Bacterial cell-surface displaying of thermo-tolerant glutamate dehydrogenase and its application in L-glutamate assay.
In this paper, glutamate dehydrogenase (Gldh) is reported to efficiently display on Escherichia coli cell surface by using N-terminal region of ice the nucleation protein as an anchoring motif. The presence of Gldh was confirmed by SDS-PAGE and enzyme activity assay. Gldh was detected mainly in the outer membrane fraction, suggesting that the Gldh was displayed on the bacterial cell surface. The optimal temperature and pH for the bacteria cell-surface displayed Gldh (bacteria-Gldh) were 70°C and 9.0, respectively. Additionally, the fusion protein retained almost 100% of its initial enzymatic activity after 1 month incubation at 4°C. Transition metal ions could inhibit the enzyme activity to different extents, while common anions had little adverse effect on enzyme activity. Importantly, the displayed Gldh is most specific to l-glutamate reported so far. The bacterial Gldh was enabled to catalyze oxidization of l-glutamate with NADP(+) as cofactor, and the resultant NADPH can be detected spectrometrically at 340nm. The bacterial-Gldh based l-glutamate assay was established, where the absorbance at 340nm increased linearly with the increasing l-glutamate concentration within the range of 10-400μM. Further, the proposed approach was successfully applied to measure l-glutamate in real samples.


Radiation therapy treatment planning using concurrent programming.
Concurrent programming can be applied to the problem of computer graphic simulation of radiation treatment of tumors (radiation treatment planning). Running several tasks or programs simultaneously on behalf of a single user provides a big improvement over the traditional sequential approach, in which editing a treatment plan and computing and displaying dose distributions are separate operations which must be invoked by explicit commands. With our system, the user sees isodose contours being updated automatically and continuously as the plan is edited; this greatly facilitates plan optimization. The complexity of parallel processing has resulted in a 'conventional wisdom' which discourages this technique. The usual approach is to have parallel processes share a common global data structure, which makes interaction hard to control and discourages modularity and data abstraction. We have developed an alternative approach based on message streams which instead enhances modularity and data abstraction while still providing the advantages of parallel processing. The system is very reliable and is used routinely in a practical clinical environment.


[Associations between glucose response and dietary fiber intakes in patients with DM].
To study the association between dietary glucose response and dietary fiber through evaluating dietary glycemic index (DGI) and diet glycemic load (DGL). Dietary intakes of 105 subjects with diabetes mellitus were measured by means of two 3-day food records through weighting. DGI and DGL were calculated according to food glycemic index and carbohydrate intakes. The content of total, insoluble and soluble dietary fiber were determined by enzymatic-gravimetric method. The values of DGI and DGL were 62.9 +/- 6.8 and 142.4 +/- 39.8 respectively. The intakes of TDF, IDF and SDF were (22.5 +/- 6.7), (16.1 +/- 5.5), (6.4 +/- 2.0) g/d respectively. The correlation coefficient between DGI and TDF were -0.407 (P < 0.01). When compared with the subjects with lowest DGI, the intakes of vegetable foods, TDF, IDF and SDF in subjects with highest DGI were decreased by 221.9 g, 6.5 g, 5.0 g, 1.4 g respectively (P < 0.01), and that of cereal foods were increased by 68.0 g (P < 0.01). Of the different sources of fiber, vegetable fiber was most strongly decreased. DGI was negtively related to TDF intakes. The diet rich in vegetables and dietary fiber may decrease the dietary glucose response.


Secular trend in the menopausal age in Finland 1997-2007 and correlation with socioeconomic, reproductive and lifestyle factors.
Only few studies have studied secular trend of menopausal age during last decade. The aim of our study is to analyze secular trend of menopausal age and to evaluate the role of socioeconomic, reproductive and lifestyle factors. National FINRISK Study sample from years 1997 and 2007 was utilized. The sample size for 1997 was 4193 and during 2007 was 4253 women. Covariance analysis included menopausal age as dependent variable and covariates (age, study year, education, occupation, parity, age at first birth, smoking, use of alcohol, physical activity, body mass index, waist circumference and waist-hip ratio) as independent variables. Median of menopausal age was 50 years in 1997 and 51 years in 2007. Differences in menopausal age by covariate were largest in smoking, education and occupation. Difference in mean menopausal age between smokers and non-smokers was larger in 2007 than in 1997 (p<0.001). Lowest educated women had lower average menopausal age during 2007 than higher educated women (p<0.001), but not in 1997. When including sociodemographic, reproductive and lifestyle factors in a model, smoking was significantly related to earlier and physical activity to later menopausal age. Study year was not significant in any model. Education, smoking and physical activity have an important role in menopausal age determination when comparing 10-year differences in menopausal age.


Brain atrophy with intracranial calcification following congenital HIV infection.
An 18-month-old girl with AIDS related complex following congenital HIV infection suffered since the first days of life from severe neurological disturbances. The extensive congenital brain atrophy and the diffuse intracranial calcification which were found were strongly reminiscent of intrauterine infection. Since the agents usually responsible for congenital infections were excluded an HIV embryopathy appears likely.


Telomerization of butadiene with glycerol: reaction control through process engineering, solvents, and additives.
Owing to the large amount of glycerol that is formed as a by-product during biodiesel production, there have been great efforts to develop new reactions and processes based on glycerol as a renewable feedstock. One example is the telomerization of butadiene with glycerol which provides an atom-economic route to amphiphilic molecules. The reaction is catalyzed by homogeneous palladium catalysts which necessitates efficient catalyst recycling. By employing an aqueous biphasic system, an increased selectivity towards the desired mono-ethers was observed in the telomerization reaction. The performance of the reaction and separation and recycling of the catalyst were optimized by the addition of organic solvents as well as cyclodextrins. By adding cyclodextrins, the conversion of glycerol could be increased and the leaching of palladium could be reduced. Leaching of palladium into the organic phase could be lowered also by using 2-octanol or 2-propanol as additional solvents. Furthermore, the catalyst system could be stabilized by adding nitriles or phosphonium salts, and radical polymerization, which leads to fouling, could be inhibited successfully.


[Molecular typing of mycobacteria].
Molecular typing is now widely used in mycobacteriology as a significant addition to conventional epidemiological studies. Several methods are available that differ with respect to their discriminating power and to the genetic marker used. Restriction fragment length polymorphism (RFLP) done using the IS6110 insertion sequence has been widely used for typing Mycobacterium tuberculosis. More recently, short repetitive DNA sequences have been used as RFLP markers, as well as for other typing methods involving gene amplification. One of these methods, the inter-direct repeat (DR)-based spoligotyping method, is rapid, directly applicable to clinical specimens, and highly discriminating.


The adhesive properties of the Staphylococcus lugdunensis multifunctional autolysin AtlL and its role in biofilm formation and internalization.
Although it belongs to the group of coagulase-negative staphylococci, Staphylococcus lugdunensis has been known to cause aggressive courses of native and prosthetic valve infective endocarditis with high mortality similar to Staphylococcus aureus. In contrast to S. aureus, only little is known about the equipment of S. lugdunensis with virulence factors including adhesins and their role in mediating attachment to extracellular matrix and plasma proteins and host cells. In this study, we show that the multifunctional autolysin/adhesin AtlL of S. lugdunensis binds to the extracellular matrix and plasma proteins fibronectin, fibrinogen, and vitronectin as well as to human EA.hy926 endothelial cells. Furthermore, we demonstrate that AtlL also plays an important role in the internalization of S. lugdunensis by eukaryotic cells: The atlL-deficient mutant Mut17 adheres to and becomes internalized by eukaryotic cells to a lesser extent than the isogenic wild-type strain Sl253 and the complemented mutant Mut17 (pCUatlL) shows an increased internalization level in comparison to Mut17. Thus, surface localized AtlL that exhibits a broad binding spectrum also mediates the internalization of S. lugdunensis by eukaryotic cells. We therefore propose an internalization pathway for S. lugdunensis, in which AtlL plays a major role. Investigating the role of AtlL in biofilm formation of S. lugdunensis, Mut17 shows a significantly reduced ability for biofilm formation, which is restored in the complemented mutant. Thus, our data provide evidence for a significant role for AtlL in adherence and internalization processes as well as in biofilm formation of S. lugdunensis.


Catalytic mechanisms of direct pyrrole synthesis via dehydrogenative coupling mediated by PNP-Ir or PNN-Ru pincer complexes: crucial role of proton-transfer shuttles in the PNP-Ir system.
Kempe et al. and Milstein et al. have recently advanced the dehydrogenative coupling methodology to synthesize pyrroles from secondary alcohols (e.g., 3) and β-amino alcohols (e.g., 4), using PNP-Ir (1) and PNN-Ru (2) pincer complexes, respectively. We herein present a DFT study to characterize the catalytic mechanism of these reactions. After precatalyst activation to give active 1A/2A, the transformation proceeds via four stages: 1A/2A-catalyzed alcohol (3) dehydrogenation to give ketone (11), base-facilitated C-N coupling of 11 and 4 to form an imine-alcohol intermediate (18), base-promoted cyclization of 18, and catalyst regeneration via H2 release from 1R/2R. For alcohol dehydrogenations, the bifunctional double hydrogen-transfer pathway is more favorable than that via β-hydride elimination. Generally, proton-transfer (H-transfer) shuttles facilitate various H-transfer processes in both systems. Notwithstanding, H-transfer shuttles play a much more crucial role in the PNP-Ir system than in the PNN-Ru system. Without H-transfer shuttles, the key barriers up to 45.9 kcal/mol in PNP-Ir system are too high to be accessible, while the corresponding barriers (<32.0 kcal/mol) in PNN-Ru system are not unreachable. Another significant difference between the two systems is that the addition of alcohol to 1A giving an alkoxo complex is endergonic by 8.1 kcal/mol, whereas the addition to 2A is exergonic by 8.9 kcal/mol. The thermodynamic difference could be the main reason for PNP-Ir system requiring lower catalyst loading than the PNN-Ru system. We discuss how the differences are resulted in terms of electronic and geometric structures of the catalysts and how to use the features in catalyst development.


Specific polymerase chain reaction for differential diagnosis of Dirofilaria immitis and Dipetalonema reconditum using primers derived from internal transcribed spacer region 2 (ITS2).
Both Dirofilaria immiti and Dipetalonema reconditum may be found in blood of infected dogs but it is not easy to distinguish D. immitis from D. reconditum in morphology. We cloned and sequenced the contiguous internal transcribed spacer (ITS) region, ITS1-5.8S-ITS2, of these two different parasites and published on GenBank as AF217800 for D. immiti and AF217801 for D. reconditum in this study. We designed two pairs of specific primers derived from ITS2 being used for polymerase chain reaction (PCR). The amplicons of ITS2 from D. immiti and D. reconditum are 302 and 348bp, respectively. Moreover, the limitation for amplifying ITS2 gene using this PCR demonstrated that 1 x 10(-2) microfilaria of each species of parasite smashed or even with mixed samples could be detected and the PCR products were predicted as the same as that described above. Thus, D. immiti and D. reconditum could be differentially diagnosed by this specific PCR. Seventeen clinical cases were evaluated and all of them were correctly identified. In this study, ITS1-5.8S-ITS2 of D. immiti or D. reconditum were the first time sequenced and analyzed. No significant similarity of ITS1 and ITS2 between D. immiti and D. reconditum could be observed.


Influence of secondary preparative parameters and aging effects on PLGA particle size distribution: a sedimentation field flow fractionation investigation.
Poly(lactic-co-glycolic acid) particles in the 200-400-nm size range were formulated through nanoprecipitation and solvent evaporation methods. Different concentrations of the polymer and stabilizer (Pluronic® F 68) were tested in order to identify the best conditions for making poly(lactic-co-glycolic acid) particles of suitable size, stable in time, and to be used as carriers for brain-targeting drugs. The particles with the best characteristics for delivery system design were those formulated by nanoprecipitation with an organic/water phase ratio of 2:30, a polymer concentration of 25 mg/mL, and a surfactant concentration of 0.83 mg/mL; their surface charge was reasonably negative (approximately -27 mV) and the average size of the almost monodisperse population was roughly 250 nm. Particle characterization was obtained through ζ-potential measurements, scanning electron microscope observations, and particle size distribution determinations; the latter achieved by both photon-correlation spectroscopy and sedimentation field flow fractionation. Sedimentation field flow fractionation, which is considered more reliable than photon-correlation spectroscopy in describing the possible particle size distribution modifications, was used to investigate the effects of 3 months of storage at 4 °C had on the lyophilized particles. Figure Particle size ditribution from the SdFFF and the PCS techniques.


Vision anomalies of Indian school children: the Lame Deer study.
The occurrence of vision anomalies among American Indian school children is measured for an entire school population, according to grade (pre-kindergarten through 12). The anomalies are identified, classified in three major groups (myopes, astigmats, all other), and each group is compared to Caucasian school children. Some conclusions and recommendations are made concerning the vision care needs of Amerind school children.


Imatinib Is Protective Against Ischemia-Reperfusion Injury in an Ex Vivo Rabbit Model of Lung Injury.
Ischemia-reperfusion injury is characterized by an increase in oxidative stress and leads to significant morbidity and death. The tyrosine kinase c-Abl is activated by oxidative stress and mediates processes that affect endothelial barrier function. We hypothesized treatment with the c-Abl inhibitor imatinib would be protective against ischemia-reperfusion injury in our ex vivo rabbit model. Heart-lung blocs were harvested from rabbits and stored in cold in Perfadex (Vitrolife, Englewood, CO) for 18 hours. Blocs were reperfused for 2 hours in an ex vivo circuit with donor rabbit blood alone (untreated group, n = 7) or donor rabbit blood and 4 mg imatinib (treatment group, n = 10). Serial clinical variables measured every 15 minutes (arterial oxygen and carbon dioxide tension and mean pulmonary artery pressures) and biochemistry of tissue samples before and after reperfusion were assessed. Compared with untreated lungs, imatinib treatment improved physiologic parameters, including oxygen, carbon dioxide, and pulmonary artery pressures. Imatinib-treated lungs had less vascular barrier dysfunction as quantified by wet-to-dry weight ratios and bronchoalveolar lavage protein concentrations. Treated lungs showed less inflammation as measured by bronchoalveolar lavage myeloperoxidase assay, less mitochondrial reactive oxygen species production, and increased antioxidant catalase levels. Finally, imatinib protected lungs from DNA damage and p53 upregulation. Imatinib treatment significantly improved the physiologic performance of reperfused lungs and biochemical indicators associated with reperfusion injury in this ex vivo model. Further study is necessary to elucidate the mechanism of tyrosine kinase inhibition in lungs exposed to ischemia and reperfusion.


Ethical safety of deep brain stimulation: A study on moral decision-making in Parkinson's disease.
The possibility that deep brain stimulation (DBS) in Parkinson's disease (PD) alters patients' decisions and actions, even temporarily, raises important clinical, ethical and legal questions. Abnormal moral decision-making can lead to ethical rules violations. Previous experiments demonstrated the subthalamic (STN) activation during moral decision-making. Here we aim to study whether STN DBS can affect moral decision-making in PD patients. Eleven patients with PD and bilateral STN DBS implant performed a computerized moral task in ON and OFF stimulation conditions. A control group of PD patients without DBS implant performed the same experimental protocol. All patients underwent motor, cognitive and psychological assessments. STN stimulation was not able to modify neither reaction times nor responses to moral task both when we compared the ON and the OFF state in the same patient (reaction times, p = .416) and when we compared DBS patients with those treated only with the best medical treatment (reaction times: p = .408, responses: p = .776). Moral judgment is the result of a complex process, requiring cognitive executive functions, problem-solving, anticipations of consequences of an action, conflict processing, emotional evaluation of context and of possible outcomes, and involving different brain areas and neural circuits. Our data show that STN DBS leaves unaffected moral decisions thus implying relevant clinical and ethical implications for DBS consequences on patients' behavior, on decision-making and on judgment ability. In conclusion, the technique can be considered safe on moral behavior.


[Ultrastructure of paired neuron contacts during the posthypoxic period].
Peculiar contacts between bodies of double neurons were revealed in the sensomotor cortex of the rat brain under ischemia and histotoxic hypoxia by means of the electron microscopic investigation. Ultrastructure of the contacting double neurons is not homogenous. In some cases, they do not differ from the normal, in other cases, certain signs of activation are revealed. In the contact area, peripheral focal chromatolysis is sometimes observed. In the double neurons the number of microtubules is often increased. In some cases, the cytoplasmic structure is altered (demonstrating certain activation in one of the cells) and combined with destructive processes in the neighbouring cell. The double neurons revealed can be divided into 3 groups: 1) pairs of morphologically identical neurons without specialized areas in the bordering membranes, 2) double neurons with specialized areas of membranes at places of their contacts, 3) double neurons with fusion of their cytoplasm at places where the cellular bodies come into contact with each other. The types of the double neurons described reveal some additional ways along which metabolism in neurons can occur.


Mechanism of PAMAM Dendrimers Internalization in Hippocampal Neurons.
Polyamidoamine (PAMAM) dendrimers are hyperbranched macromolecules which have been described as one of the most promising drug nanocarrier systems. A key process to understand is their cellular internalization mechanism because of its direct influence on their intracellular distribution, association with organelles, entry kinetics, and cargo release. Despite that internalization mechanisms of dendrimers have been studied in different cell types, in the case of neurons they are not completely described. Considering the relevance of central nervous system (CNS) diseases and neuropharmacology, the aim of this report is to describe the molecular internalization mechanism of different PAMAM-based dendrimer systems in hippocampal neurons. Four dendrimers based on fourth generation PAMAM with different surface properties were studied: unmodified G4, with a positively charged surface; PP50, with a substitution of the 50% of amino surface groups with polyethylene glycol neutral groups; PAc, with a substitution of the 30% of amino surface groups with acrylate anionic groups; and PFO, decorated with folic acid groups in a 25% of total terminal groups. Confocal images show that both G4 and PFO are able to enter the neurons, but not PP50 and PAc. Colocalization study with specific endocytosis markers and specific endocytosis inhibitor assay demonstrate that clathrin-mediated endocytosis would be the main internalization mechanism for G4, whereas clathrin- and caveolae-mediated endocytosis would be implicated in PFO internalization. These results show the existence of different internalization mechanisms for PAMAM dendrimers in neurons and the possibility to control their internalization properties with specific chemical modifications.


Conservative esthetic geriatric restoration using anhydrous glass ionomer.
A conservative technique for treatment of incisal erosion lesions using a new light-cured ionomer restorative cement is described.


Enhancing human aspects of care with young people with muscular dystrophy: Results from a participatory qualitative study with clinicians.
Most research into clinical care of Duchenne or Becker dystrophinopathies (MD) has focused on slowing progressive muscular weakness and extending lifespan. Scarce attention has been paid to the "human" aspects of care such as psychosocial health, living a fulfilling life, or dealing with disability stigma. This study partnered with clinicians to identify and address local and systemic barriers to these human aspects of care. We employed a participatory qualitative design at a multidisciplinary MD clinic using 2 methods: (a) ethnographic observations over a 6-month period of clinic visits of children with MD and families, involving 12 clinicians, and (b) 3 "dialogues" (2-way discussions) with these clinicians to collaboratively analyze practices and co-produce recommendations for change. Our methods produced rich data that, when coanalyzed with clinicians and in consultation with a family advisor, provided deep insights into the practices and underlying assumptions of a neuromuscular clinic. Staff recognized the importance of the human aspects of care but, in reviewing the observational data, identified that it was given insufficient attention in (a) routine clinical processes, (b) clinician-family patterns of interaction, and (c) staffing allocations. Although the human aspects of care were important to clinicians in the MD clinic, the routines and nature of the clinic meant these were frequently sidelined for biomedical objectives. We present collaboratively produced practical recommendations toward addressing this disjunction between ideals and practice including developing flexibility to tailor appointment frequency, composition, and length; providing time and physical space for psychosocial aspects of care; and clinician skill building to support child/family expression of "negative" emotions; and discussion of sociopolitical aspects of MD such as living with disability stigma. The study offers a set of considerations that, taking into account individual differences, offer insights for similar clinics elsewhere.


Mammary Dysbiosis and Nipple Blebs Treated With Intravenous Daptomycin and Dalbavancin.
Mammary dysbiosis, also known as subacute mastitis, may be associated with nipple blebs. These overlapping diagnoses represent a challenging clinical scenario during lactation. Little research has been published on etiology, management strategies, and outcomes of these concurrent diagnoses. We document the treatment and outcome of a patient who presented with left-breast dysbiosis and nipple blebs and whose milk culture grew multi-drug-resistant, methicillin-resistant Staphylococcus aureus. She was treated safely and effectively with intravenous daptomycin and dalbavancin. This has not been described previously in the lactation literature. The 35-year-old lactating gravida 3, para 3 patient presented at 6 months postpartum to a breast surgery clinic with a 1-week history of worsening deep left-breast pain, blebs, and recurrent plugging. She was afebrile and she had no erythema or induration on her breast exam. A culture of her milk grew multi-drug-resistant, methicillin-resistant Staphylococcus aureus, and she was referred to infectious disease for assistance with intravenous antibiotic therapy. She continued to feed expressed milk throughout treatment and demonstrated complete resolution of symptoms 8 weeks later. We report that in patients with a multi-drug-resistant, methicillin-resistant Staphylococcus aureus-positive human milk culture and a clinical presentation of mammary dysbiosis and nipple blebs, intravenous daptomycin and dalbavancin may be an effective treatment.


Frequency of aspirin non responsiveness in patients of ischemic heart disease.
Aspirin is widely used as an antiplatelet agent . Many patients have been noticed with recurrence of major ischemic events in- spite of antiplatelet therapy. The objective of this study was to determine frequency of aspirin non-responsiveness /resistance in patients of ischemic heart disease. Seventy one patients of IHD were selected from out-patient department of Punjab Institute of Cardiology Lahore. Whole Blood Platelet aggregation studies were performed on Diamed Impact R. Aspirin response assay was performed with DiaChidon (Arachidoinc Acid 16mmol/L). Non responders to aspirin were assessed on the basis of software generated results: Surface covered (SC) >2.5% was considered as response to aspirin and SC <2.5% was considered as no response (or resistance) to Aspirin. Chi-square test was applied to measure statistical significance. Non-response to Aspirin was observed in 11% (8 out of 71). There was significant association (p=0.045) between resistance to aspirin and Diabetes mellitus. Treatment resistance was also significantly associated with female gender (p=0.015). We concluded that non response to Aspirin is seen in significant number of patients of IHD. Diabetes mellitus and female gender are strong risk factors of developing failure to aspirin therapy.


Validation of the BioPRYN enzyme-linked immunosorbent assay for detection of pregnancy-specific protein-B (PSPB) and diagnosis of pregnancy in American bison (Bison bison).
This study assessed the accuracy of the commercial BioPRYN® ELISA for the detection of pregnancy-specific protein-B (PSPB) using a single blood sample to determine pregnancy status in American bison (Bison bison). A total of 49 bison cows were used in the study, and sampled at two time-points during the gestation period, fall and spring, correlating with early- to mid-term gestation (average 62.9 days post-mating) and mid- to late-term gestation (average 229.2 days post-mating), respectively. Sensitivity of the test during early- to mid-term gestation sampling period (fall) was 87.1%, while specificity was 100%; sensitivity of the test during late-term gestation sampling period (spring) was 96.3%, while specificity remained at 100%. In total, the test showed a total sensitivity of 91.4%, specificity of 100% and total accuracy of 93.8%, similar to domestic cattle. Use of the single-sample BioPRYN® ELISA in American Bison for pregnancy diagnosis is economical and practical, minimizing animal handling time, frequency and subsequent stress while providing accurate results for pregnancy diagnosis at 62 days post-mating. This method should be considered over more traditional pregnancy diagnosis methods for use in managed bison herds.


Stimulation of rat placental lactogen-II (rPL-II) secretion by cultured trophoblasts by insulin: development of a rat placental cell culture system and effects of peptide hormones on rPL-II secretion in vitro.
The purpose of this study was to develop a primary culture system using serum-free medium for rat placental trophoblast cells and to investigate the factors that control rat placental lactogen-II (rPL-II) secretion in vitro. The placentae of day 13 pregnant rats were dissociated in Medium 199 containing 0.1% collagenase and 0.002% DNAase. Dissociated cells were fractionated into five segments by centrifugation through a 40% Percoll density gradient and incubated on rat tail collagen bed in medium SFM-101 for up to 7 days. Fraction B at the Percoll gradient density of 1.05 g ml-1 was enriched with rPL-II-producing cells and the time course of rPL-II secretion was characterized by a rapid increase in the first 2 days, remaining at high values (mean: 14-16 ng micrograms-1 DNA) for the following 2-3 days and decreasing thereafter. The rPL-II-producing cells from faction B identified by immunocytochemical examination accounted for approximately 69% of total cultured cells and consisted of a few giant cells and polygonal cells. Growth factors (bovine insulin, 0.1-20 micrograms ml-1; recombinant human insulin-like growth factor (IGF)-I, IGF-II, 0.1-1.0 micrograms ml-1; murine epidermal growth factor (EGF), 0.001-10 micrograms ml-1), rat pituitary hormones (rat growth hormone, rat prolactin, 0.1-10 micrograms ml-1) and hypothalamic hormones (human growth hormone-releasing hormone (GHRH), corticotrophin-releasing hormone (CRH), LHRH, 0.1-10 micrograms ml-1) were individually added to the culture medium to investigate the putative factors that directly control rPL-II secretion by the trophoblast cells.(ABSTRACT TRUNCATED AT 250 WORDS)


Conversion of the LIMA1 tumour suppressor into an oncogenic LMO-like protein by API2-MALT1 in MALT lymphoma.
MALT1 is the only known paracaspase and is a critical mediator of B- and T-cell receptor signalling. The function of the MALT1 gene is subverted by oncogenic chimeric fusions arising from the recurrent t(11;18)(q21;q21) aberration, which is the most frequent translocation in mucosa-associated lymphoid tissue (MALT) lymphoma. API2-MALT1-positive MALT lymphomas manifest antibiotic resistance and aggressive clinical behaviour with poor clinical outcome. However, the mechanisms underlying API2-MALT1-induced MALT lymphomagenesis are not fully understood. Here we show that API2-MALT1 induces paracaspase-mediated cleavage of the tumour suppressor protein LIMA1. LIMA1 binding by API2-MALT1 is API2 dependent and proteolytic cleavage is dependent on MALT1 paracaspase activity. Intriguingly, API2-MALT1-mediated proteolysis generates a LIM domain-only (LMO)-containing fragment with oncogenic properties in vitro and in vivo. Importantly, primary MALT lymphomas harbouring the API2-MALT1 fusion uniquely demonstrate LIMA1 cleavage fragments. Our studies reveal a novel paracaspase-mediated oncogenic gain-of-function mechanism in the pathogenesis of MALT lymphoma.


Oculodermal Melanocytosis: Nevus of Ota in a Dog.
This report describes the clinical presentation, diagnosis, and histopathologic features of oculodermal melanocytosis in a young dog. A 3-year-old male neutered Labrador Retriever presented with conjunctival and scleral hyperpigmentation of the right eye, with concurrent ipsilateral cutaneous hyperpigmentation involving the right side of the face. Initial skin and conjunctival biopsies revealed an accumulation of histologically benign melanocytes within the dermis and conjunctival stroma, respectively. Enucleation was elected 19 months later by the referring veterinarian due to the progression of ocular pigmentation with concurrent marked corneal lipidosis and the suspicion of a scleral mass. On gross and histopathologic examination of the globe, there was marked panuveal melanocytosis with extension into the sclera, bulbar conjunctiva, and connective tissue surrounding the optic nerve, as well as sharply demarcated ipsilateral hyperpigmentation of the facial skin. The findings are characteristic of oculodermal melanocytosis (nevus of Ota), a dermal melanocytic hamartoma presenting as cutaneous facial hyperpigmentation that corresponds to the distribution of the ophthalmic and maxillary branches of the trigeminal nerve, often with ipsilateral ocular involvement.


The assessment of cell cycle genes in the rat mandibular condyle.
We aimed to investigate the expression profile of cell cycle genes in the mandibular condyle on mechanical strain and natural growth, and quantify their expression intensity. Three hundred and fifty 35 days old Sprague-Dawley rats were randomly divided into experimental groups fitted with bite-jumping appliances and control groups. Groups were sacrificed at days 1, 3, 7, 9, 14, 30, and 33. Then, condyles were dissected and total RNA was extracted for microarray analysis. Thirty-nine known cell cycle genes were present in the condyle, where Cyclin D1, PCNA, and Wnt5a were differentially expressed. Reverse transcriptase-PCR confirmed that Wnt5a showed a 2-fold increase on experimental day 1, Cyclin D1 showed a 2-fold increase on experimental day 1 and a 3-fold increase on experimental day 14, while PCNA shows 2.2-fold increase both on experimental days 9 and 30. PCNA, Cyclin D1, and Wnt5a were all expressed by cells in both the proliferative layer and erosive zone. Mandibular advancement leads to the expression of Cyclin D1 that accelerates entry to the S phase. The increased level of PCNA indicates increased DNA replication of MSC. Then, elevated level of Wnt5a indicates the commitment of MSC to the chondrogenic lineage.


Clinical and imaging features of pulmonary artery sling in infants without significant hemodynamic changes.
Pulmonary artery sling (PAS) is a rare congenital heart anomaly and may cause unexplained respiratory symptoms in infants. Since the non-specific respiratory symptoms of PAS may lead to misdiagnosis, the aim of this study was to clarify the clinical and imaging features of this disease for timely diagnosis and treatment. Clinical histories, physical examinations and imaging studies were retrospectively evaluated in nine infants with PAS. Chest X-ray, echocardiography and contrast-enhanced computed tomography (CT) with 3-dimensional reconstructions were performed in all patients and three of them received surgical treatment. Nine cases included six males and three females with a mean age of (4.3 ± 2.8) months ranging from 2 to 11 months old. All patients had respiratory symptoms including recurrent cough, stridor and wheezing. The onset of symptoms was within 3 months in all cases and three children had symptoms only a few days after birth. The chest X-ray showed pneumonia in all cases. Contrast-enhanced CT showed the tracheal compression at different lengths in every case. The echocardiograph findings of PAS were anomalous origins of the left pulmonary artery from the posterior aspect of the right pulmonary artery. Of the 9 cases, 8 cases were diagnosed correctly by echocardiography. Of the complicated abnormalities, there were one with secundum atrial septal defect, one with patent foramen ovale and three with persistent left superior vena cava. None of them were complicated with significant blood dynamic changes. Infants with recurrent respiratory symptoms such as chronic cough, stridor and wheezing, should be examined for the possible presence of congenital pulmonary artery sling. As a noninvasive technique, echocardiography is very helpful and should be the first-choice modality for the diagnosis of pulmonary artery sling. Contrast-enhanced CT, clearly demonstrating the anatomy of pulmonary artery sling and the position and extent of trachea compression, is necessary for the final diagnosis and pre-operation evaluation.


Prevalence of anencephaly in the region of Rijeka, Croatia.
This retrospective study determines the prevalence of anencephaly in the region of Rijeka, Croatia. Records of all spontaneous and therapeutic abortions terminated in medical institutions, all fetuses weighing more than 500 g or more than 22 weeks gestation (whether the product of abortion, therapeutic termination, stillborn or liveborn) and infants who died in the first year of life in the region of Rijeka, Croatia, during the 1963-2000 period were reviewed. There were 135,451 births; 22 of them were anencephalics (19 stillborn), which comprises 0.2% of all births and 2.1% of stillbirths. Annual prevalence of anencephaly varied in range from 0.00 to 7.42 per 10,000 births. In two cases pregnancy was electively terminated after ultrasonographic diagnosis of anencephaly. Fifteen anencephalics were female, six were male, and in one case sex was undetermined due to aplasia of genital organs. Associated congenital malformations were detected in 18 anencephalics. The importance of establishing national and international registers of congenital malformations in all countries is stressed. The authors suggested that the setting of obligatory reporting of all congenital malformations would be the first step toward this practice in Croatia, as well as in other developing countries.


[Activation and inactivation of batrachotoxin-modified sodium channels of nerve fiber membranes in the frog].
Currents through batrachotoxin (BTX)-modified sodium channels in frog myelinated nerve were measured under voltage-clamp conditions. Nonlinearity of "instantaneous" current-voltage relations was taken into account when determining steady-state parameters of channel activation. BTX induces the shift of voltage dependence of channel activation towards more negative potentials by 67 mV, without changes in its steepness. Current kinetics and effect of preceding depolarization on current size suggest that BTX-modified channels are capable for partial inactivation. High level of steady-state conductance of BTX-modified channels can be explained by suggestion that open state of the channel is energetically more profitable than inactivated one. It is concluded that effect of BTX on inactivation is different in principle from that of pronase and protein reagents.


Effect of dietary arginine on growth, intestinal enzyme activities and gene expression in muscle, hepatopancreas and intestine of juvenile Jian carp (Cyprinus carpio var. Jian).
The present study was conducted to test the hypothesis that dietary arginine promotes digestion and absorption capacity, and, thus, enhances fish growth. This improvement might be related to the target of rapamycin (TOR) and eIF4E-binding protein (4E-BP). A total of 1200 juvenile Jian carp, Cyprinus carpio var. Jian, with an average initial weight of 6.33 (SE 0.03) g, were fed with diets containing graded concentrations of arginine, namely, 9.8 (control), 12.7, 16.1, 18.5, 21.9 and 24.5 g arginine/kg diet for 9 weeks. An real-time quantitative PCR analysis was performed to determine the relative expression of TOR and 4E-BP in fish muscle, hepatopancreas and intestine. Dietary arginine increased (P < 0.05): (1) glutamate-oxaloacetate transaminase and glutamate-pyruvate transaminase activities in muscle and hepatopancreas; (2) intestine and hepatopancreas protein content, folds height, and trypsin, chymotrypsin, lipase, Na⁺/K⁺-ATPase, alkaline phosphatase, γ-glutamyl transpeptidase and creatine kinase activities in intestine; (3) Lactobacillus counts; (4) relative expression of TOR in the muscle, hepatopancreas and distal intestine (DI); (5) relative expression of 4E-BP in proximal intestine (PI) and mid-intestine (MI), as compared with the control group. In contrast, dietary arginine reduced (P < 0.05): (1) plasma ammonia content; (2) Aeromonas hydrophila and Escherichia coli counts; (3) relative expression of TOR in PI and MI; (4) relative expression of 4E-BP in the muscle, hepatopancreas and DI. The arginine requirement estimated by specific growth rate using quadratic regression analysis was found to be 18.0 g/kg diet. These results indicate that arginine improved fish growth, digestive and absorptive ability and regulated the expression of TOR and 4E-BP genes.


Clinical and metabolic response to probiotic supplementation in patients with rheumatoid arthritis: a randomized, double-blind, placebo-controlled trial.
This study was performed to determine the effects of probiotic supplementation on clinical and metabolic status of patients with rheumatoid arthritis (RA). Sixty patients with RA aged 25-70 years were assigned into two groups to receive either probiotic capsules (n = 30) or placebo (n = 30) in this randomized, double-blind, placebo-controlled trial. The patients in the probiotic group received a daily capsule that contained three viable and freeze-dried strains: Lactobacillus acidophilus (2 × 10(9) colony-forming units [CFU]/g), Lactobacillus casei (2 × 10(9) CFU/g) and Bifidobacterium bifidum (2 × 10(9) CFU/g) for 8 weeks. The placebo group took capsules filled with cellulose for the same time period. Fasting blood samples were taken at the beginning and the end of the study to quantify related markers. After 8 weeks of intervention, compared with the placebo, probiotic supplementation resulted in improved Disease Activity Score of 28 joints (DAS-28) (-0.3 ± 0.4 vs. -0.1 ± 0.4, P = 0.01). In addition, a significant decrease in serum insulin levels (-2.0 ± 4.3 vs. +0.5 ± 4.9 μIU/mL, P = 0.03), homeostatic model assessment-B cell function (HOMA-B) (-7.5 ± 18.0 vs. +4.3 ± 25.0, P = 0.03) and serum high-sensitivity C-reactive protein (hs-CRP) concentrations (-6.66 ± 2.56 vs. +3.07 ± 5.53 mg/L, P < 0.001) following the supplementation of probiotics compared with the placebo. Subjects who received probiotic capsules experienced borderline statistically significant improvement in total- (P = 0.09) and low-density lipoprotein-cholesterol levels (P = 0.07) compared with the placebo. Overall, the results of this study indicated that taking probiotic supplements for 8 weeks among patients with RA had beneficial effects on DAS-28, insulin levels, HOMA-B and hs-CRP levels.


Immunotherapy in children: report from the Reisensburg-Symposium October 20-22, 2004 and recent advances.
Immunotherapy of childhood malignancies has a magic aura of promising an easy way to cure. It is not only the public, but also the medical community, which has caused this hope. Therefore, it was worthwhile to bring together a panel of experts working in the field of immunotherapy in order to discuss ongoing projects, and to show hopes and disappointments. The result was a fascinating symposium sponsored by The Kind Philipp Leukemia Foundation (Mönchengladbach, Germany). The symposium took place in the Reissensburg near Ulm (Germany) which gives the ideal location for such an event.


Egg shell colour is affected by laying cage design.
1. When laying hens are stressed some retain their eggs in the shell gland beyond the normal time of laying and this can result in the deposition of extra-cuticular calcium which makes brown eggs appear paler. 2. Three different types of enriched modified cage were compared: the location where eggs were laid was recorded and shell colour was measured using a reflectometer. 3. In 2 types of cage with enclosed nest boxes more eggs (80%) were laid in the nests than in a design with nest hollows in the open part of the cage (41%). 4. The eggs from the cages with enclosed nests were darker (had less extraneous calcium) than those with open nest hollows. This implies that in the designs with nest boxes fewer eggs had been retained and the hens may have been less stressed. 5. The results support previous evidence that to reduce stress and improve welfare it is desirable to provide enclosed nest sites for caged laying hens.


Proteomic Comparison of Various Hepatic Cell Cultures for Preclinical Safety Pharmacology.
Experimental drugs need to be screened for safety within time constraints. Hepatotoxicity is one concerning contributor to the failure of investigational new drugs and a major rationale for postmarketing withdrawal decisions. Ethical considerations in preclinical research force the requirement for highly predictive in vitro assays using human tissue which retains functionality reflective of primary tissue. Here, the proteome of cells commonly used to assess preclinical hepatotoxicity was compared. Primary human hepatocytes (PHHs), hepatocyte-like cells (HLCs) differentiated from human pluripotent stem cells, HepG2 cell monolayers and HepG2 cell 3D spheroids were cultured and collected as whole cell lysates. Over 6000 proteins were identified and quantified in terms of relative abundance in replicate proteomic experiments using isobaric tagging methods. Comparison of these quantitative data provides biological insight into the feasibility of using HLCs, HepG2 monolayers, and HepG2 3D spheroids for hepatotoxicity testing. Collectively these data reveal how HLCs differentiated for 35 days and HepG2 cells proteomes differ from one another and that of PHHs. HepG2 cells possess a strong cancer cell signature and do not adequately express key metabolic proteins which mark the hepatic phenotype, this was not substantially altered by culturing as 3D spheroids. These data suggest that while no single hepatic model reflects the diverse array of outcomes required to mimic the in vivo liver functions, that HLCs are the most suitable investigational avenue for replacing PHHs in vitro.


Apoptotic Cancer Cells Suppress 5-Lipoxygenase in Tumor-Associated Macrophages.
The enzyme 5-lipoxygenase (5-LO) is key in the synthesis of leukotrienes, which are potent proinflammatory lipid mediators involved in chronic inflammatory diseases including cancer. 5-LO is expressed in immune cells but also found in cancer cells. Although the role of 5-LO in tumor cells is beginning to emerge, with the notion that tumor-promoting functions are attributed to its products, the function of 5-LO in the tumor microenvironment remains unclear. To understand the role of 5-LO and its products in the tumor microenvironment, we analyzed its expression and function in tumor-associated macrophages (TAMs). TAMs were generated by coculturing primary human macrophages (MΦ) with human MCF-7 breast carcinoma cells, which caused cell death of cancer cells followed by phagocytosis of cell debris by MΦ. Expression and activity of 5-LO in TAMs were reduced upon coculture with cancer cells. Downregulation of 5-LO in TAMs required tumor cell death and the direct contact between MΦ and dying cancer cells via Mer tyrosine kinase. Subsequently, upregulation of proto-oncogene c-Myb in TAMs induced a stable transcriptional repression of 5-LO. Reduced 5-LO expression in TAMs was mechanistically coupled to an attenuated T cell recruitment. In primary TAMs from human and murine breast tumors, 5-LO expression was absent or low when compared with monocyte-derived MΦ. Our data reveal that 5-LO, which is required for leukotriene production and subsequent T cell recruitment, is downregulated in TAMs through Mer tyrosine kinase-dependent recognition of apoptotic cancer cells. Mechanistically, we noticed transcriptional repression of 5-LO by proto-oncogene c-Myb and conclude that loss of stromal 5-LO expression favors tumor progression.


Impact of amino acid substitutions in the core region of HCV on multistep hepatocarcinogenesis.
The core protein of hepatitis C virus (HCV) has multiple functions for not only viral replication but also hepatocellular carcinogenesis. A significant association of the substitutions in the core region with hepatocarcinogenesis has recently been reported. In this report, we evaluated the association of the substitutions in the core region with multistep hepatocarcinogenesis. Sixty-nine non-cancerous and cancerous liver tissues were obtained from the patients with primary developed hepatocellular carcinoma (HCC) due to HCV and 17 cirrhotic liver tissues were obtained from the patients without HCC. A sequence analysis of the core protein of HCV was performed and the association between the substitution rates in the core gene and the degree of fibrosis or steatosis during the primary development of HCC and tumor differentiation was analyzed. The substitution rates of amino acid 70, 75, 91 and 147 exceeded 25% (amino acid 70, 51%; 75, 45%; 91, 36%; 147, 30%). All substitution rates were comparable among cancerous and non-cancerous region of patients with HCC and non-cancerous region without HCC. The substitution rates of these four amino acids were not associated with the degree of fibrosis, steatosis or tumor differentiation during the primary development of HCC. In addition, the substitution rates were comparable between the patients with or without HCC. The cumulative substitution numbers in the core region were also not associated with the degree of fibrosis and steatosis. It is possible that the substitutions in the core region are not associated with HCV-related multistep hepatocarcinogenesis.


[Lobar intracerebral hemorhages: a clinical study in 97 patients].
We aimed to characterize the clinical features,etiology and prognosis in lobar hemorrhages (LH). Descriptive study of 97 patients with LH included in the Sagrat Cor Hospital of Barcelona Stroke Registry over a 12 year period. The vascular risk factors and clinical profiles in a LH group and a subcortical hemorrhage group (n = 92) were compared. LH accounted for 3.9% of all acute strokes (n = 2,500) and 35.9% of all intracerebral hemorrhages. The parietal lobe was the most frequent cerebral topography (45.4%). LH were caused by arterial hypertension in 41% of patients, vascular malformations in 8.5%, bleeding diathesis in 5.5% and anticoagulation in 3%. The overall in-hospital mortality rate was 26.8%. Early seizures (odds ratio [OR] = 5.41; confidence interval [CI] 95%, 1.06-27.69), headache (OR = 2.19; CI 95%, 1.13-4.26), female sex (OR = 1.97; CI 95%, 1.04-3.75), sensory deficit (OR = 0.41; CI 95%, 0.22-0.77) and hypertension (OR = 0.34; CI 95%, 0.18-0.65) were independent predictors of LH. One in every 3 intracerebral hemorrhages is a LH. Different potentially modifiable vascular risk factors and clinical profiles were identified in LH versus subcortical hemorrhages.


Heterozygote detection in a family of Lapland dogs with a recessively inherited metabolic disease: canine glycogen storage disease type II.
The control of recessively inherited inborn errors of metabolism may benefit from quantitative biochemical screening assays enabling the identification of heterozygous individuals. Based on the principle of partial enzyme deficiency in heterozygotes, an attempt was made to identify heterozygous animals in a Lapland dog family with canine glycogen storage disease type II (acid alpha-glucosidase deficiency). Acid alpha-glucosidase activity was determined in peripheral blood leucocyte extracts of 12 related Lapland dogs, two of which were obligate heterozygotes. The use of an antiserum against acid alpha-glucosidase was necessary to increase the specificity of the assay. Twice the obligate heterozygous enzyme level was assumed to indicate the homozygous normal level. Five dogs were designated as presumptive heterozygotes, and five as presumptive normal homozygotes. The results in two dogs were inconclusive. The information obtained in this preliminary investigation may be helpful in the control of the disease in the Lapland dog breed.


National survey of the usage of lung expansion modalities for the prevention and treatment of postoperative atelectasis following abdominal and thoracic surgery.
A national survey of hospitals was conducted to evaluate the usage of lung expansion maneuvers in the prevention and management of postoperative atelectasis associated with abdominal and thoracic surgery. Equal numbers of hospitals were randomly selected from the nine American Hospital Association regions and from bed-size groups of 50 to 200 beds, 201 to 400 beds, and greater than 400 beds. Preoperative and postoperative prophylactic therapy was found to be similar in all groups except for a lower usage of intermittent positive-pressure breathing (IPPB) in the western third of the United States compared with the central third. In the treatment of postoperative atelectasis there are significant differences in the use of chest physical therapy, IPPB, and intermittent continuous positive airway pressure based on hospital size. Objective measurements of tidal volume or inspiratory capacity as a guide to therapeutic decisions are performed more frequently in the western regions. Surgical statistics relative to the number of abdominal and surgical procedures done and the incidence of postoperative atelectasis are also presented.


A comparative study of nosocomial and community-acquired strains of Escherichia coli causing bacteraemia in a Danish University Hospital.
In a previous study we found a considerably higher mortality rate in patients with nosocomial (NO) compared with community-acquired (CA) Escherichia coli bacteraemia. To establish whether this was due to host differences or to differences in the infecting bacteria, we compared 205 NO with 172 CA bacteraemic isolates of E. coli with respect to serotype, virulence factors and antimicrobial susceptibility. Overall the six most frequent O antigens were O18ac, O6, O1, O2, O15 and O75, respectively. The six most frequent capsular antigens were K1, K5, K52, K2, K7 and K34, respectively. No major differences were found regarding O-antigens, capsular antigens, production of haemolysin, P-fimbriation, serum sensitivity or antimicrobial susceptibility. Surprisingly we found 17 strains of serotype O15:K52:[H1] of both NO (eight) and CA (nine) origin with similar phenotypic characteristics to a strain causing a CA outbreak in London 1986-1987. Possibly the Danish and the English strains belong to the same clone. Our findings argue against the existence of a distinct NO flora. NO E. coli bacteraemia strains seem to originate primarily from the patients' own flora.


Assessment of vitamin K status of the newborn infant.
This article reviews our current knowledge of the assessment of vitamin K status in the human infant, where a deficiency state has been described. Laboratory measures for the functional assessment of vitamin K-dependent coagulation factors as well as the quantitative assay for vitamin K1 (phylloquinone) are reviewed. In addition, four different methods of measuring abnormal prothrombin, a protein induced by vitamin K absence (PIVKA-II), and its use in clinical pediatrics are discussed. Finally, additional methods are briefly described for assessing vitamin K status, including serum osteocalcin, urinary excretion of gamma-carboxyglutamic acid, and deficiencies of the enzymes necessary for the regeneration of phylloquinone in the so-called vitamin K cycle.


Chorioamnionitis induced by subchorionic endotoxin infusion in sheep.
This study was undertaken to determine whether subchorionic endotoxin infusion causes chorioamnionitis and preterm lung maturation, as occurs after intra-amniotic endotoxin. From day 118 of pregnancy, sheep received infusions of endotoxin (subchorionic 7.5 mg/d, n=11; intra-amniotic 2.5 mg/d, n=9) until delivery of lambs at 120 or 124 days. Other sheep received a single intra-amniotic injection of endotoxin (10 mg, n=7) at 118 days before delivery at 124 days. Controls (n=9) received equivalent saline solution treatments. Chorioamnionitis accompanied all endotoxin treatments. Lung inflammation occurred after intra-amniotic endotoxin infusion or injection but not after subchorionic endotoxin. Umbilical arterial pH was lower and Pco(2) was higher than control after subchorionic endotoxin. Lung compliance and surfactant were increased after intra-amniotic endotoxin infusion or injection but not after subchorionic endotoxin. Chorioamnionitis may result from inflammatory stimuli at various intrauterine sites, with different sites causing different fetal effects and not all cases of chorioamnionitis being accompanied by enhanced lung maturation.


Characterization of state transitions in spatially distributed, chaotic, nonlinear, dynamical systems in cerebral cortex.
The neurons of cerebral cortex are largely autonomous and generate activity that is manifested in trains of microscopic axonal action potentials. The neurons interact by sparse but numerous synaptic connections to generate macroscopic dendritic activity patterns that are observed in electroencephalographic (EEG) waves. The macroscopic patterns are constructed by the populations and they shape the output of cortical neurons in parallel arrays. Sensory cortexes receive sensory information in the form of microscopic action potentials, which induce state transitions in population dynamics. Each state transition transforms sensory information to perceptual meaning. The EEG reflects both kinds of activity. The sensory input is accessed by time ensemble averaging, whereas the perceptual output is found by spatial ensemble averaging. Spatial phase gradients in the EEG are useful for identifying EEG segments in a sequence of state transitions in response to sensory input. The rapidity and flexibility with which they take place give strong reason to postulate that the mechanism for the construction of these sequences of patterns is a dynamical system operating in a chaotic domain.


Expression of ICAM-1 and E-selectin in gingival tissues of smokers and non-smokers with periodontitis.
Tobacco smoking affects systemic concentrations of soluble intercellular adhesion molecule (ICAM)-1, but its effect on local expression of adhesion molecules in gingival tissue has not been studied previously. E-selectin and ICAM-1 expression on small blood vessel endothelia in gingival biopsies obtained from smokers (n=17) and non-smokers (n=17) with periodontitis was examined with immunohistochemistry. Blood vessels were identified with monoclonal antibody for von Willebrand's factor. A significantly larger number of vessels were observed in inflamed tissues of non-smokers than smokers (P<0.05). The number and proportion of vessels expressing both ICAM-1 and E-selectin was greater in sites with inflammation compared to non-inflamed sites in both smokers and non-smokers (P<0.05). The proportion of the total number of vessels expressing ICAM-1 in non-inflamed sites was greater in non-smokers compared with smokers (P<0.05). These results suggest that the inflammatory response in smokers with periodontitis may not be accompanied by an equivalent increase in vascularity. Reduced ICAM-1 expression in non-inflamed areas of smokers could reflect a systemic effect of tobacco smoking on ICAM-1 independent of inflammation.


[Postoperative symptoms and life quality assessment of chronic rhinosinusitis patients received endoscopic sinus surgery].
To implore the positive effects of endoscopic sinus surgery on the symptoms and quality of life (QOL) of patients with chronic rhinosinusitis (CRS) and their changing regularity of symptoms and QOL. A prospective, randomized and controlled trial was conducted to survey symptoms and QOL status of 120 CRS patients undergoing endoscopic sinus surgery, in contrast to that of 200 healthy individuals passing health examination, at the baseline and at 3- and 6-months postoperatively by visual analog score (VAS) symptom instrument and QOL instruments including medical outcome study short-form 36-items health survey (SF-36) and sino-nasal outcome test-20 (SNOT-20). In the 3 months follow-up settings, total of individual symptom severity scores and global symptom severity scores of VAS, such domains as role physical and general health of SF-36 and total of the 20 items scores and the most important 5-item scores of SNOT-20 all began to get better markedly (P < 0.05); in the 3-6 months follow-up settings, the indices above stayed fixed (P > 0.05); and in the 6-months follow-up settings, indices of symptoms and QOL status entirely improved from the baseline, with SF-36 showing no scoring difference between CRS patients and healthy individuals, whereas VAS showed that abnormal nasal drainage and decreased sense of smell were still left and SNOT-20 showed that abnormal nasal drainage and lack of good sleep left, inferior to that of healthy objects significantly (P < 0.05). On the whole CRS patients undergoing endoscopic sinus surgery recover symptomatically and come back to normal level of QOL status at the 6 months postoperatively, but such significant problems as abnormal nasal drainage, decreased sense of smell and lack of good sleep still remain and need to be treated.


Thermosensitivity of nuclear RNA polymerase during the in vitro senescence of mouse fibroblasts.
Embryonic mouse fibroblasts divide approximately twelve times in vitro prior to cessation of mitotic activity. During this period of cellular senescence the thermosensitivity of the RNA polymerase activity of isolated nuclei has been examined as a means of detecting the possible accumulation of defective enzyme molecules, as has been found by other workers for several cytoplasmic enzymes during the ageing of human fibroblasts in vitro. The total RNA polymerase activity of nuclei isolated from old (10th generation) cells is more thermoresistant than that of young (2nd generation) cells. However, the net RNA polymerase activity of nuclei from non-dividing (confluent) cells is more thermoresistant than that of exponentially growing cells of the same age. When allowance is made for the state of growth of the cultures, little difference is seens in the thermosensitivity of the activities of nuclei from old and young cells. Neither is there any difference between the thermosensitivity of the net activity of an established line of murine fibroblasts (L-cells) and cells in primary culture. Preheating nuclei increases the inhibition of their total RNA polymerase activity by alpha-amanitin, indicating that RNA polymerase II is the most heat resistance species present. There appears to be no difference between the thermosensitivity of the alpha-amanitin sensitive and resistance species of the enzyme in the nuclei of old and young cells. It is concluded that old cells resemble non-dividing young cells in containing a higher proportion of RNA polymerase II in their nuclei, resulting in greater thermoresistance of the total RNA polymerase activity over that of exponentially growing cells. However, there appears to be no increase in thermosensitivity of the enzymes with age.


The role of summating potential in the diagnosis and management of Menière's disease.
To assess the role of a negative summating potential (-SP) in the clinical diagnosis and management of Menière's disease, the relationship of -SP abnormality to clinical symptoms, hearing level and caloric test results was examined in 70 patients with unilateral Meniere's disease. The click-induced -SP and action potential (AP) were recorded by extratympanic electrocochleography. When SP/AP ratio exceeded 0.43, the -SP was considered to be abnormal. No significant relationship was found between abnormal -SP and clinical symptoms (the duration of cochlear and vestibular symptoms, recent hearing fluctuation and recent vertigo attack). Ears with abnormal -SP had a significantly worse hearing loss at high frequencies (2-8 kHz) than ears with normal -SP, whereas there was no significant difference in hearing loss at low frequencies (0.25-1 kHz) between both ears. No significant relation was found between abnormal -SP and abnormal unilateral weakness in caloric test. The results indicate that the click-induced -SP reflects the pathophysiology of Menière's disease basically in the cochlear partition at high frequencies.


Structure of the human C2 gene.
A human genomic cosmid clone, S22A, was used to characterize the structure of the human C2 gene. Eighteen exons spanning 18 kb of DNA were mapped by nucleotide sequencing of exon-containing subclones and by Southern blotting. Introns vary in length between 83 bp and 4.4 kb and all intron/exon boundaries follow the AG/GT consensus rule for splicing. Exon 1 encodes 270 bp of the 5'-untranslated region of the C2 cDNA C2HL5-3 and 15 amino acids of the leader peptide. The three short consensus repeats of C2b are encoded by the single exons 2, 3, and 4, respectively. The von Willebrand factor type A-like domain of C2a is encoded by exons 6-10 and the serine protease domain by exons 11-18. Exons 5 and 15 appear to be unique to the C2 and factor B genes.


Importance of internal mammary artery perfusion in cardiac ischemia and reperfusion.
Our purpose was to investigate the changes in antioxidant defense systems due to internal mammary artery (IMA) perfusion during coronary artery bypass graft (CABG) operations in which proximal anastomoses were completed under partial bypass with the aid of a side-biting clamp. Twenty-five patients to be studied were divided into two groups according to the criterion of whether during proximal anastomoses left internal mammary artery (LIMA) perfusion was applied (n = 15, LIMA group) or not (n = 10, non-LIMA group). The erythrocyte catalase (CAT), superoxide dismutase (SOD), serum lipid peroxidation (LPO) products and whole blood reduced glutathione (GSH) levels were measured in the blood samples taken from the coronary sinus before cardiopulmonary bypass (CPB) (t1), before declamping (t2), at the 5th min after declamping (t3), at the 5th (t4) and 15th (t5) min after removing the side-biting clamp. While erythrocyte CAT enzyme activity decreases in both groups after the basal measurements, no significant difference was detected between the groups. Although the GSH levels did not differ at t1, t2 and t3, they were found to be higher in the LIMA group at t4 (p = 0.006) and t5 (p = 0.021). The erythrocyte SOD enzyme activity decreased after the basal measurements in both groups, but this reduction was less in the LIMA group at t4 (p = 0.034) and t5 (p = 0.018) compared to the other group. LPO products rose in both groups when reperfusion started after declamping. The levels of LPO products were significantly higher at t4 and t5 in the non-LIMA group than the other group (p = 0.011 and p = 0.008). If proximal anastomoses of coronary grafts are completed under partial bypass, permitting LIMA perfusion during this procedure will be beneficial to antioxidant defense systems.


Self-adaptive genetic algorithms with simulated binary crossover.
Self-adaptation is an essential feature of natural evolution. However, in the context of function optimization, self-adaptation features of evolutionary search algorithms have been explored mainly with evolution strategy (ES) and evolutionary programming (EP). In this paper, we demonstrate the self-adaptive feature of real-parameter genetic algorithms (GAs) using a simulated binary crossover (SBX) operator and without any mutation operator. The connection between the working of self-adaptive ESs and real-parameter GAs with the SBX operator is also discussed. Thereafter, the self-adaptive behavior of real-parameter GAs is demonstrated on a number of test problems commonly used in the ES literature. The remarkable similarity in the working principle of real-parameter GAs and self-adaptive ESs shown in this study suggests the need for emphasizing further studies on self-adaptive GAs.


[Lipoatrophy after insulin injections can be treated with lipofilling].
Lipoatrophy is a known, albeit rare, side effect of insulin injections. We describe a case concerning a 17-year-old woman with insulin-dependent diabetes, who was treated with an insulin pump. In the areas surrounding the injection point subcutaneous lipoatrophy occurred. Such lipoatrophy is of cosmetic importance, and it may incur glycaemic imbalance due to altered and unpredictable absorption of insulin in the atrophied areas. With no spontaneous regeneration of the atrophied areas observed the patient was successfully treated with lipofilling in the atrophied areas.


Control of nitrification/denitrification in an onsite two-chamber intermittently aerated membrane bioreactor with alkalinity and carbon addition: Model and experiment.
Denitrifying membrane bioreactors (MBRs) are being found useful in water reuse treatment systems, including net-zero water (nearly closed-loop), non-reverse osmosis-based, direct potable reuse (DPR) systems. In such systems nitrogen may need to be controlled in the MBR to meet the nitrate drinking water standard in the finished water. To achieve efficient nitrification and denitrification, the addition of alkalinity and external carbon may be required, and control of the carbon feed rate is then important. In this work, an onsite, two-chamber aerobic nitrifying/denitrifying MBR, representing one unit process of a net-zero water, non-reverse osmosis-based DPR system, was modeled as a basis for control of the MBR internal recycling rate, aeration rate, and external carbon feed rate. Specifically, a modification of the activated sludge model ASM2dSMP was modified further to represent the rate of recycling between separate aerobic and anoxic chambers, rates of carbon and alkalinity feed, and variable aeration schedule, and was demonstrated versus field data. The optimal aeration pattern for the modeled reactor configuration and influent matrix was found to be 30 min of aeration in a 2 h cycle (104 m3 air/d per 1 m3/d average influent), to ultimately meet the nitrate drinking water standard. Optimal recycling ratios (inter-chamber flow to average daily flow) were found to be 1.5 and 3 during rest and mixing periods, respectively. The model can be used to optimize aeration pattern and recycling ratio in such MBRs, with slight modifications to reflect reactor configuration, influent matrix, and target nitrogen species concentrations, though some recalibration may be required.


Gastrointestinal behavior of orally administered radiolabeled erythromycin pellets in man as determined by gamma scintigraphy.
The behavior of single 250-mg doses of a multiparticulate form of erythromycin base (ERYC(R)), each including five pellets radiolabeled with neutron-activated samarium-153, was observed by gamma scintigraphy in seven male subjects under fasting and nonfasting conditions. The residence time and locus of radiolabeled pellets within regions of the gastrointestinal tract were determined and were correlated with plasma concentrations of erythromycin at coincident time points. Administration of food 30 minutes postdosing reduced fasting plasma erythromycin Cmax and area under the plasma erythromycin versus time curve (AUC) values by 43% and 54%, respectively. Mean peak plasma concentration of erythromycin (Cmax) in the fasting state was 1.64 micrograms/mL versus 0.94 micrograms/mL in the nonfasting state. Total oral bioavailability, as determined by mean AUC (0-infinity) of the plasma erythromycin concentration versus time curve, was 7.6 hr/micrograms/mL in the fasted state, versus 3.5 hr/micrograms/mL in the nonfasting state. Mean time to peak plasma erythromycin concentration (tmax) in the fasting state was 3.3 hours, versus 2.3 hours in the nonfasting state. Plasma concentrations of erythromycin in both fasting and nonfasting states were within acceptable therapeutic ranges. Evidence provided by this study: 1) indicates that pellet erosion and absorption of active erythromycin base begins when the enteric-coated pellets reach the highly vascular mucosa of the jejunum and proximal ileum, and is essentially completed within the ileum, with a significant portion absorbed in the medial-to-distal ileum; 2) confirms that acceptable therapeutic plasma levels of erythromycin are attained in nonfasting subjects (Cmax = 0.94 microgram/mL) and that superior plasma erythromycin concentrations (Cmax = 1.64 micrograms/mL) are achieved by administration of the dose on an empty stomach 1 to 2 hours before or after meals; 3) corroborates other comparative studies reporting greater fasting bioavailability with this multiparticulate dosage form of erythromycin base than with reference single tablet or particle-in-tablet formulations; and 4) indicates that neutron activation of stable isotopes incorporated as a normal excipient in industrially-produced formulations provides an effective means for in vivo evaluation of dosage forms through gamma scintigraphy.


Chiropractic distractive decompression in treating pelvic pain and multiple system pelvic organic dysfunction.
No data are available on the incidence of pelvic pain and organic dysfunction (PPOD) in patients suffering from low back pain. PPOD is not an uncommon finding in the low back pain patient. Women appear to be more frequently involved than men. The results of recent electrophysiologic investigations indicate that many patients with urological, bowel or anorectal dysfunction demonstrate evidence of denervation neuropathy in muscles innervated by the branches of the pudendal nerve. Six patients with low back pain meeting predetermined criteria, indicating the presence of PPOD as a result of suspected lower sacral nerve root compression (LSNRC) secondary to a mechanical disorder of the low back were treated with chiropractic distractive decompressive manipulation of the lumbar spine. Symptoms of PPOD, and indicators of LSNRC were assessed prior to the onset, and following the termination of treatment. It appears that selected indicators of LSNRC represent the most sensitive clinical signs of identifying the presence of PPOD which may respond to manipulative treatment, and may also provide the most sensitive measure of overall PPOD response. These findings, although preliminary suggest a possible etiology of PPOD in the low back pain patient with evidence of LSNRC. Further work in this area is encouraged.


Theoretical analysis of single-cycle self-compression of near infrared pulses using high-spatial modes in capillary fibers.
Soliton self-compression is demonstrated during the propagation of high spatial modes in hollow core fibers in the near-infrared spectral region, taking advantage of their negative dispersion response. We have found that there is always an optimum spatial mode to observe this phenomenon, compressing the pulses down to the single-cycle regime without needing any external compression device and with a consequent increase in the output peak power. Our result is relevant for any ultrashort laser application in which few- or single-cycle pulses are crucial.


Highly Selective Photoreduction of CO2 with Suppressing H2 Evolution over Monolayer Layered Double Hydroxide under Irradiation above 600 nm.
Although progress has been made to improve photocatalytic CO2 reduction under visible light (λ>400 nm), the development of photocatalysts that can work under a longer wavelength (λ>600 nm) remains a challenge. Now, a heterogeneous photocatalyst system consisting of a ruthenium complex and a monolayer nickel-alumina layered double hydroxide (NiAl-LDH), which act as light-harvesting and catalytic units for selective photoreduction of CO2 and H2 O into CH4 and CO under irradiation with λ>400 nm. By precisely tuning the irradiation wavelength, the selectivity of CH4 can be improved to 70.3 %, and the H2 evolution reaction can be completely suppressed under irradiation with λ>600 nm. The photogenerated electrons matching the energy levels of photosensitizer and m-NiAl-LDH only localized at the defect state, providing a driving force of 0.313 eV to overcome the Gibbs free energy barrier of CO2 reduction to CH4 (0.127 eV), rather than that for H2 evolution (0.425 eV).


The thiocarboxanilide nonnucleoside UC781 is a tight-binding inhibitor of HIV-1 reverse transcriptase.
The thiocarboxanilide nonnucleoside inhibitor (NNI) UC781 inhibited HIV-1 reverse transcriptase (RT) DNA polymerase activity at a 1:1 molar ratio of inhibitor to enzyme. Inhibition was linear uncompetitive with respect to template/primer (T/P) and mixed noncompetitive with respect to deoxynucleoside triphosphate (dNTP), typical of NNI. When the RT-T/P binary complex was incubated with UC781 and then separated from unbound inhibitor, recovery of enzyme activity was slow, with only about 60% activity recovered after 25 min. The inactivation of the RT-T/P complex was prevented by the presence of a large excess of UC84, another carboxanilide NNI that interacts with this RT mechanistic form. UC781 protected the RT-T/P-dNTP ternary complex from irreversible inactivation by a photoactivatable azido analog of nevirapine, implying that UC781 binds to the NNI pocket of this RT mechanistic form. UC781 did not photoprotect either the free enzyme or the RT-T/P binary complex; however, protein fluorescence quenching studies indicated that UC781 interacted with all RT mechanistic forms, with the order of affinity being RT-T/P-dNTP ternary complex > RT-T/P binary complex > free RT. Reaction progress curve analysis showed that the binding of UC781 to RT is rapid (k(on) approximately 1.7 x 10(6) M(-1) s(-1)), but that dissociation is slow (k(off) approximately 1.6 x 10(-3) s(-1)). UC781 is therefore a rapid tight-binding inhibitor of HIV-1 RT, the first NNI to demonstrate this property.


The effects of maternal indomethacin therapy on human fetal branch pulmonary arterial vascular impedance.
To examine whether maternal indomethacin therapy affects human fetal pulmonary arterial vascular impedance without constriction of the ductus arteriosus and to determine the changes in the pulmonary arterial vascular impedance in the presence of ductal constriction. In this cross-sectional study, 52 normal fetuses without maternal medication (control group), 33 fetuses without ductal constriction (Study group I) and 11 fetuses with ductal constriction (Study group II) during maternal indomethacin therapy between 24 and 34 weeks of gestation were examined by Doppler echo-cardiography. Blood velocity waveforms across the proximal right or left pulmonary artery were obtained and the pulsatility index (PI) of the proximal pulmonary arteries was calculated. In the control group, the proximal pulmonary artery PI was higher (p < 0.0001) at 24-25 weeks (n = 7) (3.73 +/- 0.33; mean +/- SD) than at 33-34 week of gestation (n = 11) (2.98 +/- 0.27). The PI was constantly greater (p < 0.005) in Study group I than in the control group. However, in this group the mean average weekly decrease in the PI of the proximal pulmonary arteries was similar to that in the control group. After 26 weeks of gestation, the PI values in Study group II were significantly higher than in the control group (27 weeks: 4.12 vs. 3.34 (p < 0.005); 30 weeks: 4.48 vs. 3.14 (p < 0.0001); 34 weeks: 4.96 vs. 3.00 (p < 0.0001), respectively). Human fetal pulmonary arterial vascular impedance is increased by maternal indomethacin therapy even without ductal constriction. In the presence of ductal constriction, the magnitude of the increase in the vascular impedance is related to the gestational age.


Length of surface PEG modulates nanocarrier transcytosis across brain vascular endothelial cells.
Diblock PLA-PEG nanoparticles were produced to establish the role of PEG chain length on brain vascular endothelial cell transcytosis. 100-nm nanoparticles tagged with fluorescent pyrene butanol and coated with PEG chains (Mw: 1-10 kDa), at similar PEG surface density, were used to study endocytosis and transcytosis phenomena on mouse vascular endothelial cell monolayers. The transport mechanisms were then investigated through inhibitory processes. Our results show that there is an evident correlation between PEG chain length and nanoparticle translocation. The highest transcytosis rates were obtained with PEG5000 and PEG10000 and macropinocytosis appeared to play a central role in cell uptake. This study constitutes the first systematic exploration of the role of PEG chain length on nanoparticle endocytosis and transcytosis in an in vitro model of the blood-brain barrier.


Quantum algorithms for Josephson networks.
We analyze possible implementations of quantum algorithms in a system of (macroscopic) Josephson charge qubits. System layout and parameters to realize the Deutsch algorithm with up to three qubits are provided. Special attention is paid to the necessity of entangled states in the various implementations. Further, we demonstrate explicitly that the gates to implement the Bernstein-Vazirani algorithm can be realized by using a system of uncoupled qubits.


Effect of culture conditions on the resistance of Pseudomonas aeruginosa biofilms to disinfecting agents.
The relationship between the environmental conditions of biofilm formation and resistance to disinfectants was studied. Anti-biofilm assays were performed against biofilms grown at 20, 30 and 37°C on stainless steel and polycarbonate, over 24 and 48 h. A rise in growth temperature increased the resistance of 24 h biofilms to disinfectants containing didecyldimethylammonium chloride and decreased it to a disinfectant containing alkyldimethylbenzylammonium chloride. The increase in growth temperature coupled with an incubation time of 24 h promoted increases in both matrix production and the membrane rigidity of sessile cells. An increase in incubation time also increased both matrix production and the membrane rigidity of sessile cells. Such phenomena resulted in an increased resistance to disinfectants of biofilms grown at 20 and 30°C. The resistance of 48 h biofilms to disinfectants decreased with an increase in growth temperature despite the increase in matrix production and the membrane rigidity of sessile cells.


Measurement of bubble and pellet size distributions: past and current image analysis technology.
Measurements of bubble and pellet size distributions are useful for biochemical process optimizations. The accuracy, representation, and simplicity of these measurements improve when the measurement is performed on-line and in situ rather than off-line using a sample. Historical and currently available measurement systems for photographic methods are summarized for bubble and pellet (morphology) measurement applications. Applications to cells, mycelia, and pellets measurements have driven key technological developments that have been applied for bubble measurements. Measurement trade-offs exist to maximize accuracy, extend range, and attain reasonable cycle times. Mathematical characterization of distributions using standard statistical techniques is straightforward, facilitating data presentation and analysis. For the specific application of bubble size distributions, selected bioreactor operating parameters and physicochemical conditions alter distributions. Empirical relationships have been established in some cases where sufficient data have been collected. In addition, parameters and conditions with substantial effects on bubble size distributions were identified and their relative effects quantified. This information was used to guide required accuracy and precision targets for bubble size distribution measurements from newly developed novel on-line and in situ bubble measurement devices.


Synthesis and NMR assignment of the two diastereomers of 8,6'-cyclo-2',6'-dideoxyadenosine.
We herein present the first synthesis and characterization of the two C5' diastereomers of 8,6'-cyclo-2',6'-dideoxyadenosine. Starting from commercially available 2'-deoxyadenosine, the target cyclonucleosides were synthesized in 11 linear steps. Following a zinc-mediated cyclization reaction to form the seven-membered ring, the stereochemistry of the newly formed chiral center was established using two-dimensional NOESY NMR experiments.


Genotoxicity of photoilluminated riboflavin in the presence of Cu(II).
Riboflavin is known to generate superoxide anion (O2.-) and other reactive oxygen species in the presence of Cu(II) and light as well as cause fragmentation of DNA and protein in vitro. In the present study we examined the genotoxic effects of photoilluminated riboflavin in the presence of Cu(II). Using the phage inactivation assay, a significant decline in plaque-forming unit (PFU) is seen. Results of Ames testing have suggested that probably a frameshift mutation is caused by a riboflavin-Cu(II)-mediated reaction. Using neocuproine as a Cu(I) sequestering reagent, Cu(I) has been shown to be an essential intermediate generated in the reaction between Cu(II), photoilluminated riboflavin, and DNA. Results obtained with various scavengers of active oxygen species strongly suggest that the species predominantly responsible for DNA damage is oxygen (O2) in the singlet or triplet state, together with H2O2, hydroxyl radical, and hydroxyl ion, to a lesser extent. In the case of riboflavin, a ternary complex of DNA-drug-Cu(II) is presumably formed. A redox reaction, involving riboflavin and Cu(II) in the complex, may then occur with the formation of a DNA-oxidized riboflavin-Cu(I) complex. This probably acts as a catalyst for the oxidation of Cu (I) to Cu(II), during which molecular oxygen is reduced to generate a variety of active oxygen species. The probable mechanism for the generation of these reactive oxygen species has also been proposed.


Intestinal colonization with Enterobacteriaceae in Pakistani and Swedish hospital-delivered infants.
Rectal cultures from Swedish and Pakistani hospital-delivered newborn infants were analysed regarding the early acquisition of enterobacteria. Swedish infants were delivered vaginally, Pakistani infants were delivered either vaginally or by caesarean section. The Swedish infants were all breast-fed, whereas breastfeeding was incomplete and often started late among the Pakistani infants. Both groups of Pakistani infants were more rapidly colonized with enterobacteria than were the Swedish infants. Cultures from Swedish infants seldom yielded more than one kind of enterobacteria; E. coli and Klebsiella were most frequently isolated. E. coli dominated in both Pakistani groups, but especially caesarean section delivered infants were in addition often colonized with Proteus, Klebsiella, Enterobacter or Citrobacter species. Breastfeeding from the first day of life reduced colonization with Klebsiella/Enterobacter/Citrobacter. The results suggest that environmental exposure, delivery mode and early feeding habits all influence the early intestinal colonization with enterobacteria.


Analysing georeferenced population genetics data with Geneland: a new algorithm to deal with null alleles and a friendly graphical user interface.
We introduce a new algorithm to account for the presence of null alleles in inferences of populations clusters from individual multilocus genetic data. We show by simulations that the presence of null alleles can affect the accuracy of inferences if not properly accounted for and that our algorithm improve signficantly their accuracy. This new algorithm is implemented in the program Geneland. It is freely available under GNU public license as an R package on the Comprehensive R Archive Network. It now includes a fully clickable graphical interface. Informations on how to get the software are available on folk.uio.no/gillesg/Geneland.html


Adjuvant treatment of breast cancer: impact of monoclonal antibody therapy directed against the HER2 receptor.
The use of chemotherapy and endocrine therapies as adjuncts to the treatment of early-stage breast cancer has yielded small but significant improvements in disease-free and overall survival. Increased understanding of the role of growth factor receptors enabled the rational development of agents that are capable of modulating their function. A humanised monoclonal antibody to the HER2 receptor, trastuzumab, has demonstrable single-agent activity in metastatic breast cancer and enhances the antitumour effects of chemotherapy. As a consequence, trastuzumab has been tested in the adjuvant setting the results of which have been presented recently. This review briefly summarises the use of trastuzumab in advanced breast cancer and describes recent studies of its use in the adjuvant setting.


Phosphorescence enhancement triggered by pi stacking in solid-state [Cu(N-N)(P-P)]BF4 complexes.
In this article, we report a phosphorescence enhancement phenomenon of [Cu(N-N)(P-P)]BF4 complexes in the solid state. The phosphorescence enhancement phenomenon is composed of three features: (1) an obvious emission blue shift, (2) a greatly enhanced photoluminescence quantum yield, and (3) a longer excited-state lifetime. Systematic analyses prove that the main cause of this phenomenon is nonradiative process suppression triggered by the pi stacking between [Cu(N-N)(P-P)]BF4 molecules. The phosphorescence enhancement widely exists in [Cu(N-N)(P-P)]BF4 complexes because of the pi surfaces in their diimine ligands.


Least-squares analysis of circular dichroic spectra of proteins.
It is shown that the method proposed by Baker and Isenberg [Biochemistry, 15, 629 (1976)] for estimating secondary structure composition of proteins from circular dichroic spectra is a least-squares fitting technique. Estimates obtained by this method for myoglobin, lysozyme, lactate dehydrogenase, papain, and ribonuclease are not substantively different from those obtained using unconstrained linear least squares.


Diffusion and perfusion MRI findings of the signal-intensity abnormalities of brain associated with developmental venous anomaly.
Developmental venous anomalies are the most common intracranial vascular malformation. Increased signal-intensity on T2-FLAIR images in the areas drained by developmental venous anomalies are encountered occasionally on brain imaging studies. We evaluated diffusion and perfusion MR imaging findings of the abnormally high signal intensity associated with developmental venous anomalies to describe their pathophysiologic nature. We retrospectively reviewed imaging findings of 34 subjects with signal-intensity abnormalities associated with developmental venous anomalies. All subjects underwent brain MR imaging with contrast and diffusion and perfusion MR imaging. Regions of interest were placed covering abnormally high signal intensity around developmental venous anomalies on fluid-attenuated inversion recovery imaging, and the same ROIs were drawn on the corresponding sections of the diffusion and perfusion MR imaging. We measured the apparent diffusion coefficient, relative cerebral blood volume, relative mean transit time, and time-to-peak of the signal-intensity abnormalities around developmental venous anomalies and compared them with the contralateral normal white matter. The Mann-Whitney U test was used for statistical analysis. The means of ADC, relative cerebral blood volume, relative mean transit time, and TTP of signal-intensity abnormalities around developmental venous anomalies were calculated as follows: 0.98 ± 0.13 10(-3)mm(2)/s, 195.67 ± 102.18 mL/100 g, 16.74 ± 7.38 seconds, and 11.65 ± 7.49 seconds, respectively. The values of normal WM were as follows: 0.74 ± 0.08 10(-3)mm(2)/s for ADC, 48.53 ± 22.85 mL/100 g for relative cerebral blood volume, 12.12 ± 4.27 seconds for relative mean transit time, and 8.35 ± 3.89 seconds for TTP. All values of ADC, relative cerebral blood volume, relative mean transit time, and TTP in the signal-intensity abnormalities around developmental venous anomalies were statistically higher than those of normal WM (All P < .001, respectively). The diffusion and perfusion MR imaging findings of the signal-intensity abnormalities associated with developmental venous anomaly suggest that the nature of the lesion is vasogenic edema with congestion and delayed perfusion.


Dietary patterns and vitamin B(12) status of migrant Indian preadolescent girls.
Inter-generational dietary patterns are implicated in the development of adult disease and have a critical role in cell division, growth and development. The assessment of B(12) status in six meat-eating (ME) and six non-meat-eating (NME) 9-11 years old, Tanner stage 1, migrant Indian girls. B(12) status was assessed by haematology, fasting B(12), folate, methyl malonic acid (MMA) and 7-day-diet-diary analysis. Red cell and iron measurements were within reference ranges for all girls. Reported B(12) intake correlated with serum B(12) (r=0.74, P=0.006). Serum B(12) was lower in NME girls compared with ME girls (232+/-95 vs 543+/-201 pmol l(-1), P=0.01). Two NME girls were deficient in vitamin B(12) (B(12)<170 pmol l(-1), MMA>0.26 micromol l(-1)). Overall serum B(12) was low (388+/-221 pmol l(-1)) and serum folate intake high (27+/-8 pmol l(-1); 342+/-269 microg day(-1)). Asymptomatic B(12) deficiency is present in NME migrant Indian preadolescents in the presence of high folate. Supplementation with B(12) in this group may be necessary to improve future health outcomes.


Brain pharmacokinetics of ganciclovir in rats with orthotopic BT4C glioma.
Ganciclovir (GCV) is an essential part of the Herpes simplex virus thymidine kinase (HSV-tk) gene therapy of malignant gliomas. The purpose of this study was to investigate the brain pharmacokinetics and tumor uptake of GCV in the BT4C rat glioma model. GCV's brain and tumor uptakes were investigated by in vivo microdialysis in rats with orthotopic BT4C glioma. In addition, the ability of GCV to cross the blood-brain barrier and tumor vasculature was assessed with in situ rat brain perfusion. Finally, the extent to which GCV could permeate across the BT4C glioma cell membrane was assessed in vitro. The areas under the concentration curve of unbound GCV in blood, brain extracellular fluid (ECF), and tumor ECF were 6157, 1658, and 4834 μM⋅min, respectively. The apparent maximum unbound concentrations achieved within 60 minutes were 46.9, 11.8, and 25.8 μM in blood, brain, and tumor, respectively. The unbound GCV concentrations in brain and tumor after in situ rat brain perfusion were 0.41 and 1.39 nmol/g, respectively. The highly polar GCV likely crosses the fenestrated tumor vasculature by paracellular diffusion. Thus, GCV is able to reach the extracellular space around the tumor at higher concentrations than that in healthy brain. However, GCV uptake into BT4C cells at 100 μM was only 2.1 pmol/mg of protein, and no active transporter-mediated disposition of GCV could be detected in vitro. In conclusion, the limited efficacy of HSV-tk/GCV gene therapy may be due to the poor cellular uptake and rapid elimination of GCV.


Identification of a malignant counterpart of the monocyte-dendritic cell progenitor in an acute myeloid leukemia.
Myeloblasts derived from the peripheral blood of a patient with acute myelogenous leukemia (ORL47) were found to represent the malignant counterpart of the newly elucidated monocyte-dendritic cell colony-forming unit (mono-DC-CFU). The specific cytokine conditions require to achieve intermediate and terminal maturation of DCs and monocytes from these progenitors were defined. With tumor necrosis factor (TNF) + granulocyte-macrophage colony-stimulating factor (GM-CSF) + stem cell factor treatment numerous colony-like clusters developed. In contrast with normal DC development, further advancement of mono-DC-CFU and terminal DC maturation from the leukemic cells were dependent on the addition of interleukin-6. Functional and phenotypic analysis showed that the capacity to differentiate was maintained fully in the DC compartment, but only partially in the monocyte compartment, as judged by the lack of CD14 surface expression. Cells found at intermediate stages of DC development were potent stimulators of a mixed leukocyte reaction, a function usually attributed to mature DCs. As previously shown for normal DC development, antibodies to TNF alpha and GM-CSF blocked proliferative responses and DC growth. The importance of these observations in the classification of leukemias, normal DC development, and potential clinical strategies is discussed.


Harlequin ichthyosis in association with hypothyroidism and juvenile rheumatoid arthritis.
Harlequin ichthyosis is a rare and severe congenital erythrodermic ichthyosis characterized at birth by hyperkeratotic plates covering the entire body, ectropion, eclabium, poorly developed ears, and contractures of the hands and feet. Two Chinese children, a 2-year-old boy and an 11-year-old girl, presented with these classic features as well as alopecia and loss of eyebrows and eyelashes. The boy was small for his age and was found to have hypothyroidism at the age of 18 months; he is currently on thyroxine replacement therapy. At 6 years of age, the girl developed symmetrical polyarthritis associated with positive rheumatoid factor and radiologic evidence of erosive arthritis, suggestive of juvenile rheumatoid arthritis. She received prednisolone, nonsteroidal anti-inflammatory drugs (NSAIDs), and subsequently methotrexate for her arthritis, with clinical and radiologic improvement. Early therapy with oral retinoids in both children accelerated shedding of the hyperkeratotic plates as well as improved ectropion and eclabium. There was no major adverse reaction to oral retinoids. The development of juvenile rheumatoid arthritis in survivors with harlequin ichthyosis has not been previously described. The use of prednisolone and NSAIDs in the girl did not affect the skin condition, but the addition of methotrexate led to a decrease in erythema. The association with autoimmune disease is probably coincidental. The psychosocial impact of this severe lifelong disease on the two families was enormous. Early retinoid therapy may improve the disorder and help increase survival rates. A multidisciplinary approach, including psychosocial support of the affected families, is vital in the management of this lifelong disease.


A new patient focused index for measuring quality of life in persons with severe and persistent mental illness.
The quality of life in persons with severe and persistent mental illness is often poor. Most treatment programmes have the goal of increasing quality of life. Unfortunately, existing methods to assess quality of life are cumbersome and oriented towards research rather than clinical settings. This study describes preliminary steps in the development, testing and application of a new patient focused index for measuring quality of life in persons with severe mental illness. The Quality of Life Index for Mental Health (QLI-MH) differs from existing instruments in that it is based on an easy to use, self-administered questionnaire that assesses nine separate domains that together encompass quality of life. Each domain can be individually weighted depending on its relative importance to the patient. Different parts of the instrument solicit information from the patient, the primary clinician, and, when available, the family. The instrument and its scoring system address limitations of previous approaches to quality of life measurement.