The chaperone-like sodium phenylbutyrate improves factor IX intracellular trafficking and activity impaired by the frequent p.R294Q mutation.
Essentials Missense mutations often impair protein folding, and thus intracellular trafficking and secretion. Cellular models of severe type I hemophilia B were challenged with chaperone-like compounds. Sodium phenylbutyrate improved intracellular trafficking and secretion of the frequent p.R294Q. The increased coagulant activity levels (∼3%) of p.R294Q would ameliorate the bleeding phenotype. Background Missense mutations often impair protein folding and intracellular processing, which can be improved by small compounds with chaperone-like activity. However, little has been done in coagulopathies, where even modest increases of functional levels could have therapeutic implications. Objectives To rescue the expression of factor IX (FIX) variants affected by missense mutations associated with type I hemophilia B (HB) through chaperone-like compounds. Methods Expression studies of recombinant (r)FIX variants and evaluation of secreted levels (ELISA), intracellular trafficking (immunofluorescence) and activity (coagulant assays) before and after treatment of cells with chaperone-like compounds. Results As a model we chose the most frequent HB mutation (p.R294Q, ~100 patients), compared with other recurrent mutations associated with severe/moderate type I HB. Immunofluorescence studies revealed retention of rFIX variants in the endoplasmic reticulum and negligible localization in the Golgi, thus indicating impaired intracellular trafficking. Consistently, and in agreement with coagulation phenotypes in patients, all missense mutations resulted in impaired secretion (< 1% wild-type rFIX). Sodium phenylbutyrate (NaPBA) quantitatively improved trafficking to the Golgi and dose dependently promoted secretion (from 0.3 ± 0.1% to 1.5 ± 0.3%) only of the rFIX-294Q variant. Noticeably, this variant displayed a specific coagulant activity that was higher (~2.0 fold) than that of wild-type rFIX in all treatment conditions. Importantly, coagulant activity was concurrently increased to levels (3.0 ± 0.9%) that, if achieved in patients, would ameliorate the bleeding phenotype. Conclusions Altogether, our data detail molecular mechanisms underlying type I HB and candidate NaPBA as affordable 'personalized' therapeutics for patients affected by the highly frequent p.R294Q mutation, and with reduced access to substitutive therapy.


The iliac prefabricated composite graft for dentoalveolar reconstruction: a clinical procedure.
Prefabricated composite grafts were introduced in reconstructive oral and maxillofacial surgery in the mid-1990s. The purpose of this report is to introduce a novel technique-the iliac prefabricated composite graft-for reconstruction of the alveolar ridge simultaneously with masticatory attached gingiva and integrated implants in a single procedure. Clinical and immunohistologic results are reported and reveal this technique to be an attractive alternative in dentoalveolar surgery and implant dentistry.


High plasma norepinephrine levels in patients with major affective disorder.
The authors found that patients with major affective disorder had higher levels of plasma norepinephrine and higher pulse rates (tachycardia) than healthy control subjects, but their blood pressures were normal. These measurements were similar in all three subgroups of patients with affective disorder--manic, bipolar depressed, and unipolar. Because norepinephrine is the primary neurotransmitter of the sympathetic nervous system, these data suggest sympathetic hyperactivity in the major affective disorders. This conclusion is compatible with recent speculation based on the effect of antidepressants on noradrenergic receptors and a failure of alpha-receptors to downregulate normally in patients with major affective disorder.


Further treatment of decolorization liquid of azo dye coupled with increased power production using microbial fuel cell equipped with an aerobic biocathode.
A microbial fuel cell (MFC) incorporating a recently developed aerobic biocathode is designed and demonstrated. The aerobic biocathode MFC is able to further treat the liquid containing decolorization products of active brilliant red X-3B (ABRX3), a respective azo dye, and also provides increased power production. Batch test results showed that 24.8% of COD was removed from the decolorization liquid of ABRX3 (DL) by the biocathode within 12 h. Metabolism-dependent biodegradation of aniline-like compound might be mainly responsible for the decrease of overall COD. Glucose is not necessary in this process and contributes little to the COD removal of the DL. The similar COD removal rate observed under closed circuit condition (500 Ω) and opened circuit condition indicated that the current had an insignificant effect on the degradation of the DL. Addition of the DL to the biocathode resulted in an almost 150% increase in open cycle potential (OCP) of the cathode accompanied by a 73% increase in stable voltage output from 0.33 V to 0.57 V and a 300% increase in maximum power density from 50.74 mW/m(2) to 213.93 mW/m(2). Cyclic voltammetry indicated that the decolorization products of the ABRX3 contained in the DL play a role as redox mediator for facilitating electron transfer from the cathode to the oxygen. This study demonstrated for the first time that MFC equipped with an aerobic biocathode can be successfully applied to further treatment of effluent from an anaerobic system used to decolorize azo dye, providing both cost savings and high power output.


Association of baseline neuropsychological function and progression of illness over 4 years in HIV-seropositive individuals.
The authors conducted a study to examine the association between neuropsychological markers of central nervous system impairment and systemic human immunodeficiency virus (HIV) disease progression in a sample of 64 HIV-positive asymptomatic patients who were followed for a median of 45.6 months. Patients with poorer baseline scores on the Halstead-Reitan Trail-Making A neuropsychological test developed HIV-related systemic symptoms earlier over the study period than patients with the higher scores on the same test (P < 0.05). Subclinical neuropsychological dysfunction in otherwise asymptomatic HIV-infected individuals may be a harbinger of progressive HIV-related immunologic dysfunction.


Immunoradiometric assay for human complement component C9 utilising monoclonal antibodies.
A two-site immunoradiometric assay for human C9 has been developed. The assay utilised two non-competing monoclonal antibodies to C9 in a single incubation assay protocol. The detection limit of the assay was 0.1 ng (1.4 X 10(-15) moles) in a sample volume of 100 microliters. Using this assay the C9 concentration in normal human plasma was 60.2 +/- 14.9 mg/l (mean +/- 1 standard deviation, 8.5 X 10(-10) mol/l. Significantly elevated levels were found in the plasma of patients with rheumatoid arthritis (90.4 +/- 19.9 mg/l, mean +/- 1 SD). Measurements of C9 in cerebrospinal fluid and synovial fluid were also performed. The low levels of C9 in cerebrospinal fluid (less than 1 mg/l), undetectable by previously available assay methods, were easily measurable with this highly sensitive assay.


Linewidth enhancement factor in semiconductor lasers subject to various external optical feedback conditions.
The linewidth enhancement factor α of a semiconductor laser under the influences of optical feedback with different feedback strengths, external cavity lengths, and feedback phases are studied both experimentally and theoretically. The value of α is determined from the minimum of the Hopf bifurcation curve when the laser is subject to both optical feedback and optical injection. In the experiment, a pellicle beamsplitter mounted on a PZT stage placed on a linear translation stage is used as the reflector, where the external cavity length can be adjusted continuously from the long cavity regime to the short cavity regime with phase accuracy. With a moderate feedback strength, α is found to increase as the feedback strength increases. Moreover, while α is insensitive to the feedback phase in the long cavity regime, it can be tuned continuously in the short cavity regime when varying the phase. A normalized variation range of 21.59% is obtained experimentally at an external cavity length of 1.5 cm, which can be further enhanced by shortening the external cavity. To the best of our knowledge, this is the first detailed study of α from the long to the short cavity regime in a semiconductor laser subject to optical feedback. More particularly, the continuous tuning of α under phase variation is demonstrated the first time.


Nonfebrile mitral valve endocarditis due to Neisseria subflava.
Native valve endocarditis normally presents with fever and only later in its course demonstrates dysfunction of the affected valve. We describe a case of endocarditis due to Neisseria subflava, a Gram-negative diplococcal saprophyte of the oral cavity, which was unsuspected clinically and found unexpectedly during a mitral valve operation performed for symptomatic prolapse with regurgitation.


Comparative proteomic analysis of the short-term responses of rice roots and leaves to cadmium.
Cadmium (Cd) is a non-essential heavy metal that is recognized as a major environmental pollutant. While Cd responses and toxicities in some plant species have been well established, there are few reports about the effects of short-term exposure to Cd on rice, a model monocotyledonous plant, at the proteome level. To investigate the effect of Cd in rice, we monitored the influence of Cd exposure on root and leaf proteomes. After Cd treatment, root and leaf tissues were separately collected and leaf proteins were fractionated with polyethylene glycol. Differentially regulated proteins were selected after image analysis and identified using MALDI-TOF MS. A total of 36 proteins were up- or down-regulated following Cd treatment. As expected, total glutathione levels were significantly decreased in Cd-treated roots, and approximately half of the up-regulated proteins in roots were involved in responses to oxidative stress. These results suggested that prompt antioxidative responses might be necessary for the reduction of Cd-induced oxidative stress in roots but not in leaves. In addition, RNA gel blot analysis showed that the proteins identified in the proteomic analysis were also differentially regulated at the transcriptional level. Collectively, our study provides insights into the integrated molecular mechanisms of early responses to Cd in rice.


Possible involvement of dopamine in the long-term serotonin depletion by p-chloroamphetamine and beta,beta-difluoro-p-chloroamphetamine in rats.
The role of dopamine in the long-term depletion of serotonin in rat brain by p-chloroamphetamine (PCA) and related compounds was investigated by comparing effects of beta,beta-difluoro-p-chloroamphetamine (beta,beta-difluoro-PCA) and 4-methyl-alpha-ethyl-meta-tyramine (H75/12), reported to cause only short-term serotonin depletion, with those of PCA. A single dose of beta,beta-difluoro-PCA had no long-term effects on serotonin in whole rat brain, even after pretreatment with proadifen which decreased the rate at which beta,beta-difluoro-PCA disappeared from brain. The possibility that proadifen might antagonize serotonin depletion was ruled out; proadifen did not prevent long-term serotonin depletion by PCA. Long-term depletion of brain serotonin was found after repeated injections of beta,beta-difluoro-PCA (five injections 4 hr apart) and was prevented by fluoxetine pretreatment. beta,beta-Difluoro-PCA given after the monoamine oxidase inhibitor pargyline or after carbidopa/L-dopa also caused long-term serotonin depletion, although H75/12 did not. At early times after single doses producing the same initial depletion of serotonin, PCA caused a large increase in dopamine and a large decrease in the metabolite 3,4-dihydroxyphenylacetic acid in whole brain, thereby increasing the ratio dopamine/3,4-dihydroxyphenylacetic, and the other two drugs caused smaller effects. Extracellular dopamine was increased markedly by PCA, less by beta,beta-difluoro-PCA, and not at all by H75/12. These results suggest an association between dopamine release and long-term depletion of serotonin and add to evidence that dopamine release by PCA may be essential to its neurotoxic actions on brain serotonin neurons.


Evidence for specific TRPM8 expression in human prostate secretory epithelial cells: functional androgen receptor requirement.
TRPM8 (melastatine-related transient receptor potential member 8), a member of the transient receptor potential (TRP) superfamily of cation channels, has been shown to be a calcium-channel protein. TRPM8 mRNA has also been shown to be overexpressed in prostate cancer and is considered to play an important role in prostate physiology. This study was designed to determine the androgen-regulation mechanisms for TRPM8 mRNA expression and to identify the phenotype of TRPM8-expressing cells in the human prostate. Our findings show that trpm8 gene expression requires a functional androgen receptor. Furthermore, this article argues strongly in favour of the fact that the trpm8 gene is a primary androgen-responsive gene. Single-cell reverse transcriptase PCR and immunohistochemical experiments also showed that the trpm8 gene was mainly expressed in the apical secretory epithelial cells of the human prostate and trpm8 down-regulation occurred during the loss of the apical differentiated phenotype of the primary cultured human prostate epithelial cells. The androgen-regulated trpm8 expression mechanisms are important in understanding the progression of prostate cancer to androgen-independence. These findings may contribute to design a strategy to predict prostate cancer status from the TRPM8 mRNA level. Furthermore, as the TRPM8 channel is localized in human prostate cells, it will be interesting to understand its physiological function in the normal prostate and its potential role in prostate cancer development.


[Cytomegalovirus infection in transplant recipients].
Cytomegalovirus (CMV) is the most important infectious agent in transplant recipients. The critical step in the pathogenesis of CMV infection is the reactivation of latent virus, which is affected by the immunosuppressive therapy and/or alloantigenic stimulation. The clinical effects of CMV infection include CMV disease (syndrome), an immunosuppressed state, and allograft injury. Recently, the incidence of serious CMV diseases after transplant has been decreased, probably due to the advance in the method for rapid diagnosis, the ganciclovir administration, and the effective prevention of CMV diseases. Seronegative or filtered blood products, CMV immune globulin, and prophylactic or preemptive therapy with ganciclovir appear contribute to the improvement in the prophylaxis for CMV diseases after transplant. Antigenemia-guided early treatment may be promising for the effective prevention of CMV diseases after transplant.


Assessing zinc from a nail clipping using mono-energetic portable X-ray fluorescence.
A mono-energetic X-ray beam from a portable X-ray fluorescence device was used to excite characteristic X-rays from zinc in a series of nail clipping phantoms. Twenty nail clipping phantoms having equal zinc concentrations of ~40 µg/g, but with different physical characteristics, were measured individually for 300 s using a small diameter (~1 mm) X-ray beam. Energy spectra obtained from the measurements were analyzed using PyMca software. Zinc signal size varied widely between the different clippings, with a relative standard deviation of 41% observed in the combined signal from zinc Kα and Kβ characteristic X-rays. Three different normalization approaches were introduced to account for variation in the amounts of sample interrogated by the X-ray beam. All three approaches produced similar results, and successfully reduced the relative standard deviation to between 12% and 13%. A clear trend was still observed, however, between the normalized zinc signal and the thickness of clipping measured. To account for this effect, normalized signals were adjusted to calculate "thickness-corrected" values. The relative standard deviation of these thickness-corrected values was 6.2%. Reproducibility of measurement from individual clippings was excellent, with relative standard deviations on the order of 1%, with or without normalization. Overall, this new method of measuring zinc in nail shows promise for the assessment of zinc status in humans using a portable device. The method is sensitive, rapid, and requires only a single nail clipping.


The impact of proton pump inhibitors on bone regeneration and implant osseointegration.
Proton pump inhibitors (PPIs) have become known for the treatment of gastric-acid related disorders. Similar to any other drugs, PPIs have possible adverse reactions, being associated with bone fractures, infections, kidney disease, mineral deficiency, dementia, and pneumonia. Multiple analyses have stated that PPIs therapy may affect bone regeneration and osseointegration process, causing an increased risk of bone fracture, deterioration of bone metabolism and impaired bone healing. In this review, we emphasized the current literature regarding the influence of proton pump inhibitors in the bone regeneration process. Results from the studies suggest a link between PPIs intake and bone regeneration, but several concerns are raised regarding inadequate recipient bone, surgical trauma, limitations on the titanium surface, comorbidities or interference with other pharmacological agents. Further studies are needed to determine whether the impaired bone regeneration process is due to PPI or coexisting factors.


Topical application of tetracycline in regenerative periodontal surgery in beagles.
This study was designed to test the effect of tetracycline on healing subsequent to periodontal surgery. Aqueous solutions of tetracyclines are highly acidic and may therefore represent a suitable substitute for citric acid. Furthermore, tetracyclines react with dental hard tissues to from long-lasting antimicrobial compounds, and they have a retarding effect on pellicle and plaque formation and an antienzymatic effect. The alveolar bone around mandibular premolars was surgically reduced up to 6 mm from the cementoenamel junction in two beagles. The denuded root surfaces were exposed to the oral environment during 3 months without plaque control. Regenerative surgery was then carried out, using root surface conditioning with 1% tetracycline and coronally repositioned flaps. Six months later, histologic evaluation showed connective tissue attachment extending to the cementoenamel junction in most of the specimens. Superficial root resorption was prevalent in the cervical region, below which a collar of replacement resorption partly surrounded the roots in a characteristic manner. Morphometric analysis showed that attachment gain was similar to that obtained with citric acid in a preceding series of seven dogs. These preliminary results indicate that connective tissue attachment gain after topical use of tetracycline is similar to that obtained with citric acid. In addition, the antibacterial capacity and biological effects of tetracycline warrant further study of its possible clinical use in periodontal reconstructive surgery.


Charged Porous Polymers using a Solid C-O Cross-Coupling Reaction.
Herein, we report a green, fast, efficient mechanochemical strategy for charged porous polymers (CPPs). A cationic CPP with basic anions and an anionic CPP with Li(+) cations were fabricated by solid grinding under solvent-free conditions. Compared with solution-based synthesis, mechanochemical grinding can shorten the reaction time from dozens of hours to several minutes (60-90 min) to form polymers possessing a high molecular mass and low polydispersity. During the construction of CPPs, a Pd-catalyzed solid polycondensation based on unactivated organic linkers was introduced. In particular, CPPs with basic phenolic or proline anions showed good activity and stability in SO2 capture, and Li(+) -functionalized CPPs can be post-modified to CPPs with other metal ions by ion exchange, highlighting the tailorable feature of ionic-modified CPPs.


The route of transmission of hemagglutinating encephalomyelitis virus (HEV) 67N strain in 4-week-old rats.
Four-week-old Wistar rats were inoculated with HEV by different routes. Animals died of encephalitis after intraperitoneal (i.p.), subcutaneous (s.c.) and intravenous (i.v.) as well as intracerebral (i.c.) and intranasal (i.n.) inoculation. However when inoculated subcutaneously, rats died a few days earlier than those inoculated i.p. and i.v., suggesting that the virus might be transmitted to the central nervous system (CNS) by the neuronal route rather than by blood stream. Rats which were inoculated subcutaneously at the site of the neck (group A) began to die on day 4 p.i., a few days earlier than animals inoculated in the foot pad of the right leg (group B). On day 2 and 3 after inoculation, the virus titer in the brain was higher in group A, but group B animals showed higher virus titers in the lumber region of spinal cord than group A animals. In order to follow the virus spread from the peripheral nerve to the brain, the virus was inoculated into the sciatic nerve of rats. The inoculated rats developed clinical signs on day 4 and began to die on day 6. On day 2, virus was detected in the posterior half of the spinal cord and migrated toward the anterior half and in the brain where it was present on day 3. The highest virus titers in the brain were recorded on day 4 to 6, meanwhile the virus titers in the spinal cord tend to decrease. By immunohistochemical study, antigen positive neurons were found in the spinal cord and brain on day 4.(ABSTRACT TRUNCATED AT 250 WORDS)


Optometric supply and demand in Australia: 2011-2036.
The effective size of the optometric workforce is dependent on graduate numbers, retention rates and immigration and is influenced by age, gender and working hours of optometrists. This paper presents modelling results of the relationship between the projected Australian optometric workforce and projected demand for optometric services for the period 2011 to 2036. Nine hypothetical optometric supply-side and demand-side scenarios are presented. Data from the Australian Bureau of Statistics on age and gender of people listing optometry as their major qualification in the 2011 census were projected over a 25-year period, accounting for factors such as concordance with Health Workforce Australia figures for registered optometrists in Australia in 2011, ageing, attrition, hours worked, new graduates and immigration. Data were compared to the numbers of optometrists calculated as necessary to meet the demand for services of the Australian population to 2036 using nine different scenarios. It was estimated that there would be a surplus of over 1,200 equivalent full-time optometrists (EFTO) in 2036 for the highest service demand scenario of 13.8 million Medicare services, where 21 hours of a 38-hour week per EFTO were allowed for the provision of optometric services under Medicare. Substantial surpluses were predicted in all states and territories except Queensland, Tasmania and the Northern Territory where predicted supply was within six EFTO of predicted demand. Projections using current weightings for mortality, attrition, proportion of optometrists in active practice, working hours, immigration, new graduates and 21 hours per EFTO per week available for Medicare services indicate that in 2036, there will be excess optometrists in relation to projected demand for services, if service utilisation is maintained at current levels or increased by 10 or 20 per cent. Substantially greater excesses result if each EFTO has 28 or 35 hours per week available for Medicare services.


Closed, locked intramedullary nailing of pediatric femoral shaft fractures through the tip of the greater trochanter.
Closed femoral nailing is universally accepted as the treatment of choice in almost all diaphyseal femoral fractures in adults. Numerous authors reported favorable results applying the same surgical technique in the adolescent patient group. Nevertheless, reports of complications such as avascular necrosis and alteration of the proximal femoral anatomy have dampened the initial enthusiasm. The purpose of this paper was to evaluate the possible effect of closed intramedullary nailing through the greater trochanter on the proximal femoral anatomy. We report the results of intramedullary nailing in 20 skeletally immature patients (13 men and 7 women) with a mean age of 14.4 years (range, 11-16 years). All were treated with closed, reamed, percutaneously performed nailing, using the tip of the greater trochanter as the nail insertion point. The patients were followed for 29 months in average (range, 19-37 months). No major complication (limb length discrepancy, avascular necrosis, coxa valga) occurred during the observation period. All fractures healed clinically and radiographically within 9 weeks in average (8-13 weeks) and all patients returned to the preinjury activity level. The mean ATD difference was 1.10 +/- 3.51 (range, -5-7 mm, 95% CI -0,54/2,74, p = 0.177). The mean LTA distance difference was 0.3 mm (range, -6-5 mm, p = 0.158), the mean femoral length difference was 1.9 mm (-9-12 mm, p = 0.122) and the overall limb length difference was 1.4 mm (-25-20 mm, p = 0.178). The mean neck-shaft angle difference was 0.20 +/- 1.74 (range, -3-4, p = 0.612) and the mean neck width was 0.60 +/- 1.50 (range, -3-3, p = 0.09). Fourteen nails (70%) were removed within 13 months in average (range, 10-18 months) without any complications. This study showed that with strict adherence to a surgical technique that respects the growing proximal femur and its vascular anatomy, using the tip of the greater trochanter as an entry point to the femoral canal, the proven advantages of closed, intramedullary nailing can safely be offered to the adolescent patient population as well.


Crystal structure of human prostatic acid phosphatase .
Prostatic acid phosphatase (hPAP) is a major product of the human prostate gland, yet its physiological substrate remains unknown. Human PAP, purified from semen, was crystallized using polyethylene glycol as the precipitant and its crystal structure was determined using X-ray diffraction. The structure was refined at 3.1 A resolution to R = 16% and R(free) = 27%. The structure of hPAP is similar to that of other known histidine phosphatases, and the positions of its catalytic residues are conserved. N-linked carbohydrates are present at each of the possible glycosylation sites. It appears that high-mannose chains are attached to Asn 62 and Asp 301, while complex chains are at Asn 188. The similarity of the three-dimensional structures of rat PAP and human PAP indicates that the mechanistic analyses of the catalytic mechanism proposed for the rat enzyme should be extended to the human enzyme without reservations. The crystallographic data allowed the correlation of attachment sites of N-linked carbohydrate chains with a given carbohydrate type. The carbohydrates of the protein produced in the prostate cells and in the baculovirus expression system appear to differ at the site of complex carbohydrates attachment.


The CHESS score: a simple tool for early prediction of shunt dependency after aneurysmal subarachnoid hemorrhage.
Acute hydrocephalus is an early and common complication of aneurysmal subarachnoid hemorrhage (SAH). However, considerably fewer patients develop chronic hydrocephalus requiring shunt placement. Our aim was to develop a risk score for early identification of patients with shunt dependency after SAH. Two hundred and forty-two SAH individuals who were treated in our institution between January 2008 and December 2013 and survived the initial impact were retrospectively analyzed. Clinical parameters within 72 h after the ictus were correlated with shunt dependency. Independent predictors were summarized into a new risk score which was validated in a subsequent SAH cohort treated between January and December 2014. Seventy-five patients (31%) underwent shunt placement. Of 23 evaluated variables, only the following five showed independent associations with shunt dependency and were subsequently used to establish the Chronic Hydrocephalus Ensuing from SAH Score (CHESS, 0-8 points): Hunt and Hess grade ≥IV (1 point), location of the ruptured aneurysm in the posterior circulation (1 point), acute hydrocephalus (4 points), the presence of intraventricular hemorrhage (1 point) and early cerebral infarction on follow-up computed tomography scan (1 point). The CHESS showed strong correlation with shunt dependency (P = 0.0007) and could be successfully validated in both internal SAH cohorts tested. Patients scoring ≥6 CHESS points had significantly higher risk of shunt dependency (P < 0.0001) than other patients. The CHESS may become a valuable diagnostic tool for early estimation of shunt dependency after SAH. Further evaluation and external validation will be required in prospective studies.


Maternal adherence with recommendations for folic acid and iodine supplements: A cross-sectional survey.
We conducted a survey of 535 New Zealand women to determine supplement use pre-pregnancy, during pregnancy and while breastfeeding to evaluate adherence with national recommendations for folic acid and iodine supplementation. Our findings suggest that despite these women being well-educated and motivated to complete an online survey, there was low adherence to recommendations for folic acid and iodine supplements. Only 38% of women reported supplement intake that met both folic acid and iodine recommendations. This cross-sectional survey provides information useful to policy makers, clinicians and researchers seeking to protect and promote maternal and infant health.


Adaptive plasticity of autophagic proteins to denervation in aging skeletal muscle.
Aging muscle exhibits a progressive decline in mass and strength, known as sarcopenia, and a decrease in the adaptive response to contractile activity. The molecular mechanisms mediating this reduced plasticity have yet to be elucidated. The purposes of this study were 1) to determine whether denervation-induced muscle disuse would increase the expression of autophagy genes and 2) to examine whether selective autophagy pathways (mitophagy) are altered in aged animals. Denervation reduced muscle mass in young and aged animals by 24 and 16%, respectively. Moreover, young animals showed a 50% decrease in mitochondrial content following denervation, an adaptation that was not matched by aged animals. Basal autophagy protein expression was higher in aged animals, whereas young animals exhibited a greater induction of autophagy proteins following denervation. Localization of LC3II, Parkin, and p62 was significantly increased in the mitochondrial fraction of young and aged animals following denervation. Moreover, the unfolded protein response marker CHOP and the mitochondrial dynamics protein Fis1 were increased by 17- and 2.5-fold, respectively, in aged animals. Lipofuscin granules within lysosomes were evident with aging and denervation. Thus reductions in the adaptive plasticity of aged muscle are associated with decreases in disuse-induced autophagy. These data indicate that the expression of autophagy proteins and their localization to mitochondria are not decreased in aged muscle; however, the induction of autophagy in response to disuse, along with downstream events such as lysosome function, is impaired. This may contribute to an accumulation of dysfunctional mitochondria in aged muscle.


Multitriggered Shape-Memory Acrylamide-DNA Hydrogels.
Acrylamide-acrylamide nucleic acids are cross-linked by two cooperative functional motives to form shaped acrylamide-DNA hydrogels. One of the cross-linking motives responds to an external trigger, leading to the dissociation of one of the stimuli-responsive bridges, and to the transition of the stiff shaped hydrogels into soft shapeless states, where the residual bridging units, due to the chains entanglement, provide an intrinsic memory for the reshaping of the hydrogels. Subjecting the shapeless states to counter stimuli restores the dissociated bridges, and regenerates the original shape of the hydrogels. By the cyclic dissociation and reassembly of the stimuli-responsive bridges, the reversible switchable transitions of the hydrogels between stiff shaped hydrogel structures and soft shapeless states are demonstrated. Shaped hydrogels bridged by K(+)-stabilized G-quadruplexes/duplex units, by i-motif/duplex units, or by two different duplex bridges are described. The cyclic transitions of the hydrogels between shaped and shapeless states are stimulated, in the presence of appropriate triggers and counter triggers (K(+) ion/crown ether; pH = 5.0/8.0; fuel/antifuel strands). The shape-memory hydrogels are integrated into shaped two-hydrogel or three-hydrogel hybrid structures. The cyclic programmed transitions of selective domains of the hybrid structures between shaped hydrogel and shapeless states are demonstrated. The possible applications of the shape-memory hydrogels for sensing, inscription of information, and controlled release of loads are discussed.


Protocadherin 10 suppresses tumorigenesis and metastasis in colorectal cancer and its genetic loss predicts adverse prognosis.
Protocadherin 10 (PCDH10), a novel tumor suppressor gene in human cancers, is located in a common deleted region at chromosome 4q28 in colorectal cancer (CRC). This study aimed to ascertain the genetic loss of PCDH10 and its clinical relevance in CRC and to explore the tumor suppressor function of PCDH10. The genetic deletion of PCDH10 was determined in 171 pairs of primary tumors and corresponding normal mucosae by loss of heterozygosity study. In total, 53 carcinomas were positive for allelic loss of PCDH10. The genetic aberration was significantly associated with tumor progression and distant metastasis (p = 0.021 and p = 0.018, respectively) and was an independent predictor of poor survival for CRC patients (p = 0.005). Expression of PCDH10 gene was silenced or markedly down-regulated in all of 12 CRC cell lines tested and in 41 of 53 colorectal carcinomas compared with their matched normal mucosae. Ectopic expression of PCDH10 suppressed cancer cell proliferation, anchorage-independent growth, migration and invasion in vitro. Subcutaneous injection of PCDH10-expressing CRC cells into SCID mice revealed the reduction of tumor growth compared with that observed in mock-inoculated mice. Furthermore, through intrasplenic implantation, the re-expression of PCDH10 in silenced cells restrained liver metastasis and improved survival in SCID mice. In conclusion, PCDH10 is a pivotal tumor suppressor in CRC, and the loss of its function promotes not only tumor progression but also liver metastasis. In addition, the genetic deletion of PCDH10 represents an adverse prognostic marker for the survival of patients with CRC.


Molecular frame image restoration and partial wave analysis of photoionization dynamics of NO by time-energy mapping of photoelectron angular distribution.
The benchmark system of molecular photoionization dynamics, the (1+1{'}) two-photon ionization of NO via the A state, is investigated using the time-energy mapping of the photoelectron angular distribution in a laboratory frame. The molecular frame photoelectron angular distribution and partial wave composition are determined from time-energy maps and compared with those obtained by Schwinger variational calculation (SVC) and state-to-state photoelectron spectroscopy. Good agreement is found with SVC. By comparison of the phase shifts of the scattering waves and the quantum defects of the Rydberg states, the l hybridization of p waves is identified.


[Diagnosis of membrane disorders in acute pancreatitis].
In 110 patients with acute pancreatitis the authors studied the activity of intracellular enzymes in the blood (LDH, CPC, ALaT, ASaT, transamidinase) in a complex with indices characterizing the condition of the membrane cell systems, in particular: activity of acid phosphatase, content of 17-ketosteroids, activity of plasma chemiluminescence, concentration of beta-lipoproteins in blood. Complex study of these indices makes it possible to make a sufficiently objective judgement of the severity of the membrane disorders in acute pancreatitis and verity the form and control the treatment of the disease.


Regulation of renin gene expression in hypertensive rats.
A carboxy terminal renin complementary DNA (cDNA) clone from rat kidney was isolated, characterized, and used as a probe for renin messenger RNA (mRNA) quantification in normotensive and hypertensive rats. RNA blotting analysis detected renin mRNA in control kidney and brain. Deoxycorticosterone acetate (DOCA) and high salt (1%) treatment of experimental animals resulted in a greater than 95% decrease in the content of renin mRNA in the kidney, as compared with values in control rats receiving 0.4% NaCl in their diet. In contrast, high salt (1%) treatment alone caused only a twofold decrease in kidney renin mRNA content, as compared with values in controls. DOCA and low salt (0.04%) or low salt (0.04%) treatment alone caused a 1.5-fold increase in the kidney renin mRNA content, as compared with values in control rats. These results indicate that DOCA and salt have a synergistic effect in depressing renin mRNA levels in kidney. Clipping of the left renal artery caused a threefold increase in the steady state level of renin mRNA in the ischemic kidney and a 0.5-fold decrease in the hypertrophied kidney. The data are consistent with the hypothesis that blood pressure and other stimuli regulate the expression of the renin gene in vivo.


Boronic Acid functionalized core-shell polymer nanoparticles prepared by distillation precipitation polymerization for glycopeptide enrichment.
The boronic acid-functionalized core-shell polymer nanoparticles, poly(N,N-methylenebisacrylamide-co-methacrylic acid)@4-vinylphenylboronic acid (poly(MBA-co-MAA)@VPBA), were successfully synthesized for enriching glycosylated peptides. Such nanoparticles were composed of a hydrophilic polymer core prepared by distillation precipitation polymerization (DPP) and a boronic acid-functionalized shell designed for capturing glycopeptides. Owing to the relatively large amount of residual vinyl groups introduced by DPP on the core surface, the VPBA monomer was coated with high efficiency, working as the shell. Moreover, the overall polymerization route, especially the use of DPP, made the synthesis of nanoparticles facile and time-saving. With the poly(MBA-co-MAA)@VPBA nanoparticles, 18 glycopeptides from horseradish peroxidase (HRP) digest were captured and identified by MALDI-TOF mass spectrometric analysis, relative to eight glycopeptides enriched by using commercially available meta-aminophenylboronic acid agarose under the same conditions. When the concentration of the HRP digest was decreased to as low as 5 nmol, glycopeptides could still be selectively isolated by the prepared nanoparticles. Our results demonstrated that the synthetic poly(MBA-co-MAA)@VPBA nanoparticles might be a promising selective enrichment material for glycoproteome analysis.


Anti-inflammatory activity of Camellia japonica oil.
Camellia japonica oil (CJ oil) has been used traditionally in East Asia to nourish and soothe the skin as well as help restore the elasticity of skin. CJ oil has also been used on all types of bleeding instances. However, little is known about its anti-inflammatory effects. Therefore, the anti-inflammatory effects of CJ oil and its mechanisms of action were investigated. CJ oil inhibited LPS-induced production of NO, PGE(2), and TNF-α in RAW264.7 cells. In addition, expression of COX-2 and iNOS genes was reduced. To evaluate the mechanism of the anti-inflammatory activity of CJ oil, LPS-induced activation of AP-1 and NF-κB promoters was found to be significantly reduced by CJ oil. LPS-induced phosphorylation of IκBα, ERK, p38, and JNK was also attenuated. Our results indicate that CJ oil exerts anti-inflammatory effects by downregulating the expression of iNOS and COX-2 genes through inhibition of NF-κB and AP-1 signaling. [BMB reports 2012; 45(3): 177-182].


Functions of LiBH4 in the hydrogen sorption reactions of the 2LiH-Mg(NH2)2 system.
LiBH(4) is an effective additive in enhancing hydrogen desorption and absorption properties for the 2LiH-Mg(NH(2))(2) hydrogen storage system. The improving factors brought by LiBH(4) were studied experimentally. Although a minimum in activation energy was observed in the middle stage of hydrogen desorption for the samples with and without LiBH(4) doping, a reduction in the activation energy was manifest in the overall conversion range due to the LiBH(4) introduction. In contrast to the pure 2LiH-Mg(NH(2))(2), the system component Mg(NH(2))(2) crystallized in the presence of LiBH(4) at about 150 °C in situ from the post-milled amorphous mixture prior to hydrogen desorption. We believe the formation of the N-atom matrix by the crystalline Mg(NH(2))(2) is relevant to the subsequent dehydrogenation as it shares a similar sublattice structure with the desorption product. The formation and disappearance of Li(4)(BH(4))(NH(2))(3) in the dehydrogenation step witnessed the capturing and releasing of LiNH(2) that is generated from the dehydrogenation of 2LiH-Mg(NH(2))(2), which alters not only the kinetics but also thermodynamics in favor of an improved hydrogen sorption performance. The reaction mechanism could be further clarified by the refinement of powder neutron diffraction data of the intermediate compound Li(4)(BD(4))(NH(2))(3).


Level of alimentation and line of breeding on oxygen uptake by ovine jejunal mucosa.
Targhee rams from a selected and control line of breeding were provided with the same diet at either 100 or 40% of ad libitum feed intake levels for 28 days. The rams were killed in a serial fashion over a 35-day period. A sample of the jejunum mucosa was taken, and O2 consumption rates with and without ouabain were measured. Total O2 consumption and ouabain-sensitive respiration rates of the jejunal mucosa taken from rams of the select line were greater than for samples taken from the control rams. Line of breeding did not influence the percent of the total O2 consumption that was ouabain sensitive and the proportion of the jejunum that was mucosa. Differences in rates of total and ouabain-sensitive O2 consumption attributed to level of feed intake were not detected as significant. Both the percentage of the total respiration rates that was ouabain sensitive and the proportion of the jejunum that was mucosa were greater for rams fed at high intake levels compared with rams fed at low intake levels. Thus both breeding sheep on the basis of improved growth potential and level of alimentation influenced aspects of jejunal mucosa respiration that was Na+-K+-adenosinetriphosphatase dependent.


Conventional and molecular characterization of coagulase-negative staphylococcus in hospital isolates.
During the last decade, coagulase-negative staphylococci (CoNS) have emerged as a major cause of nosocomial infections. They constitute a major component of the normal skin and mucosal microflora, and are particularly responsible for catheter- and medical device-related sepsis. They present unique problems in diagnosis and treatment of infections. The present study has been designed to evaluate phenotypic and genotypic characteristics of CoNS among nosocomial isolates. This study was carried out in a tertiary care hospital. Data from 150 samples collected from 73 hospitalized patients and 15 healthy volunteers between October 2003 and May 2005 were analyzed. A total of 100 CoNS strains responsible for sepsis or implant-associated infections and 50 saprophytic strains were studied. Invasive CoNS strains were selected on the basis of different colony morphologies, drug resistance patterns, and biofilm formation. The same criteria were used to select saprophytic isolates. Multiplex PCR was used to explore the ica, mecA, and atlE genes, which might contribute to the pathogenicity of CoNS and the formation of biofilms. Most of the invasive strains that formed the biofilm were resistant to multiple antibiotics, with more than 80% resistant to methicillin. ica and mecA genes were detected significantly in pathogenic strains (chi-square test, P<0.0001) whereas atlE was ubiquitously amplified in all the strains. All those strains which had ica and mecA genes were resistant to multiple antibiotics and were positive for biofilm formation. These genetic markers thus appear to discriminate between potential invasive virulent and saprophytic strains of CoNS.


Measurement error and dietary intake.
This chapter reviews work of Carroll, Freedman, Kipnis, and Li (1998) on the statistical analysis of the relationship between dietary intake and health outcomes. In the area of nutritional epidemiology, there is some evidence from biomarker studies that the usual statistical model for dietary measurements may break down due to two causes: (a) systematic biases depending on a person's body mass index; and (b) an additional random component of bias, so that the error structure is the same as a one-way random effects model. We investigate this problem, in the context of (1) the estimation of the distribution of usual nutrient intake; (2) estimating the correlation between a nutrient instrument and usual nutrient intake; and (3) estimating the true relative risk from an estimated relative risk using the error-prone covariate. While systematic bias due to body mass index appears to have little effect, the additional random effect in the variance structure is shown to have a potentially important impact on overall results, both on corrections for relative risk estimates and in estimating the distribution usual of nutrient intake. Our results point to a need for new experiments aimed at estimation of a crucial parameter.


Intracellular protozoan infection in small intestinal biopsies of patients with AIDS. Light and electron microscopic evaluation.
Small intestinal biopsies of 21 patients with acquired immunodeficiency syndrome (AIDS) with light microscopic findings diagnostic or suspicious for parasite infection were investigated by transmission electron microscopy (TEM). TEM allowed us to identify and specify the genus and species of involved parasites in 16 out of the 21 cases: 7 Cryptosporidium parvum, 5 Enterocytozoon bieneusi and 4 Isospora belli. Cryptosporidium was easily identified on light microscopy (LM), and only slightly influenced by parasite burden in all the 7 cases; TEM confirmed LM diagnosis and made it possible to characterize the parasites as C. parvum. The identification of Microsporidium on LM in our cases was related to the burden of parasite; its presence was certainty identified in 2 cases and suspected in 3. TEM allowed to identify these parasites as E. bieneusi. Intracytoplasmic coccidia could be detected with certainly in semithin sections in all 4 cases, but TEM was always needed to specify the infectious agent as I. belli. In 5 cases the suspicious of protozoan infection on LM (3 microsporidia, 1 intracytoplasmic coccidia and 1 Cryptosporidium) was not confirmed by TEM. Our data suggest that TEM is an appropriate diagnostic tool in this field of pathology and necessary in most of the cases.


Utility of interphase FISH to stratify patients into cytogenetic risk categories at diagnosis of AML in an Eastern Cooperative Oncology Group (ECOG) clinical trial (E1900).
We evaluated the efficacy of FISH to detect chromosome anomalies in the evaluation of young (<60 years) patients with AML. Patients were enrolled in E1900, an ECOG clinical trial for AML. The protocol was designed to collect bone marrow or blood for both cytogenetic and FISH studies at study entry (diagnosis). FISH for each patient was performed and utilized eight probe sets to detect t(8;21), t(9;22), t(11;var), t(15;17), inv(16), +8, -5/5q, and -7/7q. We analyzed 237 specimens with complete cytogenetic and FISH results. Results for each specimen were classified by probe set into one of six categories. The concordance rate between cytogenetic and FISH results ranged from 98 to 100% for all probe sets and kappa analysis for concordance had a p-value of <0.0001. The high level of agreement between cytogenetic and FISH results demonstrate the accuracy of a panel of eight FISH probe sets for the detection of significant abnormalities in AML. Data from this investigation support the use of FISH as an adjunct method to increase the yield of useful cytogenetic results in large cooperative trials and demonstrate the potential of FISH as a follow-up study of minimal residual disease in ECOG trials.


Results from a comparative dietary assessment in Europe: II. Feasibility of pooling individual-based dietary data between countries.
Dietary investigations in four central European survey populations carried out in the German Democratic Republic, Poland and Denmark between 1982 and 1984 using different methodologies were analysed in order to assess the possibilities of characterizing the dietary habits of individual survey participants in a comparable fashion. This was done with the view of assessing the feasibility of a pooled cancer cohort study. For this purpose a method has been devised to combine dietary information derived by food frequency questionnaires and quantitative recording methods into a quantitative characterization of individuals' habits. A comparable characterization between different cultural settings could be demonstrated for a selected list of food items. The selection was determined by the food items considered in common in the different food frequency questionnaires and yielding sufficient and comparable variation as well as absolute amounts of intake. This was more clearly found for food items such as 'fruit' which experience a concise role in dietary habits. However, the observed discrepancies of the different dietary methods within the countries, and, most importantly, between the countries, result in distributions of average daily consumption values which are not deemed to be comparable.


An empirical assessment of the relationship between Official Development Aid and child mortality, 2000-2015.
To analyze the effect of Official Development Aid (ODA) dollars on child mortality over the course of the United Nation's Millennium Development Goals initiative. The relationship between child mortality and Official Development Aid over the duration of the Millennium Development Goals (2000-2015) is examined here using a longitudinal panel of country-level data from the World Bank and the United Nations. An Ordinary Least Squares regression approach was used with country-level fixed effects. Models were estimated for the full sample and by Human Development Index development strata (high, medium, and low developed countries) with clustered standard errors. ODA appears to be most strongly associated with decreases in child mortality in Medium Developed Countries. Every one dollar per capita increase in ODA is associated with a 0.035 decrease in child deaths per 1000 births. Significant gains were made in decreasing child mortality over the last 15 years. The need for more progress remains. Allocation of ODA to developing countries can be an effective policy tool in achieving public health goals.


Luciferase Advisor: High-Accuracy Model To Flag False Positive Hits in Luciferase HTS Assays.
Firefly luciferase is an enzyme that has found ubiquitous use in biological assays in high-throughput screening (HTS) campaigns. The inhibition of luciferase in such assays could lead to a false positive result. This issue has been known for a long time, and there have been significant efforts to identify luciferase inhibitors in order to enhance recognition of false positives in screening assays. However, although a large amount of publicly accessible luciferase counterscreen data is available, to date little effort has been devoted to building a chemoinformatic model that can identify such molecules in a given data set. In this study we developed models to identify these molecules using various methods, such as molecular docking, SMARTS screening, pharmacophores, and machine learning methods. Among the structure-based methods, the pharmacophore-based method showed promising results, with a balanced accuracy of 74.2%. However, machine-learning approaches using associative neural networks outperformed all of the other methods explored, producing a final model with a balanced accuracy of 89.7%. The high predictive accuracy of this model is expected to be useful for advising which compounds are potential luciferase inhibitors present in luciferase HTS assays. The models developed in this work are freely available at the OCHEM platform at http://ochem.eu .


Exceptional thermal stability of Pd@CeO2 core-shell catalyst nanostructures grafted onto an oxide surface.
Monolayer films of highly catalytically active Pd@CeO2 core-shell nanocomposites were grafted onto a planar YSZ(100) (yttria-stabilized zirconia, YSZ) single crystal support that was functionalized with a CVD-deposited layer of triethoxy(octyl)silane (TEOOS). The resulting monolayer films were found to exhibit exceptionally high thermal stability compared to bare Pd nanoparticles with the Pd@CeO2 nanostructures remaining intact and highly dispersed upon calcining in air at temperatures in excess of 1000 K. The CeO2 shells were also shown to be more easily reduced than bulk CeO2, which may partially explain their unique activity as oxidation catalysts. The use of both TEOOS and tetradecylphosphonic acid (TDPA) as coupling agents for dispersing Pd@CeO2 core-shell nanocomposites onto a high surface area γ-Al2O3 support is also demonstrated.


HLA-A3 supermotif defined by quantitative structure-activity relationship analysis.
Activation of a cytotoxic T cell requires specific binding of antigenic peptides to major histocompatibility complex (MHC) molecules. This paper reports a study of peptides binding to members of the HLA-A3 superfamily using a recently developed 2D-QSAR method, called the additive method. Four alleles with high phenotype frequency were included in the study: A*0301, A*1101, A*3101 and A*6801. The influence of each of the 20 amino acids at each position of the peptide on binding was studied. A refined A3 supertype motif was defined in the study.


Depletion of Gr-1+, but not Ly6G+, immune cells exacerbates virus replication and disease in an intranasal model of herpes simplex virus type 1 infection.
In the absence of a viable 'knockout' mouse, researchers have relied extensively on monoclonal antibody (mAb) RB6-8C5 [anti-granulocyte receptor 1 (Gr-1)] to deplete neutrophils in murine models of inflammation and infection. Using an intranasal model of herpes simplex virus type 1 (HSV-1) infection, we demonstrate that mAb RB6-8C5 also binds to plasmacytoid dendritic cells, F4/80(+) macrophages/monocytes and CD8(+) T cells recovered from the airways of HSV-1-infected mice. In contrast, mAb 1A8 (anti-Ly6G) bound specifically to Ly6G(high) neutrophils. Following intranasal infection of C57BL/6 mice with HSV-1, few Ly6G(high) neutrophils were recruited to the airways and treatment of mice with purified mAb 1A8 induced systemic neutropenia, but did not alter virus replication or disease progression. In contrast, treatment of HSV-1-infected mice with mAb RB6-8C5 led to exacerbated virus replication, disease severity and mortality. These findings highlight the limitations associated with widespread use of antibody-mediated depletion of Gr-1(+) cells to define the role of neutrophils in vivo. Furthermore, we use mAb 1A8 to demonstrate that specific depletion of neutrophils does not modulate disease or alter virus replication following intranasal infection with HSV-1.


Initial fermentation of sea sludge using aerobic and thermophilic microorganisms in a mangrove soil.
As sea sludge has the potential to cause eutrophication and red tide resulting in the death of shellfish and offensive odors, an effective method to treat it is needed. It was found that adding soil taken from a mangrove swamp to sea sludge promoted an initial fermentation of the sludge constituents. The result suggested that certain microorganisms that were thought to inhabit the sub-tropical mangrove soil had the potential to play a significant role in the fermentation and that the use of the microorganisms in the mangrove soil might be useful for composting sea sludge.


High serum level of procollagen type III amino-terminal peptide contributes to the efficacy of spironolactone and angiotensin-converting enzyme inhibitor therapy on left ventricular hypertrophy in essential hypertensive patients.
We recently demonstrated that spironolactone may have beneficial effects on left ventricular hypertrophy in selected patients with essential hypertension undergoing treatment with an angiotensin-converting enzyme (ACE) inhibitor. To clarify the possible mechanisms by which spironolactone improves cardiac hypertrophy, we investigated the change in serum procollagen type III amino-terminal peptide (PIIINP) in 11 patients with essential hypertension treated with spironolactone and an ACE inhibitor for 24 weeks. Both blood pressure and serum PIIINP levels were significantly decreased by treatment. There was a statistical significant correlation between the changes in LVMI and those in PIIINP. The reduction in PIIINP was significant in patients whose initial serum PIIINP levels were above the normal range. Before treatment, there were no statistically significant correlations between serum PIIINP levels and either LVMI, blood pressure, or plasma aldosterone concentration. Essential hypertensive patients matched in terms of duration of therapy, blood pressure and LVMI and treated with an ACE inhibitor alone showed no change in serum PIIINP levels. In conclusion, the results of the present study demonstrate that patients with essential hypertension and high serum levels of PIIINP are particularly responsive to MR blockade in terms of left ventricular hypertrophy. Moreover, these results suggest that spironolactone limits cardiac collagen turnover in such patients. Larger studies may provide definitive evidence for the involvement of aldosterone in left ventricular hypertrophy in patients with abnormally high PIIINP levels.


Cadmium exposure in relation to insulin production, insulin sensitivity and type 2 diabetes: a cross-sectional and prospective study in women.
Cadmium is a wide-spread pollutant. Observational studies suggest associations between cadmium and prevalence of type 2 diabetes. Experimental studies indicate that cadmium may cause impaired insulin production. To examine whether cadmium exposure is associated with impaired glucose tolerance and type 2 diabetes and impaired pancreatic beta-cell function. Oral glucose tolerance tests were used in a screening examination of 64-year old women (n=2595) to identify all with diabetes, impaired (IGT) and normal (NGT) glucose tolerance. Random samples of women with type 2 diabetes (n=215), IGT (n=207) and NGT (n=194) were recruited in a cross-sectional examination including measurement of pancreatic insulin production (acute insulin response) and insulin sensitivity (homeostasis model assessment). Cadmium concentrations were measured in blood and urine. A follow-up examination was performed after 5.4 years with assessment of new cases with diabetes or impaired glucose tolerance. At baseline, neither blood nor urinary cadmium concentrations showed any statistically significant differences between women with type 2 diabetes, IGT or NGT. The prospective analysis included 68 cases with incident diabetes, 58 with IGT and 118 women with NGT and no associations with cadmium levels at baseline were observed. Blood and urinary cadmium at baseline were not associated with insulin production, blood glucose, HbA1c, or changes in HbA1c during follow-up. This is the first study of cadmium and diabetes with detailed data on pancreatic beta-cell function, insulin sensitivity and glucose tolerance. Cadmium exposure was not associated with increased risk of type 2 diabetes or IGT.


Cognitive dysfunction and symptoms of movement disorders in adult-onset leukoencephalopathy with axonal spheroids and pigmented glia.
Adult-onset leukoencephalopathies are clinically and genetically heterogeneous disorders that affect predominantly the cerebral white matter of the central nervous system. Clinical and neuroimaging-based approaches have been developed to improve diagnostic processes for adult-onset leukoencephalopathies. However, the differential diagnosis is often challenging. Recently, knowledge of the genetic basis of leukoencephalopathies has been accumulated rapidly, which provides powerful diagnostic approaches. The article provides an overview of adult-onset leukoencephalopathy with axonal spheroids and pigmented glia (ALSP), focusing on the clinical presentations of cognitive impairment and symptoms of movement disorders. ALSP is a subtype of dominantly inherited leukoencephalopathy caused by CSF1R mutations. ALSP typically develop in adulthood, with cognitive decline, psychiatric symptoms, and motor symptoms of movement disorders. Cognitive symptoms in ALSP are characterized by frontal lobe dysfunctions such as executive dysfunction, attention deficits and indifference. The cardinal motor symptoms of movement disorders ALSP were gait disturbance and bradykinesia, which may appear as the initial symptoms. Thus, ALSP should be recognized as both a cognitive disorder and a movement disorder.


Assembly of homogeneous norovirus-like particles accomplished by amino acid substitution.
Infection of insect cells with recombinant baculoviruses carrying the VP1 gene from Chiba strain norovirus resulted in the production of 57 and 50 kDa proteins, and the assembly of a smaller, 23 nm form of the virus-like particles (VLPs), together with the normal, 38 nm form of the VLPs. The N-terminal residues of the 57 and 50 kDa proteins were Ala4 and Thr45, respectively. When the tripeptide Leu43-Ala44-Thr45 was changed to Ala-Pro-Val, only 38 nm VLPs were assembled. The 38 nm VLPs showed essentially the same pattern of carbohydrate binding as the 23 nm VLPs, despite the significant difference in the degree of Lewis b antigen binding.


[The incidence of multiple ovulations in Dutch warmblood mares].
In the period 1994-1996 1030 oestruses of Dutch Warmblood mares were evaluated on the incidence of multiple ovulations diagnosed by rectal palpation. Ultrasonography was not used. All follicles > 25 mm were noted. The examination was repeated every 48 hours. The incidence of multiple follicle development was 7.3%. In a group of 50 ultrasonographically diagnosed twin-pregnant mares only 14 (28%) had been detected by rectal palpation as a multiple ovulation. This suggests an incidence of multiple ovulations in Dutch Warmblood mares of 7.3 x 1/28 x 100 = 26%. Restrictions of rectal palpation are discussed. Ultrasonography is essential for the diagnosis of multiple ovulations. The importance of a good twin-prevention programme is emphasized.


Static magnetic field as biological modifier: a study on temperature dependent influence.
Magnetic fields seemingly alter a number of physiological indicators in intact animals and influence cellular metabolism. We have studied the magnetic field effects on the membrane and receptors of the reticulo-endothelial cells of bone marrow which are mainly responsible for the phagocytic activity of nanocolloid particles of human serum albumin tagged with Tc99m. A series of experiments carried out on immobilized mice exposed to a static 1.4 T SMF for 60 min at 27 degrees C or 37 degrees C body temperature showed an increased phagocytic activity at 37 degrees C and decreased activity at 27 degrees C.


Rapid determination of hydrogen peroxide in pulp bleaching effluents by headspace gas chromatography.
A headspace gas chromatographic (HS-GC) method has been developed for the determination of residual hydrogen peroxide in pulp bleaching effluents. The method is based on the reaction of hydrogen peroxide and permanganate in an acidic medium (0.1 mol/L), in which hydrogen peroxide is quantitatively converted to oxygen within 10 min at 60°C in a sealed headspace sample vial. The released oxygen is then determined by GC equipped with a thermal conductivity detector. The method is robust, sensitive, and accurate, with reproducibility characterized by a relative standard deviation of <0.5%, a sensitivity whose limit of quantification (LOQ) is 0.96 μmol, and a demonstrated recovery ranging from 98 to 103%. Further, the method is simple, rapid, and automated.


Cutaneous extraskeletal mesenchymal chondrosarcoma in a cat.
A 4-year-old, male cat was presented with a fixed, subcutaneous mass in the lumbosacral region. A histopathological examination revealed a well-defined but nonencapsulated neoplasm characterized by a proliferation of predominantly spindle cells, with high mitotic activity. Interspersed between these cells were single cellular elements with chondroid differentiation. Large areas of cartilaginous tissue with foci of endochondral ossification, necrosis and myxoid tissue were also observed within the neoplastic parenchyma. A diagnosis of extraskeletal mesenchymal chondrosarcoma was made based on the histological pattern - characterized by the coexistence of cartilaginous islands and undifferentiated mesenchymal cells, results of Alcian blue staining at various pH, immunohistochemical reactivity against vimentin and S-100, and the absence of skeletal involvement or other primary tumour sites. Clinical history of the cat excluded traumas, vaccinations or other types of subcutaneous inoculation. Six months on from surgical treatment, neither recurrence nor metastases have been detected.


Membrane properties related to the firing behavior of zebrafish motoneurons.
The physiological and pharmacological properties of the motoneuron membrane and action potential were investigated in larval zebrafish using whole cell patch current-clamp recording techniques. Action potentials were eliminated in tetrodotoxin, repolarized by tetraethylammonium (TEA) and 3,4-diaminopyridine (3,4-AP)-sensitive potassium conductances, and had a cobalt-sensitive, high-threshold calcium component. Depolarizing current injection evoked a brief (approximately 10-30 ms) burst of action potentials that was terminated by strong, outwardly rectifying voltage-activated potassium and calcium-dependent conductances. In the presence of intracellular cesium ions, a prolonged plateau potential often followed brief depolarizations. During larval development (hatching to free-swimming), the resting membrane conductance increased in a population of motoneurons, which tended to reduce the apparent outward rectification of the membrane. The conductances contributing to action potential burst termination are hypothesized to play a role in patterning the synaptically driven motoneuron output in these rapidly swimming fish.


The role of Peyer's patches in the age-related incidence of Crohn's disease.
Recently researchers have suggested that clinical subsets of Crohn's disease occur, which are variously described as inflammatory, fibrostenotic, and fistulizing. In addition, it has been observed that within families with multiple cases, often there is concordance of the site and type of disease. The lesions of Crohn's disease occur in segments that suggest that distribution of Peyer's patches. When the age-related incidence of Crohn's disease was plotted for all countries from which such data were available, the peaks of greatest case frequency occurred at ages 15 to 25 years and paralleled a similar peak representing the number of Peyer's patches as a function of age. This correlation suggests that Crohn's disease may develop as an inflammatory process specifically targeting these important lymphoid structures. Similar peaks of activity in the adolescent to early adult years occur for appendicitis and tonsillitis.


Cerebral hemodynamics in ischemic cerebrovascular disease.
During the past decade, technological advances have made it possible to measure regional cerebral hemodynamics in individual patients. Studies performed with these techniques have demonstrated that the degree of carotid stenosis correlates poorly with the hemodynamic status of the ipsilateral cerebral circulation. The primary determinant of cerebral perfusion pressure and blood flow under these circumstances is the adequacy of collateral circulatory pathways. Since collateral circulation varies from patient to patient, there is no critical degree of carotid stenosis that consistently produces hemodynamic compromise of the cerebral circulation. It is, thus, time to abandon the concept of the hemodynamically significant carotid stenosis as it relates to the pathogenesis and treatment of cerebrovascular disease. Measurements of regional cerebral hemodynamics have provided new insight into the pathogenesis of transient ischemic attacks and generated some preliminary data on the prognostic and therapeutic importance of chronic reductions in regional cerebral perfusion pressure. Further investigations into the importance of hemodynamic factors in ischemic stroke can now be based on accurate assessment of cerebral (not carotid or vertebrobasilar) hemodynamics in the context of other coexisting epidemiological, clinical, hematological, and angiographic risk factors.


Age-associated elongation of the ascending aorta in adults.
To determine whether human aorta lengthens with aging and to evaluate the impact of the hypothesized aortic elongation on pulse wave velocity (PWV) measurements. Although it is generally thought that the aorta becomes tortuous with aging, there has been no systematic study to date in healthy adults to determine if this is so. Such age-related aortic elongation may be a confounding factor for the PWV measurement in elderly people. Arterial lengths were computed by the 3-dimensional transverse magnetic resonance image arterial tracing of the aorta and carotid and iliac arteries in 256 apparently healthy adults (age 19 to 79 years). The ascending aorta was greater with advancing age (r = 0.72), whereas the lengths of the descending aorta and carotid and iliac arteries were not associated with age. The elongation of the ascending aorta was associated with the corresponding increases in aortic PWV (beta = 0.50) and brachial/aortic pulse pressure ratio (beta = 0.24), which is an index of pulse wave amplification. The straight distance between carotid and femoral sites (car-fem), the most popular arterial length measurement, overestimated the aortic length measured with the magnetic resonance image by approximately 25%. The most accurate arterial length estimation was the distance obtained by subtracting carotid length from the car-fem, with <5% difference from the magnetic resonance image-measured length. Because the ascending aorta was omitted or subtracted from the length estimation in PWV, the impact of age-related elongation of the aorta on PWV was small. The aorta lengthens with age, even in healthy humans, due primarily to the elongation of the ascending aorta. Age-related aortic elongation has little impact on PWV measurements, as the ascending aorta, which undergoes lengthening with age, is not included in the arterial length measurements.


Investigation of the interaction between calreticulin and immunoglobulin G by capillary electrophoresis and computer modeling.
Calreticulin (CRT), a highly conserved ubiquitous multifunctional protein, has immense implications in physiological, developmental, and pathological processes due to its interaction with many proteins. In this work, we utilized various capillary electrophoresis (CE) modes for CRT analysis and its interaction with IgG both qualitatively and quantitatively. The interaction was evaluated by capillary zone electrophoresis (CZE) and affinity capillary electrophoresis (ACE) modes that took only 4-6 min for each run. The nanomolar level Kd was determined between CRT and IgG by CZE-LIF and ACE-UV, whereas, CRT did not exhibit any interaction with HSA (as the control protein). By computer modeling, we obtained modeled structures of the CRT-IgG complex which mapped six hot spots on the CRT and IgG interacting interface. Our work is the first report which establishes CE methods for qualitative and quantitative analysis of CRT and IgG interaction. Moreover, the results of computer modeling provide the first molecular insights into the CRT binding sites for IgG that may act as the key anchors and contribute to the interaction studies between CRT and its interacting protein partners.


Involvement of CD45 in dexamethasone- and heat shock-induced apoptosis of rat thymocytes.
CD45 is a transmembrane tyrosine-specific phosphatase which participates in lymphoid cell signal transduction during T cell activation, as well as in intrathymic negative and positive selection. In mammals, this molecule exhibits a variety of isoforms of different molecular weight, whose roles have still to be fully elucidated. We report here that apoptosis of rat thymocytes after in vitro dexamethasone and heat shock treatment was accompanied by an early significative increase of cells expressing CD45RC, the high molecular weight isoform of CD45 molecule. The same phenomenon was observed in thymocytes derived from in vivo dexamethasone-treated rats. However, the increase of CD45RC+ cells was not apparently characteristic of cells undergoing apoptosis, as the same phenomenon was also observed in rat thymocytes induced to proliferate by Concanavalin A. On the whole, these results suggest that CD45 modulation can be added to the list of early molecular events, such as the increased expression of genes (ornithine decarboxylase), proto-oncogenes (c-fos, c-jun, c-myc) and activation of transcription factors (AP-1, NFkB), we previously demonstrated in the same experimental model to occur and to be shared by these two apparently opposite biological processes, i.e., cell proliferation and apoptosis, both likely depending on a complex balance of protein phosphorylation and dephosphorylation.


Catalytic mechanism of SHCHC synthase in the menaquinone biosynthesis of Escherichia coli: identification and mutational analysis of the active site residues.
(1R,6R)-2-Succinyl-6-hydroxy-2,4-cyclohexadiene-1-carboxylate (SHCHC) synthase (MenH) is an alpha/beta fold enzyme containing a catalytically essential serine-histidine-aspartate triad typical of serine proteases but catalyzes a pyruvate elimination reaction initiated by alpha-proton abstraction in the menaquinone biosynthetic pathway of Escherichia coli. In this study, we identify the active site residues in the synthase through sequence analysis and structural modeling and study their mechanistic roles in MenH catalysis. Steady-state kinetic characterization of site-directed mutants of the active site residues shows that three conserved arginine residues (Arg-90, Arg-124, and Arg-168) likely form ionic salt bridges with three carboxylate groups of the substrate in the Michaelis complex and that the side-chain polar groups of the conserved tyrosine (Tyr-85) and tryptophan (Trp-147) residues likely donate hydrogen bonds to form an "oxyanion hole". In addition, the pH dependence of the MenH kinetic properties reveals a catalytic base with a pK(a) highly dependent on the hydroxyl group of the triad serine residue in the enzymatic reaction. Moreover, proton inventory experiments demonstrate that the SHCHC synthase adopts one-proton catalysis like many serine proteases. These results allow the proposal of a mechanism in which the histidine residue of the MenH triad serves as a general base catalyst to deprotonate the triad seryl hydroxyl group in the alpha-proton abstraction from the substrate. As such, the MenH triad performs a simple and fundamental proton transfer reaction occurring repeatedly in the reactions catalyzed by serine proteases and alpha/beta fold hydrolases, suggesting a common evolutionary origin for all serine-histidine-aspartate triads serving different catalytic functions.


Alternative methods of treating pain.
The range of non-invasive methods of treatment of chronic pain which are currently available has been discussed. They fall into three categories; those dependent on counter stimulation, those which influence the higher centres and those, like acupuncture, which possibly combine both mechanisms. The empirical, but frequently successful nature of such treatments has been emphasised.


Mutations of the HIV type 1 V3 loop under selection pressure with neutralizing monoclonal antibody NM-01.
Variants of human immunodeficiency virus type 1 (HIV-1) were selected for resistance to the neutralizing monoclonal antibody (MAb) NM-01. MAb NM-01 recognizes the center of the third hypervariable domain (V3 loop) of the envelope gp120, and neutralizes diverse HIV-1 strains. In the continuous presence of MAb NM-01, transmission and propagation of molecularly cloned HIV-1 were performed in vitro to isolate escape variants. The polymerase chain reaction-single-strand conformation polymorphism and sequence analyses of these variants indicated that the antigenic change against MAb NM-01 is due to a single base substitution resulting in one amino acid interchange within the recognition site of MAb NM-01 in the V3 loop. Mutational analyses also demonstrated a nonrandom event of variability and the existence of mutational hot spots in the V3 loop. The bias of variability could be interpreted by the specificity of error-prone replication by HIV-1 reverse transcriptase. Furthermore, the results suggest that distribution of mutability might correlate closely with the stability of the secondary structure of RNA encoding the V3 loop region.


Direct observation of the spatial distribution of charges on a polypropylene fiber via Electrostatic Force Microscopy.
The spatial distribution of electrical charges along the longitudinal axes of a polypropylene electret fiber was determined using Electrostatic Force Microscopy (EFM). EFM mapping on highly curved surfaces, such as those of polymeric fibers, is a challenging endeavour and most work reported in the scientific literature has been limited to single line-scan analysis or flat specimens. Charged polymeric fibers, electrets, are extensively used in high performance filtration applications and methods to determine the amount and magnitude of the charges on these fibers remain elusive. Electrical charge maps of individual fibers were obtained by biasing the tip to -10 V and maintaining a constant tip-sample distance of 100 nm. Spatially dependant EFM phase and magnitude gradients were determined and the developed technique may provide a unique understanding into the heterogeneous charge distribution on electrets fibers. Direct mapping of the charge distribution in electrets fibers can offer new insights in the development of antistatic additives, new means to facilitate electrostatic self-assembly of nano-moieties on the surface of fibrous materials and a quantitative metrics capable of determining discharge dynamics and predicting the shelf-life of filtration media.


HEMET: mathematical model of biochemical pathways for simulation and prediction of HEpatocyte METabolism.
Many computer studies and models have been developed in order to simulate cell biochemical pathways. The difficulty of integrating all the biochemical reactions that occur in a cell in a single model is the main reason for the poor results in the prediction and simulation of cell behaviour under different chemical and physical stimuli. In this paper we have translated biochemical reactions into differential equations for the development of modular model of metabolism of a hepatocyte cultured in static and standard conditions (in a plastic multiwell placed in an incubator at 37 degrees C with 5% of CO(2)). Using biochemical equations and energetic considerations a set of non-linear differential equations has been derived and implemented in Simulink. This set of equations mimics some of the principal metabolic pathways of biomolecules present in the culture medium. The software platform developed is subdivided into separate modules, each one describing a different metabolic pathway; they constitute a library which can be used for developing new modules and models to project, predict and validate cell behaviour in vitro.


The obesity epidemic: metabolic imprinting on genetically susceptible neural circuits.
The apparent obesity epidemic in the industrialized world is not explained completely by increased food intake or decreased energy expenditure. Once obesity develops in genetically predisposed individuals, their obese body weight is avidly defended against chronic caloric restriction. In animals genetically predisposed toward obesity, there are multiple abnormalities of neural function that prime them to become obese when dietary caloric density and quantity are raised. Once obesity is fully developed, these abnormalities largely disappear. This suggests that obesity might be the normal state for such individuals. Formation of new neural circuits involved in energy homeostasis might underlie the near permanence of the obese body weight. Such neural plasticity can occur during both nervous system development and in adult life. Maternal diabetes, obesity, and undernutrition have all been associated with obesity in the offspring of such mothers, especially in genetically predisposed individuals. Altered brain neural circuitry and function often accompanies such obesity. This enhanced obesity may then be passed on to subsequent generations in a feed-forward, upward spiral of increasing body weight across generations. Such findings suggest a form of "metabolic imprinting" upon genetically predisposed neural circuits involved in energy homeostasis. Centrally acting drugs used for obesity treatment lower the defended body weight and alter the function of neural pathways involved in energy homeostasis. But they generally have no permanent effect on body weight or neural function. Thus, early identification of obesity-prone mothers, infants, and adults and treatment of early obesity may be the only way to prevent the formation of permanent neural connections that promote and perpetuate obesity in genetically predisposed individuals.


EDD, the human hyperplastic discs protein, has a role in progesterone receptor coactivation and potential involvement in DNA damage response.
The ubiquitin-protein ligase EDD encodes an orthologue of the hyperplastic discs tumor suppressor gene, which has a critical role in Drosophila development. Frequent allelic imbalance at the EDD chromosomal locus in human cancers suggests a role in tumorigenesis. In addition to a HECT (homologous to E6-AP carboxyl terminus) domain, the EDD protein contains a UBR1 zinc finger motif and ubiquitin-associated domain, each of which indicates involvement in ubiquitinylation pathways. This study shows that EDD interacts with importin alpha 5 through consensus basic nuclear localization signals and is localized in cell nuclei. EDD also binds progesterone receptor (PR) and potentiates progestin-mediated gene transactivation. This activity is comparable with that of the coactivator SRC-1, but, in contrast, the interaction between EDD and PR does not appear to involve an LXXLL receptor-binding motif. EDD also binds calcium- and integrin-binding protein/DNA-dependent protein kinase-interacting protein, a potential target of ubiquitin-mediated proteolysis, and an altered association is found between EDD and calcium- and integrin-binding protein/DNA-dependent protein kinase-interacting protein in response to DNA damage. The data presented here demonstrate a role for EDD in PR signaling but also suggest a link to cancer through DNA damage response pathways.


DNA-templated silver nanoclusters for fluorescence turn-on assay of acetylcholinesterase activity.
We have developed a fluorescence turn-on assay using DNA-templated silver nanoclusters (Ag NCs) (i.e., 12 polycytosine-templated silver nanoclusters, dC12-Ag NCs), which is amenable to rapid, ultrasensitive assay of acetylcholinesterase (AChE). The detection mechanism is based on the concept, that is, AChE hydrolyzes the acetylthiocholine (ATCh) chloride to produce thiocholine (TCh). Subsequently, TCh sensitively and rapidly reacts with dC12-Ag NCs via Ag-S bond forming and enhances the fluorescence of dC12-Ag NCs. Using dC12-Ag NCs, detection of TCh has a linear concentration range of 2.0 nM to 16.0 nM and a detection limit of 0.3 nM. Due to the sensitive and rapid fluorescence turn-on response of dC12-Ag NCs to TCh, AChE with activity as low as 0.5 × 10(-4) U/mL (signal/noise = 3) can be analyzed with a dynamic range of 0.1 to 1.25 × 10(-3) U/mL. The promising application of the proposed method in AChE inhibitor screening was demonstrated. AChE concentrations were determined in human blood red cell (RBC) membranes from clinical specimens using dC12-Ag NCs, and the quantitative results were validated with Ellman's method. Aside from the ease of manufacture, reduction of matrix effect, and low background noise, the continuous detection format and detection sensitivity can open up to wider applications to AChE activity assay in neurobiology, toxicology, and pharmacology, among other fields.


Staged cranial reconstruction after epidural abscess associated with dural substitute exposure.
Cadaveric dura mater was widely used for a long time but has recently been associated with the transmission of Creutzfelt-Jakob disease. Expanded polytetrafluoethylene sheet has been suggested as a suitable alternative in Japan. Recently epidural abscesses associated with exposure to Expanded polytetrafluoethylene have been on the increase in Japan. We experienced one case of staged cranial reconstruction for chronic ulcer of the head associated with MRSA epidural abscess. the exposed expanded polytetrafluoethylene must be completely removed along with the skin. Reconstruction of dura mater should be made watertight with the application of autologous free fascia lata collected from the paralyzed thigh. Autologous free fascia lata is reconstructed with the rectus abdominis muscle so that there is no dead space remaining. Calvarial bone should be reconstructed in a separate operation as at a second stage.


Greater immediate gastric acid suppression with lansoprazole 30 mg administered as a 2-minute intravenous bolus injection versus a 30-minute infusion.
To compare the pharmacokinetics, pharmacodynamics, and safety of lansoprazole administered as a 2-minute intravenous bolus injection versus a 30-minute continuous infusion. Phase I, open-label, randomized, crossover, single-center trial. Clinical research facility. Thirty-eight healthy volunteers aged 18-55 years. Each subject received one of three regimens. Each regimen contained the same three treatments but in a different sequence: an intravenous injection of lansoprazole 30 mg/10 ml of normal saline over 2 minutes, an infusion of lansoprazole 30 mg/60 ml of normal saline over 30 minutes, and an intravenous injection of 10 ml of normal saline (placebo) over 2 minutes. Each treatment was administered once/day for 7 days, with a washout period of at least 5 days separating each one. Blood samples were assayed for lansoprazole concentrations by using liquid chromatography with tandem mass spectrometry. After the 2-minute administration, mean peak lansoprazole concentrations were about 2-fold higher than those after the 30-minute administration on days 1 and 7. Lansoprazole area under the concentration versus time curve met criteria for bioequivalence on both days. Two-minute administration resulted in a greater percentage of time that the 24-hour pH was above 4 compared with the 30-minute administration (53% vs 47%, p=0.045), with comparable 24-hour integrated gastric acidity (114.4 vs 91.6 mmol*hr/L for 2-min vs 30-min, p=0.335). Significantly greater acid suppression occurred during the first hour after the 2-minute administration compared with that after the 30-minute administration (p < or = 0.001). Safety profiles were similar among the regimens. Greater immediate gastric acid suppression occurred after administration of lansoprazole 30 mg over 2 minutes than over 30 minutes, with other pharmacokinetic, pharmacodynamic, and safety profiles being similar.


Evaluating limited specificity of drug pumps reduced relative resistance in human MDR phenotypes.
In the parallel paper, we developed a property to characterize drug efflux pumps, i.e. the reduced relative resistance (RRR). Using this RRR, we here investigate whether the observed diversity in human multidrug resistance (MDR) phenotypes might be due to variable levels of P-glycoprotein encoded by MDR1. We analyzed resistance phenotypes of various human cell lines in which either one, or both, classical human multidrug resistance genes, MDR1 and MDR3, are overexpressed. In addition, RRR values were calculated for MDR phenotypes presented in the literature. The results suggest that more than a single mechanism is required to account for the observed phenotypic diversity of classical multidrug resistance. This diversity is only partly due to differences in plasma membrane permeabilities between cell line families. It is discussed whether the alternative MDR phenotypes might be MDR1 phenotypes modified by other factors that do not themselves cause MDR. The method we here apply may also be useful for other nonspecific enzymes or pumps.


Suprarenal clamping is a safe method of aortic control when infrarenal clamping is not desirable.
We evaluated the safety of suprarenal aortic clamping in patients with abdominal aortic aneurysm (AAA) treated by open aortic replacement by retrospectively reviewing all patients who underwent elective AAA replacement at a university hospital from 1993 until 2003. We reviewed 249 patient charts and divided them into three groups according to the clamp location during aortic replacement: group 1, infrarenal clamp group (n = 185); group 2, suprarenal clamp group (n = 52); and group 3, supraceliac clamp group (n = 12). Groups 1 and 2 were compared with respect to risk factors, intraoperative events, and postoperative events. Statistical analysis was done using Wilcoxon's rank-sum test, chi-squared test, and Fisher's exact test. Risk factors were comparable in groups 1 and 2 except for weight, which was higher in group 1. Intraoperative urine output, hypotensive episodes, and use of renal protective drugs were comparable in the two groups. Operation time, blood loss, and use of IV fluids were all significantly higher in group 2, while total aortic clamp time was higher in group 1. Postoperative events were comparable except for postoperative peak creatinine, intensive care unit length of stay, and postoperative length of stay, which were higher in group 2; however, discharge creatinine was comparable without a significant difference. Suprarenal clamping is a safe method of aortic control during open AAA replacement surgery. The selection of clamping site should be individualized according to the intraoperative anatomy. Supraceliac clamping is not necessarily the preferable method of aortic control when the infrarenal location is not suitable for clamping.


Pulmonary edema induced by intravenous administration of contrast media: experimental study in rats.
An experimental study of changes in the lung induced by the intravenous administration of iodinated contrast media was carried out using rats. The increase in the relative lung weight and histological findings, especially pulmonary edema, were evaluated in control, physiological saline, diatrizoate sodium meglumine, iohexol, and ioxaglate groups. The relative lung weight increased significantly in all contrast media groups. Proximal and distal perivascular edema were confirmed microscopically. No statistically significant differences among the contrast media were noted in the increase in relative lung weight or in the degree of perivascular edema. There was a close correlation between the increase in relative lung weight and the degree of perivascular edema. These physiological and histological changes in the lung occurred irrespective of the osmolality of the contrast materials. We considered these phenomena to be induced by non-osmolality related toxicity of the contrast medium.


Anti-TNFα agents curb platelet activation in patients with rheumatoid arthritis.
Cardiovascular disease is important in rheumatoid arthritis (RA). Tissue factor (TF) is expressed upon platelet activation and initiates coagulation. Anti-tumour necrosis factor-α (TNFα) agents seem to decrease RA-associated cardiovascular events. We investigated whether (1) TNFα activates human platelets and (2) TNFα pharmacological blockade modulates the platelet-leucocyte reciprocal activation in RA. The expression of platelet TNFα receptors has been assessed by flow cytometry and immunogold electron microscopy. Platelet and leucocyte activation has been assessed also in the presence of antibodies against the TNFα receptors 1 and 2 and of infliximab. TF expression, binding to fibrinogen and phosphatidylserine exposure, has been assessed by flow cytometry, TF activity by coagulation time and by endogenous thrombin generation. Markers of platelet and leucocyte activation have been assessed in 161 subjects: 42 patients with RA, 12 with osteoarthritis, 37 age-matched and sex-matched patients with chronic stable angina and 70 age-matched and sex-matched healthy subjects. TNFα elicited the platelet activation and the expression of TF, which in turn prompted thrombin generation and clot formation. Inhibition of the TNFα-induced activation restricted platelet ability to activate leucocytes and to induce leucocyte TF. TNFα inhibition did not influence platelet activation induced by collagen, ADP or thrombin receptor activating peptide-6. Platelets of patients with RA were more activated than those of controls. Activation was reduced in patients treated with TNFα inhibitors. TNFα-dependent pathways control platelet activation and TF expression in RA. Further studies will verify whether the protective effect of TNFα inhibitors on cardiovascular events involves their ability to modulate platelet function.


Neither MafA/L-Maf nor MafB is essential for lens development in mice.
The importance of the large Maf transcription factor family has been investigated in lens development in the chick, Xenopus and mammals. Previously we reported that c-maf-deficient mice exhibit severe defects in lens fibre cells. Here, we report the roles of other large Mafs, MafA/L-Maf and MafB, during mouse lens development. MafA/L-Maf and MafB were expressed in lens epithelial cells and fibre cells at E12.5 but had largely disappeared from the lens at E18.5. The lens of mafA-, mafB-deficient and mafA::mafB double-deficient mice developed normally. In c-maf-deficient mice, the pattern of expression of MafA and MafB differed from their expression in wild-type mice. Moreover, the expression of crystallin genes was unchanged in mafA-, mafB- and mafA::mafB double-deficient lens. These results indicate that c-Maf alone is essential for lens development, and that MafA/L-Maf and MafB are dispensable in mice.


Accurate visual guidance despite severe neglect.
Unilateral inactivation of the superior colliculus causes profound neglect. In cats, this neglect has been studied previously using tasks that require gaze orientation to, or detection of, a stimulus appearing somewhere in the visual field of an attentive animal. We investigated how neglect affects a completely different kind of task, visually guided foot placement while walking across a cluttered surface. We made muscimol injections into one superior colliculus, and performed perimetry to gauge the extent of the cat's neglect. Cats then walked repeatedly through a cluttered test alley. Most of the time, their gaze was deviated towards the side of the injection, so that they saw the alley floor ahead of them in their neglected hemifield. Surprisingly, they accurately avoided stepping on the densely scattered objects, just as they normally do. We surmise that cats process 'neglected' visual stimuli to guide foot placement even when they are unable to consciously perceive these stimuli.


Sibutramine and L-carnitine compared to sibutramine alone on insulin resistance in diabetic patients.
To evaluate the effects of one year of treatment with sibutramine plus L-carnitine compared to sibutramine on body weight, glycemic control, and insulin resistance state in type 2 diabetic patients. Two hundred and fifty-four patients with uncontrolled type 2 diabetes mellitus (T2DM) [glycated hemoglobin (HbA(1c)) >8.0%] in therapy with different oral hypoglycemic agents or insulin were enrolled in this study and randomised to take sibutramine 10 mg plus L-carnitine 2 g or sibutramine 10 mg in monotherapy. We evaluated at baseline, and after 3, 6, 9, and 12 months these parameters: body weight, body mass index (BMI), glycated hemoglobin (HbA(1c)), fasting plasma glucose (FPG), post-prandial plasma glucose (PPG), fasting plasma insulin (FPI), homeostasis model assessment insulin resistance index (HOMA-IR), total cholesterol (TC), low density lipoprotein-cholesterol (LDL-C), high density lipoprotein-cholesterol (HDL-C), triglycerides (Tg), retinol binding protein-4 (RBP-4), resistin, visfatin, high sensitivity-C reactive protein (Hs-CRP). There was a decrease in body weight, BMI, HbA(1c), FPI, HOMA-IR, and RBP-4 in both groups, even when the values obtained with sibutramine plus L-carnitine were lower than the values obtained in sibutramine group. There was a faster decrease of FPG, PPG, TC, LDL-C, resistin and Hs-CRP with sibutramine plus L-carnitine even when no differences between the two groups were obtained. Furthermore, only sibutramine plus L-carnitine improved Tg, and visfatin. Sibutramine plus L-carnitine gave a faster improvement of lipid profile, insulin resistance parameters, glycemic control, and body weight compared to sibutramine.