Mycoplasma hyopneumoniae and Mycoplasma flocculare differential domains from orthologous surface proteins induce distinct cellular immune responses in mice.
Mycoplasma hyopneumoniae and Mycoplasma flocculare are two genetically close species found in the swine respiratory tract. Despite their similarities, while M. hyopneumoniae is the causative agent of porcine enzootic pneumonia, M. flocculare is a commensal bacterium. Genomic and transcriptional comparative analyses so far failed to explain the difference in pathogenicity between these two species. We then hypothesized that such difference might be, at least in part, explained by amino acid sequence and immunological or functional differences between ortholog surface proteins. In line with that, it was verified that approximately 85% of the ortholog surface proteins from M. hyopneumoniae 7448 and M. flocculare present one or more differential domains. To experimentally assess possible immunological implications of this kind of difference, the extracellular differential domains from one pair of orthologous surface proteins (MHP7448_0612, from M. hyopneumoniae, and MF_00357, from M. flocculare) were expressed in E. coli and used to immunize mice. The recombinant polypeptides (rMHP61267-169 and rMF35767-196, respectively) induced distinct cellular immune responses. While, rMHP61267-169 induced both Th1 and Th2 responses, rMF35767-196 induced just an early pro-inflammatory response. These results indicate that immunological properties determined by differential domains in orthologous surface protein might play a role in pathogenicity, contributing to elicit specific and differential immune responses against each species.


Identification of patterns of transmission of Salmonella within swine production systems using pulsed field gel electrophoresis (PFGE) and repetitive sequence polymerase chain reaction (REP-PCR): a quantitative analysis.
Pulsed field gel electrophoresis (PFGE) using 3 enzymes (Spe I, Xba I, Avr II) and repetitive sequence polymerase chain reaction (REP-PCR) with 3 primers (BOX, ERIC, REP) were compared with respect to their validity as a method for identifying transmission of Salmonella on swine farms. Sixty-eight isolates of Salmonella were obtained from feces of swine, cats, mice, and birds, insect body parts, water and floor samples, and boot scrapings collected on 9 swine farms in Illinois USA. Genetic distances between isolates were calculated using the Dice matching coefficient. Cluster analysis of distance matrices was conducted using the UPG-MA algorithm. There was no significant difference between PFGE and REP-PCR in the genetic diversity detected; however, REP-PCR differentiated between 14 pairs of isolates which PFGE identified as identical. There were no significant differences between PFGE and REP-PCR in identifying all or most close genetic links as isolates from the same farm, the same building, and from the same sampling visit, suggesting ecological validity for both methods. Thus, REP-PCR should be considered as an acceptable and perhaps preferable alternative to PFGE as a genotyping method for studies of Salmonella transmission.


Pharmacokinetics of sodium borocaptate: a critical assessment of dosing paradigms for boron neutron capture therapy.
The pharmacokinetics of sodium borocaptate (BSH), a drug that has been used clinically for boron neutron capture therapy (BNCT) of malignant brain tumors, have been characterized by measuring boron concentrations by direct current plasma-atomic emission spectroscopy (DCP-AES) in a group of 23 patients with high-grade gliomas. The disposition of BSH following intravenous (i.v.) infusion, which was determined by measuring plasma boron concentrations by DCP-AES, was consistent with a three-compartment open model with zero-order input and first-order elimination from the central compartment. Boron disposition was linear over the dose range of 26.5-88.2 mg BSH/kg body weight (b.w.), corresponding to 15-50 mg boron/kg b.w. Mean total body boron plasma clearance was 14.4 +/- 3.5 ml/min and the harmonic mean half-lives (range) were 0.6 (0.3-3.7), 6.5 (4.8-10.1) and 77.8 (49.6-172.0) h for the alpha, beta, and gamma disposition phases, respectively. Using an empirically determined plasma: blood boron concentration ratio of 1.3 +/- 0.2, the calculated total body boron blood clearance was 18.5 +/- 4.5 ml/min. In order to develop a model for selecting the optimum dosing paradigm, a pharmacokinetic correlation was established between the boron content of normal brain, solid tumor, and infiltrating tumor to the shallow tissue pharmacokinetic compartment (C2). Based on our model, it was concluded that although multiple i.v. infusions of BSH might increase absolute tumor boron concentrations, they will not improve the tumor: plasma boron concentration ratios over those attainable by a single i.v. infusion. The results from our study are confirmatory of those previously reported by others when blood sampling has been carried out for a sufficient period of time to adequately characterize the pharmacokinetics.


Multi-institutional validation of the predictive value of Ki-67 in patients with high grade urothelial carcinoma of the upper urinary tract.
We validate the independent predictive value of Ki-67 in patients with high grade upper tract urothelial carcinoma. A total of 475 patients from the international Upper Tract Urothelial Carcinoma Collaboration who underwent extirpative surgery for high grade upper tract urothelial carcinoma were included in this study. Immunohistochemical staining for Ki-67 was performed on tissue microarray formed from this patient cohort. Ki-67 expression was assessed in a semiquantitative fashion and considered over expressed at a cutoff of 20%. Multivariate analyses were performed to assess independent predictors of oncologic outcomes and Harrell's C indices were calculated for predictive models. The median age of the cohort was 69.7 years and 55.2% of patients were male. Ki-67 was over expressed in 25.9% of patients. Ki-67 over expression was significantly associated with ureteral tumor location, higher pT-stage, lymphovascular invasion, sessile tumor architecture, tumor necrosis, concomitant carcinoma in situ and regional lymph node metastases. On Kaplan-Meier analyses over expressed Ki-67 was associated with worse recurrence-free survival (HR 12.6, p <0.001) and cancer specific survival (HR 15.8, p <0.001). On multivariate analysis Ki-67 was an independent predictor of recurrence-free survival (HR 1.6, 95% CI 1.07-2.30, p=0.021) and cancer specific survival (HR 1.9, 95% CI 1.29-2.90, p=0.001). Ki-67 improved Harrell's C index from 0.66 to 0.70 (p <0.0001) for recurrence-free survival as well as cancer specific survival in our preoperative model, and from 0.81 to 0.82 (p=0.0018) for recurrence-free survival and 0.81 to 0.83 (p=0.005) for cancer specific survival in our postoperative model. Ki-67 was validated as an independent predictor of recurrence-free survival and cancer specific survival in patients treated with extirpative surgery for high grade upper tract urothelial carcinoma in a large, multi-institutional cohort.


Allergy and brain tumors in the INTERPHONE study: pooled results from Australia, Canada, France, Israel, and New Zealand.
A history of allergy has been inversely associated with several types of cancer although the evidence is not entirely consistent. We examined the association between allergy history and risk of glioma, meningioma, acoustic neuroma, and parotid gland tumors using data on a large number of cases and controls from five INTERPHONE study countries (Australia, Canada, France, Israel, New Zealand), to better understand potential sources of bias in brain tumor case-control studies and to examine associations between allergy and tumor sites where few studies exist. A total of 793 glioma, 832 meningioma, 394 acoustic neuroma, and 84 parotid gland tumor cases were analyzed with 2,520 controls recruited during 2000-2004. Conditional logistic regression models were used to obtain odds ratios (ORs) and 95 % confidence intervals (CIs) for associations between self-reported allergy and tumor risk. A significant inverse association was observed between a history of any allergy and glioma (OR = 0.73, 95 % CI 0.60-0.88), meningioma (OR = 0.77, 95 % CI 0.63-0.93), and acoustic neuroma (OR = 0.64, 95 % CI 0.49-0.83). Inverse associations were also observed with specific allergic conditions. However, inverse associations with asthma and hay fever strengthened with increasing age of allergy onset and weakened with longer time since onset. No overall association was observed for parotid gland tumors (OR = 1.21, 95 % CI 0.73-2.02). While allergy history might influence glioma, meningioma, and acoustic neuroma risk, the observed associations could be due to information or selection bias or reverse causality.


Modulation of TRPV4 by diverse mechanisms.
Transient receptor potential ion channels (TRP) are a superfamily of non-selective ion channels which are opened in response to a diverse range of stimuli. The TRP vanilloid 4 (TRPV4) ion channel is opened in response to heat, mechanical stimuli, hypo-osmolarity and arachidonic acid metabolites. However, recently TRPV4 has been identified as an ion channel that is modulated by, and opened by intracellular signalling cascades from other receptors and signalling pathways. Although TRPV4 knockout mice show relatively mild phenotypes, some mutations in TRPV4 cause severe developmental abnormalities, such as the skeletal dyplasia and arthropathy. Regulated TRPV4 function is also essential for healthy cardiovascular system function as a potent agonist compromises endothelial cell function, leading to vascular collapse. A better understanding of the signalling mechanisms that modulate TRPV4 function is necessary to understand its physiological roles. Post translational modification of TRPV4 by kinases and other signalling molecules can modulate TRPV4 opening in response to stimuli such as mechanical and hyposmolarity and there is an emerging area of research implicating TRPV4 as a transducer of these signals as opposed to a direct sensor of the stimuli. Due to its wide expression profile, TRPV4 is implicated in multiple pathophysiological states. TRPV4 contributes to the sensation of pain due to hypo-osmotic stimuli and inflammatory mechanical hyperalsgesia, where TRPV4 sensitizaton by intracellular signalling leads to pain behaviors in mice. In the vasculature, TRPV4 is a regulator of vessel tone and is implicated in hypertension and diabetes due to endothelial dysfunction. TRPV4 is a key regulator of epithelial and endothelial barrier function and signalling to and opening of TRPV4 can disrupt these critical protective barriers. In respiratory function, TRPV4 is involved in cystic fibrosis, cilary beat frequency, bronchoconstriction, chronic obstructive pulmonary disease, pulmonary hypertension, acute lung injury, acute respiratory distress syndrome and cough.In this review we highlight how modulation of TRPV4 opening is a vital signalling component in a range of tissues and why understanding of TRPV4 regulation in the body may lead to novel therapeutic approaches to treating a range of disease states.


Influenza A virus potentiates bacteria-induced histamine release. Examination of normal individuals and patients allergic to bacteria.
Influenza A virus was found to enhance basophil histamine release induced by Escherichia coli, Salmonella enteritidis, Staphylococcus aureus, Streptococcus pneumoniae and Streptococcus sanguis, but did not per se release histamine. This potentiating effect of the virus was seen both when the bacteria-induced mediator release was IgE-dependent (i.e. patient allergic to bacterium) and when the bacterium caused histamine release by a non-immunological mechanism independent of IgE (putative sugar-lectin mediated). Also histamine release induced by other immunological and non-immunological stimuli, such as anti-IgE, calcium ionophore or agarose beads was enhanced in the presence of the virus. The potentiating effect of the virus on bacteria-induced mediator release might be of importance for the conversion from latent to manifest asthma in upper respiratory tract infections.


Microcystin production in benthic mats of cyanobacteria in the Nile River and irrigation canals, Egypt.
The present study describes for the first time the species composition and toxicity of benthic cyanobacteria forming mats on the Nile River and irrigation canal sediments in Egypt. A total of 19 species of cyanobacteria were isolated from these mats during this study. The toxicity of the extracts of these species was investigated using Artemia salina assay, mouse bioassay and enzyme linked immunosorbent assay (ELISA). The results showed that all the 19 benthic species isolated from cyanobacterial mats, were toxic to A. salina. Two of these species, namely Calothrix parietina and Phormidium tenue, caused toxicity to mice with neurotoxic signs appeared within 12 h after injection. Whereas, five species showed hepatotoxic effects to mice within 6 h after injection. The results of ELISA showed that all the extracts which had hepatotoxic effects to mice, contained high levels of microcystins with concentrations ranging from 1.6 to 4.1 mg g(-1) dry weight. HPLC analysis for heptotoxic extracts revealed that these extracts contained two peaks corresponding to microcystin-YR and -LR with different proportions. This study suggests that benthic species should be considered along with planktonic species during monitoring of toxic cyanobacteria in water sources, particularly the Nile river which is the main source of drinking water in Egypt.


[Celiac disease].
Celiac disease is an enteropathy due to gluten intake in genetically predisposed individuals (HLA DQ2/DQ8). Celiac disease occurs in adults and children at rates approaching 1% of population in Europe and USA. Clinical features observed in celiac disease are extremely various and anaemia, oral aphthous stomatis, amenorrhea or articular symptoms may be the only presenting manifestations. Diagnosis relies on the evidence of histological villous atrophy in proximal small bowel and the presence of specific serum antibodies. Treatment relies on eviction of gluten (wheat, barley, rye) from diet. Gluten free diet allows prevention of malignant complications such as small bowel adenocarcinoma and lymphoma, and osteopenia. The main cause of resistance to gluten free diet is its poor observance. If not the case, serious complications of celiac disease, such as clonal refractory celiac sprue and intestinal T-cell lymphoma should be suspected. Current therapeutic challenges concern alternative to gluten free diet and new efficient treatments of lymphomatous complications.


The kinetics of the removal of nitrogen and organic matter from parboiled rice effluent by cyanobacteria in a stirred batch reactor.
The aim of this research was to evaluate the kinetics of the removal of nitrogen and organic matter from parboiled rice effluent by the cyanobacteria Aphanothece microscopica Nägeli. From the results obtained, it was verified that maximum efficiency for the removal of organic matter expressed as COD and total nitrogen (N-TKN) occurred after 15 h of cultivation, being 83.44% and 72.74, respectively. The scale-up process indicated that the volume of the estimated reactor would be 100.2m(3), containing parboiled rice effluent and biomass.


Seasonal variation in lipoprotein lipase and plasma lipids in physically active, normal weight humans.
Adipose tissue lipoprotein lipase (ATLPL) provides free fatty acids (FFA) for storage in adipocytes, whereas in skeletal muscle LPL (SMLPL) provides FFA for oxidation. In hibernating animals, the level of SMLPL is relatively higher in summer than winter (promoting fat oxidation), whereas the opposite is seen with ATLPL. A patient-controlled study was designed to determine whether such seasonal variation occurs in normal weight humans. Eighteen subjects were studied in the summer and winter. After 2 days of a standardized diet, they underwent muscle and adipose biopsies for LPL activity, assessment of fitness by VO2 max, and determination of body composition by hydrostatic weighing. The percentages of body fat, body mass index, VO2 max, insulin, glucose, FFA, glycerol, and leptin were not affected by the season. Total cholesterol was higher in the winter than in the summer (157 +/- 5.5 vs. 148 +/- 4.2 mg/dL respectively; P = 0.03). The ATLPL activity was also higher in the winter than in the summer (4.4 +/- 0.8 vs. 2.3 +/- 0.6 nmol FFA/10(6) cells-min; P = 0.04). SMLPL activity trended to be higher in the winter than in the summer (1.9 +/- 0.5 vs. 1.0 +/- 0.1 nmol FFA/g x min; P = 0.06). In summary, ATLPL is seasonally regulated. It appears that SMLPL is similarly regulated by season. For physically active lean subjects, this increase in SMLPL may be a compensatory mechanism to help protect from seasonal weight gain.


In vivo intracellular characteristics of inferior colliculus neurons in guinea pigs.
Intracellular in vivo recordings of physiologically identified inferior colliculus central nucleus (ICc) auditory neurons (n = 71) were carried out in anesthetized guinea pigs. The neuronal membrane characteristics are described showing mainly quantitative differences with a previous report [Nelson, P.G. and Erulkar, S.D., J. Neurophysiol., 26 (1963) 908-923]. The spontaneous spike activity was consistent with the discharge pattern of most extracellularly recorded units. The action potentials showed different spike durations, short and long, and some of them exhibited hyperpolarizing post-potentials. There were also differences in firing rate. The ICc neurons exhibited irregular activity producing spike trains as well as long silent periods (without spikes). Intracellular current injection revealed membrane potential adaptation and shifts that outlasted the electrical stimuli by 20-30 ms. Both evoked synaptic potentials and the spike activity in response to click and tone-burst stimulation were analyzed. Depolarizing-hyperpolarizing synaptic potentials were found in response to contralateral and binaural sound stimulation that far outlasted the stimulus (up to 90 ms). When ipsilaterally stimulated, inhibitory responses and no-responses were also recorded. Although few cells were studied, a similar phenomenon was observed using tone-burst stimulation; moreover, a good correlation was obtained between membrane potential shifts and the triggered spikes (input-output relationship). These in vivo results demonstrate the synaptic activity underlying many of the extracellularly recorded discharge patterns. The data are consistent with the known multi-synaptic ascending pathway by which signals arrive at the ICc as well as the descending corticofugal input that may contribute to the generation of long duration post-synaptic potentials.


Evidence of Hepatitis E virus breaking through the blood-brain barrier and replicating in the central nervous system.
Neurologic dysfunctions such as Guillain-Barre' syndrome, encephalitis, meningitis and transverse myelitis occur frequently in patients with hepatitis E virus (HEV) infection, and this study was conducted to better characterize the role of HEV in the pathogenesis of neurologic disorders. Genotype 4 strain of swine HEV was used to inoculate Mongolian gerbils. Reverse transcription-nested polymerase chain reaction (RT-nPCR), ELISA, histopathology, ultrastructural pathology and enzyme immunohistochemistry method were conducted to investigate the replication and localization of HEV in the central nervous system (CNS) and the consequent pathological changes. Both positive- and negative-strand HEV RNA was detectable in brain and spinal cord from 7 to 28 dpi (days postinoculation) via RT-nPCR. Various pathological changes such as perineural invasion, neuron necrosis, microglia nodule, lymphocyte infiltration, perivascular cuff and myelin degeneration were observed in HEV-positive brains and spinal cords. Immunohistochemical (IHC) staining targeting on HEV ORF2 protein revealed positive signals concentrated mainly in the cytoplasm of neuron, ependymal epithelium and choroid plexus area. Positive area density of ZO-1 (zonula occludens-1) in brain of HEV-positive gerbils decreased, while the GFAP (glial fibrillary acidic protein) expression was upregulated compared with control groups. These results provide strong evidence that HEV is able to damage the blood-brain barrier (BBB), replicate in brain and spinal cord, and hammer the causative role of HEV in the pathogenesis of neurologic disorders.


Iron bound to the lipophilic iron chelator, 8-hydroxyquinoline, causes DNA strand breakage in cultured lung cells.
The formation of DNA-strand breaks was studied in cultured human lung cells (A 549) subjected to iron, either in the form of iron(III) citrate or in combination with the metal chelators ethylene diamine tetra-acetic acid (EDTA), nitrilo triacetic acid (NTA), or 8-hydroxyquinoline (8HQ). After 15 min exposure to 5 microM iron(III) citrate or iron chelate, the cellular levels of iron were found to be three times higher in cells subjected to iron-8HQ than in cells subjected to iron(III) citrate, iron-EDTA or iron-NTA. Exposure to iron-8HQ caused extensive DNA-strand breakage, whereas no such breakage was found in cells exposed to iron-EDTA or iron-NTA. The DNA damage caused by iron-8HQ increased with time and dose, and DNA-strand breakage was clearly demonstrable in cells after 15 min exposure to as little as 0.1 microM iron-8HQ. Moreover, iron-8HQ was strongly toxic to the cells and inhibited their growth after exposure. Along with the formation of DNA-strand breaks, the concentration of cellular malondialdehyde increased four-fold after exposure to iron-8HQ and two-fold after exposure to iron-EDTA or iron-NTA, suggesting that reactive oxygen metabolites might be involved in the toxic action. Moreover, both iron-EDTA and iron-NTA caused a considerable hydroxylation of deoxyguanosine (dG) residues in DNA in vitro, whereas iron(III) citrate and iron-8HQ only caused a minor hydroxylation of dG. This points to the possibility that iron-8HQ-mediated DNA-strand breakage in cells might be due to the action of a metal-bound oxyl radical formed from the iron-8HQ complex rather than to the formation of hydroxyl radicals. Altogether, these findings indicate that iron bound to the lipophilic chelator, 8HQ, has strong toxic properties and that it may cause substantial DNA-strand breakage and lipid peroxidation in living cells.


Enhancement of 5-aminolevulinic acid phototoxicity by encapsulation in polysaccharides based nanocomplexes for photodynamic therapy application.
Polysaccharides based nanocomplexes have been developed for encapsulation, controlled delivery and to enhance the phototoxicity of the photosensitizer 5-aminolevulinic acid for application in photodynamic therapy. The nanocomplexes were prepared by coacervation in a solvent free environment using chitosan as polycation while alginic and polygalacturonic acid as polyanions. The complexes showed average dimension in the range 90-120nm, good stability in simulated physiological media and high drug encapsulation efficiency, up to 800μg per mg of carrier. Release studies demonstrate the possibility to tune the overall release rate and the intensity of the initial burst by changing the external pH. Cytotoxicity and photocytotoxicity tests confirmed the not toxicity of the used polysaccharides. Cell viability results confirmed the improvement of 5-aminolevulinic acid phototoxicity when loaded into the carrier compared to the free form. No effect of the irradiation on the nanocomplexes structure and on the release kinetics of the drug was observed. The results demonstrate that the prepared formulations have suitable properties for future application in photodynamic therapy and to ameliorate the therapeutic efficacy and overcome the side-effects related to the use of the photosensitizer 5-aminolevulinic acid.


Formation of Adventitious Buds in Stem Internodes of Populus spp. Cultured in vitro on Basal Medium: Influence of Endogenous Properties of Explants.
Stem-internodes of the following poplar species formed adventitious buds when cultured in vivo on basal medium in the absence of exogenous growth regulators: Populus nigra, P. deltoides, P. tacamahaca, P. trichocarpa, P. alba, P. wilsonii and Populus hybrid TT32 (P. trichocarpa × P. tacamahaca). The number of buds formed per internodal explant varied with each genotype. Adventitious buds differentiated in tissue produced by cambium and phloem cells. Capacity for bud formation was highest at the physiological apical end of explants. Morphogenesis was reduced in explants cultured with an axillary bud. Formation of adventitious buds was greatest during the initial period of explant culture; bud-formation was reduced or absent in re-cultured explants. Bud-forming capacity was related to length of explant; explants 2mm in length were unresponsive whereas a mean of eight buds were obtained per 8mm explant. Internodes close to the shoot apex failed to produce buds and morphogenic competence increased with increasing distance from the apex.


Corporate colonization of health activism? Irish health advocacy organizations' modes of engagement with pharmaceutical corporations.
This article is based on a study that aimed to shed light on the "cultures of action" of Irish health advocacy organizations, and particularly their modes of engagement with pharmaceutical corporations. Debates about what some interpret as the "corporate colonization" of health activism provide the backdrop for the analysis. The empirical dimension of the study involved a survey of 112 organizations and in-depth study of a small number of organizations that manifest diverse modes of engagement with the pharmaceutical industry. The varying modes of interaction are plotted along a continuum and characterized as corporatist, cautious cooperation, and confrontational. Evidence is presented of a strong and growing cultural tendency in Irish health advocacy organizations to frame pharmaceutical corporations as allies in their quests for better health. The analysis of four constitutive dimensions of organizations' cultures of action can reveal the legitimating logics underlying their diverging positions around pharmaceutical industry sponsorship. While the research shows that pharmaceutical corporations have largely succeeded in defining themselves as a philanthropic force and rightful players in Irish health activism, it cautions against a simplistic conclusion that this is evidence of corporate colonization.


[Mitral valve replacement after previous coronary artrey bypass grafting( CABG) with functioning left internal thoracic artery( LITA) grafts in an elderly patient; report of a case].
An 85-year-old woman had a history of coronary artery bypass grafting (CABG) performed 7 years ago, and dyspnea on effort had been worsening recently. Since echocardiography showed severe mitral valve regurgitation( MR), mitral valve repair was suggested. Preoperative enhanced computed tomography (CT) showed the patent functioning left internal thoracic artery (LITA) graft. Mitral valve replacement (MVR) using a 25 mm CEP bioprosthesis was performed successfully via resternotomy without any intraoperative injury of the heart. Myocardial protection without clamping of functioning LITA was done by both antegrade and retrograde continuous coronary perfusion (RCCP) under mild hypothermia. The postoperative clinical course was uneventful without any hemodynamic compromise. She was discharged on postoperative day 21 without any cardiac events following early introduction of cardiac rehabilitation. From these results, mitral valve reoperation by RCCP under mild hypothermia without control of functioning internal thoracic artery( ITA) grafts could be a safe option in some cases.


A randomized controlled trial comparing ceftriaxone with cefazolin for antibiotic prophylaxis in abdominal hysterectomy.
To compare the effectiveness of ceftriaxone versus cefazolin for the prevention of febrile morbidity and postoperative infections among patients after abdominal hysterectomy. In a double-blind, randomized, controlled trial in Bangkok, Thailand, 320 patients undergoing abdominal hysterectomy between July 2008 and July 2009 were randomly assigned to receive 1g of either ceftriaxone or cefazolin intravenously in a single dose before surgery. The participants were evaluated for postoperative fever and infection for up to 4 weeks. χ(2) or Fisher exact tests were used for statistical analysis. There was no significant difference between the ceftriaxone and cefazolin groups in incidence of febrile morbidity (9.4% versus 11.2%), wound infection (3.8% versus 1.9%), vaginal cuff infection (3.8% versus 1.9%), or urinary tract infection (1.9% versus 1.9%). There was no difference between the use of single-dose preoperative ceftriaxone and cefazolin in preventing infectious morbidity among patients undergoing hysterectomy.


Yield enhancement strategies for artemisinin production by suspension cultures of Artemisia annua.
Artemisinin, isolated from the shrub-Artemisia annua, is a sesquiterpene lactone used to treat multi-drug resistant strains of falciparum malaria. It is also effective against a wide variety of cancers such as leukemia and colon cancer. To counter the present low content in leaves and uneconomical chemical synthesis, alternate ways to produce artemisinin have been sought. But this compound remains elusive in cell cultures of A. annua despite the extensive studies undertaken. This work reports the first successful approach for production of artemisinin by cell cultures of Indian variety of A. annua. In the present study, an integrated yield enhancement strategy, developed by addition of selected precursor (mevalonic acid lactone) and elicitor (methyl jasmonate) at optimized concentrations, resulted in 15.2g/l biomass and 110.2mg/l artemisinin, which was 5.93 times higher in productivity in comparison to control cultures.


Straight-wire appliances: standard versus individual prescription.
In this article the individual patient (IP) appliance is described. It consists of 250 options of bracket and band variations as the straight wire appliances. Increasing the bracket capabilities means using an increasing number of brackets, each with a specific design created for a treatment situation. The objective of IP appliance is to eliminate wire bending from orthodontic treatment and improve the treatment results. To manage this technique, a computer software is needed. Internet offers significant possibilities in managing each patient by patient basis. The clinician is required to make the diagnosis and treatment plan before ordering the appliance. Two clinical cases are described with the aim to present the advantages of this technique.


[60]Fullerene-fused lactones: manganese(III) acetate-mediated synthesis and novel reductive ring opening.
[reaction: see text] [60]Fullerene-fused lactones were prepared by the manganese(III) acetate-mediated reactions of [60]fullerene with carboxylic acids, carboxylic anhydrides, or malonic acids. Novel reductive ring opening of the lactones with Grignard reagents was observed.


Reliability and validity of a simulation-based acute care skills assessment for medical students and residents.
Medical students and residents are expected to be able to manage a variety of critical events after training, but many of these individuals have limited clinical experiences in the diagnosis and treatment of these conditions. Life-sized mannequins that model critical events can be used to evaluate the skills required to manage and treat acute medical conditions. The purpose of this study was to develop and test simulation exercises and associated scoring methods that could be used to evaluate the acute care skills of final-year medical students and first-year residents. The authors developed and tested 10 simulated acute care situations that clinical faculty at a major medical school expects graduating physicians to be able to recognize and treat at the conclusion of training. Forty medical students and residents participated in the evaluation of the exercises. Four faculty members scored the students/residents. The reliability of the simulation scores was moderate and was most strongly influenced by the choice and number of simulated encounters. The validity of the simulation scores was supported through comparisons of students'/residents' performances in relation to their clinical backgrounds and experience. Acute care skills can be validly and reliably measured using a simulation technology. However, multiple simulated encounters, covering a broad domain, are needed to effectively and accurately estimate student/resident abilities in acute care settings.


Activity of paclitaxel in advanced or recurrent squamous cell cancer of the cervix.
Twenty-six patients with squamous cell cancer of the cervix were treated with i.v. paclitaxel, 250 mg/m2 over 3 h every 21 days. They received steroid, H1 and H2 blocker premedications, and granulocyte-colony-stimulating factor (G-CSF) support (5 microgram/kg/day). No prior chemotherapy, except as a radiation sensitizer, was allowed. The median age was 50 (range, 36-81) years, and performance status Zubrod was 1 (range, 0-2). Eight (33%) patients had prior surgery, and 22 (92%) had prior radiation therapy. Twenty-four patients were evaluable for response; 2 were later found to be ineligible. Five patients had partial responses (21%; 95% confidence interval, 6-40%), and 14 (58%; 95% confidence interval, 35-78%) had stable disease. The median duration of response was 10 (range, 3-27+) weeks. The responses were within the radiation port (four responses) and outside of it (one response). The median interval from the start of irradiation to the start of paclitaxel in responding patients was 94 weeks, whereas in patients with stable disease it was 68 weeks, and in patients whose disease progressed it was 46 weeks. Eighty-eight percent of the 105 cycles of paclitaxel were administered at a dose of 250 mg/m2 or higher. Granulocytopenia was brief and noncumulative, with grades 3 and 4 experienced by 5 and 3 patients, respectively. G-CSF was used for a median of 7 (range, 2-14) days/cycle. Anemia was mild, with G3 noted in 3 patients, and thrombocytopenia was not significant. Infections and musculoskeletal pain were mild and infrequent. Sensory (14 patients G1 or G2 and 2 patients G3) and motor (4 patients G1 or G2 and 1 patient G3) neurotoxicity was noted. There was no significant cardiovascular toxicity. Paclitaxel is active in patients with squamous cell cancer of the cervix and is well tolerated at this dose schedule with G-CSF support.


Novel two-dimensional reversed-phase liquid chromatography/hydrophilic interaction chromatography, an excellent orthogonal system for practical analysis.
A novel two-dimensional reversed-phase liquid chromatography/hydrophilic interaction chromatography (2D-RPLC/HILIC) system is developed with the introduction of the click beta-cyclodextrin (beta-CD) stationary phase. The offline system shows excellent abilities for the separation of polar and medium-polarity components in traditional Chinese medicine (TCM). It facilitates us not only to separate components that cannot be resolved by uni-dimensional chromatography, but also to achieve much more efficient detection of components with low abundance. The orthogonality of this new system is excellent. A simple geometric approach is developed to characterize the practical orthogonality of 2D-LC in the analysis of complex unknown samples. The mathematical characterization results are in good accordance with the experimental findings, and both demonstrate that the proposed 2D-RPLC/HILIC system is a powerful tool for the separation of polar and medium-polarity complex samples. This system may have a great potential for the separation of peptides and metabolomic compounds with similar polarity.


Histopathological and capillaroscopical features of the cuticles and bleeding clots in ring or middle fingers of systemic scleroderma patients.
Sixty-three patients with systemic scleroderma (SSc) (Barnett I, 41; Barnett II, 17; Barnett III, 5), 14 with systemic lupus erythematosus (SLE), 9 with dermatomyositis (DM) and 10 healthy controls (HC) were subjected to histopathological examinations of the cuticles of ring or middle fingers. The sex ratios (male/female) in the patients with SSc, SLE, DM and HC were 7:56, 5:9, 5:4 and 5:5, and the ages were 22-74, 19-78, 45-70 and 13-78 years old, respectively. Biopsy samples were taken from the central portion of the cuticles, which showed the most severe change of elongation with or without bleeding clots of cuticle-proximal nailfolds (BC). Histopathologically, 61 (96.8%) cuticles of SSc patients consisted of the upper (U), middle (M) and lower (L) layers, which represent obliquely stacked, parabolic, and parallel stacked layers, respectively. The middle parabolic layer appeared to discharge homogenous eosinophilic globular deposits (ED). On the other hand, this typical three-layer-nail pattern was seen only in 9 (64.3%) of SLE, 3 (33.3%) of DM and none of HC, in total 12 (36.4%) of the non-SSc group, which included SLE, DM and HC. In SSc, there were statistical correlations (R2) between ED and BC, ED and cuticle-elongation, cuticle-layer and cuticle-elongation, ED and cuticle-layer, BC and cuticle-elongation. Capillaroscopically, bleeding clots located in the middle layer with ED of the cuticles in eight patients with SSc were transported rapidly within 1-2 weeks.(ABSTRACT TRUNCATED AT 250 WORDS)


Constituents of the Egyptian Centaurea scoparia; Chlorinated Guaianolides of the Aerial Parts.
A new chlorine-containing guaianolide, chloroscoparin, was isolated from the ethanol extract of the aerial parts of CENTAUREA SCOPARIA Sieb. together with three known chlorine-containing guaianolides. The structures of the isolated compounds were elucidated by spectroscopic methods including (1)H-NMR-, 2D (1)H- (1)H COSY-, (13)C-NMR-, APT-, DEPT-, HETCOR-, long-range-HETCOR-, 2D INADEQUATE-, and mass spectra. The relative configuration was determined by 2D-NOESY studies. New NMR data for the known compounds are reported.


Bisphosphonate-related osteonecrosis of the jaws: a review of 34 cases and evaluation of risk.
The purpose of this study was to identify factors that influence bisphosphonate-related osteonecrosis of the jaws (BRONJ). Patients undergoing treatment for BRONJ (n=34) were evaluated. Sex, age, underlying diagnosis, type of bisphosphonate (BP), duration and route of administration, location of osteonecrosis, clinical symptoms, Actinomyces colonisation, treatment and outcome were recorded. Symptom onset was analysed with respect to BP potency and cumulative dose. Underlying diagnoses indicating BP-treatment included multiple myeloma, breast carcinoma, prostate carcinoma and osteoporosis. In 31 patients, BRONJ was preceded by tooth extraction, root apicotomy, ill-fitting dentures, cystenucleation, implant insertion or trauma; in 3 patients, the precipitating event was not identified. Actinomyces colonisation was observed in 18 patients (53%). The occurrence of BRONJ was not directly related to BP dose or potency. More women with multiple myeloma had BRONJ than did males. BRONJ was observed in osteoporotic patients treated with both corticosteroids and BPs. BRONJ was not primarily associated with BP potency or dose. Factors that increased the risk of osteonecrosis were female sex, oral surgery and corticosteroids plus intravenous or oral BP administration. BP deposition in the jaw bones might enhance BRONJ by promoting bacterial colonisation; however, this hypothesis requires more study.


A study on small-world brain functional networks altered by postherpetic neuralgia.
Understanding the effect of postherpetic neuralgia (PHN) pain on brain activity is important for clinical strategies. This is the first study, to our knowledge, to relate PHN pain to small-world properties of brain functional networks. Functional magnetic resonance imaging (fMRI) was used to construct functional brain networks of the subjects during the resting state. Sixteen patients with PHN pain and 16 (8 males, 8 females for both groups) age-matched controls were studied. The PHN patients exhibited decreased local efficiency along with non-significant changes of global efficiency in comparison with the healthy controls. Moreover, regional nodal efficiency was found to be significantly affected by PHN pain in the areas related to sense (postcentral gyrus, inferior parietal gyrus and thalamus), memory/affective processes (parahippocampal gyrus) and emotional activities (putamen). Significant correlation (p<0.05) was also found between the nodal efficiency of putamen and pain intensity in PHN patients. Our results suggest that PHN modulates the local efficiency, and the small-world properties of brain networks may have potentials to objectively evaluate pain information in clinic.


Amyotrophic lateral sclerosis mortality in Italy, 1958 to 1987: a cross-sectional and cohort study.
We analyzed the mortality rates of amyotrophic lateral sclerosis (ALS) in Italy from 1958 to 1987. The overall mean annual mortality rate, adjusted to the 1981 Italian population, was 0.68/100,000 (95% confidence interval, 0.64 to 0.72); 0.86 for men; and 0.62 for women. During that period, mortality increased for both sexes: from 1958/1962 to 1983/1987, mortality increased by 60% for women and 24% for men. The slope for women was 0.07, and for men, 0.02. Moreover, the mortality rate increased in older age groups and was stable for subjects under 45 years old. There was no clear birth-cohort effect, but in any cohort the age-specific mortality rate increased with advancing age, with no decline for the oldest age. Based on these data, the increase of ALS mortality rates is probably due to the effect of changing methodologic and demographic variables and the decrease of mortality of some competing diseases, rather than a real rise of the risk in the whole population or in specific birth cohorts.


Women's and men's negative experience of child birth-A cross-sectional survey.
A negative birth experience may influence both women and men and can limit their process of becoming a parent. This study aimed to analyze and describe women's and men's perceptions and experiences of childbirth. A cross-sectional study of women and their partners living in one Swedish county were recruited in mid pregnancy and followed up two months after birth. Women (n=928) and men (n=818) completed the same questionnaire that investigated new parents' birth experiences in relation to socio-demographic background and birth related variables. Women (6%) and men (3%) with a negative birth experiences, experienced longer labours and more often emergency caesarean section compared to women (94%) and men (97%) with a positive birth experience. The obstetric factors that contributed most strongly to a negative birth experience were emergency caesarean and was found in women (OR 4.7, 95% CI 2.0-10.8) and men (OR 4.5, Cl 95% 1.4-17.3). In addition, pain intensity and elective caesarean section were also associated with a negative birth experiences in women. Feelings during birth such as agreeing with the statement; 'It was a pain to give birth' were a strong contributing factor for both women and men. A negative birth experience is associated with obstetric factors such as emergency caesarean section and negative feelings. The content of negative feelings differed between women and men. It is important to take into account that their feelings differ in order to facilitate the processing of the negative birth experience for both partners.


Serum selenium levels in individuals on PKU diets.
Serum selenium levels in 73 patients with phenylketonuria were significantly lower than in controls. The phenylketonuric and hyperphenylalaninaemic individuals taking the non-supplemented amino acid mixture generally had lower levels: 36% were below the normal range as defined by our laboratory, compared with 19% in the supplemented group. The low levels were present even in those on diet, who had a greater phenylalanine tolerance--that is, a tolerance for more than 9 x 50 mg phenylalanine exchanges per day, in other words a higher intake of natural protein. Individuals on long-term synthetic diets may be at risk for selenium deficiency even on selenium supplements. In areas where the soil may be low in selenium, the deficiency may be aggravated. Long-term low levels may impair health but the required amount of selenium supplementation remains uncertain.


Two-dimensional agarose gel electrophoresis as a tool to isolate genus- and species-specific repetitive DNA sequences.
Two-dimensional electrophoresis in agarose gels separates DNA-restriction fragments not only by molecular weight but also according to their AT-cluster content. The method produced genus-specific spot patterns of multicopy DNA fragments of grains as well as spot patterns of highly repetitive DNA fragments of ciliates, demonstrated for barley, spelt, and Tetrahymena. Further investigations in regard to their specificity by hybridization with three other grain species (wheat, oat, and rye) and three ciliate species (Tetrahymena thermophila, Tetrahymena pigmentosa, and Tetrahymena borealis) were performed. The DNA samples from spelt and Tetrahymena were demonstrated to be genus specific for Triticum and species specific for Tetrahymena pyriformis, respectively.


[Rxtrinsic determinants for using complementary medical therapies].
Numerous studies have explored intrinsic factors that influence the use of complementary and alternative medicine (CAM). Extrinsic determinants of patients' choice of therapy have seldom been explored. This study analyzes the effects of extrinsic determinants on the first-time use of CAM. Furthermore, it analyzes how extrinsic determinants influence the choice of particular CAM methods and the frequency of CAM use. The analysis bases on a data set generated by a survey using standardized questionnaires. 31.8% out of a1,000-patient sample responded. 86.6% of the respondents were aged 21-60 years, 79.5% were female. The analysis employs descriptive statistics, univariate tests and multivariate fractional logit models. Recommendations by friends and family are the strongest determinant of first time use of CAM. The strongest determinant of the choice ofa particular therapy is its perceived efficacy. Patients with as trong preference for effective therapies use CAM more frequently(OR 1.687; 90% CI 1.040-3.378). In particular, these patients use phytotherapy (1.787; 1.040-3.372), homeopathy(2.282; 1.161-10.806) and traditional Chinese medicine(TCM) (2,386; 1,421-4,396) more frequently. Patients with a strong preference for physicians' recommendations use manipulative therapies more frequently (1,610; 1,177-2,374),and energetic therapies less frequently (0,451; 0.261-0,693). Patients valuing high-profile therapies (0,714; 0.585-0,883)and expert recommendations (0,608; 0.508-0,732) use CAM less frequently. Socio-demographic characteristics do not affect the total usage of CAM. The usage pattern of CAM does not only depend on intrinsic factors. It is also determined by extrinsic factors.


Vertical transmission of HIV in Ghanaian women diagnosed in cord blood and post-natal samples.
HIV RNA detection in the newborn is the main diagnostic tool for vertical transmission. Most infections are thought to occur peri- or post-natally, hence preventive antiviral therapy administered days before and during delivery. This study used cord blood for molecular diagnosis, examined viral load and HIV-1 subtypes as determinants of transmission, and compared molecular variability of maternal, cord blood, and post-natal quasispecies. Ninety-seven seropositive mother-cord blood paired plasmas from Ghana were tested for HIV RNA. Viral load was quantified and a subgroup of 45 random women samples was typed and subtyped. HIV-1 from infected pairs was cloned, sequenced, and analyzed phylogenetically. The prevalence of HIV infection in pregnant women was 3.3%. 13/97 cord blood samples (13.5%) contained HIV RNA. No correlation between either viral load at labor (range 10(3)-10(7)) or HIV-1 subtype and in utero transmission was found. In both transmitting and non-transmitting mothers, 56% of HIV-1 strains were CRF02_AG. In three pairs, maternal and cord blood quasispecies were closely related, suggesting late pregnancy or perinatal transmission, while in four pairs, genetic distances suggested transmission earlier during gestation. Maternal viral load and genotype did not correlate with HIV-1 pre-natal transmission. HIV infection during gestation appears relatively frequent.


Tracking children from poverty at risk for specific language impairment: a 3-year longitudinal study.
A 3-year longitudinal study of the language performance of children from poverty was designed to address the problem of separating children with a specific language impairment (SLI) from low-scoring normal children in the borderline area on the continuum of language performance where normal ends and abnormal begins. Two approaches to definition were compared: an experimental approach (using story-retelling, rote-memory ability, and invented-morpheme learning) and a traditional approach (using standardized-test discrepancy scores). Results indicated that 6 of 34 children tracked from kindergarten through second grade appeared to be SLI at the end of the study. The best kindergarten predictor for the outcome status of these 6 children was a combination of the score on the Oral Vocabulary subtest of the TOLD-2P and the score on a combination of the experimental tasks. The best single kindergarten predictor of the academic status of the 15 children in the study who received academic remediation was story-retelling. Children's scores on the experimental and standardized tests of language performance and nonverbal intelligence were profiled over the 3 years of the study, and patterns of change in many instances reveal the lifting of the early influences of poverty.


Cost-effectiveness of five strategies for gonorrhea and chlamydia control among female and male emergency department patients.
Previous studies have shown screening for gonorrhea and chlamydia to be cost-effective for limiting the sequelae of infection and the associated costs of management. To evaluate the cost-effectiveness of enhanced screening for gonorrhea and chlamydia in an emergency department (ED) setting. Five strategies were compared with use of decision analysis for theoretical cohorts of 10,000 female and 10,000 male ED patients aged 18 years to 31 years: standard ED practice, three enhanced screening strategies, and mass treatment. Main outcome measures were untreated gonorrhea or chlamydia cases and their sequelae, transmission to a partner, congenital outcomes, and cost to prevent a case. This analysis, from the perspective of the healthcare sector, included medical case costs expressed in US dollars (1999), discounted at an annual rate of 3%. Mass treatment was the most cost-effective strategy among women and men. Of the screening strategies for women, universal screening combined with standard practice was the most cost-effective; it was used for treating 499 more cases of gonorrhea and chlamydia than was standard practice, saving $95.70 per case treated. Standard ED practice remained the most cost-effective strategy for men under a variety of circumstances. The authors recommend urine ligase chain reaction screening for gonorrhea and chlamydia in women aged 18 years to 31 years in the ED, in conjunction with standard ED practice, to decrease the occurrence of the sequelae and costs associated with infection.


Long-term effects of local ionizing radiation treatment on Langerhans cells in mouse footpad epidermis.
Langerhans cells (LC) were studied with ADPase histochemistry in sheets of hind footpad epidermis from groups of CBA/H mice. Single, local 20-Gy doses of 250-kV x-rays were administered to the right hind feet of the mice when they were 3-4 months old, and LC were counted at intervals ranging from 2 to 24 months later. In unirradiated mice, aged 5-19 months, the mean density of LC in footpad was 1521-1617 cells/mm2. It dropped to 1137 +/- 86 cells/mm2 (mean +/- SE) in untreated 28-month-old mice. At times from 2-15 months after irradiation, normal mean densities of LC were present in footpad epidermis. On average, LC numbers were subsequently reduced to 1078 +/- 65/mm2 by 19 months after irradiation (71% of the cells in age-matched controls) and to 789 +/- 53/mm2 by 24 months (59% of the cells in age-matched controls). Loss of cells was focal. Chronic radiation-induced fibrosis and damage to circulatory function in skin may have contributed to impaired replacement of LC from bone marrow precursors. The possibility that late radiation-related depletion of the LC population permits development of skin tumors as a delayed consequence of exposure to ionizing radiation is discussed.


Pharmacokinetics and drug interactions with zonisamide.
Polypharmacy is a widely employed treatment strategy in epilepsy, particularly for individuals with poorly controlled seizures. Drug combinations should be carefully considered to minimize the potential for unfavorable interactions. Older-generation antiepileptic drugs (AEDs) are well known for their pharmacokinetic interaction potential, which generally results from alterations in the metabolism of concomitant drugs due to effects on the cytochrome P450 (CYP) and uridine glucuronyl transferase enzyme systems. Newer agents, such as zonisamide, are less likely to cause adverse drug interactions. A series of interaction studies has revealed zonisamide to be without effect on the steady-state pharmacokinetics of carbamazepine, phenytoin, sodium valproate, or lamotrigine. However, zonisamide is principally inactivate by CY3A4-dependent reduction. Consequently, carbamazepine, phenytoin, and phenobarbital all increase its clearance, an interaction that may necessitate a dosage increase, but which will also permit more rapid attainment of steady-state zonisamide concentrations. Otherwise, zonisamide is essentially devoid of clinically significant interactions with other AEDs, oral contraceptives and, indeed, all other classes of therapeutic agents investigated to date. As a result, it is reasonable to conclude that zonisamide has a favorable pharmacokinetic profile and that it may be a useful and uncomplicated agent when employed as adjunctive therapy in refractory epilepsy.


Intraoperative Intratumoral Embolization of a Complex Recurrent Hemangiopericytoma: Technical Report and Review of the Literature.
Objective Recurrent brain tumors represent a challenge for neurosurgeons because of the extensive blood loss and the time needed for surgical resection. Only a few hemostatic agents are useful to prevent the bleeding and thus facilitate the surgical resection. Fibrin sealant can be used to achieve sealing, tissue adherence, or hemostasis when other means of hemostasis are inadequate or inappropriate. We report the feasibility and positive effects of direct intratumoral injection of fibrin sealant during resection of a recurrent hemangiopericytoma. Material and Methods The intraoperative intratumoral injection of fibrin sealant changed the tumor properties of a recurrent hemangiopericytoma of the tentorium with infra- and supratentorial extension. From a loose friable briskly bleeding tumor, this complex lesion became a nonbleeding well-demarcated soft-firm tumor that could easily be dissected off the pial surface and totally resected without extensive bleeding. Results There are several benefits of intratumoral injection of fibrin sealant in hemangiopericytomas: (1) the extensive bleeding is diminished and blood loss minimized; (2) the restriction of the surgical view by the venous oozing is diminished, making the microsurgical dissection of the tumor capsule off the pial surface easier and safer; (3) the loose consistency of the tumor becomes firmer and facilitates the manipulation of the tumor and leads to a safer resection; and (4) a shorter operating time is needed. Conclusion The use of intratumoral fibrin glue injection is a safe and useful technique that could be used for hemostasis of highly vascularized tumors to facilitate a safer resection and to reduce blood loss.


Role of a bound ubiquinone on reactions of the Escherichia coli cytochrome bo with ubiquinol and dioxygen.
To probe the functional role of a bound ubiquinone-8 in cytochrome bo-type ubiquinol oxidase from Escherichia coli, we examined reactions with ubiquinol-1 and dioxygen. Stopped-flow studies showed that anaerobic reduction of the wild-type and the bound ubiquinone-free (delta UbiA) enzymes with ubiquinol-1 immediately takes place with four kinetic phases. Replacement of the bound ubiquinone with 2,6-dibromo-4-cyanophenol (PC32) suppressed the anaerobic reduction of the hemes with ubiquinol-1 by eliminating the fast phase. Flow-flash studies in the reaction of the fully reduced enzyme with dioxygen showed that the heme b to heme o electron transfer occurs with a rate constant of approximately 10(4) s-1 in all three preparations. These results support our previous proposal that the bound ubiquinone is involved in facile oxidation of substrates in subunit II and subsequent intramolecular electron transfer to low-spin heme b in subunit I.


Propionic acid-induced calcium mobilization in human neutrophils.
The ability of propionic acid to elicit an increase in the level of cytoplasmic free calcium in human neutrophils was examined in detail. Propionic acid induced a rapid and dose-dependent mobilization of calcium that relied on both internal and external sources of calcium. The effects of propionic acid on the mobilization of calcium were inhibited by pertussis toxin, but not cholera toxin, implicating a guanine nucleotide binding protein. Furthermore, preincubation of the neutrophils with phorbol 12-myristate 13-acetate resulted in a decreased mobilization of calcium. This inhibitory activity of phorbol myristate acetate was antagonized by the protein kinase C inhibitor H-7. Preincubation of the cells with the synthetic chemotactic factor fMet-Leu-Phe caused a reduction in the magnitude of the calcium transient elicited by propionic acid. However, the calcium response to propionic acid was not affected by antagonists of fMet-Leu-Phe and platelet-activating factor binding or by an inhibitor of leukotriene synthesis. Propionic acid did not elicit a mobilization of calcium in monocytes, platelets, lymphocytes, or undifferentiated HL-60 cells. However, the treatment of the HL-60 cells with dimethylsulfoxide resulted in the appearance of a calcium response to propionic acid. The potential physiological significance of these findings are discussed.


Combined internal and external fixation of distal radius fractures.
Combined internal and external fixation of distal radius fractures is used most commonly to treat injuries with joint surface or metaphyseal comminution. External fixation aids reduction intraoperatively and facilitates arthroscopic, per-cutaneous, or open manipulation of the fracture. Internal fixation maintains precise reduction of critical anatomy, principally the contour and orientation of the articular surface. Postoperatively the fixator functions as a neutralization device, preventing fracture collapse and decreasing the biomechanical demands on the internal fixation hardware. The combined technique exploits the benefits of both forms of fixation, allowing each to be used to full advantage in the treatment of complex distal radius fractures.


Subject Profile Assessment of Cases Recommended with Nutritional Supplement Containing Red Yeast Rice, Grape Seed Extract and Black Pepper Extract: A Prospective, Multicentric, Post-marketing Study.
To assess the profile and general health indices in subjects prescribed with a nutritional supplement containing red yeast rice, grape seed extract and black pepper extract (NS-YGP; PreLipid®; Abbott Healthcare Pvt. Ltd.) in routine clinical setting in India. In this prospective, postmarketing observational study, consecutive subjects recommended NS-YG at their outpatient department were recruited from 39 sites in India. The primary outcome of interest was to identify the profiles of subjects recommended with NS-YGP. The secondary outcome measures were to evaluate the changes in lipid profile from baseline to 3 and 6 months and assess the safety of NS-YGP. A total of 573 subjects were enrolled, of which 527 (92%) completed the study. Majority of subjects receiving NS-YGP were males (76.6%), non-obese (85.9%) and had borderline dyslipidemia (87.3%) at the time of enrolment. There was a significant improvement in lipid profile over a period of 6 months; with a significant decline in the levels of total cholesterol, low-density lipoprotein cholesterol, non-high-density lipoprotein cholesterol (non-HDL-C) and triglycerides from baseline to month 3 and 6 (p < 0.0001 for each parameters). Furthermore, HDL-C level significantly improved from baseline till month 6 (p = 0.0007). There was a significant change in levels of other laboratory parameters, including creatine phosphokinase, serum creatinine, hemoglobin and white blood cells from baseline to 3 and 6 months (p< 0.05). No adverse events were reported in the study. Most of the subjects prescribed with NS-YG had borderline elevated lipid levels. NS-YGP significantly improved lipid profile and was safe, well-tolerated and can be recommended in the primordial prevention of borderline dyslipidemia.


Universal method facilitating the amplification of extremely GC-rich DNA fragments from genomic DNA.
Polymerase chain reaction (PCR) is a basic technique with wide applications in molecular biology. Despite the development of different methods with various modifications, the amplification of GC-rich DNA fragments is frequently troublesome due to the formation of complex secondary structure and poor denaturation. Given the fact that GC-rich genes are closely related to transcriptional regulation, transcriptional silencing, and disease progression, we developed a PCR method combining a stepwise procedure and a mixture of additives in the present work. Our study demonstrated that the PCR method could successfully amplify targeted DNA fragments up to 1.2 Kb with GC content as high as 83.5% from different species. Compared to all currently available methods, our work showed satisfactory, adaptable, fast and efficient (SAFE) results on the amplification of GC-rich targets, which provides a versatile and valuable tool for the diagnosis of genetic disorders and for the study of functions and regulations of various genes.


Prevalence and Outcomes of Patients Meeting Palliative Care Consultation Triggers in Neurological Intensive Care Units.
Palliative care needs among patients in the neurological intensive care unit (neuroICU) are poorly characterized. Our aim was to explore the prevalence and type of potential palliative care consultation triggers in neuroICUs. We conducted a retrospective cohort study of neuroICU admissions in Project IMPACT from 2001 to 2008. We assessed the prevalence of neuroICU admissions meeting one or more of five validated palliative care consultation triggers and compared the percentage of admissions meeting these triggers in other ICUs from the same hospitals. Among 1268 admissions to 2 neuroICUs, 200 (15.8 %) met one or more triggers for palliative care consultation. Among 13,694 admissions to non-neuroICUs in the same hospitals, 1909 (13.9 %) met one or more palliative care triggers (p = 0.44). The most common trigger in the neuroICU was intracerebral hemorrhage with mechanical ventilation (n = 92; 7.3 %). The most common trigger in non-neuroICUs was ICU admission following ≥10-day hospital stay (n = 805; 5.9 %). Although ICU mortality was not significantly higher in neuroICU vs. non-neuroICU patients meeting triggers (23.4 vs 19.9 %, p = 0.46), neuroICU patients were significantly more likely to have withdrawal of life-sustaining therapies (19.4 vs 8.0 %, p < 0.001). Among neuroICU patients, 15.8 % met triggers for palliative care consultation. Although prevalence of admissions meeting any trigger was similar amongst all ICUs, neuroICU admissions met different types of triggers and were more likely to have withdrawal of life-sustaining therapy. These data suggest that palliative care needs are common among neuroICU patients and discussions with patients and families regarding limitation of life-sustaining therapy may differ in this setting.


[Lateral suboccipital puncture of the cisterna magna (author's transl)].
Lateral puncture of the cisterna magna cerebellomedullaris offers a more efficient conception of the lateral approach for the use of positive and negative contract medium in myelography and cisternography. The extracranial part of the cisterna magna cerebellomedullaris gives enough space to ensure the definite fixation of the needle without any major risk after puncture through the atlanto-occipital membrane. Positioning of the needles tip is possible without x-Ray control. Injury of the spinal cord is most unlikely to occur.


Hb Santa Clara (beta 97His-->Asn), a human haemoglobin variant: functional characterization and structure modelling.
This study examines the functional and structural effects of amino acid substitution at alpha(1)beta(2) interface of Hb Santa Clara (beta 97His-->Asn). We have characterized the variation by a combination of electrospray ionisation mass spectrometry and DNA sequence analysis followed by oxygen-binding experiments. Functional studies outlined an increased oxygen affinity, reduced effect of organic phosphates and a reduced Bohr effect with respect to HbA. In view of the primary role of this interface in the cooperative quaternary transition from the T to R conformational state, a theoretical three-dimensional model of Hb Santa Clara was generated. Structural investigations suggest that replacement of Asn for His beta 97 results in a significant stabilization of the high affinity R-state of the haemoglobin molecule with respect to the low affinity T-state. The role of beta FG4 position has been further examined by computational models of known beta FG4 variants, namely Hb Malmö (beta 97His-->Gln), Hb Wood (beta 97His-->Leu), Hb Nagoya (beta 97His-->Pro) and Hb Moriguchi (beta 97His-->Tyr). These findings demonstrate that, among the various residues at the alpha(1)beta(2) (and alpha(2)beta(1)) intersubunit interface, His beta FG4 contributes significantly to the quaternary constraints that are responsible for the low oxygen affinity of human deoxyhaemoglobin.


Tripeptidyl peptidase II is dispensable for the generation of both proteasome-dependent and proteasome-independent ligands of HLA-B27 and other class I molecules.
A significant fraction of the HLA-B27-bound peptide repertoire is resistant to proteasome inhibitors. The possible implication of tripeptidyl peptidase II (TPPII) in generating this subset was analyzed by quantifying the surface re-expression of HLA-B*2705 after acid stripping in the presence of two TPPII inhibitors, butabindide and Ala-Ala-Phe-chloromethylketone. Neither decreased HLA-B27 re-expression under conditions in which TPPII activity was largely inhibited. This was in contrast to a significant effect of the proteasome inhibitor epoxomicin. The failure of TPPII inhibition to decrease surface re-expression was not limited to HLA-B27, since it was also observed in several HLA-B27-negative cell lines, including Mel JuSo. Actually, HLA class I re-expression in Mel JuSo cells increased as a function of butabindide concentration, which is consistent with an involvement of TPPII in destroying HLA class I ligands. Inhibition of TPPII with small interfering RNA also failed to decrease the surface expression of HLA class I molecules on 143B cells. Our results indicate that TPPII is dispensable for the generation of proteasome-dependent HLA class I ligands and, without excluding its role in producing some individual epitopes, this enzyme is not involved to any quantitatively significant extent, in generating the proteasome-independent HLA-B27-bound peptide repertoire.


Pituitary-adrenal axis function in systemic inflammatory response syndrome.
This study characterizes the hypothalamic-pituitary-adrenal axis function in patients with sepsis syndrome now known as systemic inflammatory response syndrome (SIRS). One hundred and thirteen patients with SIRS had their pituitary-adrenal axis tested with the use of a 250 µg IV ACTH stimulation test. No patient received corticosteroids prior to the ACTH stimulation test. Serum cortisol concentrations were measured prior to and 30 and 60 min after ACTH administration. 26% of the patients had bacteremia, 22% bacteruria, 22% AIDS, 17% renal failure, 15% diabetes, 13% severe liver disease, 8% GI bleed, 4% pancreatitis, 3% trauma and 1% classical Addison's disease. Several patients had more than one disorder. The overall mortality was 28%. Multivariate analysis identified that both the baseline cortisol concentration and delta cortisol concentration were significant indicators of mortality. Despite the fact that mortality was double in the bacteremic patients, the baseline cortisol concentrations were similar to the non-bacteremic patients (25.4 ± 1.9vs 25.1 ± 2.5 µg/dl). The only cortisol abnormality noted in the bacteremic patients was a significantly smaller delta cortisol response to ACTH (14.7 ± 2.2vs 18.9 ± 1.2 µg/dl;P<0.05). These data imply that bacteremic processes may alter the adrenal responsiveness to intravenous ACTH administration. Understanding the pathophysiological disturbances responsible for an impaired adrenal reserve may identify new treatment strategies for patients with bacteremia.


The present and future of cardiac arrest care: international experts reach out to caregivers and healthcare authorities.
The purpose of this review is to describe the epidemiology of out-of-hospital cardiac arrest (OHCA), disparities in organisation and outcome, recent advances in treatment and ongoing controversies. We also outline the standard of care that should be provided by the critical care specialist and propose future directions for cardiac arrest research. Narrative review with contributions from international resuscitation experts. Although it is recognised that survival rates from OHCA are increasing there is considerable scope for improvement and many countries have implemented national strategies in an attempt to achieve this goal. More resources are required to enable high-quality randomised trials in resuscitation. Increasing international collaboration should facilitate resuscitation research and knowledge translation. The International Liaison Committee on Resuscitation (ILCOR) has adopted a continuous evidence review process, which facilitate the implementation of resuscitation interventions proven to improve patient outcomes.


A bi-monomeric, nonlinear Becker-Döring-type system to capture oscillatory aggregation kinetics in prion dynamics.
In this article, in order to understand the appearance of oscillations observed in protein aggregation experiments, we propose, motivate and analyse mathematically the differential system describing the kinetics of the following reactions: [Formula: see text] with n finite or infinite. This system may be viewed as a variant of the seminal Becker-Döring system, and is capable of displaying sustained though damped oscillations.


Surgical treatment of complicated pilonidal sinus with a fasciocutaneous V-Y advancement flap.
In this study, we investigated the value of fasciocutaneous V-Y advancement flap reconstruction in the treatment of complicated or recurrent sacrococcygeal pilonidal sinus. Patients with complicated pilonidal sinus treated with a V-Y advancement flap from July 2002 through December 2005 were studied. The duration of inpatient stay, early wound complications, time of return to active work, recurrence, and patient satisfaction with the operation were recorded. Forty-three patients were treated for recurrent or complicated sacrococcygeal pilonidal disease. All flaps remained viable without necrosis. However, 4 patients (9.3%) had wound infections, and 3 patients (7%) had hematoma. The mean hospital stay was 3 (range, 2-5) days, and time to return to active life was 17 (range, 13-25) days. The mean duration of follow-up was 48 (range, 27 to 69) months. Recurrence was detected in only one patient in the early period. Seven patients (16.3%), including 4 bilateral repairs, had numbness over the operative site. Thirty-nine patients (90.7%) were not satisfied with scars; 4 reported that the scar was not important. In selected patients, V-Y advancement flaps provide tension-free repair with minimal recurrence rates and reliable skin coverage while flattening the natal cleft that predisposes to recurrence. Because patient dissatisfaction with cosmetic results is high, potential treatment outcomes should be discussed with patients in detail before the operation.


Mechanisms of leucocyte sodium influx in essential hypertension.
1. Leucocyte Na+ influx in media containing 10 mmol/l Na+ was studied directly using a triple-isotope method for measuring initial 22Na uptake rates in 20 normal and 18 untreated hypertensive subjects. The effects of 1 mmol/l amiloride (a Na+-H+-antiport inhibitor) and 0.1 mmol/l bumetanide (a Na+,K+,Cl-symport inhibitor) were also examined. 2. The total, amiloride-sensitive and bumetanide-sensitive influx rates were raised in hypertensive compared with normotensive subjects [median (range): total 0.63 (0.25-1.82) vs 0.40 (0.09-0.65) mmol min-1 l-1, P less than 0.002; amiloride-sensitive 0.43 (0.18-1.56) vs 0.26 (0.04-0.48) mmol min-1 l-1, P less than 0.002; bumetanide-sensitive 0.12 (-0.03 to 0.83) vs 0.02 (-0.25 to 0.21) mmol min-1 l-1, P less than 0.005]. 3. We conclude that hypertensive patients have a raised leucocyte total Na+ influx when measured in media containing 10 mmol/l Na+ and that this is contributed mainly by amiloride-sensitive and bumetanide-sensitive Na+ influx mechanisms.


Secreted Frizzled related protein-4 (sFRP4) promotes epidermal differentiation and apoptosis.
The skin provides vital protection from infection and dehydration. Maintenance of the skin is through a constant program of proliferation, differentiation and apoptosis of epidermal cells, whereby proliferating cells in the basal layer differentiating to form the keratinized, anucleated stratum corneum. The WNT signalling pathway is known to be important in the skin. WNT signalling has been shown to be important both in epidermal development and in the maintenance and cycling of hair follicles and epidermal stem cells. However, the precise role for this pathway in epidermal differentiation remains unknown. We investigated the role of the WNT signalling inhibitor sFRP4 in epidermal differentiation. sFRP4 is expressed in both normal skin and keratinocytes in culture. Expression of sFRP4 mRNA and protein increases with keratinocyte differentiation and apoptosis, whilst exposure of keratinocytes to exogenous sFRP4 promotes apoptosis and expression of the terminal differentiation marker Involucrin. These data suggest sFRP4 promotes epidermal differentiation.


Correlates of HIV vaccine trial participation: an Indian perspective.
Successful conduct of HIV vaccine trials in a population of great cultural diversity like India could be a challenge. Concerns, knowledge gaps and willingness to participate in future HIV vaccine trials were studied among 349 patients attending three sexually transmitted infections clinics and one reproductive tract infections clinic. Overall willingness to volunteer for HIV vaccine trials was 48%. Women and men at risk of HIV infection were willing to participate in the HIV vaccine trials. Factors associated with increased willingness to participate in these trials were awareness of current HIV vaccine efforts, realization of importance of vaccine for self, concern about adverse events and altruism.


Computer handling of occupational exposure data.
A system has been developed for the computer handling of industrial hygiene data in the Michigan Division of Dow Chemical U.S.A. With the implementation of this system, future epidemiological studies might be completed in a few weeks as opposed to months when done menually. The system is capable of providing lists, either current or historical, of the chemicals used in a particular building; and all the available exposure data for specific employees can be obtained throughout their entire periods of employment. The system is also capable of summarizing all available exposure information for a given chemical. The heart of the chemical exposure data system lies in the record content. Each record contains: 1. Chemical identification number; 2. Employee number; 3. Job classification number; 4. Department index number; 5. Data type code; 6. Exposure data; 7. Units code; 8. Date; 9. Building number and 10. Reference. A detailed discussion of each of these parameters is contained in the text.


Role of 5-lipoxygenase pathway in the regulation of RAW 264.7 macrophage proliferation.
Arachidonic acid (AA) metabolites control cell proliferation, among other physiologic functions. RAW 264.7 macrophages can metabolise AA through the cyclooxygenase and lipoxygenase (LOX) pathways. We aimed to study the role of AA-metabolites derived from 5-LOX in the control of RAW 264.7 macrophage growth. Our results show that zileuton, a specific 5-LOX inhibitor, and nordihydroguaiaretic acid (NDGA), a non-specific LOX inhibitor, inhibit cell proliferation and [(3)H]-thymidine incorporation in a concentration-dependent fashion. Growth inhibition induced by NDGA can be explained by an apoptotic process, while zileuton does not seem to induce apoptosis. Moreover, these treatments delay the cell cycle, as analysed by flow cytometry. On the other hand, the leukotriene (LT) B(4) receptor antagonist U-75302, the LTD(4) receptor antagonists LY-171883 and MK-571, and the cysteinyl-LT receptor antagonist REV-5901 also inhibit cell proliferation and [(3)H]-thymidine incorporation in a concentration-dependent manner, and delay the RAW 264.7 cell cycle. However, these antagonists did not induce annexin V staining, caspase activation or DNA fragmentation. Furthermore, we demonstrated that exogenous addition of LTB(4) or LTD(4) revert the cell growth inhibition induced by zileuton or the leukotriene receptor antagonists mentioned above. Finally, we observed that LTB(4) and LTD(4), in the absence of growth factors, have pro-proliferative effects on macrophages, and we obtained preliminary evidences that this effect could be through mitogen-activated protein kinase (MAPK) and phosphatidylinositol 3-kinase (PI3K) pathways. In conclusion, our results show that the interaction between LTB(4) and LTD(4) with its respective receptor is involved in the control of RAW 264.7 macrophage growth.


Kidney donation--where some of the problems lie.
If a viable kidney transplant programme is to be continued on a national basis, many of the problems at present surrounding organ transplantation must be tackled urgently. This paper discusses the psychological and emotional problems which are presently seen to hinder a successful renal transplantation policy, and considers the relative advantages and disadvantages of the kidney donor card scheme.


[A future image of clinical inspection from health economics].
Do you let medical costs increase in proportion to the growth rate of GDP? A way of thinking of the Council on Economic and Fiscal Policy. Should we exclude public medical insurance? It is not a problem, it is an absolute sum if you are effective. If there is no insurance, and individuals pay the total amount, there is no problem, but it is impossible. Economic development will cease if there is no insurance. As medical personnel, to offer good medical care with an appropriate cost. An appeal to the nation is necessary. Economic technical evaluation to identify a cheap method for each clinical inspection. Does medical insurance have a deficit? I. Japanese Health insurance system. (1) Health insurance union. When you look at the contribution money, it is originally 2,479,800,000,000 yen, with premium income and a profit of 45%. (2) Government management health insurance. When you look at the contribution money, it is originally 2,163,300,000,000 yen, with premium income and a profit of 36%. (1) + (2) Employed insurance meter. (3) Mutual aid. (4) National Health Insurance. II. A clinical economic method. III. Expense of medical care and its effect. A. Expense. B. A medical economic technical evaluation method. 1. Cost-effectiveness analysis CEA. 2. Cost utility analysis CUA. 3. Cost-benefit analysis CBA. 4. Expense minimization analysis.


Ethnic variations in risk factor profile, pattern and recurrence of non-cardioembolic ischemic stroke.
Ischemic stroke is influenced by ethnic and geographical variations. The aim of this study was to identify the risk factor profiles, subtypes and recurrence of non-cardioembolic ischemic stroke for the two largest subpopulations, the Arab and South Asians, at the only stroke-admitting hospital in Qatar. Data on stroke patients admitted to Hamad Medical Corporation from January through December 2001 were reviewed in January 2005. Only patients with non-cardioembolic ischemic stroke and complete work-up were included. A total of 303 patients with ischemic non-cardioembolic stroke fit the entry criteria. Sixty seven percent of the overall patient population (sex ratio 2:6, M/F) was of Arab origin, and 32% were South Asians. Hypertension was the most commonly encountered risk factor followed by dyslipidemia, diabetes mellitus, and obesity. Significant differences between the Arab and South Asian subgroup of patients were observed with respect to number of risk factors and occurrence of obesity and diabetes. Carotid artery stenotic lesions, ventricular wall motion abnormalities and stroke recurrence were observed with a higher frequency in the Arab subgroup of patients compared with the South Asians. The majority of strokes were lacunar hemispheric strokes (68%), followed by lacunar brainstem strokes (15%) and large-vessel hemispheric infarctions (10%). Patients with a previous history of stroke had a higher frequency of carotid artery stenosis (p = 0.05) and risk of stroke recurrence (p = 0.04). Unlike in other studies originating from the Arabian Gulf, lacunar stroke is the most common subtype of non-cardioembolic ischemic stroke in both the Arabs and South Asians in Qatar. Significant ethnic differences in age of occurrence, risk factor profile, and cardiovascular variables were observed.


Dorsoventral patterning and oligodendroglial specification in the developing central nervous system.
While the bulk of oligodendendrocytes are generated postnatally in rodents, it is now clear that the first oligodendrocytes are born during midembryonic development. Recent studies imply that the first oligodendrocytes to appear are specified concurrently with certain neuronal subtypes. In addition, patterning molecules known to confer positional information on neural tissues during development, such as sonic hedgehog and bone morphogenetic proteins, have also been implicated in the specification of glial fate. This review discusses some of the recent advances in our knowledge of how oligodendrocytes are generated and the mechanisms by which this might occur in the developing brain and spinal cord.


Total synthesis of (-)-morphine.
We have developed an efficient total synthesis of (-)-morphine in 5% overall yield with the longest linear sequence consisting of 17 steps from 2-cyclohexen-1-one. The cyclohexenol unit was prepared by means of an enzymatic resolution and a Suzuki-Miyaura coupling as key steps. Construction of the morphinan core features an intramolecular aldol reaction and an intramolecular 1,6-addition. Furthermore, mild deprotection conditions to remove the 2,4-dinitrobenzenesulfonyl (DNs) group enabled the facile construction of the morphinan skeleton. We have also established an efficient synthetic route to a cyclohexenol unit containing an N-methyl-DNs-amide moiety.


Role of reverse transcription in the generation of extrachromosomal copia mobile genetic elements.
The Drosophila genetic element copia is one of the best studied eukaryotic transposable sequences. Copia shares structural features with a wide variety of mobile elements in Drosophila, Lepidoptera, yeast and vertebrates, the last class being the retrovirus proviruses. Furthermore, retrovirus-like particles containing copia RNA have been isolated from Drosophila cells and extrachromosomal circular copias with structures closely resembling circular retrovirus proviruses have been isolated and cloned. Therefore, copia-like elements and retroviruses may be members of a class of mobile genetic elements existing throughout the eukaryotic kingdom. Consequently, there has been speculation that retroviruses evolved from transposable elements and, conversely, that copia-like elements transpose as retroviruses or retrovirus-like particles. To date, however, there has been no demonstration that copia RNA is reverse transcribed into copia DNA. The present report describes the isolation of linear extrachromosomal copias whose structure closely resembles the analogous retrovirus provirus linears and whose synthesis is unaffected by inhibitors of the cellular DNA polymerase responsible for chromosomal DNA replication.


Late responses after transcranial magnetic stimulation in stroke.
In addition to early responses after transcranial magnetic stimulation, usually designated as Motor Evoked Potentials (MEP), several late responses have been reported in intervals up to 500 ms following stimulus. Our attention in this work was focused on the response in interval 150-300 ms after stimulus output, which is also designated as S > 150 response. We monitored longitudinally late S > 150 response in group of 19 stroke patient with verified hemispherical ischemic lesion. Our results have shown that the presence of late S > 150 response on the first day after stroke predicts an excellent improvement of clinical deficit and that late S > 150 response is more sensitive to hemispheric lesion than early MEPs. From this point of view, recording of late responses can supplement early MEP recording. Our study demonstrated that supratentorial structures are involved in the origin of the S > 150 response. According to the observation that in some patients a normal early response was present and an S > 150 response was absent on the hemiparetic side, and both responses were present on the unaffected side, we hypothesize that cortical structures play an important role in S > 150 response.


A tailored approach to asthma management: Arg(16) holds the key?
Asthma is heterogeneous with respect to clinical presentation, underlying disease mechanisms and response to existing drugs making tailored therapy desirable. Pharmacogenetics, the study of the influence of genetic polymorphisms on drug efficacy and/or adverse effects, is relatively advanced in asthma with replicated genetic associations identified in the main drug classes. In the present issue of Clinical Science, Lipworth and co-workers report a proof-of-concept study and demonstrate that, in asthmatic children carrying the β(2)-adrenergic receptor gene Arg(16) polymorphism, a combination of corticosteroid plus leukotriene receptor antagonist provides superior asthma control (e.g. quality of life scores) compared with corticosteroid plus a long-acting β(2)-adrenergic receptor agonist as add-on therapy. The basis of these observations is well founded, as it has been demonstrated previously that the Arg(16) polymorphism may confer an increased risk of exacerbation following prolonged β(2)--adrenergic receptor agonist use. These results suggest Gly16Arg genotyping in Caucasian asthma patients may have a role in the clinical management of asthma by influencing the decision of which add-on therapy to prescribe; however, larger studies are required to provide definitive conclusions regarding the clinical utility of this approach.


Digital amputations.
Digital amputations may be indicated for trauma, tumor, transplantation, infection, peripheral vascular disease, congenital deformities, and chronic pain. This article reviews indications, general principles of amputation, surgical procedures, complications, postoperative course and rehabilitation, and clinical cases.


Narrative text analysis of accident reports with tractors, self-propelled harvesting machinery and materials handling machinery in Austrian agriculture from 2008 to 2010 - a comparison.
The aim of this study was the identification of accident scenarios and causes by analysing existing accident reports of recognized agricultural occupational accidents with tractors, self-propelled harvesting machinery and materials handling machinery from 2008 to 2010. As a result of a literature-based evaluation of past accident analyses, the narrative text analysis was chosen as an appropriate method. A narrative analysis of the text fields of accident reports that farmers used to report accidents to insurers was conducted to obtain detailed information about the scenarios and causes of accidents. This narrative analysis of reports was made the first time and yielded first insights for identifying antecedents of accidents and potential opportunities for technical based intervention. A literature and internet search was done to discuss and confirm the findings. The narrative text analysis showed that in more than one third of the accidents with tractors and materials handling machinery the vehicle rolled or tipped over. The most relevant accident scenarios with harvesting machinery were being trapped and falling down. The direct comparison of the analysed machinery categories showed that more than 10% of the accidents in each category were caused by technical faults, slippery or muddy terrain and incorrect or inappropriate operation of the vehicle. Accidents with tractors, harvesting machinery and materials handling machinery showed similarities in terms of causes, circumstances and consequences. Certain technical and communicative measures for accident prevention could be used for all three machinery categories. Nevertheless, some individual solutions for accident prevention, which suit each specific machine type, would be necessary.


Living cell manipulation, manageable sampling, and shotgun picoliter electrospray mass spectrometry for profiling metabolites.
A modified cell pressure probe and an online Orbitrap mass spectrometer were used to sample in situ plant single cells without any additional manipulation. The cell pressure probe, a quartz capillary tip filled with an oil mixture, was penetrated to various depths into parenchyma cells of tulip bulb scale, followed by a hydraulic continuity test to determine the exact location of the tip inside target cells. The operation was conducted under a digital microscope, and the capillary tip was photographed to calculate the volume of the cell sap sucked. The cell sap sample was then directly nebulized/ionized under high-voltage conditions at the entrance of the mass spectrometer. Several sugars, amino acids, organic acids, vitamins, fatty acids, and secondary metabolites were detected. Because picoliter solutions can be accurately handled and measured, known volumes of standard solutions can be added to cell sap samples inside the capillary tip to be used as references for metabolite characterization and relative quantitation. The high precision and sensitivity of the cell pressure probe and Orbitrap mass spectrometer allow for the manipulation and analysis of both femtoliter cell sap samples and standard solutions.


[Field epidemiological study on news reports that related to public health emergencies].
All news reports (NR) that were related to public health emergency (PHE) were collected from the Southern Metropolis Daily (SMD) to explore the characteristics of epidemiology in the fields. Based on the theory of communication that including both case and text analysis, qualitative analysis on all the NR regarding PHE published in SMD from the years of 2008 to 2012, was carried out and input to database using the EpiData. Numbers of articles as indicators were compared to show the yearly change of different types of events. Various features of the NR including coverage, source of information, location of the incident, style and size of news, with or without editorials etc. were statistically analyzed by SPSS version 18.0. Among all the 998 reports related to PHE, higher proportion was found in the events of Infectious diseases (35.3%) and food safety (34.1%)respectively. Events on vaccines and drugs used for disease prevention and control (8.9%), environmental pollution caused incidents (8.0%)appeared to be less frequent. Events related to occupational disease, poisoning, bioterrorism and biochemical events were rare. Looking at the monthly distribution of reports, we noticed that the peaks occurred in 2008 and in 2009, which were caused by the Melamine-contamination events and the 2009 H1N1 pandemic. Between 2010 and 2012, figures of monthly reports were smooth, including some critical events from the interests of the media. Most events took place in Guangdong province (34.3%) and other provinces (50.9%), with some were from Hong Kong, Macao and Taiwan regions (9.5%). However, international events (5.2%)were less seen. Extensive coverage accounted for 17.6% of all of reports, and 11.5% allotted the editorials or other forms of in-depth reports. Most of the source of reports on infectious diseases and food safety were from the official release, however. The main sources of occupational diseases and poisoning, vaccines and drug incidents, environmental pollution related incidents were reported by active journalists through interview. Reports on hand, foot and mouth disease, influenza, milk safety, AIDS and lead pollution showed continued concern in the past five years by SMD. NR on public health emergencies by SM had encompassed all 10 categories-related events formulated by the Ministry of Health. Sustained and in-depth coverage were more commonly seen. Field-epidemiologists should learn interdisciplinary sciences on the theory and methodology of communication. They also need to interact with media people during the whole processes of public health emergency preparedness and responses.


Functional characterization of calcium-signaling pathways of human skin-derived mesenchymal stem cells.
Mesenchymal stem cells (MSCs) derived from adult human tissues are able to differentiate into various specialized cell types. In research, they can therefore be used like embryonic cells but without the ethical restrictions. Among the various human tissues, skin as a source is characterized by great accessibility and availability using noninvasive procedures and is without the risk of oncogenesis after transplantation. The recent isolation of MSCs has shown the lack of knowledge regarding their specific features, including the calcium-signaling pathways. In this study, we isolated MSCs from human skin biopsies (S-MSCs) and characterized them phenotypically and their calcium-signaling pathways by the means of Ca2+ imaging and video microscopic experiments. The cytofluorimetric analysis of the expression of surface markers on S-MSCs revealed that they express the normal pattern present on MSCs. Interestingly, these cells appeared to be successfully cryopreserved at early passages. Calcium imaging on single S-MSCs shows that these cells did not display significant spontaneous activity or a response to a depolarizing agent. However, ATP or acetylcholine-induced intracellular calcium increase via ionotropic or metabotropic receptors, respectively. The results presented here reveal that S-MSCs show morphological and functional features that make them useful as an in vitro model to study cell differentiation.


Proton and iodine-127 nuclear magnetic resonance studies on the binding of iodide by lactoperoxidase.
Interaction of an iodide ion with lactoperoxidase was studied by the use of 1H NMR, 127I NMR, and optical difference spectrum techniques. 1H NMR spectra demonstrated that a major broad hyperfine-shifted signal at about 60 ppm, which is ascribed to the heme peripheral methyl protons, was shifted toward high field by adding KI, indicating the binding of iodide to the active site of the enzyme; the dissociation constant was estimated to be 38 mM at pH 6.1. The binding was further detected by 127I NMR, showing no competition with cyanide. Both 1H NMR and 127I NMR revealed that the binding of iodide to the enzyme is facilitated by the protonation of an ionizable group with a pKa value of 6.0-6.8, which is presumably the distal histidyl residue. Optical difference spectra showed that the binding of an aromatic donor molecule to the enzyme is slightly but distinctly affected by adding KI. On the basis of these results, it was suggested that an iodide ion binds to lactoperoxidase outside the heme crevice but at the position close enough to interact with the distal histidyl residue which possibly mediates electron transport in the iodide oxidation reaction.


Delayed recurrent massive thromboembolism through a vena caval clip after pulmonary embolectomy.
A case of recurrent massive pulmonary embolization through a modified Miles' clip two weeks after successful emergency pulmonary embolectomy is reported. Vena caval ligation is probably a safer alternative in these critically ill patients.