Real-World Effectiveness of the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) mRNA Vaccines in Preventing Confirmed Infection in Patients on Chronic Hemodialysis.
Persons on chronic hemodialysis have a significantly diminished humoral immune response to SARS-CoV-2 vaccines. Whether this translates to reduced vaccine effectiveness (VE) is unknown. We used the US Department of Veterans Affairs COVID-19 Shared Data Resource to identify all veterans who were tested for SARS-CoV-2 between 26 January and 31 August 2021. Using International Classification of Diseases, 10th edition, codes and attendance at a dialysis clinic/center, we identified those who were on chronic hemodialysis. We used a test-negative, case-control design using a doubly robust logistic regression model to determine the VE of the BNT-162b2 (Pfizer) or mRNA-1273 (Moderna) vaccines in preventing confirmed SARS-CoV-2 infection. Among 847 199 veterans tested for SARS-CoV-2 between 26 January and 31 August 2021, there were 6076 veterans on chronic hemodialysis. Among those, we identified 1270 cases (580 fully vaccinated) and 2959 controls (2120 fully vaccinated). The overall VE >14 days after the second dose in preventing documented infection was 68.2% (95% CI: 62.6-72.9%). VE was 68.9% (95% CI: 61.9-74.7%) for Pfizer BNT-162b2 and 66.7% (95% CI: 58.9-73.0%) for Moderna mRNA-1273 vaccine. There was no difference in VE by age (<70 vs >70 years), race, or sex. There were no events recorded in persons with a Charlson's comorbidity index score <2. VE of 2 doses of current mRNA vaccines in preventing SARS-CoV-2 infection in persons on chronic hemodialysis is lower than historic VE rates in the general population. Effects of additional doses in improving VE in this special population need further study.


Spin-polarized Fermi surface, hole-doping and band gap in graphene with boron impurities.
Embedding foreign atoms in graphene and interchanging the underlying substrate are proved to be efficient methods for manipulating the properties of graphene. Combining ARPES experiments with DFT calculations we show that boron-doped graphene (B-graphene) grown on a Co(0001) substrate by chemical vapor deposition (CVD) becomes hole doped and its Fermi surface near the K-point reveals strongly spin-polarized states. The latter stems from the spin-polarized mini Dirac cone that is an intrinsic two-dimensional feature of the graphene/Co(0001) interface and is formed by a mixture of C 2pz and Co 3d states. Since the CVD method allows the achievement of up to 20 at% of incorporated B atoms, this provides a certain flexibility for handling the spin-polarized properties of the system. We also show that the bonding of the B-graphene layer to the Co(0001) substrate can be released by intercalation of Li into the interface. This allows the exploration of the doping effect in detail. Finally, our ARPES data indicate a gap opening in the Dirac cone as a result of the highly unbalanced boron concentrations in the two graphene sublattices.


Who does the numbers? The role of third-party technology assessment to inform health systems' decision-making about the funding of health technologies.
There is an increasing number of health-care systems using economic evaluations to inform decisions about the reimbursement of health technologies. There are usually two separate elements of this process: assembling relevant evidence and undertaking analyses (technology assessment), and decision-making. In most systems, technology assessment is undertaken by the manufacturer of the technology. In a few, "third-party" assessment is used. In the United Kingdom, the National Institute for Health and Clinical Excellence used a combination of third-party and manufacturer assessments between 1999 and 2005. After this point, a Single Technology Appraisal program (using manufacturer-based assessment) was instituted for some technologies. Here the role of third-party assessment is considered in this from of decision-making. The article reviews the requirements of economic evaluation to support decision-making, and considers the extent to which each type of assessment is likely to meet these requirements. It also attempts to address whether the two forms of assessment differ in their impact on decision-making using a comparison of the decisions made by National Institute for Health and Clinical Excellence (NICE) (under its multiple-technology appraisal system) and the Scottish Medicines Consortium (SMC), which relies on manufacturer assessment. The comparison is limited by the small number of technologies considered by both bodies. Nevertheless, it suggests that there are potentially important differences between the two bodies, with NICE generally placing more restrictions of the use of technologies. The article concludes that there are potential advantages to third-party assessment, but its cost and timing may preclude its use for all new technologies. A hybrid arrangement is suggested where third-party assessment is used in particular circumstances.


Noncontact anterior cruciate ligament injuries: mechanisms and risk factors.
Significant advances have recently been made in understanding the mechanisms involved in noncontact anterior cruciate ligament (ACL) injury. Most ACL injuries involve minimal to no contact. Female athletes sustain a two- to eightfold greater rate of injury than do their male counterparts. Recent videotape analyses demonstrate significant differences in average leg and trunk positions during injury compared with control subjects. These findings as well as those of cadaveric and MRI studies indicate that axial compressive forces are a critical component in noncontact ACL injury. A complete understanding of the forces and risk factors associated with noncontact ACL injury should lead to the development of improved preventive strategies for this devastating injury.


Quality assessment of morphine hydrochloride solutions.
The therapeutic substances in solution prepared in pharmaceutical laboratories (prescribed drugs) must preserve their activity. Therefore, they must be stable throughout the period of storage in home conditions. The maintenance of stability is particularly difficult for morphine hydrochloride solutions administered orally to cancer patients at the last stage of the disease being at home. This study, aiming at the assessment of stability of morphine hydrochloride solutions, was performed on samples of 0.5% water solutions of the drug alone, 0.25% and 0.5% solutions of the drug in water with chloroform as well as injection solutions (Morphinum hydrochloricum, 20 mg, Polfa Warsaw). All the samples were kept at 20 degrees C for six months. Throughout this time observations were made to detect changes in their appearence and pH values. Their qualitative composition was determined by TLC and the content of morphine was checked by UV spectrophotometry in an environment of 0.1 mol/l of hydrochloric acid at 285 nm. Results of the kinetic study permitted drawing conclusions as to the mechanism of the decomposition of morphine hydrochloride in the solutions studied - according to a simple first order reaction and determination of the rate constants (k, s(-1)) of the process. Results of the chromatographic and spectrophotometric study did not show differences in the stability of water and chloroform/water solutions of morphine hydrochloride studied after 4 weeks and 6 months. After that time the decrease of morphine content was 10 and 25%, respectively.


The molecular immunology of mucositis: implications for evidence-based research in alternative and complementary palliative treatments.
The terms 'mucositis' and 'stomatitis' are often used interchangeably. Mucositis, however, pertains to pharyngeal-esophago-gastrointestinal inflammation that manifests as red, burn-like sores or ulcerations throughout the mouth. Stomatitis is an inflammation of the oral tissues proper, which can present with or without sores, and is made worse by poor dental hygiene. Mucositis is observed in a variety of immunosuppressed patients, but is most often consequential to cancer therapy. It appears as early as the third day of intervention, and is usually established by Day 7 of treatment. Mucositis increases mortality and morbidity and contributes to rising health care costs. The precise immune components involved in the etiology of mucositis are unclear, but evidence-based research (EBR) data has shown that applications of granulocyte-macrophage-colony stimulating factor prevent the onset or the exacerbation of oropharyngeal mucositis. The molecular implications of this observation are discussed from the perspective of future developments of complementary and alternative treatments for this condition. It must be emphasized that this article is meant to be neither a review on mucositis and the various treatments for it, nor a discussion paper on its underlying molecular immunology. It is a statement of the implications of EBR for CAM-based interventions for mucositis. It explores and discusses the specific domain of molecular immunology in the context of mucositis and its direct implications for EBR research in CAM-based treatments for mucositis.


The contribution of withholding or withdrawing care to newborn mortality.
We sought to determine the contribution of withholding or withdrawing care to neonatal mortality in our hospital over a 10-year period from 1993 through 2002. Although not initiating or withholding intensive care for certain newborns has been practiced openly for the past 25 years, little information has been published concerning the results of these practices on neonatal mortality. All infants who were born in our hospital and remained in the hospital until their death were evaluated for the care they received at the time of birth and near the time of their death. The contribution of not initiating care or withdrawing care to the neonatal mortality rates in our hospital was determined. Information was obtained directly from the patients' charts as well as the neonatal database and monthly neonatal mortality and morbidity review. Other information, if needed, was obtained from the monthly ethics committee reviews of all nursery deaths. Information was collected relating to birth weight, gestational age, diagnosis, time of death, and year of death. Hospital and neonatal unit protocols were evaluated to determine how closely they were followed. During the 10-year period, 380 deaths (0.8%) of a total of 47820 live births occurred in our hospital. Care was not initiated or was withdrawn in close to 72% of those deaths; total care until death occurred in 28%. Total care for infants who died over the 10-year period decreased markedly as care not initiated or care withdrawn increased. Most of this increase in not initiating care and in withdrawal of care was in the smaller of the extremely low birth weight infants. The majority of nursery deaths of infants born in our hospital occurred as the result of selected noninitiating of care or as a result of withdrawing care in infants not responding or considered to have a futile outcome. Only slightly more than one quarter of the infants received total care until the time of death.


Genetic, molecular and clinical features of familial isolated pituitary adenomas.
Pituitary adenomas occur in a familial setting in about 5% of all cases, and over half of these are due to multiple endocrine neoplasia type 1 (MEN1) and Carney's complex (CNC). Non-MEN1/CNC familial pituitary tumours of all tumour phenotypes, known as familial isolated pituitary adenomas (FIPA), were first described in the late 1990s. Clinical features of FIPA differ from those of sporadic pituitary adenomas, as patients with FIPA have a younger age at diagnosis and larger tumours. About 15% of patients with FIPA have mutations in the aryl hydrocarbon receptor-interacting protein gene (AIP), which indicates that FIPA may have a diverse genetic pathophysiology. This article describes the clinical features of FIPA, the tumour pathologies found in this setting and the genetic/molecular data that have recently been reported in FIPA.


Calcium and vitamin d supplementation in men.
Calcium and vitamin D supplements reverse secondary hyperparathyroidism and are widely prescribed to prevent osteoporotic fractures, with proven antifracture efficacy when targeted to individuals with documented insufficiencies. Men who should particularly be considered for calcium and vitamin D supplements include elderly or institutionalized individuals, patients with documented osteoporosis on antiresorptive or anabolic medication, and individuals receiving glucocorticoids. Benefits are most apparent when a daily dose of 1000-1200 mg calcium is complemented with 800 IU vitamin D. Compliance is the key to optimizing clinical efficacy. While (conventionally dosed) vitamin D has not been associated with safety concerns, recent meta-analytic data have provided evidence to suggest that calcium supplements (without coadministered vitamin D) may potentially be associated with cardiovascular risks.


Isoenzyme profiles of Trypanosoma cruzi stocks from different areas of Paraguay.
Twenty one Trypanosoma cruzi stocks from humans, domiciliary triatomines and one sylvatic animal of different areas of Paraguay were subjected to isoenzyme analysis. Thirteen enzyme systems (15 loci in total) were studied. MN cl2 (clonets 39) and SO34 cl4 (clonets 20) were used as references. Relationships between stocks were depicted by an UPGMA dendrogram constructed using the Jaccard's distances matrix. Among the Paraguayan stocks 14 zymodemes were identified (Par1 to Par14), Par 5 being the most frequent. Polymorphism rate and clonal diversity were 0.73 and 0.93, respectively. Average number of alleles per polymorphic locus was 2.5 (range 2-4). These measurements show a high diversity, which is confirmed by the dendrogram topology. All stocks belong to the same lineage, as MN cl2 reference strain (T. cruzi II). Moreover three distinct subgroups were identified and two of them correspond to Brazilian and Bolivian zymodemes, respectively. The third subgroup, the most common in Paraguay, is related to Tulahuen stock. The large geographical distribution of some zymodemes agrees with the hypothesis of clonality for T. cruzi populations. However sample size was not adequate to detect genetic recombination in any single locality.


Application of Bacteriophages to Limit Campylobacter in Poultry Production.
Campylobacter is a major foodborne pathogen with over a million United States cases a year and is typically acquired through the consumption of poultry products. The common occurrence of Campylobacter as a member of the poultry gastrointestinal tract microbial community remains a challenge for optimizing intervention strategies. Simultaneously, increasing demand for antibiotic-free products has led to the development of several alternative control measures both at the farm and in processing operations. Bacteriophages administered to reduce foodborne pathogens are one of the alternatives that have received renewed interest. Campylobacter phages have been isolated from both conventionally and organically raised poultry. Isolated and cultivated Campylobacter bacteriophages have been used as an intervention in live birds to target colonized Campylobacter in the gastrointestinal tract. Application of Campylobacter phages to poultry carcasses has also been explored as a strategy to reduce Campylobacter levels during poultry processing. This review will focus on the biology and ecology of Campylobacter bacteriophages in poultry production followed by discussion on current and potential applications as an intervention strategy to reduce Campylobacter occurrence in poultry production.


Two-Channel Portable Biopotential Recording System Can Detect REM Sleep Behavioral Disorder: Validation Study with a Comparison of Polysomnography.
Sleep disorders are frequent nonmotor symptoms of Parkinson's disease (PD). Polysomnography (PSG) has been the gold standard for its assessment. However, it requires patients to stay overnight in a hospital or sleep center. The mobile two-channel electroencephalography (EEG)/electrooculography (EOG) recording system is a self-applicable and affordable method to objectively assess sleep at home. We aimed at evaluating patients with PD to confirm the difference in sleep parameters between the portable recording system and PSG. PSG and the portable recording system were simultaneously performed on a similar night in eight patients with PD. We compared the difference in sleep parameters between them using nonparametric tests. All patients displayed a score of both PDSS -2 ≥ 15 and PSQI ≥ 5, respectively, which revealed poor sleep quality. There was no difference in the sleep parameters between the portable recording system and PSG, except for the percentage of sleep stage N3. Regarding the detection of REM sleep without atonia, we observed accordance between the portable recording system and PSG in six patients (P=0.686). The portable EEG/EOG recording system may gain an advantage from home-based evaluations for habitual sleep at home. Our study on device validation may contribute to measuring natural sleep, including rapid eye movement (REM) sleep behavioral disorder (RBD), in an outpatient care setting.


Characterization of polysaccharides from the flowers of Nerium indicum and their neuroprotective effects.
Degeneration of neurons is a key problem in Alzheimer's disease (AD) and neuroprotection is a possible way to safeguard neurons from neurodegeneration. Polysaccharides isolated from Chinese medicinal herbs have been investigated extensively for their anti-tumor and immune stimulating effects. Yet, little is known about the effects of polysaccharides in neurons. Recently, two pure polysaccharides isolated from the flowers of Nerium indicum were shown to stimulate proliferation and differentiation of PC12 pheochromocytoma cells, an effect similar to that observed from nerve growth factor. In this notion, it is hypothesized that polysaccharides isolated from the flowers of N. indicum could exhibit beneficial effects in neurons. In this study, we isolated, characterized and investigated two new polysaccharides from the flowers of N. indicum for their neuroprotective effects on neurons against serum-deprivation and beta-amyloid (Abeta) peptide toxicity in primary rat cortical neuronal cultures. Pretreatment of the polysaccharides significantly reduced the number of apoptotic neurons revealed by DAPI staining when neurons were exposed to serum-free medium. Besides, the polysaccharides could also decrease the activity of caspase-3 triggered by Abeta peptides. Western blot analysis indicated that polysaccharides stimulated the phosphorylation of PDK-1 (Serine 241) and Akt (Threonine 308). In conclusion, the polysaccharides J2, J3 and J4 isolated from N. indicum provide a lead for future development of neuroprotective agent against neuronal death in neurodegenerative diseases and the neuroprotective mechanism may primarily rely on activation of Akt survival signaling pathway.


Coupled Mn(II) and Cr(III) Oxidation Mediated by Ascomycete Fungi.
Manganese (Mn) oxides are considered as the primary oxidant of trivalent chromium [Cr(III)] in the environment. Microbial activities are responsible for the majority of Mn oxide formation in nature, thus likely influencing Cr(III) oxidation. Previous studies have been limited to Cr(III) oxidation by bacterial Mn oxides. Herein, we report coupled Mn(II) and Cr(III) oxidation in the presence of three Mn(II)-oxidizing Ascomycete fungi. In contrast to the previously reported inhibitory effect of Cr(III) on bacterial Mn(II) oxidation, varying effects of Cr(III) on fungal Mn(II) oxidation were observed, which may be linked to their Mn(II)-oxidation mechanisms. Under the concentrations of Mn(II) and Cr(III) applied in this study, Cr(III) promoted Mn(II) oxidation if it was mediated by hyphae-associated processes, but inhibited Mn(II) oxidation if it was achieved via extracellular enzymes/metabolites. The Cr(III) oxidation rate and extent were affected by Cr(III) speciation, Cr(VI) removal capacity (i.e., adsorption/reduction) of fungi, and organic content. The morphology and spatial relationship of Mn oxides with fungi varied, depending on fungal species and Cr(III) presence. Our findings highlight the importance of Mn(II)-oxidizing fungi in biogeochemical cycles of Mn and Cr and have significant implications for the origin of geogenic Cr(VI) and stability of reduced chromium in contaminated environments.


Adult respiratory distress syndrome: an unrecognized premortem event in systemic lupus erythematosus.
A retrospective study of the medical records of 46 patients, comprising our SLE population over a 6-year period, noted that mortality (7 of 46:15.2%) was directly related to the adult respiratory distress syndrome. In five, a concomitant severe infection was detected. Overall, 17 of our 46 patients presented with infectious episodes. The only statistically significant factor predisposing to both infection and the adult respiratory distress syndrome was high dose corticosteroid administration. Our results suggest that adult respiratory distress syndrome is a clinical condition related to the death of lupus patients and commonly complicates infection.


Discovery and genomic characterization of a novel ovine partetravirus and a new genotype of bovine partetravirus.
Partetravirus is a recently described group of animal parvoviruses which include the human partetravirus, bovine partetravirus and porcine partetravirus (previously known as human parvovirus 4, bovine hokovirus and porcine hokovirus respectively). In this report, we describe the discovery and genomic characterization of partetraviruses in bovine and ovine samples from China. These partetraviruses were detected by PCR in 1.8% of bovine liver samples, 66.7% of ovine liver samples and 71.4% of ovine spleen samples. One of the bovine partetraviruses detected in the present samples is phylogenetically distinct from previously reported bovine partetraviruses and likely represents a novel genotype. The ovine partetravirus is a novel partetravirus and phylogenetically most related to the bovine partetraviruses. The genome organization is conserved amongst these viruses, including the presence of a putative transmembrane protein encoded by an overlapping reading frame in ORF2. Results from the present study provide further support to the classification of partetraviruses as a separate genus in Parvovirinae.


An easy-to-use primer design tool to address paralogous loci and T-DNA insertion sites in the genome of Arabidopsis thaliana.
More than 90% of the Arabidopsis thaliana genes are members of multigene families. DNA sequence similarities present in such related genes can cause trouble, e.g. when molecularly analysing mutant alleles of these genes. Also, flanking-sequence-tag (FST) based predictions of T-DNA insertion positions are often located within paralogous regions of the genome. In such cases, the prediction of the correct insertion site must include careful sequence analyses on the one hand and a paralog specific primer design for experimental confirmation of the prediction on the other hand. GABI-Kat is a large A. thaliana insertion line resource, which uses in-house confirmation to provide highly reliable access to T-DNA insertion alleles. To offer trustworthy mutant alleles of paralogous loci, we considered multiple insertion site predictions for single FSTs and implemented this 1-to-N relation in our database. The resulting paralogous predictions were addressed experimentally and the correct insertion locus was identified in most cases, including cases in which there were multiple predictions with identical prediction scores. A newly developed primer design tool that takes paralogous regions into account was developed to streamline the confirmation process for paralogs. The tool is suitable for all parts of the genome and is freely available at the GABI-Kat website. Although the tool was initially designed for the analysis of T-DNA insertion mutants, it can be used for any experiment that requires locus-specific primers for the A. thaliana genome. It is easy to use and also able to design amplimers with two genome-specific primers as required for genotyping segregating families of insertion mutants when looking for homozygous offspring. The paralog-aware confirmation process significantly improved the reliability of the insertion site assignment when paralogous regions of the genome were affected. An automatic online primer design tool that incorporates experience from the in-house confirmation of T-DNA insertion lines has been made available. It provides easy access to primers for the analysis of T-DNA insertion alleles, but it is also beneficial for other applications as well.


[Diagnosis of iron overload syndrome].
The article presents a classification of conditions, accompanied with increasing of iron accumulation of the liver cells, normal iron metabolism in the human body, etiology, epidemiology, multifactorial structure of the pathogenesis of HFE homozygous hemochromatosis, clinic of iron overload syndrome. The modern algorithms of diagnostic and tactics of patients with the use of genotyping and correction of iron content.


[Current aspects of imported drug-resistant Plasmodium falciparum].
24 cases of Plasmodium falciparum malaria in a group of 73 non-immune subjects coming mostly from central Africa have been tested in vitro for chloroquine sensitivity: 5 are sensitive, 19 are resistant among whom one is also quinine-resistant. The comparison of the major clinical and biological features in the two groups by methods of statistical calculation shows that the parasitological diagnosis is retarded and that hemolysis is more frequent in the strains of Plasmodium falciparum resistant to chloroquine. Thrombopenia is more frequent with chemosensitives strains. In any case, the delay between the first symptoms and the parasitological diagnosis is increased with an average of 17 days in comparison to what was observed in the 1970's. Under the influence of several complex factors including human ones, the clinical aspects are changed in comparison to forms usually observed. In this context of an extension of the emergence of the chemoresistance and in the aim of efficiency, only the quinine I.V., with an average dose of 16.5 mg/kg per day of quinine-base, has been chosen as a curative treatment during 5 days allowing a rapid recovery without after-effects for all the cases.


Lugol chromoendoscopic screening for esophageal dysplasia/early squamous cell carcinoma in patients with esophageal symptoms in low-risk region in China.
Detection of esophageal dysplasia/early esophageal squamous cell carcinoma (ESCC) is essential for improving 5-year survival. The aim of this prospective study was to evaluate whether Lugol chromoendoscopy improves the detection of esophageal dysplasia/early ESCC in patients with esophageal symptoms in a low-incidence region in China. Eligible patients were randomly assigned into two groups who received routine endoscopy or Lugol chromoendoscopy. During endoscopy, between one and five biopsies were taken from visible lesions for routine endoscopy, or unstained areas of >0.5 cm in diameter for Lugol chromoendoscopy. In total, 812 patients were enrolled, 395 for routine endoscopy and 417 for Lugol chromoendoscopy. The overall detection rate of esophageal dysplasia/early ESCC was 10.6% (86/812), the detection rates were 7.3% (29/395) and 13.7% (57/417) in routine and chromoendoscopy groups, respectively (χ2=8.58, P=0.003). The detection rates were 8.3% (48/580), 17.2% (17/99) and 16.5% (22/133), respectively, in patients with reflux, dysphagia and globus sensation symptoms. In the chromoendoscopy group, 213 patients had unstained lesions of >0.5 cm, the detection rates of dysplasia/early carcinoma were 5.3% (4/76) in those with lesions of 0.5-1.0 cm, and 37.2% (51/137) in those with lesions >1.0 cm (χ2=21.46, P<0.001). These results indicate that Lugol chromoendoscopy improves the detection rate of esophageal dysplasia/early carcinoma in patients with esophageal symptoms compared with routine endoscopy. We propose that Lugol chromoendoscopy must therefore be considered in addition to routine endoscopy in patients with esophageal symptoms.


Combined treatment with bone marrow mesenchymal stem cells and methylprednisolone in paraquat-induced acute lung injury.
To evaluate the efficacy of combined treatment with bone marrow mesenchymal stem cell (BMSC) transplantation and methylprednisolone (MP) to treat paraquat (PQ)-induced acute lung injury. A total of 102 female rats were randomly divided into five groups: PQ, BMSC, MP, BMSC + MP and normal control. After 14 days of PQ poisoning, the survival of rats, wet/dry weight ratio of lung tissue, serum levels of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, IL-10, malondialdehyde (MDA) and superoxidase dismutase (SOD), and the expression of nuclear factor (NF)-кB p65 in lung tissue were determined. Rats in BMSC and BMSC + MP groups survived. BMSC transplantation significantly decreased the wet/dry weight ratio of lung tissue, down-regulated NF-кB p65 expression in lung tissue, lowered serum levels of TNF-α, IL-1β, IL-6 and MDA, and increased serum levels of IL-10 and SOD. These changes were particularly significant on days 7-14 after PQ poisoning. The above changes were more significant in the MP group on days 1-3 after PQ poisoning, compared with those of the BMSC group. However, the BMSC + MP group showed more significant changes on days 1-14 after PQ poisoning than those of both BMSC and MP groups. MP inhibits the inflammatory response, reduces the products of lipid peroxidation and promotes survival of transplanted BMSC, thus improving the intermediate and longer term efficacy of BMSC transplantation for treatment of PQ-induced lung injury.


Evaluation of four molecular methods for the diagnosis of tuberculosis in pulmonary and blood samples from immunocompromised patients.
The present study analysed the concordance among four different molecular diagnostic methods for tuberculosis (TB) in pulmonary and blood samples from immunocompromised patients. A total of 165 blood and 194 sputum samples were collected from 181 human immunodeficiency virus (HIV)-infected patients with upper respiratory complaints, regardless of suspicious for TB. The samples were submitted for smear microscopy, culture and molecular tests: a laboratory-developed conventional polymerase chain reaction (PCR) and real-time quantitative PCR (qPCR) and the Gen-Probe and Detect-TB Ampligenix kits. The samples were handled blindly by all the technicians involved, from sample processing to results analysis. For sputum, the sensitivity and specificity were 100% and 96.7% for qPCR, 81.8% and 94.5% for Gen-Probe and 100% and 66.3% for Detect-TB, respectively. qPCR presented the best concordance with sputum culture [kappa (k) = 0.864)], followed by Gen-Probe (k = 0.682). For blood samples, qPCR showed 100% sensitivity and 92.3% specificity, with a substantial correlation with sputum culture (k = 0.754) and with the qPCR results obtained from sputum of the corresponding patient (k = 0.630). Conventional PCR demonstrated the worst results for sputa and blood, with a sensitivity of 100% vs. 88.9% and a specificity of 46.3% vs. 32%, respectively. Commercial or laboratory-developed molecular assays can overcome the difficulties in the diagnosis of TB in paucibacillary patients using conventional methods available in most laboratories.


Design of Fuzzy Aided Cognition System for International Economic and Trade Information under Supply Chain Management.
The existing fuzzy assistant cognition system has the problem of imperfect retrieval function, which leads to long execution time. This paper designs a fuzzy assistant cognition system of international economic and trade information under supply chain management. Hardware Part. Optimize wireless sensor and connect power interface and related accessories. In the software part, the dynamic characteristics of international economic and trade information elements are extracted, the flexible operation model of supply chain management is constructed, the demand-oriented organizational structure is established, the output set is determined by the product reasoning, and the retrieval function of fuzzy assistant cognitive system is designed by using the fuzzy association algorithm. Experimental Results. The average execution time of the fuzzy assistant cognitive system and the other two systems is 123.593 s, 165.733 s, and 169.506 s, which proves that the cognitive system integrated with supply chain management has a higher practical application value.


The host defense system in the human newborn: the role of interferon and the natural killer cell.
The human newborn is particularly susceptible to infections in the neonatal period, due in part to the immaturity and naive state of the immune system. The role of interferon and natural killer cells in host defense in this age group is poorly defined. We have studied natural killer cells and the activation of the interferon system in cord blood. Most newborns (75%) had low natural killer cell activity, and this was unrelated to levels of the intracellular interferon-associated enzyme, 2'-5' oligoadenylate synthetase (2-5A synthetase). Newborn cells responded to interferon in vitro with an increase in natural killer activity, suggesting that this low natural killer cell activity is the result of fewer numbers of effector cells rather than a functional immaturity. Circulating serum interferon was detected in greater than 50% of maternal samples tested; however, this did not correlate with 2-5A synthetase levels in maternal or the corresponding cord blood mononuclear cells. Natural killer cell activity was low in the newborn in spite of activation of the interferon system in 39% of these individuals. This may be an important factor in susceptibility to infections in this period.


Prediction Model for COVID-19 Vaccination Intention among the Mobile Population in China: Validation and Stability.
Since China's launch of the COVID-19 vaccination, the situation of the public, especially the mobile population, has not been optimistic. We investigated 782 factory workers for whether they would get a COVID-19 vaccine within the next 6 months. The participants were divided into a training set and a testing set for external validation conformed to a ratio of 3:1 with R software. The variables were screened by the Lead Absolute Shrinkage and Selection Operator (LASSO) regression analysis. Then, the prediction model, including important variables, used a multivariate logistic regression analysis and presented as a nomogram. The Receiver Operating Characteristic (ROC) curve, Kolmogorov-Smirnov (K-S) test, Lift test and Population Stability Index (PSI) were performed to test the validity and stability of the model and summarize the validation results. Only 45.54% of the participants had vaccination intentions, while 339 (43.35%) were unsure. Four of the 16 screened variables-self-efficacy, risk perception, perceived support and capability-were included in the prediction model. The results indicated that the model has a high predictive power and is highly stable. The government should be in the leading position, and the whole society should be mobilized and also make full use of peer education during vaccination initiatives.


2-Aminoisobutyric acid in Co(II) and Co(II)/Ln(III) chemistry: homometallic and heterometallic clusters.
The synthesis and magnetic properties of 13 new homo- and heterometallic Co(II) complexes containing the artificial amino acid 2-amino-isobutyric acid, aibH, are reported: [Co(II)(4)(aib)(3)(aibH)(3)(NO(3))](NO(3))(4)·2.8CH(3)OH·0.2H(2)O (1·2.8CH(3)OH·0.2H(2)O), {Na(2)[Co(II)(2)(aib)(2)(N(3))(4)(CH(3)OH)(4)]}(n) (2), [Co(II)(6)La(III)(aib)(6)(OH)(3)(NO(3))(2)(H(2)O)(4)(CH(3)CN)(2)]·0.5[La(NO(3))(6)]·0.75(ClO(4))·1.75(NO(3))·3.2CH(3)CN·5.9H(2)O (3·3.2CH(3)CN·5.9H(2)O), [Co(II)(6)Pr(III)(aib)(6)(OH)(3)(NO(3))(3)(CH(3)CN)(6)]·[Pr(NO(3))(5)]·0.41[Pr(NO(3))(3)(ClO(4))(0.5)(H(2)O)(1.5)]·0.59[Co(NO(3))(3)(H(2)O)]·0.2(ClO(4))·0.25H(2)O (4·0.25H(2)O), [Co(II)(6)Nd(III)(aib)(6)(OH)(3)(NO(3))(2.8)(CH(3)OH)(4.7)(H(2)O)(1.5)]·2.7(ClO(4))·0.5(NO(3))·2.26CH(3)OH·0.24H(2)O (5·2.26CH(3)OH·0.24H(2)O), [Co(II)(6)Sm(III)(aib)(6)(OH)(3)(NO(3))(3)(CH(3)CN)(6)]·[Sm(NO(3))(5)]·0.44[Sm(NO(3))(3)(ClO(4))(0.5)(H(2)O)(1.5)]·0.56[Co(NO(3))(3)(H(2)O)]·0.22(ClO(4))·0.3H(2)O (6·0.3H(2)O), [Co(II)(6)Eu(III)(aib)(6)(OH)(3)(NO(3))(3)(CH(3)OH)(4.87)(H(2)O)(1.13)](ClO(4))(2.5)(NO(3))(0.5)·2.43CH(3)OH·0.92H(2)O (7·2.43CH(3)OH·0.92H(2)O), [Co(II)(6)Gd(III)(aib)(6)(OH)(3)(NO(3))(2.9)(CH(3)OH)(4.9)(H(2)O)(1.2)]·2.6(ClO(4))·0.5(NO(3))·2.58CH(3)OH·0.47H(2)O (8·2.58CH(3)OH·0.47H(2)O), [Co(II)(6)Tb(III)(aib)(6)(OH)(3)(NO(3))(3)(CH(3)CN)(6)]·[Tb(NO(3))(5)]·0.034[Tb(NO(3))(3)(ClO(4))(0.5)(H(2)O)(0.5)]·0.656[Co(NO(3))(3)(H(2)O)]·0.343(ClO(4))·0.3H(2)O (9·0.3H(2)O), [Co(II)(6)Dy(III)(aib)(6)(OH)(3)(NO(3))(2.9)(CH(3)OH)(4.92)(H(2)O)(1.18)](ClO(4))(2.6)(NO(3))(0.5)·2.5CH(3)OH·0.5H(2)O (10·2.5CH(3)OH·0.5H(2)O), [Co(II)(6)Ho(III)(aib)(6)(OH)(3)(NO(3))(3)(CH(3)CN)(6)]·0.27[Ho(NO(3))(3)(ClO(4))(0.35)(H(2)O)(0.15)]·0.656[Co(NO(3))(3)(H(2)O)]·0.171(ClO(4)) (11), [Co(II)(6)Er(III)(aib)(6)(OH)(4)(NO(3))(2)(CH(3)CN)(2.5)(H(2)O)(3.5)](ClO(4))(3)·CH(3)CN·0.75H(2)O (12·CH(3)CN·0.75H(2)O), and [Co(II)(6)Tm(III)(aib)(6)(OH)(3)(NO(3))(3)(H(2)O)(6)]·1.48(ClO(4))·1.52(NO(3))·3H(2)O (13·3H(2)O). Complex 1 describes a distorted tetrahedral metallic cluster, while complex 2 can be considered to be a 2-D coordination polymer. Complexes 3-13 can all be regarded as metallo-cryptand encapsulated lanthanides in which the central lanthanide ion is captivated within a [Co(II)(6)] trigonal prism. dc and ac magnetic susceptibility studies have been carried out in the 2-300 K range for complexes 1, 3, 5, 7, 8, 10, 12, and 13, revealing the possibility of single molecule magnetism behavior for complex 10.


Inflammation stimulates niacin receptor (GPR109A/HCA2) expression in adipose tissue and macrophages.
Many of the beneficial and adverse effects of niacin are mediated via a G protein receptor, G protein-coupled receptor 109A/hydroxycarboxylic acid 2 receptor (GPR109A/HCA2), which is highly expressed in adipose tissue and macrophages. Here we demonstrate that immune activation increases GPR109A/HCA2 expression. Lipopolysaccharide (LPS), TNF, and interleukin (IL) 1 increase GPR109A/HCA2 expression 3- to 5-fold in adipose tissue. LPS also increased GPR109A/HCA2 mRNA levels 5.6-fold in spleen, a tissue rich in macrophages. In peritoneal macrophages and RAW cells, LPS increased GPR109A/HCA2 mRNA levels 20- to 80-fold. Zymosan, lipoteichoic acid, and polyinosine-polycytidylic acid, other Toll-like receptor activators, and TNF and IL-1 also increased GPR109A/HCA2 in macrophages. Inhibition of the myeloid differentiation factor 88 or TIR-domain-containing adaptor protein inducing IFNβ pathways both resulted in partial inhibition of LPS stimulation of GPR109A/HCA2, suggesting that LPS signals an increase in GPR109A/HCA2 expression by both pathways. Additionally, inhibition of NF-κB reduced the ability of LPS to increase GPR109A/HCA2 expression by ∼50% suggesting that both NF-κB and non-NF-κB pathways mediate the LPS effect. Finally, preventing the LPS-induced increase in GPR109A/HCA2 resulted in an increase in TG accumulation and the expression of enzymes that catalyze TG synthesis. These studies demonstrate that inflammation stimulates GPR109A/HCA2 and there are multiple intracellular signaling pathways that mediate this effect. The increase in GPR109A/HCA2 that accompanies macrophage activation inhibits the TG accumulation stimulated by macrophage activation.


Influence of fingertip contact with a wall on postural sway and electromyographyic activity of the soleus muscle.
Standing and walking balance decreases in many kinds of neuromuscular, musculoskeletal and sensory disorders, and in elderly people because of age-related physiologic diminution. Recent studies have shown that contact cues from a cane or a fingertip provide information that leads to reduced postural sway in subjects. The first purpose of this study was to investigate the influence of light and forceful fingertip contacts with a wall on the postural sway while standing upright. The second purpose was to investigate the influence of fingertip contact on the EMG activity of the soleus muscle. Twelve healthy male volunteers (20.8 +/- 0.7 years old) participated in this study. Standing balance or postural sway was measured by sway of the center of pressure (COP) under three conditions: (1) standing without touching a wall, (2) standing with the right index fingertip lightly touching (< 1N) to a wall (3), and standing with the right index fingertip forcefully touching (5 approximately 10N) to a wall. Bipolar surface electrodes were placed over the right soleus muscle to measure EMG activity associated with anterior-posterior sway. More significant length, and rectangular and environmental areas of the COP were observed while standing without touching a wall than while standing while lightly or forcefully touching a wall, but there was no significant difference between the two touching conditions. No significant difference in the EMG activity of the soleus muscle was observed among the three conditions. The results of this study suggested that the fingertip touch contact with a wall decreased sway during static standing but did not influence the EMG activity of the soleus muscle.


Trends of height-for-age Z-scores according to age among Brazilian children under 5 years old from 2006 to 2019.
This study compared the distribution of stunting and height-for-age (HAZ) Z-scores among age groups in data from the Brazilian National Survey on Demography and Health of Women and Children (PNDS 2006) and the Brazilian National Survey on Child Nutrition (ENANI-2019). The final sample comprised 4,408 and 14,553 children < 59 months of age in the PNDS 2006 and ENANI-2019, respectively. Children with HAZ scores < -2 according to the World Health Organization (WHO) growth standard were classified as stunted. Prevalence, 95% confidence intervals (95%CI), means, and standard deviations were estimated for Brazil and according to age. The distribution of HAZ scores at each age (in months) was estimated using the svysmooth function of the R survey package. Analyses considered the complex sampling design of the studies. Statistical differences were determined by analyzing the 95%CI of the overlap of point estimates. From 2006 to 2019, the prevalence of stunting for children < 12 months of age increased from 4.7% to 9%. As expected, the smoothed curves showed a higher mean HAZ score for children < 24 months of age in 2006 than in 2019 with no overlap of 95%CI among children aged 6-12 months. For children ≥ 24 months of age, we observed a higher mean HAZ score in 2019. Although the prevalence of stunting among children < 59 months of age was similar between 2006 and 2019, mean HAZ scores among children ≥ 24 months of age increased, whereas the mean HAZ score among children < 24 months of age decreased. Considering the deterioration in living conditions and the potential impact of the COVID-19 pandemic, we expect a greater prevalence of stunting in Brazil in the near future.


Isolation, Characterization and Transcriptome Analysis of a Cytokinin Receptor Mutant Osckt1 in Rice.
Cytokinins play important roles in regulating plant development, including shoot and root meristems, leaf longevity, and grain yield. However, the in planta functions of rice cytokinin receptors have not been genetically characterized yet. Here we isolated a rice mutant, Osckt1, with enhanced tolerance to cytokinin treatment. Further analysis showed that Osckt1 was insensitive to aromatic cytokinins but responded normally to isoprenoid and phenylurea-type cytokinins. Map-based cloning revealed that the mutation occurred in a putative cytokinin receptor gene, histidine kinase 6 (OsHK6). OsCKT1 was found to be expressed in various tissues throughout the plant and the protein was located in the endoplasmic reticulum. In addition, whole-genome gene expression profiling analysis showed that OsCKT1 was involved in cytokinin regulation of a number of biological processes, including secondary metabolism, sucrose and starch metabolism, chlorophyll synthesis, and photosynthesis. Our results demonstrate that OsCKT1 plays important roles in cytokinin perception and control of root development in rice.


Diagnostic accuracy of procalcitonin and interleukin-6 for postoperative infection in major gastrointestinal surgery: a systematic review and meta-analysis.
We aim to assess the diagnostic accuracy of procalcitonin (PCT) and interleukin-6 (IL-6) as diagnostic biomarkers for postoperative infection/sepsis following major abdominal surgery. Postoperative infection is an important cause for morbidity and mortality in major surgery. Early diagnosis and antimicrobial treatment improves outcomes, and high-performing biomarkers could guide clinical decision making. A systematic database search was conducted for studies reporting diagnostic performance of biomarkers (including PCT and IL-6) for infection/sepsis following major abdominal surgery. Studies were assessed for reporting of diagnostic accuracy, relevance and quality. Data were extracted for meta-analysis. Ten studies with 1,611 participants reported the diagnostic accuracy of PCT, with pooled sensitivity, specificity and summary receiver operator curve of 72% (95% CI 66-78), 62% (95% CI 59-64) and 0.766, respectively. Four studies with 175 participants reported the diagnostic accuracy of IL-6, with pooled sensitivity, specificity and summary receiver operator curve of 84% (95% CI 72-92), 76% (95% CI 68-84) and 0.878, respectively There was variability in the timing of sampling and cut-off values and significant heterogeneity and inconsistency between studies (I2 diagnostic odds ratio (DOR)= 43.2% for PCT, I2 DOR=0% for IL-6). PCT performs only moderately well as a diagnostic test for postoperative infection/sepsis in major abdominal surgery, demonstrating limited sensitivity and specificity. Heterogeneity between studies is a limitation of the meta-analysis. There is an ongoing need for a rapid, accurate biomarker for postoperative infection or sepsis.


Does female education affect fertility behaviour in Pakistan?
The study explores the relationship between female education and fertility in Pakistan and is based on data from the Pakistan Fertility Survey 1975. Only slight differentials were identified between women with no education and those who had primary or less schooling. However, women with more than primary education had notably lower fertility. Also the role of the intermediate variables such as proportions married, length of breastfeeding, and contraceptive use had significant associations with female education.


Real-World Clinical Data of Palbociclib in Asian Metastatic Breast Cancer Patients: Experiences from Eight Institutions.
Use of cyclin-dependent kinase 4/6 inhibitors improved survival outcome of hormone receptor (HR) positive metastatic breast cancer (MBC) patients, including Asian population. However, Asian real-world data of palbociclib is limited. We analyzed the real-world clinical practice patterns and outcome in HR-positive, MBC Asian patients treated with palbociclib. Between April 2017 to November 2019, 169 HR-positive, human epidermal growth factor-2-negative MBC patients treated with letrozole or fulvestrant plus palbocilib were enrolled from eight institutions. Survival outcome (progression-free survival [PFS]), treatment response and toxicity profiles were analyzed. Median age of letrozole plus palbociclib (145 patients, 85.8%) and fulvestrant plus palbociclib (24 patients, 14.2%) was 58 and 53.5 years, with median follow-up duration of 14.63 months (range 0.2 to 33.9 months). Median PFS (mPFS) of letrozole plus palbociclib and fulvestrant plus palbociclib was 25.6 (95% confidence interval [CI], 19.1 to not reached) and 6.37 months (95% CI, 5.33 to not reached), comparable to previous phase 3 trials. In letrozole plus palbociclib arm, luminal A (hazard ratio, 2.86; 95% CI, 1.20 to 6.80; p=0.017) and patients with good performance (Eastern Cooperative Oncology Group 0-1 [hazard ratio, 3.68; 95% CI, 1.70 to 7.96]) showed better mPFS. In fulvestrant plus palbociclib group, chemotherapy naïve patients showed better mPFS (hazard ratio, 12.51, 95% CI, 1.59 to 99.17; p=0.017). The most common grade 3 or 4 adverse event was neutropenia (letrozole 86.3%, fulvestrant 88.3%). To our knowledge, this is the first real-world data of palbociclib reported in Asia. Palbociclib showed comparable benefit to previous phase 3 trials in Asian patients during daily clinical practice.


Growth requirements and characterization of rat cervical epithelial cells in culture.
The extended culture of rat cervical epithelial cells can be achieved in the absence of a fibroblast feeder layer by utilizing collagen gels and a complex growth medium. The medium contains a 1:1 mixture of RPMI-1640 and Ham's F12 supplemented with 7.5% porcine serum and epidermal growth factor, cholera toxin, transferrin, insulin, and hydrocortisone. Under these culture conditions the cells show rapid log-phase growth and high saturation densities while retaining the ultrastructural characteristics of immature squamous metaplastic cells of the rat uterine cervix even after extended passage. In a manner similar to epithelial cells from a variety of sources, rat cervical epithelial cells form hemicysts at confluence in vitro when cultured on impermeable substrates. The development of these methods for culturing cervical epithelial cells provides an experimental system for the study of factors important in regulating the growth and differentiation of metaplastic squamous epithelial cells.


Case Report: Rapid Recognition and Immune Modulation of Secondary HLH Due to Disseminated HSV Infection.
We describe the case of a newborn who presented with multiple organ dysfunction syndrome (MODS) and hyperferritinemia, who eventually met criteria for hemophagocytic lymphohistiocytosis (HLH) due to disseminated herpes simplex virus 1 (HSV-1). While the cytokine storm abated after administration of multiple immune modulatory therapies including dexamethasone, etoposide, intravenous immune globulin, anakinra, as well as the interferon gamma antagonist emapalumab, multiple organ dysfunction syndrome progressed. Care was withdrawn after 5 days. Subsequent genetic testing did not reveal any mutations associated with familial HLH. This case highlights that even with appropriate antiviral treatment and immune suppression, disseminated HSV is often fatal. Further study is warranted to determine whether early immune modulatory therapy including interferon gamma blockade can interrupt the HLH inflammatory cascade and prevent progression of MODS.


A polymeric protein anchors the chromosomal origin/ParB complex at a bacterial cell pole.
Bacterial replication origins move towards opposite ends of the cell during DNA segregation. We have identified a proline-rich polar protein, PopZ, required to anchor the separated Caulobacter crescentus chromosome origins at the cell poles, a function that is essential for maintaining chromosome organization and normal cell division. PopZ interacts directly with the ParB protein bound to specific DNA sequences near the replication origin. As the origin/ParB complex is being replicated and moved across the cell, PopZ accumulates at the cell pole and tethers the origin in place upon arrival. The polar accumulation of PopZ occurs by a diffusion/capture mechanism that requires the MreB cytoskeleton. High molecular weight oligomers of PopZ assemble in vitro into a filamentous network with trimer junctions, suggesting that the PopZ network and ParB-bound DNA interact in an adhesive complex, fixing the chromosome origin at the cell pole.


[Interactions of mononuclear cells of peripheral blood with islet cells in vitro].
Peripheral blood mononuclear cells from Type I diabetic patients health donors or Wistar rats were cultured with rat islet cells for 18 h. Then TNF content of the medium and basal and stimulated insulin release were determined. Mononuclear cells from both healthy donors and diabetic patients could inhibit the insulin release with no correlation to TNF content. Addition of lipopolysaccharide resulted in a 5-7 times increase of TNF content of the medium followed by a more pronounced inhibition of insulin release. Rat mononuclear cells inhibited the beta-cell function almost completely and initially produced large amounts of TNF. The data indicate that inhibition of insulin release by blood mononuclear cells in vitro does not reflect anti beta-cell specific cellular immunity, involves cytokines and, probably depends upon the initial properties of the cells.


Interlimb Dynamic after Unilateral Focal Lesion of the Cervical Dorsal Corticospinal Tract with Endothelin-1.
Handedness is one of the most recognized lateralized behavior in humans. Usually, it is associated with manual superiority regarding performance proficiency. For instance, more than 90% of the human population is considered more skilled with the right hand, which is controlled by the left hemisphere, than with the left. However, during the performance of bimanual tasks, the two hands usually assume asymmetric roles, with one hand acting on objects while the other provides support, stabilizing the object. Traditionally, the role of the two hands is viewed as fixed. However, several studies support an alternate view with flexible assignments for the two hands depending on the task. The supporting role of the hand depends on a closed loop pathway based on proprioceptive inputs from the periphery. The circuit's efferent arm courses through the dorsal corticospinal tract (dCST) in rodents and terminate on spinal cord interneurons which modulate the excitability of motoneurons in the ventral horn. In the present work, we developed an experimental model of unilateral lesion targeting the cervical dCST with microinjections of the vasoconstrictor endothelin-1 (ET-1) to evaluate the degree of flexibility of forelimb assignment during a food manipulation task. Our results show that just 3 days after unilateral corticospinal tract (CST) injury in the cervical region, rats display severe motor impairment of the ipsilateral forepaw together with a remarkable reversal of motor assignment between the forelimbs.


Traditional and commercial turkeys show similar susceptibility to foot pad dermatitis and behavioural evidence of pain.
1. An experiment was conducted to compare the development and clinical significance of foot pad dermatitis (FPD) in 4 lines of turkey poults kept on wet or dry litter for 6 d. 2. Turkeys kept on wet litter were less active and expressed fewer unique sequences of behaviour than poults kept on dry litter, suggesting that high FPD scores are associated with pain during movement. 3. Heavier lines of turkeys had higher mean foot scores that developed earlier, than those in a traditional line, but the effect was relatively small and not statistically significant after 6 d, whereas there were large differences between individuals within line. 4. Body weight gains were greater, and feed intake was lower in the commercial lines kept on dry litter compared with wet litter. There were no differences in water intake between poults kept on wet or dry litter.


Comparison of the TestPack Strep A enzyme immunoassay system with anaerobically incubated cultures for detection of group A streptococci from oropharyngeal swabs.
To find a rapid and sensitive test for detection of Group A streptococci (GABS), the performance of the TestPack Strep A (TPSA; Abbott) was compared with culture with the use of rayon-tipped throat swabs from symptomatic patients six months to 90 years of age. Each swab was first inoculated to a 5% sheep blood agar plate and then tested for GABS antigen with the use of the TPSA and the manufacturer's instructions. Cultures were incubated anaerobically at 35 degrees C for 36-48 hours unless positive results were obtained after one night. GABS were identified with a fluorescent antibody method or a latex antibody test. From 1,616 throat swabs, 296 (18.3%) of the cultures contained GABS. The sensitivity and specificity of the TPSA were 73.3% and 94.8%, respectively, whereas the predictive values of positive and negative results were 75.9% and 94.1%, respectively. Results varied significantly, however, with different production lots of TPSA. The TPSA does not appear to provide a sensitive alternative to an anaerobic culture for detection of GABS.


Impact of physiological factors on longitudinal structural MRI measures of the brain.
Longitudinal MRI is used in clinical research studies to examine illness progression, neurodevelopment, and the effect of medical interventions. Such studies typically report changes in brain volume of less than 5%. However, there is a concern that these findings could be obscured or confounded by small changes in brain volume estimates caused by physiological factors such as, dehydration, blood pressure, caffeine levels, and circadian rhythm. In this study, MRI scans using the ADNI-III protocol were acquired from 20 participants (11 female) at two time points (mean interval = 20.3 days). Hydration, systolic and diastolic blood pressure, caffeine intake, and time of day were recorded at both visits. Images were processed using FreeSurfer. Three a priori hypothesised brain regions (hippocampus, lateral ventricles, and total brain) were selected, and an exploratory analysis was conducted on FreeSurfer's auto-segmented brain regions. There was no significant effect of the physiological factors on changes in the hypothesised brain regions. We provide estimates for the maximum percentage change in regional brain volumes that could be expected to occur from normal variation in each of the physiological measures. In this study, normal variations in physiological parameters did not have a detectable effect on longitudinal changes in brain volume.


Intimate partner violence during infancy and cognitive outcomes in middle childhood: Results from an Australian community-based mother and child cohort study.
The cognitive functioning of children who experience intimate partner violence (IPV) has received less attention than their emotional-behavioral outcomes. Drawing upon data from 615 (48.4% female) 10-year-old Australian-born children and their mothers (9.6% of mothers born in non-English speaking countries) participating in a community-based longitudinal study between 2004 and 2016, this study examined the associations between IPV in infancy and cognition in middle childhood (at age 10). Results showed that IPV in the first 12 months of life was associated with lower general cognitive ability and poorer executive attention but not working memory skills. IPV in middle childhood (in the 10th year postpartum) was not associated with cognition. This study provides evidence for the long-term impact of early life exposure to IPV on children's cognition, and points to the importance of early intervention to optimize development.


Development of the My Positive Health dialogue tool for children: a qualitative study on children's views of health.
Children's views of health were explored in order to develop a health dialogue tool for children. A qualitative research design was used as part of a codesign process. Based on semi-structured interviews with both healthy children and children with a chronic condition (aged 8-18). Two approaches were applied. The first was an open exploration of children's views on health, which was then thematically analysed. Subsequently, a framework was used, based on the six-dimensional My Positive Health (MPH) dialogue tool for adults, to guide the second part of the interviews, focusing on reviewing the children's view on health within the context of the framework. For the final draft of the dialogue tool, a framework analysis was conducted and then validated by members of the 'children's council' of the Wilhelmina Children's Hospital. We interviewed 65 children, 45 of whom had a chronic condition and 20 were healthy. The children described a broad concept of health with the central themes of 'feeling good about yourself' and 'being able to participate'. Based on the subsequent framework analysis, the wording of two of the six dimensions of the MPH dialogue tool was adjusted and the related aspects were adapted for better alignment with the children's concept of health. After these modifications, the tool fully matched the children's concept of health. The MPH dialogue tool for children was developed for children with and without a chronic condition, to help them open up about what they consider important for their health and well-being, and to improve directorship over decisions and actions that would affect their health. The MPH dialogue tool aims to support healthcare professionals in providing the type of care and treatment that is in line with the needs of their young patients/clients.


Early epigenetic events regulate the adaptive immune response gene CIITA.
Precise regulation of Major Histocompatibility class II (MHC II) genes plays important roles in initiation, propagation and termination of adaptive immune responses by controlling antigen presentation to CD4+ T cells. MHC II genes are constitutively expressed in only a few cell types and are inducibly expressed by the inflammatory response cytokine interferon gamma (IFNγ) in all nucleated cells. The regulation of MHC II is tightly controlled by a Master Regulator, the class II transactivator (CIITA), which is a general regulator of both constitutive and inducible MHC II expression. Although much is known about the transcription factors necessary for CIITA expression, less is known about the epigenetic modifications and the requisite enzymes needed to provide these transcription factors access to DNA. We show here that multiple epigenetic changes occur at the IFNγ inducible CIITA promoter within 20' of IFNγ stimulation and that these changes correlate with the opening of the promoter and the initiation of transcription. Our study links these rapidly occurring epigenetic events at the inducible CIITA promoter to decreased promoter binding of the histone methyltransferase EZH2, and shows that decreased promoter binding of EZH2 transforms this previously tightly regulated and cytokine inducible promoter into a constitutively active and dysregulated gene.


Proximal tibial fracture following total knee arthroplasty.
The patient was a 74-year-old man, with a history of total knee arthoplasty 14 years earlier, after having sustained a pathological fracture of the proximal diaphysis of the left tibia following a fall. Given the unstable nature of the fracture and the severe osteolysis noted below the total knee arthroplasty, surgical management 1 day after the fall entailed packing cancellous bone graft into the defect and realigning the fracture.


E6AP/UBE3A catalyzes encephalomyocarditis virus 3C protease polyubiquitylation and promotes its concentration reduction in virus-infected cells.
The encephalomyocarditis virus (EMCV) 3C protease (3Cpro) is one of a small number of viral proteins whose concentration is known to be regulated by the cellular ubiquitin-proteasome system. Here we report that the ubiquitin-conjugating enzyme UbcH7/UBE2L3 and the ubiquitin-protein ligase E6AP/UBE3A are components of a previously unknown EMCV 3Cpro-polyubiquitylating pathway. Following the identification of UbcH7/UBE2L3 as a participant in 3Cpro ubiquitylation, we purified a UbcH7-dependent 3Cpro-ubiquitylating activity from mouse cells, which we identified as E6AP. In vitro reconstitution assays demonstrated that E6AP catalyzes the synthesis of 3Cpro-attached Lys48-linked ubiquitin chains, known to be recognized by the 26S proteasome. We found that the 3Cpro accumulates to higher levels in EMCV-infected E6AP knockdown cells than in control cells, indicating a role for E6AP in in vivo 3Cpro concentration regulation. We also discovered that ARIH1 functions with UbcH7 to catalyze EMCV 3Cpro monoubiquitylation, but this activity does not influence the in vivo 3Cpro concentration.


Beneficial autoimmunity and maladaptive inflammation shape epidemiological links between cancer and immune-inflammatory diseases.
Chronic inflammation drives proliferative responses, hence increasing cellular multiplication with the consequent risk of malignant transformation. Autoimmune responses against self-antigens drive chronic inflammation but may also enhance cancer immunosurveillance with the consequent reduction of tumor incidence and progression. These notions, which have been well established at the preclinical level, may explain the generally positive associations between immune-inflammatory diseases but also some negative associations, for example between breast cancer and rheumatoid arthritis or systemic lupus erythematosus, which have recently been confirmed in a study enrolling close to half a million participants from the UK Biobank.


Success against lymphatic filariasis.
A campaign against lymphatic filariasis began in China's Shandong Province during 1956. Epidemiological surveillance since 1984 indicates that the transmission of infection has been interrupted. The factors accounting for this are discussed and the continuing challenges presented by the disease are outlined.


A positive correlation between GC content and growth temperature in prokaryotes.
GC pairs are generally more stable than AT pairs; GC-rich genomes were proposed to be more adapted to high temperatures than AT-rich genomes. Previous studies consistently showed positive correlations between growth temperature and the GC contents of structural RNA genes. However, for the whole genome sequences and the silent sites of the codons in protein-coding genes, the relationship between GC content and growth temperature is in a long-lasting debate. With a dataset much larger than previous studies (681 bacteria and 155 archaea with completely assembled genomes), our phylogenetic comparative analyses showed positive correlations between optimal growth temperature (Topt) and GC content both in bacterial and archaeal structural RNA genes and in bacterial whole genome sequences, chromosomal sequences, plasmid sequences, core genes, and accessory genes. However, in the 155 archaea, we did not observe a significant positive correlation of Topt with whole-genome GC content (GCw) or GC content at four-fold degenerate sites. We randomly drew 155 samples from the 681 bacteria for 1000 rounds. In most cases (> 95%), the positive correlations between Topt and genomic GC contents became statistically nonsignificant (P > 0.05). This result suggested that the small sample sizes might account for the lack of positive correlations between growth temperature and genomic GC content in the 155 archaea and the bacterial samples of previous studies. Comparing the GC content among four categories (psychrophiles/psychrotrophiles, mesophiles, thermophiles, and hyperthermophiles) also revealed a positive correlation between GCw and growth temperature in bacteria. By including the GCw of incompletely assembled genomes, we expanded the sample size of archaea to 303. Positive correlations between GCw and Topt appear especially after excluding the halophilic archaea whose GC contents might be strongly shaped by intense UV radiation. This study explains the previous contradictory observations and ends a long debate. Prokaryotes growing in high temperatures have higher GC contents. Thermal adaptation is one possible explanation for the positive association. Meanwhile, we propose that the elevated efficiency of DNA repair in response to heat mutagenesis might have the by-product of increasing GC content like that happens in intracellular symbionts and marine bacterioplankton.


Evidence for contagious behaviors in budgerigars (Melopsittacus undulatus): an observational study of yawning and stretching.
Yawning is contagious in humans and some non-human primates. If there are social functions to contagious behaviors, such as yawning, they might occur in other highly social vertebrates. To investigate this possibility, we conducted an observational study of yawning and an associated behavior, stretching, in budgerigars (Melopsittacus undulatus), a social, flock-living parrot. Flock-housed budgerigars were videotaped for 1.5h at three time-blocks during the day (early morning, afternoon and early evening), and the times of all yawns and stretches for each bird were recorded. Both yawning and stretching were temporally clumped within sessions, but were uniformly distributed across the trials of a particular time-block. This suggests that clumping was not a result of circadian patterning and that both behaviors could be contagious. There was additional evidence of contagion in stretching, which occurred in two forms - a posterior-dorsal extension of either one foot or both feet. Birds that could have observed a conspecific stretch, and that then stretched themselves within 20s, replicated the form of the earlier stretch significantly more often than expected by chance. This study provides the first detailed description of temporal patterns of yawning under social conditions in a flock-living species as well as the first support for contagious yawning and stretching in a non-primate species in a natural context. Experimental evidence will be necessary to confirm the extent of contagion in either behavior.


Association of eNOS Gene Polymorphisms G894T and T-786C with Risk of Hepatorenal Syndrome.
Background. There are no studies investigating the relationship between endothelial nitric oxide synthase (eNOS) gene polymorphisms and hepatorenal syndrome (HRS). Aim. The purpose of this study is to elucidate whether eNOS gene polymorphisms (G894T and T-786C) play a role in the development of type-2 HRS. Methods. This study was carried out in a group of 92 patients with cirrhosis (44 patients with type-2 HRS and 48 without HRS) and 50 healthy controls. Polymorphisms were determined by polymerase chain reaction (PCR) and melting curve analysis. Results. We did not find any significant difference in allele and genotype distributions of the eNOS -T-786C polymorphism among the groups (p = 0.440). However, the frequency of GT (40.9%) and TT (13.6%) genotypes and mutant allele T (34.1%) for the eNOS G894T polymorphism were significantly higher (p < 0.001 and p < 0.001, resp.) in the HRS group than in both the stable cirrhosis (14.6%, 4.2%, and 11.5%, resp.) and the control (22.0%, 2.0%, and 13.0%, resp.) groups. Conclusion. The occurrence of mutant genotypes (GT/TT) and mutant allele T in eNOS -G894T polymorphisms should be considered as a potential risk factor in cirrhotic patients with HRS.


Translation and Cultural Adaptation to European Portuguese of the "Measure of Moral Distress - Healthcare Professionals" Scale: A Cross-Sectional Multicenter Study.
Moral distress occurs when one knows the morally correct action to take but is constrained from taking that action. The aims of this study were to translate into European Portuguese and culturally adapt the "Measure of Moral Distress - Healthcare Professionals" questionnaire to the context of the Portuguese healthcare system and to explore the frequency and intensity of moral distress occurring among medical students. The "Measure of Moral Distress - Healthcare Professionals" questionnaire was translated and culturally adapted to European Portuguese, following the internationally accepted "COnsensus-based Standards for the selection of health Measurement INstruments". Afterwards, a web-based survey was conducted, following the "Checklist for Reporting Results of Internet E-Surveys" guidelines. Medical students were asked to rate potentially morally distressing situations on frequency and intensity. Of approximately 4300 medical students, 939 (22%) completed the survey. Participants experienced, on average, 16 morally distressing situations. Median of composite score of moral stress was 79 (IQR 44 - 118). Only 31% of the students felt well prepared to handle a morally distressing situation, 26% considered leaving medical school and 28% thought about choosing a non-clinical specialty due to moral distress. Despite a plethora of studies on this topic, the results suggested that moral distress is still a common phenomenon among medical students with a cumulative effect over time. These results emphasize the importance of a critical review of medical education, reducing the harmful effects of preventable psychological phenomena in clinical practice and in the lives of future healthcare professionals.


Defunctioning loop ileostomy for pelvic anastomoses: predictors of morbidity and nonclosure.
The aim of this study was to determine the morbidity of a defunctioning loop ileostomy and the subsequent closure rate, and to identify the predictors of complications and nonclosure of stoma. This study is a retrospective review of a single-institution experience. All patients who underwent a planned temporary defunctioning loop ileostomy performed synchronously with a pelvic anastomosis during a 6-year period were included. The primary outcome measures were the ileostomy complication rate for the entire spectrum of care, readmission and reoperation rates to treat ileostomy complications, and subsequent closure rate. Patient and treatment factors were evaluated for their independent effect on complications and closure rate with the use of multivariable logistic regression. One hundred twenty-three patients were identified (median age, 51 years). Of these patients, 64.2% developed ≥1 minor or major ileostomy complications (13.8% during index hospitalization, 52.8% as outpatient, and 23.4% after closure). Readmitted for dehydration following ileostomy formation were 11.4% of patients. The ileostomy was closed in 76.4% of patients with 8.6% requiring a midline laparotomy. The overall ileostomy-related reoperation rate was 10.4% (2.4% during index hospitalization, 1.6% at readmission, and 6.4% following ileostomy closure). Obesity (BMI ≥30 kg/m) was associated with a higher overall ileostomy complication rate (OR 8.56, 95% CI 1.64-44.74) and outpatient complication rate (OR 7.69, 95% CI 2.48-23.81). Age >65 years (OR 53.34, 95% CI 4.21-676.14) and hypertension (OR 8.36, 95% CI 1.09-64.43) increased the risks of high ileostomy output and dehydration. Obesity (OR 4.61, 95% CI 1.14-18.54) and smoking (4.47, 95% CI 1.43-13.98) decreased the likelihood of ileostomy closure. This study was limited by its retrospective nature. The morbidity of a defunctioning loop ileostomy remains significant. Obesity is an independent predictor of ileostomy complications. Older age and hypertension increase the risks of high-output stoma and dehydration. Almost one quarter of patients never have the ileostomy closed. Obesity and smoking are associated with less likelihood of a subsequent ileostomy closure.


[Interaction of topotecan, DNA topoisomerase I inhibitor, with double-stranded polydeoxyribonucleotides. III. Binding at the minor groove].
Interaction of topotecan (TPT) with calf thymus DNA, coliphage T4 DNA, and poly(dG-dC). poly(dG-dC) was studied by optical (linear flow dichroism, UV-vis spectroscopy) and quantum chemical methods. The linear dichroism (LD) signal of TPT bound to DNA was shown to have positive sign in the range 260-295 nm. This means that the plane of quinoline fragment (rings A and B) of TPT molecule form an angle lower 54 degrees with the long axis of DNA, and hence TPT molecule can not intercalate between DNA base pairs. TPT was established to bind to calf thymus DNA as readily as to coliphage T4 DNA whose all cytosines in the major groove were glycosylated at the 5th position. Consequently, the DNA major groove does not participate in TPT binding. TPT molecule was shown to compete with distamycin for binding sites in the minor groove of DNA and poly(dG-dC). poly(dG-dC). Thus, it was demonstrated for the first time that TPT binds to DNA at its minor groove.


Morphology, microstructure, crystallography, and chemistry of distinct CaCO3 deposits formed by early recruits of the scleractinian coral Pocillopora damicornis.
Scleractinian corals begin their biomineralization process shortly after larval settlement with the formation of calcium carbonate (CaCO(3)) structures at the interface between the larval tissues and the substrate. The newly settled larvae exert variable degrees of control over this skeleton formation, providing an opportunity to study a range of biocarbonate structures, some of which are transient and not observed in adult coral skeletons. Here we present a morphological, structural, crystallographic, and chemical comparison between two types of aragonite deposits observed during the skeletal development of 2-days old recruits of Pocillopora damicornis: (1) Primary septum and (2) Abundant, dumbbell-like structures, quasi-randomly distributed between initial deposits of the basal plate and not present in adult corals-At the mesoscale level, initial septa structures are formed by superimposed fan-shaped fasciculi consisting of bundles of fibers, as also observed in adult corals. This organization is not observed in the dumbbell-like structures. However, at the ultrastructural level there is great similarity between septa and dumbbell components. Both are composed of <100 nm granular units arranged into larger single-crystal domains.Chemically, a small difference is observed between the septae with an average Mg/Ca ratio around 11 mmol/mol and the dumbbell-like structures with ca. 7 mmol/mol; Sr/Ca ratios are similar in the two structures at around 8 mmol/mol-Overall, the observed differences in distribution, morphology, and chemistry between septa, which are highly conserved structures fundamental to the architecture of the skeleton, and the transient, dumbbell-like structures, suggest that the latter might be formed through less controlled biomineralization processes. Our observations emphasize the inherent difficulties involved in distinguishing different biomineralization pathways based on ultrastructural and crystallographical observations.


Novel bis-cyclic guanidines as potent membrane-active antibacterial agents with therapeutic potential.
We designed a class of small dimeric cyclic guanidine derivatives which display potent antibacterial activity against both multidrug-resistant Gram-negative and Gram-positive bacteria. They could compromise bacterial membranes without developing resistance, inhibit biofilms formed by E. coli, and exhibit excellent in vivo activity in the MRSA-infected thigh burden mouse model.


Influence of a biofilm bioreactor on water quality and microbial communities in a hypereutrophic urban river.
Biofilms play an important role in degradation, transformation and assimilation of anthropogenic pollutants in aquatic ecosystems. In this study, we assembled a tubular bioreactor containing a biofilm substrate and aeration device, which was introduced into mesocosms to explore the effects of bioreactor on physicochemical and microbial characteristics of a hypereutrophic urban river. The biofilm bioreactor greatly improved water quality, especially by decreasing dissolved inorganic nitrogen (DIN) concentrations, suggesting that biofilms were the major sites of nitrification and denitrification with an oxygen concentration gradient. The biofilm bioreactor increased the abundance of planktonic bacteria, whereas diversity of the planktonic microbial community decreased. Sequencing revealed that Proteobacteria, Bacteroidetes, Planctomycetes, and Actinobacteria were the four predominant phyla in the planktonic microbial community, and the presence of the biofilm bioreactor increased the relative abundance of Proteobacteria. Variations in microbial communities were most strongly affected by the presence of the biofilm bioreactor, as indicated by principal component analysis (PCA) and redundancy analysis (RDA). This study provides valuable insights into changes in ecological characteristics associated with self-purification processes in hypereutrophic urban rivers, and may be of important for the application of biofilm bioreactor in natural urban river.


Fatherhood reduces the survival of adult-generated cells and affects various types of behavior in the prairie vole (Microtus ochrogaster ).
Motherhood has profound effects on physiology, neuronal plasticity, and behavior. We conducted a series of experiments to test the hypothesis that fatherhood, similarly to motherhood, affects brain plasticity (such as cell proliferation and survival) and various behaviors in the highly social prairie vole (Microtus ochrogaster). In Experiment 1, adult males were housed with their same-sex cage mate (control), single-housed (isolation), or housed with a receptive female to mate and produce offspring (father) for 6 weeks. Fatherhood significantly reduced cell survival (assessed by bromodeoxyuridine labeling), but not cell proliferation (assessed by Ki67-labeling), in the amygdala, dentate gyrus of the hippocampus, and ventromedial hypothalamus, suggesting that fatherhood affects brain plasticity. In Experiment 2, neither acute (20 min) nor chronic (20 min daily for 10 consecutive days) pup exposure altered cell proliferation or survival in the brain, but chronic pup exposure increased circulating corticosterone levels. These data suggest that reduced cell survival in the brain of prairie vole fathers was unlikely to be due to the level of pup exposure and display of paternal behavior, and may not be mediated by circulating corticosterone. The effects of fatherhood on various behaviors (including anxiety-like, depression-like, and social behaviors) were examined in Experiment 3. The data indicated that fatherhood increased anxiety- and depression-like behaviors as well as altered aggression and social recognition memory in male prairie voles. These results warrant further investigation of a possible link between brain plasticity and behavioral changes observed due to fatherhood.


Diet Quality and Cardiovascular Disease Risk in Postmenopausal Women With Type 2 Diabetes Mellitus: The Women's Health Initiative.
Background Dietary patterns are associated with cardiovascular disease (CVD) risk in the general population, but diet-CVD association in populations with diabetes mellitus is limited. Our objective was to examine the association between diet quality and CVD risk in a population with type 2 diabetes mellitus. Methods and Results We analyzed prospective data from 5809 women with prevalent type 2 diabetes mellitus at baseline from the Women's Health Initiative. Diet quality was defined using alternate Mediterranean, Dietary Approach to Stop Hypertension, Paleolithic, and American Diabetes Association dietary pattern scores calculated from a validated food frequency questionnaire. Multivariable Cox's proportional hazard regression was used to analyze the risk of incident CVD. During mean 12.4 years of follow-up, 1454 (25%) incident CVD cases were documented. Women with higher alternate Mediterranean, Dietary Approach to Stop Hypertension, and American Diabetes Association dietary pattern scores had a lower risk of CVD compared with women with lower scores (Q5 v Q1) (hazard ratio [HR]aMed 0.77, 95% CI 0.65-0.93; HRDASH 0.69, 95% CI 0.58-0.83; HRADA 0.71, 95% CI 0.59-0.86). No association was observed between the Paleolithic score and CVD risk. Conclusions Dietary patterns that emphasize higher intake of fruits, vegetables, whole grains, nuts/seeds, legumes, a high unsaturated:saturated fat ratio, and lower intake of red and processed meats, added sugars, and sodium are associated with lower CVD risk in postmenopausal women with type 2 diabetes mellitus.


Creatine metabolism at the uterine-conceptus interface during early gestation in sheep†.
Ruminant conceptuses that elongate and attach to the uterine luminal epithelium (LE) to establish pregnancy require a large amount of adenosine triphosphate (ATP). The creatine (Cr)-creatine kinase (CK)-phosphocreatine (PCr) system re-generates ATP in dividing and migrating cells such as the conceptus trophectoderm cells. However, little is known about metabolism of Cr within uterine and conceptus tissues in livestock species during early gestation. In this study, Suffolk ewes were ovariohysterectomized on Days 9, 12, 15, 16, 17, 18, 20, or 21 of pregnancy (n = 2-5 animals/per day) to investigate metabolites, mRNAs, and proteins of the Cr-CK-PCr system at uterine-conceptus interface. Amounts of Cr and guanidinoacetate (GA) in uterine flushings increased between Days 12 and 17 of pregnancy. Endometrial expression of mRNAs for GA formation (AGAT), Cr synthesis (GAMT), and Cr/PCr utilization (CKB) was greater on Days 17 and 21 than on Days 9 and 12 of pregnancy. Immunoreactive AGAT was detected in uteri only on Day 21 but not in uteri or conceptuses at earlier days of pregnancy. GAMT, SLC6A8, and CKs were expressed in uterine luminal and glandular epithelia. Immunoreactive CKs (CKB, CKM, and CKMT1) appeared greater on Day 9 than Day 17 of pregnancy. Immunoreactive GAMT and CKs appeared greater in trophectoderm of conceptuses on Day 20 than on Day 15 of pregnancy, whereas the opposite was observed for that of SLC6A8. This study provides insights into cell-, tissue-, and time-specific metabolism of Cr at the uterine-conceptus interface suggesting a role for the Cr-CK-PCr system in ovine conceptus development and implantation.


Bases of variation in resistance to beta-lactams in Klebsiella oxytoca isolates hyperproducing K1 beta-lactamase.
Nineteen isolates of Klebsiella oxytoca were examined, representing 18 distinct strains. All were from a 1994 survey of resistance amongst klebsiellae in intensive care units in Europe, and all had reduced susceptibility, or were resistant, to cefuroxime, ceftriaxone and aztreonam, suggesting hyperproduction of the chromosomal K1 beta-lactamase. We sought to confirm this mechanism and to identify why the levels of resistance varied between isolates. Possible reasons for variation were differences in the quantity or subtype of the K1 enzyme or differences in this enzyme's interplay with permeability. Spectrophotometric assays showed that all 19 isolates had K1-like beta-lactamases and that these were present at > or = 15-fold higher levels than in beta-lactam-sensitive K. oxytoca isolates. Fourteen of the 19 isolates had the OXY-2 form of K1 enzyme, while the remaining five had the OXY-1 form, as determined by isoelectric focusing and PCR amplification. Most isolates with the OXY-2 enzyme were more resistant than those with the OXY-1 subtype, but this difference partly reflected enzyme quantity rather than subtype. More generally, and irrespective of enzyme subtype, levels of resistance were broadly related to beta-lactamase specific activity, and the degree of hyperproduction was a major determinant of the level of resistance. Nevertheless, other factors had a role too: several isolates had reduced susceptibility or were resistant to cefoxitin, which is not a substrate for K1 enzyme, and examination of outer membrane protein profiles revealed considerable strain-to-strain diversity in the molecular weight range typical of the major enterobacterial porins (40-48 kDa).


Palliative care curriculum development: a model for a content and process-based approach.
To ensure its success, a new curriculum has to meet the needs of learners, patients, and the institution. A review of the literature indicates that despite a tremendous need for palliative care services and a lack of appropriate knowledge and attitudes among physicians, few palliative care curricula for medical residents have been developed. Most are developed by national organizations, and as a result can not meet the individual needs of different institutions. This paper outlines the process of developing a palliative care curriculum in the context of available institutional resources that meets the learners' needs. The development of a curriculum can be divided into four phases: needs assessment, curricular design, implementation, and evaluation. Content (curricular content, instructional strategies and available resources for the curriculum and the developmental process) and process (methods through which the curriculum is developed and institutional issues are addressed) issues that are pertinent to the successful completion of each phase are discussed. Two hypothetical institutions are used to illustrate relevant issues. Methods that have been successfully used to develop residency curricula are discussed.


Dissipation of insecticidal Cry1Ac protein and its toxicity to nontarget aquatic organisms.
The widespread cultivation of Bacillus thuringiensis crops has raised public concerns on their risk to nontarget organisms. Persistence of Cry1Ac protein in soil, sediment and water and its toxicity to nontarget aquatic organisms were determined. The dissipation of Cry1Ac toxin was well described using first order kinetics, with the half-lives (DT50) ranging from 0.8 to 3.2, 2.1 to 7.6 and 11.0 to 15.8 d in soil, sediment and water, respectively. Microbial degradation played a key role in the dissipation of Cry1Ac toxin and high temperature accelerated the processes. Cry1Ac toxin was more toxic to the midge Chironomus dilutus than the amphipod Hyalella azteca, with the median lethal concentration (LC50) of C. dilutus being 155 ng/g dry weight and 201 ng/mL in 10-d sediment and 4-d water bioassays, respectively. While Cry1Ac toxin showed toxicity to the midges, risk of Bt proteins to aquatic nontarget organisms was limited because their environmentally relevant concentrations were much lower than the LC50s.


New monoclonal antibodies in CD44 and CD58: their use to quantify CD44 and CD58 on normal human erythrocytes and to compare the distribution of CD44 and CD58 in human tissues.
The cell-surface glycoproteins CD44 and CD58 are involved in cell adhesion reactions. In this paper 12 monoclonal antibodies in CD44 and two in CD58 are described. Competitive binding assays using CD44 antibodies identified three distinct epitope groups. Antibodies in Group 1 and, with one exception (BRIC 214), antibodies in group 2, but not antibodies in Group 3, recognized epitopes that are sensitive to reduction and to trypsin or chymotrypsin treatment of intact erythrocytes, and so these epitopes probably reside on the N-terminal disulphide-bonded domain of CD44. Antibodies in CD44 did not inhibit the binding of CD58 antibodies to erythrocytes or vice versa. Quantitative binding studies using radioiodinated IgG measured 1888-5592 copies of CD44 and 1772-3290 copies of CD58 on normal erythrocytes. Similar measurements with radioiodinated Fab fragments gave values of 6508-10,450 (CD44) and 3457-7622 (CD58). Immunocytochemical studies indicated that CD44 is much more widely expressed in non-haemopoietic tissues than CD58. Comparison with previously described CD44 antibodies suggests that antibodies in our Group 1 encompass Hermes 2 and that those in Group 2 encompass Hermes 1. All the CD44 antibodies gave weakened reactions with Lu(a-b-) erythrocytes of the In(Lu) type by one or more methods. BRIC 214 and antibodies in epitope Group 3 were used to demonstrate that CD44 on these variant cells gives membrane-bound trypsin and chymotrypsin cleavage fragments of similar molecular weight to those obtained with normal erythrocytes.


[Prognostic value of bone marrow hematogones in childhood B-lineage acute lymphoblastic leukemia].
To study the prognostic value of hematogones (HGs) for childhood B-lineage acute lymphoblastic leukemia (B-ALL) during consolidation chemotherapy. A retrospective analysis was conducted for 196 children with newly-diagnosed B-ALL. They were divided into high-risk group (n=55), intermediate-risk group (n=69), and low-risk group (n=72) by risk stratification, and into complete remission group (n=165) and relapse group (n=31) by clinical outcome. The European BIOMED-1 standard flow cytometry for minimal residual disease (MRD) was used to determine the number of HGs during consolidation chemotherapy. The Kaplan-Meier survival curve was used to assess event-free survival (EFS). The high-risk group had a significantly lower number of HGs than the intermediate-risk and low-risk groups (P<0.05). The number of HGs in the complete remission group was significantly higher than in the relapse group (P<0.05). The children with HGs ≤1.0% had a significantly lower EFS than those with HGs <1.0% (P<0.05). HGs can be used to assess the treatment outcome and prognosis in children with B-ALL, and proliferation of HGs reflects the good effect of chemotherapy in such children.


Molecular mechanisms of inherited demyelinating neuropathies.
The past 15 years have witnessed the identification of more than 25 genes responsible for inherited neuropathies in humans, many associated with primary alterations of the myelin sheath. A remarkable body of work in patients, as well as animal and cellular models, has defined the clinical and molecular genetics of these illnesses and shed light on how mutations in associated genes produce the heterogeneity of dysmyelinating and demyelinating phenotypes. Here, we review selected recent developments from work on the molecular mechanisms of these disorders and their implications for treatment strategies.


Immunohistochemical analysis of forensic evidence from a double homicide.
We report the use of immunohistochemical staining for analysis of forensic evidence from a double homicide. A 38-year-old woman and her 7-year-old daughter were murdered by multiple blows to the head and face with a tomahawk, resulting in multiple fragments of brain tissue scattered about the murder scene. The victims' husband and father was the main suspect, who maintained that he was out of town on business during the evening of the murders. However, a shirt taken from the suspect's car on the morning after the murders (secured by the police before the suspect visited the murder scene) was found to have two small stains. DNA analysis on the stains showed the presence of the deceased wife's DNA, and immunohistochemical stains on shirt fragments conclusively documented the presence of deep central nervous system tissue, providing the critical piece of evidence needed to arrest and prosecute the suspect. This report demonstrates that shirt or similar cloth fragments can be processed into paraffin blocks and subsequently immunostained to search for and classify types of tissue fragments that may be present on the fabric.


Autopsy diagnoses can increase DRG reimbursement.
To determine how missed clinical diagnoses revealed at autopsy might affect third-party payment through DRGs, the authors performed a retrospective comparison of clinical and pathologic diagnoses in 100 consecutive cases that came to autopsy. In 13 of these cases, the autopsy findings resulted in a change in DRG assignment, which could have produced an additional $14,235 for the hospital.


Evaluation of the systemic acute phase response and endometrial gene expression of serum amyloid A and pro- and anti-inflammatory cytokines in mares with experimentally induced endometritis.
Infectious infertility in the mare is clinically well described, little is however known about the systemic acute phase reaction (APR) and local immunological responses accompanying equine endometritis. The aim of this study was to monitor selected markers of the APR in the systemic circulation and to correlate them to the local innate immune response in the uterus during infectious endometritis. Six adult standard bred mares received an intrauterine infusion of 10(9)CFU Escherichia coli. Blood samples were obtained before (0 h) and 3, 6, 12, 24, 36, 48, 72, 96 and 120 h post inoculation (pi), and endometrial biopsies were sampled before, and 3, 12, 24, 48 and 72 h pi. The infectious endometritis elicited a systemic APR with significantly increased concentrations of the acute phase proteins (APPs) serum amyloid A (SAA) and fibrinogen. Relative gene expression analyses were performed on extracted RNA from endometrial biopsies using quantitative real-time PCR and specific primers for SAA and pro- and anti-inflammatory cytokines. Expression of SAA was significantly up-regulated at 3 and 12h pi, and a significant up-regulated expression of IL-1β, TNFα, IL-8 and IL-10 was observed at 3h pi. Plasma concentration of SAA was significantly correlated to endometrial SAA expression. The results of the present study demonstrate that endometritis gives rise to a systemic APR and an up-regulated endometrial gene expression of SAA and several pro-and anti-inflammatory cytokines. Understanding endometrial expression of acute phase proteins and selected cytokines contributing to uterine immunity in equine endometritis could improve understanding of events leading to infertility in the mare and help identify candidate genes of mediators/markers for diagnostic use.