Transport of glucose and galactose in kidney-cortex cells.
1. The aerobic transport of d-glucose and d-galactose in rabbit kidney tissue at 25 degrees was studied. 2. In slices forming glucose from added substrates an accumulation of glucose against its concentration gradient was found. The apparent ratio of intracellular ([S](i)) and extracellular ([S](o)) glucose concentrations was increased by 0.4mm-phlorrhizin and 0.3mm-ouabain. 3. Slices and isolated renal tubules actively accumulated glucose from the saline; the apparent [S](i)/[S](o) fell below 1.0 only at [S](o) higher than 0.5mm. 4. The rate of glucose oxidation by slices was characterized by the following parameters: K(m) 1.16mm; V(max.) 4.5mumoles/g. wet wt./hr. 5. The active accumulation of glucose from the saline was decreased by 0.1mm-2,4-dinitrophenol, 0.4mm-phlorrhizin and by the absence of external Na(+). 6. The kinetic parameters of galactose entry into the cells were: K(m) 1.5mm; V(max) 10mumoles/g. wet wt./hr. 7. The efflux kinetics from slices indicated two intracellular compartments for d-galactose. The galactose efflux was greatly diminished at 0 degrees , was inhibited by 0.4mm-phlorrhizin, but was insensitive to ouabain. 8. The following mechanism of glucose and galactose transport in renal tubular cells is suggested: (a) at the tubular membrane, these sugars are actively transported into the cells by a metabolically- and Na(+)-dependent phlorrhizin-sensitive mechanism; (b) at the basal cell membrane, these sugars are transported in accordance with their concentration gradient by a phlorrhizin-sensitive Na(+)-independent facilitated diffusion. The steady-state intracellular sugar concentration is determined by the kinetic parameters of active entry, passive outflow and intracellular utilization.


The research team, calibration, and quality assurance in clinical trials in periodontics.
The present report provides an overview of practical issues related to training, calibrating, and monitoring the individuals involved in conducting a clinical trial. These aspects are considered in relation to a theoretical multi-center clinical trial designed to test the safety and efficacy of a new periodontal treatment. Many factors need to be controlled in order to reduce inherent variability and provide standardization across the study population and study centers in a large-scale multi-center clinical trial. Components incorporated to control these factors include identification and recruitment of study centers, pre-study training and organization of the teams at the different research centers, quality assurance procedures and monitoring of the study for compliance with protocol and safety requirements, and management of databases. Details are given regarding the use of pre-study survey questionnaires to assess research center capabilities, methods for training and calibrating examiners and therapists and methods for integrating these aspects prior to actual study commencement, and quality assurance after the study has begun. The review considers the critical practical details regarding all of these aspects of study organization and execution.


Pseudohypoparathyroidism. Variable manifestations within a family.
A wide variety of clinical and biochemical manifestations have been reported in association with pseudohypoparathyroidism and associated disorders. This is illustrated by a family study in which the affected members show widely differing characteristics.


Biomarkers for cardiovascular risk assessment in autoimmune diseases.
Autoimmune diseases, such as antiphospholipid syndrome, systemic lupus erythematosus, and rheumatoid arthritis, are characterized by a high prevalence of cardiovascular (CV) disease (CVD), which constitutes the leading causes of morbidity and mortality among such patients. Although such effects are partly explained by a higher prevalence of traditional CV risk factors, many studies indicate that such factors do not fully explain the enhanced CV risk in these patients. In addition, risk stratification algorithms based upon traditional CV risk factors are not as predictive in autoimmune diseases as in the general population. For these reasons, the timely and accurate assessment of CV risk in these high-risk populations still remains an unmet clinical need. An enhanced contribution of different inflammatory components of the immune response, as well as autoimmune elements (e.g. autoantibodies, autoantigens, and cellular response), has been proposed to underlie the incremental CV risk observed in these populations. Recent advances in proteomic tools have contributed to the discovery of proteins involved in CVDs, including some that may be suitable to be used as biological markers. In this review we summarize the main markers in the field of CVDs associated with autoimmunity, as well as the recent advances in proteomic technology and their application for biomarker discovery in autoimmune disease.


Regulation of adipokine production in human adipose tissue by propionic acid.
Dietary fibre (DF) has been shown to be protective for the development of obesity, insulin resistance and type 2 diabetes. Short-chain fatty acids, produced by colonic fermentation of DF might mediate this beneficial effect. Adipose tissue plays a key role in the regulation of energy homeostasis, therefore, we investigated the influence of the short-chain fatty acid propionic acid (PA) on leptin, adiponectin and resistin production by human omental (OAT) and subcutaneous adipose tissue (SAT). As PA has been shown to be a ligand for G-protein coupled receptor (GPCR) 41 and 43, we investigated the role of GPCR's in PA signalling. Human OAT and SAT explants were obtained from gynaecological patients who underwent surgery. Explants were incubated for 24 h with PA. Adipokine secretion and mRNA expression were determined using ELISA and RT-PCR respectively. We found that PA significantly stimulated leptin mRNA expression and secretion by OAT and SAT, whereas it had no effect on adiponectin. Furthermore, PA reduced resistin mRNA expression. Leptin induction, but not resistin reduction, was abolished by inhibition of Gi/o-coupled GPCR signalling. Moreover, GPCR41 and GPCR43 mRNA levels were considerably higher in SAT than in OAT. We demonstrate that PA stimulates expression of the anorexigenic hormone leptin and reduces the pro-inflammatory factor resistin in human adipose tissue depots. This suggests that PA is involved in regulation of human energy metabolism and inflammation and in this way may influence the development of obesity and type 2 diabetes.


Using Virtual Reality in the Inference-Based Treatment of Compulsive Hoarding.
The present study evaluated the efficacy of adding a virtual reality (VR) component to the treatment of compulsive hoarding (CH), following inference-based therapy (IBT). Participants were randomly assigned to either an experimental or a control condition. Seven participants received the experimental and seven received the control condition. Five sessions of 1 h were administered weekly. A significant difference indicated that the level of clutter in the bedroom tended to diminish more in the experimental group as compared to the control group F(2,24) = 2.28, p = 0.10. In addition, the results demonstrated that both groups were immersed and present in the environment. The results on posttreatment measures of CH (Saving Inventory revised, Saving Cognition Inventory and Clutter Image Rating scale) demonstrate the efficacy of IBT in terms of symptom reduction. Overall, these results suggest that the creation of a virtual environment may be effective in the treatment of CH by helping the compulsive hoarders take action over their clutter.


Multifaceted roles and regulation of nucleotide-binding oligomerization domain containing proteins.
Nucleotide-binding oligomerization domain-containing proteins, NOD1 and NOD2, are cytosolic receptors that recognize dipeptides and tripeptides derived from the bacterial cell wall component peptidoglycan (PGN). During the past two decades, studies have revealed several roles for NODs beyond detecting PGN fragments, including activation of an innate immune anti-viral response, NOD-mediated autophagy, and ER stress induced inflammation. Recent studies have also clarified the dynamic regulation of NODs at cellular membranes to generate specific and balanced immune responses. This review will describe how NOD1 and NOD2 detect microbes and cellular stress and detail the molecular mechanisms that regulate activation and signaling while highlighting new evidence and the impact on inflammatory disease pathogenesis.


Calculation of rotational partition functions by an efficient Monte Carlo importance sampling technique.
The evaluation of the classical rotational partition function represented by a configuration integral over all external and internal rotational degrees of freedom of nonrigid chain polyatomic molecules is described. The method of Pitzer and Gwinn is used to correct the classical partition function for quantum mechanical effects at low temperatures. The internal rotor hindrance and all coupling arising from the external and internal rotational degrees of freedom are explicitly taken into account. Importance sampling Monte Carlo based on the adaptive VEGAS algorithm to perform multidimensional integration is implemented within the TINKER program package. A multidimensional potential energy hypersurface is calculated with the MM3(2000) molecular mechanics force field. Numerical tests are performed on a number of small n-alkanes (from ethane to octane), for which the absolute entropies calculated at three different temperatures are compared both with the experimental values and with the previous theoretical results. The application of a more efficient importance sampling technique developed here results in a substantial reduction of statistical errors in the evaluation of the configuration integral for a given number of Monte Carlo steps. Error estimates for the calculated entropies are given, and possible sources of systematic errors, and their importance for a reliable prediction of the absolute entropy, are discussed.


Microbial metabolism of pseudoprotodioscin.
Microbial transformation of the furostanol saponin pseudoprotodioscin ( 1) using Aspergillus fumigatus resulted in the isolation of two new steroidal metabolites, 3- O-[bis- alpha- L-rhamnopyranosyl-(1-->2 and 1-->4)- beta- D-glucopyranosyl]-22 R,25 R-spirost-5-ene-3 beta,20 alpha-diol ( 2) and 3- O-[bis- alpha- L-rhamnopyranosyl-(1-->2 and 1-->4)- beta- D-glucopyranosyl]-25 R-furost-5-ene-3 beta,22 alpha,26-triol ( 3), in addition to the previously reported steroidal saponins: dioscin ( 4) and progenin II ( 5). The structure elucidation of these metabolites was based primarily on 1D and 2D NMR analyses. Metabolites 2 - 5 showed significant cytotoxicity against cancer cell lines A375, L929, and HeLa with IC (50) values ranging from 1.18 microM to 17.88 microM.


Novel luminescent benzopyranothiophene- and BODIPY-derived aroylhydrazonic ligands and their dicopper(II) complexes: syntheses, antiproliferative activity and cellular uptake studies.
Two novel unsymmetrical binucleating aroylhydrazonic ligands and four dicopper(II) complexes carrying fluorescent benzopyranothiophene (BPT) or boron dipyrromethene (BODIPY) entities were synthesized and fully characterized. Complex 1, derived from the BPT-containing ligand H3L1, had its crystal structure elucidated through X-ray diffraction measurements. The absorption and fluorescence profiles of all the compounds obtained were discussed. Additionally, the stability of the ligands and complexes was monitored by UV-vis spectroscopy in DMSO and biologically relevant media. All the compounds showed moderate to high cytotoxicity towards the triple negative human breast cancer cell line MDA-MB-231. BPT derivatives were the most cytotoxic, specially H3L1, reaching an IC50 value up to the nanomolar range. Finally, fluorescence microscopy imaging studies employing mitochondria- and nucleus-staining dyes showed that the BODIPY-carrying ligand H3L2 was highly cell permeant and suggested that the compound preferentially accumulates in the mitochondria.


Alterations in cellular differentiation, mitogenesis, cytoskeleton and growth characteristics during Syrian hamster embryo cell multistep in vitro transformation.
In vitro Syrian hamster embryo (SHE) cell transformation is a neoplastic process that proceeds through several identifiable consecutive stages including in vitro morphological transformation (mt), acquisition of immortality (I+), acquisition of tumorigenicity (T+) and tumor-derived cells (I'TD). Eight transformed lineages consisting of cells at the mt, I+, T+ and I'TD stages were assayed for alterations in general markers of cell differentiation, mitogenic signaling pathways, cytoskeleton and cellular growth in 3D matrix. Alterations in cellular differentiation markers included a decrease in H19 gene expression and placental alkaline phosphatase enzymatic activity at the mt stage in all lineages examined with a complete absence of H19 gene expression and placental alkaline phosphatase enzymatic activity by the I'TD stage in a majority of transformed lineages. Changes in mitogenic signaling pathways included the production of autocrine growth factors and alterations in growth factor-induced immediate early gene expression by the I'TD stage of transformation in the majority of transformed lineages investigated. By the I'TD stage of transformation in most lineages, changes in both the cytoskeleton (including a decrease in tropomyosin-I gene expression) and the Matrigel growth characteristics of SHE cells were observed.


Low-dose controlled release of mTOR inhibitors maintains T cell plasticity and promotes central memory T cells.
An important goal for improving vaccine and immunotherapy technologies is the ability to provide further control over the specific phenotypes of T cells arising from these agents. Along these lines, frequent administration of rapamycin (Rapa), a small molecule inhibitor of the mammalian target of rapamycin (mTOR), exhibits a striking ability to polarize T cells toward central memory phenotypes (TCM), or to suppress immune function, depending on the concentrations and other signals present during administration. TCM exhibit greater plasticity and proliferative capacity than effector memory T cells (TEFF) and, therefore, polarizing vaccine-induced T cells toward TCM is an intriguing strategy to enhance T cell expansion and function against pathogens or tumors. Here we combined biodegradable microparticles encapsulating Rapa (Rapa MPs) with vaccines composed of soluble peptide antigens and molecular adjuvants to test if this approach allows polarization of differentiating T cells toward TCM. We show Rapa MPs modulate DC function, enhancing secretion of inflammatory cytokines at very low doses, and suppressing function at high doses. While Rapa MP treatment reduced - but did not stop - T cell proliferation in both CD4+ and CD8+ transgenic T cell co-cultures, the expanding CD8+ T cells differentiated to higher frequencies of TCM at low doses of MP Rapa MPs. Lastly, we show in mice that local delivery of Rapa MPs to lymph nodes during vaccination either suppresses or enhances T cell function in response to melanoma antigens, depending on the dose of drug in the depots. In particular, at low Rapa MP doses, vaccines increased antigen-specific TCM, resulting in enhanced T cell expansion measured during subsequent booster injections over at least 100days.


Severe Strongyloidiasis in Solid Organ Transplant Recipients: Should We Preventively Treat the Recipient, the Donor, or Both?
Strongyloidiasis is caused by a soil-transmitted helminth that is endemic in tropical and subtropical countries. The parasite can complete its life cycle without leaving the host, allowing autoinfection and persistence. The risk of infection in travelers is low, but the disease may become lethal following immunosuppression. In case of solid organ transplantation, the risk of donor transmission has been suspected for several years. However, the management of live donors in this context has only recently been considered, and no guidelines exist for the management of deceased donors. To highlight the complexity of diagnosing, treating, and preventing strongyloidiasis donor transmission, we describe a case of possible transmission of severe strongyloidiasis to a kidney transplant recipient with limited travel history. Taking into account the difficulty of diagnosing chronic strongyloidiasis infection and the increase in travel and immunosuppressive treatments, we recommend pragmatic management guidelines to limit the risks of infection.


FOOD TEXTURE MEASUREMENTS IN CANADA.
A survey of the food industry, government agencies and educational institutions dealing with foods has been conducted to establish the nature and extent to which instrumental texture measurements are employed in Canada. Two hundred and fifteen questionnaires were sent out and 123 (57%) returned. Of those replying, 52% were using texture evaluation instruments. Within the food industry the use of texture measuring instruments was as follows: meat 36%; fish 20%; canner/freezer 79%; dairy 42%; confectionery 73%; baking 50%; fats and oils 78%; multi product 77%; beverage 0%. Most widely used instruments in the quality control area were rotational viscometers and penetrometers. The more complex instruments, such as the Universal Testing Machine and the Shear Press, were used to a larger extent in the research area. The use of some instruments appeared to be restricted to particular commodity groups. The questionnaire dealt among other things with the use of taste panels, statistical evaluation of results, and the need for expanded use of instruments. There was general agreement that more standardization of methods is desirable.


Long non-coding RNA HOTAIR promotes glioblastoma cell cycle progression in an EZH2 dependent manner.
The long non-coding RNA Hox transcript antisense intergenic RNA (HOTAIR) was recently implicated in breast cancer metastasis and is predictive of poor prognosis in colorectal and pancreatic cancers. We recently discovered that HOTAIR is a cell cycle-related lncRNA in human glioma, and its expression is closely associated with glioma staging and poor prognosis. Although lysine specific demethylase 1 (LSD1) and polycomb repressive complex 2 (PRC2) have been demonstrated to be functional targets of HOTAIR, how HOTAIR regulates glioma cell cycle progression remains largely unknown. In this study, we found that EZH2 (predominant PRC2 complex component) inhibition blocked cell cycle progression in glioma cells, consistent with the effects elicited by HOTAIR siRNA. However, the inhibition of LSD1 did not affect cell cycle progression in glioma cells. These results suggest that HOTAIR might regulate cell cycle progression through EZH2. Our intracranial mice model also revealed delayed tumor growth in HOTAIR siRNA- and EZH2 inhibitor-treated groups. Moreover, in HOTAIR knock-down cell lines, the expression of the PRC2-binding domain of HOTAIR (5' domain) but not of the LSD1-binding domain of HOTAIR (3' domain) resulted in accelerated cell cycle progression. In conclusion, HOTAIR promotes cell cycle progression in glioma as a result of the binding of its 5' domain to the PRC2 complex.


Cryptococcal meningitis: diagnostic value of cryptococcal antigen in cerebrospinal fluid.
In three previously reported cases of cryptococcal meningitis, the only laboratory evidence for this diagnosis was the presence of cryptococcal antigen in the cerebrospinal fluid (CSF). Three additional patients had chronic meningitis and repeatedly negative CSF cultures and had cryptococcal antigen demonstrated in the CSF. In our patients, the diagnosis was further supported by the complete recovery after amphotericin B therapy in two and the demonstration of Cryptococcus neoformans in the meninges at autopsy in the third. In certain patients with chronic meningitis, the detection of cryptococcal antigen in the CSF may be the only means of establishing a diagnosis during life. In such patients, if cryptococcal antigen is present in the CSF in a titer of larger than or equal to 1:8, antifungal therapy should be initiated, pending results of other diagnostic studies.


Are child-centric aspects in newborn and child health systematic review and meta-analysis protocols and reports adequately reported?-two systematic reviews.
Evidence suggests that newborn and child health systematic reviews and meta-analyses exhibit poor quality in reporting. The "Preferred Reporting Items in Systematic Review and Meta-Analysis" (PRISMA) and PRISMA-Protocols (PRISMA-P) checklists have been developed to improve the reporting of systematic review results and protocols, respectively. We aimed to evaluate the clarity and transparency in reporting of child-centric items in child health systematic reviews (SRs) and SR protocols and to identify areas where reporting could be strengthened. Two preliminary lists of potential child-centric reporting items were used to examine current reporting. The Cochrane, DARE, MEDLINE, and EMBASE libraries were searched from 2010 to 2014 for systematic reviews that included children. Each report and protocol that met the inclusion criteria had their quality of reporting assessed by their reporting of child-centric items. Quality of reporting was assessed per whether one third, one to two thirds, or more than two thirds of papers complied with potential child-centric potential modifications/extensions to PRISMA and were analyzed by the following: (i) paper type (i.e., report vs. protocol), (ii) publication type (i.e., Cochrane vs. non-Cochrane), and (iii) population type (i.e., child-only vs. mixed populations vs. family/maternal). Of the 414 eligible articles, 248 reports and 76 protocols were included. In 21 of 24 potential SR reporting items and 13 of 14 potential SR protocol reporting items, less than two thirds of papers met the child-centric reporting item requirements. Mixed population studies displayed significantly poorer reporting in comparison to child-only and family/maternal intervention studies for 11 potential SR reporting items (p < 0.05) and five potential SR protocol items (p < 0.05). When comparing non-Cochrane to Cochrane reports and protocols, five items in both lists were found to perform significantly poorer in non-Cochrane reports (p < 0.05). Significant differences in reporting quality were found in three of 14 items shared between the potential SR reporting items and potential SR protocol reporting items (p < 0.05). Newborn and child health systematic reviews and meta-analyses exhibit incomplete reporting, thereby hindering prudent decision-making by healthcare providers and policy makers. These results provide a rationale for the implementation of child-centric extensions and modifications to current PRISMA and PRISMA-P, such as to improve reporting in this population.


Effect of alcoholic beverages on surface roughness and microhardness of dental composites.
The aim of this study was to evaluate the microhardness and surface roughness of composite resins immersed in alcoholic beverages. Three composite resins were used: Durafill (Heraeus Kulzer), Z250 (3M-ESPE) and Z350 XT (3M-ESPE). The inital surface roughness and microhardness were measured. The samples were divided into four groups (n=30): G1-artificial saliva; G2-beer; G3-vodka; G4-whisky. The samples were immersed in the beverages 3× a day for 15 min and 30 days. The surface roughness and microhardness assays were repeated after immersion period. The data were statistically analyzed by two-way ANOVA and Tukey-HSD test (p<0.05). Surface roughness increased for all composite resins immersed in beer and whisky. Microhardness of all groups decreased after immersion in alcoholic beverages. The effect of these beverages on dental composites is depended upon the chemical composition, immersion time, alcohol content and pH of solutions.


Automated Feature Mining for Two-Dimensional Liquid Chromatography Applied to Polymers Enabled by Mass Remainder Analysis.
A fast algorithm for automated feature mining of synthetic (industrial) homopolymers or perfectly alternating copolymers was developed. Comprehensive two-dimensional liquid chromatography-mass spectrometry data (LC × LC-MS) was utilized, undergoing four distinct parts within the algorithm. Initially, the data is reduced by selecting regions of interest within the data. Then, all regions of interest are clustered on the time and mass-to-charge domain to obtain isotopic distributions. Afterward, single-value clusters and background signals are removed from the data structure. In the second part of the algorithm, the isotopic distributions are employed to define the charge state of the polymeric units and the charge-state reduced masses of the units are calculated. In the third part, the mass of the repeating unit (i.e., the monomer) is automatically selected by comparing all mass differences within the data structure. Using the mass of the repeating unit, mass remainder analysis can be performed on the data. This results in groups sharing the same end-group compositions. Lastly, combining information from the clustering step in the first part and the mass remainder analysis results in the creation of compositional series, which are mapped on the chromatogram. Series with similar chromatographic behavior are separated in the mass-remainder domain, whereas series with an overlapping mass remainder are separated in the chromatographic domain. These series were extracted within a calculation time of 3 min. The false positives were then assessed within a reasonable time. The algorithm is verified with LC × LC-MS data of an industrial hexahydrophthalic anhydride-derivatized propylene glycol-terephthalic acid copolyester. Afterward, a chemical structure proposal has been made for each compositional series found within the data.


CircTLK1 alleviates oxygen-glucose deprivation/reperfusion induced apoptosis in HK-2 cells through miR-136-5p/Bcl2 signal axis.
The biological functions of circTLK1 in acute kidney injury (AKI), which mainly results from renal ischemia-reperfusion (IR), remain largely unknown. HK-2 cell treatment with oxygen and glucose deprivation, reoxygenation, and glucose (OGD/R) was used to simulate an AKI model that was mainly caused by renal IR. Then, the circTLK1 expression level in HK-2 cells treated with OGD/R was assessed by quantitative reverse transcription polymerase chain reaction (RT-qPCR). Functional experiments were performed with circTLK1 knockdown of HK-2 cells via Cell Counting Kit-8 (CCK8), flow cytometry (FCM), RT-qPCR, and western blotting. The circTLK1-miRNAs-mRNAs network was constructed following the ceRNA mechanism and visualized by Cytoscape software to investigate the mechanism of circTLK1 in AKI. RT-qPCR was performed to verify the relationship between circTLK1, miR-136-5p, and Bcl2. The level of miR-136-5p was knocked down to ensure its function in OGD/R-triggered apoptosis through experiments, including CCK8, FCM, RT-qPCR, and western blotting. CircTLK1 was downregulated in HK-2 cells subjected to OGD/R treatment and in mouse kidney tissues after renal IR, but the expression of miR-136-5p was the opposite. Interference with circTLK1 expression accelerated HK-2 cell apoptosis, which was overturned by miR-136-5p inhibitors. CircTLK1 targets miR-136-5p to upregulate Bcl2 expression and attenuate apoptosis in HK-2 cells. These data revealed the possible role of circTLK1 as a new biomarker for diagnosis as well as a target in AKI through the miR-136-5p/Bcl2 signaling axis.


Transcriptome profiling reveals insertional mutagenesis suppressed the expression of candidate pathogenicity genes in honeybee fungal pathogen, Ascosphaera apis.
Chalkbrood disease is caused by Ascosphaera apis which severely affects honeybee brood. Spore inoculation experiments shown pathogenicity varies among different strains and mutants, however, the molecular mechanism of pathogenicity is unclear. We sequenced, assembled and annotated the transcriptomes of wild type (SPE1) and three mutants (SPE2, SPE3 and SPE4) with reduced pathogenicity that were constructed in our previous study. Illumina sequencing generated a total of 394,910,604 clean reads and de novo Trinity-based assembled into 12,989 unigenes, among these, 9,598 genes were successfully annotated to known proteins in UniProt database. A total of 172, 3,996, and 650 genes were up-regulated and 4,403, 2,845, and 3,016 genes were down-regulated between SPE2-SPE1, SPE3-SPE1, and SPE4-SPE1, respectively. Overall, several genes with a potential role in fungal pathogenicity were detected down-regulated in mutants including 100 hydrolytic enzymes, 117 transcriptional factors, and 47 cell wall related genes. KEGG pathway enrichment analysis reveals 216 genes involved in nine pathways were down-regulated in mutants compared to wild type. The down-regulation of more pathways involved in pathogenicity in SPE2 and SPE4 than SPE3 supports their lower pathogenicity during in-vitro bioassay experiment. Expression of 12 down-regulated genes in mutants was validated by quantitative real time PCR. This study provides valuable information on transcriptome variation caused by mutation for further functional validation of candidate pathogenicity genes in A. apis.


Survey on attitudes of Italian pediatricians toward cough.
Children's cough is a daily concern for most pediatricians. The management of both acute and chronic cough requires a systematic and comprehensive approach. Despite the approved protocols for management, the pediatric assessment of cough and the corresponding prescribing attitude frequently do not fit these protocols, which can be affected by parental suggestions - sometimes substantially. The objective of this study was to investigate both the perception and the behavior of a representative sample of Italian pediatricians toward cough in real life. A specific questionnaire consisting of 18 questions was prepared. The questionnaire was completed by 300 pediatricians (all members of PAIDOSS: Italian National Observatory on Health of Childhood and Adolescence) who represented ~300,000 children. A vast majority of children have cough throughout the year (99.3% of respondents have cough during autumn/winter and 64.7% in spring/summer). Allergic disease is the most frequent suspected cause of chronic cough in children (53%), and this is supported by the high demand for consultations: 73% seek the opinion of allergologists, 62% of otorhinolaryngologists and only 33% of pulmonologists. The majority of pediatricians (92%) reported that they prescribe therapy in acute cough regardless of cough guidelines. Moreover, the survey pointed out the abuse of aerosol therapy (26% in acute cough and 38% in chronic cough) and of antibiotics prescription (22% in acute cough and 42% in chronic cough). Our survey suggests that some Italian pediatricians' therapeutic attitudes should be substantially improved in order to achieve better management of cough in children and to minimize the burden of cough.


Human herpesvirus-6B active infection associated with relapsing bilateral anterior optic neuritis.
Human herpesvirus-6 (HHV-6) is the causative agent of exanthem subitum. Both HHV-6 variants, A and B, have been associated with central nervous system (CNS) diseases, suggesting a wide neuropathogenic potential. We describe a case of recurrent bilateral anterior optic neuritis with HHV-6 active infection associated with clinical relapses. A 23-year old woman presented with progressive visual impairment, bilateral papillitis and painful ocular movements. Nested polymerase chain reaction (PCR) for DNA viruses, HHV-6 variant specific real time quantitative PCR, serological analysis and retrotranscription PCR (RT-PCR) for HHV-6 mRNA transcripts were performed. Nested PCR in PBMC and CSF samples was negative for all viruses but positive for HHV-6 DNA, subtyped as HHV-6B. The disease had a relapsing/remitting course. During relapses PBMC samples remained positive for HHV-6 DNA, and HHV-6 active infection was confirmed by the presence of anti-HHV-6 IgM and of HHV-6 U27 mRNA transcript. High viremia levels and relapses were overlapping. After the last relapse, the patient was successfully treated with gancyclovir. The case reported here suggests a possible association of HHV-6 in bilateral optic neuritis. HHV-6 could be monitored when bilateral optic neuritis is identified, in order to establish an appropriate antiviral therapy.


[Feasibility of a three-sided encapsulation procedure based on fascia anatomy in laparoscopic lateral lymph node dissection for middle and low rectal cancer].
Objective: To explore the feasibility and value of performing a three-sided encapsulation procedure based on fascia anatomy in laparoscopic lateral lymph node dissection (LLND) for middle and low rectal cancer. Methods: This was a retrospective review. The study cohort comprised patients who met the diagnostic criteria for rectal cancer according to the Chinese Guidelines for the Diagnosis and Treatment of Colorectal Cancer, had a short lymph node diameter of >5 mm on the lateral side within the 15 days before surgery, were evaluated as feasible candidates for laparoscopic total mesorectal excision+LLND surgery, had been diagnosed with low or intermediate level rectal cancer, and whose tumor was less than 8 cm away from the anal verge according to pathological examination of the operative specimen. Patients with a history of other malignant tumors of the abdomen or with incomplete follow-up data were excluded. Forty-two patients with middle and low rectal cancer who had undergone lateral lymph node dissection in diagnosis and treatment center of Gastrointestinal Cancer of Guangdong Hospital of Chinese Medicine from Jan.2018 to Dec.2022 were enrolled. There were 24 men (57.1%) and 18 women (42.9%) aged 58.4±11.8 years and the median BMI was 22.5 (19.3-24.1) kg/m2. The main point of the three-sided encapsulation procedure is to expand the external side medial to the external iliac artery and vein, narrowing the range of exterior side dissection. The anterior-medial side is designed to expand the vesical fascia to define the range of anterior-medial side extension. The internal side is fully extended to the ureterohypogastric nerve fascia; the distal point of the caudal extension reaches the level of the Alcock canal and the bottom reaches the piriformis, enabling dissection of the obturator nerve and No.283 lymph nodes. No.263D lymph nodes are dissected by exposing the internal iliac artery and its branches, dissecting the group No.263P lymph nodes, and severing the inferior vesical artery. Finally, the lateral lymphatic tissue is completely resected. Relevant variables were recorded, including the number of lateral lymph nodes detected, the rate of lymph node metastasis, operation duration, intraoperative blood loss, postoperative complications, postoperative hospital stay, and 3-year overall survival rate. Results: Laparoscopic surgery was successfully completed in all patients with no conversions to open surgery and no intraoperative complications. Twenty-seven (64.3%) of the study patients underwent left-sided LLND, 10 (23.8%) right-sided LLND, and five (11.9%) bilateral LLND, with lymph nodes cleared on both sides. All patients' lymph nodes were examined pathologically. A median of 17.0 (11.7, 26.0) lymph nodes was detected, the median of lateral lymph nodes being 5.0 (2.0, 10.2). The median operation time was 254.5 (199.0, 325.2) minutes. The median intra-operative blood loss was 50.0 (30.0, 100.0) mL. All patients were diagnosed with adenocarcinoma by pathological examination of the operative specimen. Two patients developed postoperative intestinal obstruction, one lymphatic leakage, and one a perineal incision infection. There were no cases of anastomotic leakage. The median postoperative hospital stay was 6.0 (5.0, 7.0) days and the median follow-up time 23.5 (9.0, 36.7) months. During follow-up, three patients (7.1%) died of tumor recurrence and metastasis. Two (4.8%) experienced mild urinary dysfunction, and one (2.4%) had moderate postoperative erectile dysfunction. One patient (2.4%) was found to have prostate and lung metastases 3 month after surgery. The 3-year overall survival rate was 74.4%. Conclusions: Three sided encapsulation is a safe and feasible procedure for LLND, achieving accurate and complete clearance of lateral lymphatic tissue.


The "second stalk" of Escherichia coli ATP synthase: structure of the isolated dimerization domain.
The b subunit of E. coli F(0)F(1)-ATPase links the peripheral F(1) subunits to the membrane-integral F(0) portion and functions as a "stator", preventing rotation of F(1). The b subunit is present as a dimer in ATP synthase, and residues 62-122 are required to mediate dimerization. To understand how the b subunit dimer is formed, we have studied the structure of the isolated dimerization domain, b(62-122). Analytical ultracentrifugation and solution small-angle X-ray scattering (SAXS) indicate that the b(62-122) dimer is extremely elongated, with a frictional ratio of 1.60, a maximal dimension of 95 A, and a radius of gyration of 27 A, values that are consistent with an alpha-helical coiled-coil structure. The crystal structure of b(62-122) has been solved and refined to 1.55 A. The protein crystallized as an isolated, monomeric alpha helix with a length of 90 A. Combining the crystal structure of monomeric b(62-122) with SAXS data from the dimer in solution, we have constructed a model for the b(62-122) dimer in which the two helices form a coiled coil with a right-handed superhelical twist. Analysis of b sequences from E. coli and other prokaryotes indicates conservation of an undecad repeat, which is characteristic of a right-handed coiled coil and consistent with our structural model. Mutation of residue Arg-83, which interrupts the undecad pattern, to alanine markedly stabilized the dimer, as expected for the proposed two-stranded, right-handed coiled-coil structure.


Syntheses, characterization, and catalytic ability in alkane oxygenation of chloro(dimethyl sulfoxide)ruthenium(II) complexes with tris(2-pyridylmethyl)amine and its derivatives.
New ruthenium(II) complexes having a tetradentate ligand such as tris(2-pyridylmethyl)amine (TPA), tris[2-(5-methoxycarbonyl)pyridylmethyl]amine [5-(MeOCO)3-TPA], tris(2-quinolylmethyl)amine (TQA), or bis(2-pyridylmethyl)glycinate (BPG) have been prepared. The reaction of the ligand with [RuCl2(Me2SO)4] resulted in a mixture of trans and cis isomers of the chloro(dimethyl sulfoxide-kappaS)ruthenium(II) complexes containing a TPA or a BPG, whereas a trans(Cl,N(amino)) isomer was selectively obtained for 5-(MeOCO)3-TPA and TQA. The trans and cis isomers of the [RuCl(TPA)(Me2SO)]+ complex were easily separated by fractional recrystallization. The molecular structures of trans- and cis(Cl,N(amino))-[RuCl(TPA)(Me2SO)]+ complexes and the trans(Cl,N(amino))-[RuCl{5-(MeOCO)3-TPA}(Me2SO)]+ complex have been determined by X-ray structural analyses. The reaction of TPA with [RuCl2(PhCN)4] gave a single isomer of the chloro(benzonitrile)ruthenium(II) complex, whereas the bis(benzonitrile)ruthenium(II) complex was obtained with BPG. The cis(Cl,N(amino))-[RuCl(TPA)(Me2SO)]+ complex is thermodynamically much less stable than the trans isomer and isomerizes in dimethyl sulfoxide at 65-100 degrees C. Oxygenation of alkanes catalyzed by these ruthenium(II) complexes has been examined. The chloro(dimethyl sulfoxide-kappaS)ruthenium(II) complexes with TPA and its derivatives using m-chloroperbenzoic acid as a cooxidant showed high catalytic ability. Adamantane was efficiently and selectively oxidized to give 1-adamantanol up to 88%. The chloro(dimethyl sulfoxide-kappaS)ruthenium(II) complex with 5-(MeOCO)3-TPA was found to be the most active catalyst among the complexes examined.


NCLscan: accurate identification of non-co-linear transcripts (fusion, trans-splicing and circular RNA) with a good balance between sensitivity and precision.
Analysis of RNA-seq data often detects numerous 'non-co-linear' (NCL) transcripts, which comprised sequence segments that are topologically inconsistent with their corresponding DNA sequences in the reference genome. However, detection of NCL transcripts involves two major challenges: removal of false positives arising from alignment artifacts and discrimination between different types of NCL transcripts (trans-spliced, circular or fusion transcripts). Here, we developed a new NCL-transcript-detecting method ('NCLscan'), which utilized a stepwise alignment strategy to almost completely eliminate false calls (>98% precision) without sacrificing true positives, enabling NCLscan outperform 18 other publicly-available tools (including fusion- and circular-RNA-detecting tools) in terms of sensitivity and precision, regardless of the generation strategy of simulated dataset, type of intragenic or intergenic NCL event, read depth of coverage, read length or expression level of NCL transcript. With the high accuracy, NCLscan was applied to distinguishing between trans-spliced, circular and fusion transcripts on the basis of poly(A)- and nonpoly(A)-selected RNA-seq data. We showed that circular RNAs were expressed more ubiquitously, more abundantly and less cell type-specifically than trans-spliced and fusion transcripts. Our study thus describes a robust pipeline for the discovery of NCL transcripts, and sheds light on the fundamental biology of these non-canonical RNA events in human transcriptome.


A Cre Transcription Fidelity Reporter Identifies GreA as a Major RNA Proofreading Factor in Escherichia coli.
We made a coupled genetic reporter that detects rare transcription misincorporation errors to measure RNA polymerase transcription fidelity in Escherichia coli Using this reporter, we demonstrated in vivo that the transcript cleavage factor GreA, but not GreB, is essential for proofreading of a transcription error where a riboA has been misincorporated instead of a riboG. A greA mutant strain had more than a 100-fold increase in transcription errors relative to wild-type or a greB mutant. However, overexpression of GreB in ΔgreA cells reduced the misincorporation errors to wild-type levels, demonstrating that GreB at high concentration could substitute for GreA in RNA proofreading activity in vivo.


[Accelerated phase of idiopathic pulmonary fibrosis].
The natural history of idiopathic pulmonary fibrosis is characterized by a slow progression resulting in respiratory failure and death. The progression to the fulminant form is rapid in a small percentage of cases, however. Within weeks or months, patients develop respiratory distress, and extensive ground-glass patterns can be seen in computed tomography scans and hyaline membranes in biopsy samples. This is described as an accelerated phase of idiopathic pulmonary fibrosis, in which elevated levels of acute-phase reactants and tumor markers have been reported. To date, the monoclonal tumor marker, CA 15/3 has not been associated with the accelerated phase.


Porcine and human insulin absorption from subcutaneous tissues in normal and insulin-dependent diabetic subjects: a deconvolution-based approach.
The mechanisms of sc insulin absorption are not understood, and models for interpreting in vivo data cannot be developed without gross simplification. To overcome this difficulty we developed a new approach which makes use of deconvolution analysis and does not require any model of the sc tissue. In five normal subjects and seven insulin-dependent diabetic (IDDM) patients endogenous insulin secretion was suppressed by means of a hypoglycemic glucose clamp procedure (approximately 2.8 mmol/L) sustained by a continuous insulin infusion (approximately 4 pmol/min.kg). A bolus injection of insulin (5.4 nmol) was administered iv, and plasma insulin concentrations were measured frequently for 2 h to assess iv insulin kinetics. Insulin then was injected sc in the abdominal region, and plasma insulin concentrations were measured for 8 h. Each subject was studied twice, with porcine and semisynthetic human insulin (Actrapid, Novo). The rate of insulin absorption was reconstructed by deconvolution from the plasma concentrations and iv insulin kinetic data. Linearity of the iv insulin kinetics, essential for deconvolution analysis, was confirmed by a dose-response study in the range of the measured concentrations (150-1800 pmol/L). In most instances, a two-compartment model was adequate to describe the iv response. The mean plasma insulin clearance rates were 15.5 +/- 1.9 (+/- SD) mL/min.kg (porcine) and 17.2 +/- 6.0 (human) in normal subjects and 20.7 +/- 8.8 (porcine) and 20.9 +/- 9.1 (human) in the IDDM patients. The rate of appearance of human insulin from sc tissue was faster than that of porcine insulin in both normal and IDDM subjects, but no significant differences were found in bioavailability, which was 55 +/- 12% (+/- SD; porcine) and 61 +/- 34% (human) in the normal subjects, and 84 +/- 28% (porcine) and 86 +/- 23% (human) in the IDDM patients. The rate of absorption and bioavailability were higher in the IDDM patients than in the normal subjects, a difference possibly related to increased sc blood flow in the IDDM patients. No differences were found with regard to glucose requirement values, normalized to plasma insulin concentrations, in agreement with the finding that the bioavailability of the two insulin species was similar.


The dissociative processing style:a cognitive organization activated by perceived or actual threat in clinical dissociators.
This paper proposes a cognitive organization that operates during times of perceived or actual threat in individuals with dissociative psychopathology. This organization, referred to as the dissociative processing style (DPS), serves as a threat monitoring system. It is characterized by (1) a shift from selective attention processing to multiple streams of information processing, (2) weakened cognitive inhibitory functioning which allows these streams to be operational and (3) the directing of awareness towards some and away from other information streams. Whilst DPS activation has the potential for adaptive and protective functions, it also heightens the likelihood of dissociative symptom experience and dissociation itself. Dissociation is understood as a failure to integrate encoded information from multiple input streams. The DPS is argued to be activated by top-down processes which signal danger, such as the appraisal of contextual cues. A clinical example is used to highlight the characteristics of the DPS.


A clinical evaluation of transcatheter arterial embolization on hepatocellular carcinoma.
TAE was performed in 49 cases of HCC for clinical assessment of the usefulness. This procedure was aimed to subside intraperitoneal hemorrhage in 8 cases among them and to provide a possible antitumor effect in the remaining 41 cases. The prognosis in the 8 cases of intraperitoneal hemorrhage was 54.9 +/- 41.1 days including one case survived as long as 116 days. The prognosis in the remaining 41 cases was 12.6 +/- 8.3 months in group A, 7.8 +/- 6.1 months in group B and 1.6 +/- 1.3 months in group C according to Child's classification, while it was 2.8 +/- 2.5 months in 1st branch occluded, 9.4 +/- 6.1 months in IInd branch occluded and 19.2 +/- 6.7 months in IIIrd branch occluded group according to the portal vein occlusion due to tumor thrombi suggesting that a more prolonged survival was attained with more favorable degree of Child's classification and less affected portal embolization. The cumulative survival time (by Kaplan-Meier's methods) was 6 months in 89% of the cases examined, 1 year in 59%, 2 years in 34% and 3 years or more in 11%, indicating significantly higher survival as compared to our TAI group. Angiographic re-opening of tumor vessel within 3 months was observed in 46.7%. TAE on HCC was useful both for the purpose of antitumor effect and of hemostasis. The degree of Child's classification and severity of portal occlusion at the initial examination may closely relate to the prognosis. Thus, angiography should be repeated within 3 months following the first TAE at least.


Innate and adaptive immune response to pneumonia virus of mice in a resistant and a susceptible mouse strain.
Respiratory syncytial virus (RSV) is the leading cause of infant bronchiolitis. The closely related pneumonia virus of mice (PVM) causes a similar immune-mediated disease in mice, which allows an analysis of host factors that lead to severe illness. This project was designed to compare the immune responses to lethal and sublethal doses of PVM strain 15 in Balb/c and C57Bl/6 mice. Balb/c mice responded to PVM infection with an earlier and stronger innate response that failed to control viral replication. Production of inflammatory cyto- and chemokines, as well as infiltration of neutrophils and IFN-γ secreting natural killer cells into the lungs, was more predominant in Balb/c mice. In contrast, C57Bl/6 mice were capable of suppressing both viral replication and innate inflammatory responses. After a sublethal infection, PVM-induced IFN-γ production by splenocytes was stronger early during infection and weaker at late time points in C57Bl/6 mice when compared to Balb/c mice. Furthermore, although the IgG levels were similar and the mucosal IgA titres lower, the virus neutralizing antibody titres were higher in C57Bl/6 mice than in Balb/c mice. Overall, the difference in susceptibility of these two strains appeared to be related not to an inherent T helper bias, but to the capacity of the C57Bl/6 mice to control both viral replication and the immune response elicited by PVM.


Factors Influencing the Placebo Effect in Patients with Primary Open-Angle Glaucoma or Ocular Hypertension: An Analysis of Two Randomized Clinical Trials.
To explore factors related to the placebo effect in patients with primary open-angle glaucoma (POAG) or ocular hypertension (OH). This was a retrospective cohort study of patients with POAG and patients with OH who were treated with placebo. The patients' data were extracted from two randomized, double-masked, parallel, multicenter clinical trials (trial 1 and trial 2) in Japan. We explored the baseline factors that were associated with the intraocular pressure (IOP)-lowering effect of placebo ophthalmic solution after 4 weeks of instillation treatment at two time points by using multivariable models. The time points were Hour 0 (between 08:30 and 10:30 before instillation) and Hour 2 (within 1.5 to 2.5 h after instillation and by 12:30) at the baseline date and after 4 weeks. The changes in IOP from baseline to 4 weeks at the two time points were evaluated for the IOP-lowering effect induced by placebo instillation. Of the 330 patients included in the two trials, 89 patients were eligible for the analysis. The results of the multivariable analysis for Hour 0 indicated a high IOP at the baseline date (coefficient: 0.24, 95% confidence interval (CI): 0.02 to 0.46, P = 0.03), and the magnitude of the IOP fluctuation at the baseline date (coefficient: 0.57, 95% CI: 0.24 to 0.90, P = 0.001) was associated with the IOP-lowering effect after 4 weeks. With respect to Hour 2, the trial type was associated with the IOP-lowering effect (coefficient: -1.15, 95% CI: -2.14 to -0.16, P = 0.02). A large fluctuation in IOP during the day is associated with the IOP-lowering effect induced by placebo in patients with POAG or OH. This finding would be helpful to researchers when designing studies related to glaucoma in the early stages of clinical development of drugs.


Cell surface glycosaminoglycans do not serve as ligands for PECAM-1. PECAM-1 is not a heparin-binding protein.
Previous studies have suggested that PECAM-1 mediates cellular interactions via both homophilic and heterophilic adhesive mechanisms. Cell surface glycoaminoglycans have been implicated as one of the heterophilic ligands for PECAM-1. To determine whether PECAM-1 is capable of interacting directly with glycosaminoglycans, we examined the adhesive properties of multiple monovalent and multivalent forms of this adhesion molecule. We found that the binding of a bivalent PECAM-1/IgG chimeric protein or multivalent PECAM-1-containing proteoliposomes to multiple different cell lines was 1) strictly dependent upon cell surface expression of PECAM-1 and 2) unaffected by the presence of excess heparin or heparan sulfate. The extracellular domain of PECAM-1 failed to interact specifically with heparin-Sepharose, 3H-labeled heparin, or a heparin-bovine serum albumin conjugate. In addition, an amino acid sequence motif inadvertently created by the juxtaposition of PECAM-1 and IgG sequences within the hinge region of certain PECAM-1/IgG chimeric constructs was found to confer glycosaminoglycan binding properties not normally present within the extracellular domain of the native molecule. Together, these data suggest that the mechanism by which heparin is able to affect PECAM-1-dependent cell-cell adhesion is indirect and occurs via inhibition of events that occur downstream from PECAM-1 engagement.


Characterization and natural course of cardiac autonomic nervous dysfunction in HIV-infected patients.
To examine the degree, pattern, and natural history of cardiac autonomic nervous dysfunction in patients infected with HIV. Cross-sectional and prospective longitudinal cohort study. Primary care and tertiary referral university centre. Thirty-five consecutive HIV-infected patients who had either not yet developed AIDS (15 pre-AIDS patients) or who were at the Centers for Disease Control and Prevention (CDC) AIDS stage (n = 20), and 29 healthy age- and sex-matched HIV-negative controls. Computer-aided power spectral analysis of 15 standardized parameters of heart-rate variability (HRV). Pre-AIDS patients as a group did not exhibit any HRV parameters to be significantly different from healthy controls (P > 0.017), whereas AIDS patients demonstrated reduced HRV in 14 parameters (93.3%) compared with healthy subjects (p > 0.017). Median proportion of abnormal HRV parameters (< 10th percentile of controls) per individual was 9.1% in pre-AIDS patients and 61.3% in AIDS patients (P = 0.0347). Progressive CDC stages inversely correlated to 10 HRV parameters (66.7%; -0.50 < or = r < or = -0.36; P < 0.05). Follow-up testing in 10 pre-AIDS and six AIDS patients after 6-16 months (median, 12.5 months) did not reveal deterioration of HRV (P < 0.05). A dysautonomia symptom score correlated to 10 HRV parameters (66.7%; -0.14 < r < -0.55; P < 0.05). Cardiac autonomic nervous dysfunction is severe in AIDS patients, although not significant in pre-AIDS patients. Cardiac autonomic nervous dysfunction proceeds with HIV disease progression, although its individual course is slow.


Ursolic acid improves the bacterial community mapping of the intestinal tract in liver fibrosis mice.
Liver fibrosis often appears in chronic liver disease, with extracellular matrix (ECM) deposition as the main feature. Due to the presence of the liver-gut axis, the destruction of intestinal homeostasis is often accompanied by the development of liver fibrosis. The inconsistent ecological environment of different intestinal sites may lead to differences in the microbiota. The traditional Chinese medicine ursolic acid (UA) has been proven to protect the liver from fibrosis. We investigated the changes in the microbiota of different parts of the intestine during liver fibrosis and the effect of UA on these changes based on high-throughput sequencing technology. Sequencing results suggest that the diversity and abundance of intestinal microbiota decline and the composition of the microbiota is disordered, the potentially beneficial Firmicutes bacteria are reduced, and the pathways for functional prediction are changed in the ilea and anal faeces of liver fibrosis mice compared with normal mice. However, in UA-treated liver fibrosis mice, these disorders improved. It is worth noting that the bacterial changes in the ilea and anal faeces are not consistent. In conclusion, in liver fibrosis, the microbiota of different parts of the intestines have different degrees of disorder, and UA can improve this disorder. This may be a potential mechanism for UA to achieve anti-fibrosis. This study provides theoretical guidance for the UA targeting of intestinal microbiota for the treatment of liver fibrosis.


Is acute fibrinous and organizing pneumonia the expression of immune dysregulation?
Acute fibrinous and organizing pneumonia (AFOP) is a recently described histologic pattern of diffuse pulmonary disease. In children, all cases reported to date have been fatal. In this study, we describe the first nonfatal AFOP in a child and review the literature. A 10-year-old boy developed very severe aplastic anemia (VSAA) after being admitted to our hospital with a fulminant hepatic failure of unknown origin. A chest computed tomography scan revealed multiple lung nodules and a biopsy of a pulmonary lesion showed all the signs of AFOP. Infectious workup remained negative. We started immunosuppressive therapy with antithymocyte globulin and cyclosporine to treat VSAA. Subsequent chest computed tomography scans showed a considerable diminution of the lung lesions but the VSAA did not improve until we performed hematopoietic stem cell transplantation 5 months later. Aplastic anemia is associated with a variety of autoimmune syndromes. The sequence of events in our patient suggests that the hepatic failure, AFOP, and the VSAA may all have been part of an autoimmune syndrome. AFOP could be the result of immune dysregulation in this pediatric case with favorable outcome after immunosuppressive therapy and hematopoietic stem cell transplantation.


Impact of a publicly-funded pharmacare program policy on benzodiazepine dispensing among children and youth: a population-based natural experiment.
In January 2018, the Government of Ontario, Canada, initiated a universal pharmacare program (OHIP+) for all individuals aged 24 years and younger. In April 2019, the program was amended to cover only children and youth without private insurance. Because benzodiazepines are commonly prescribed to children and youth despite their potential hazards, we examined whether changes in publicly-funded drug coverage influenced benzodiazepine dispensing trends in this demographic. We conducted a population-based natural experiment study of benzodiazepine dispensing to children and youth in Ontario between January 2013 and March 2020. We used interventional autoregressive integrated moving average models to estimate the impact of OHIP + and its subsequent modification on these trends. The implementation of OHIP + was associated with an immediate increase in the monthly rate of benzodiazepine dispensing of 12.9 individuals per 100,000 population (95% confidence interval [CI]; 7.5 to 18.3 per 100,000). Benzodiazepine dispensing rates rose from 214.2 to 241.5 per 100,000 from December 2017 to March 2019, a 12.8% (95% CI 9.6-16.0%) increase. In stratified analyses, increases were most pronounced among females, children and youth living in the lowest income neighbourhoods and individuals aged 20 to 24. The April 2019 modification to OHIP + was not associated with changes in monthly benzodiazepine dispensing trends (0.39 individuals per 100,000; 95% CI -1.3 to 2.1 per 100,000). However, rates remained elevated relative to the period preceding OHIP + implementation. Implementation of a publicly-funded pharmacare program resulted in more children and youth being prescribed benzodiazepines.


Ultrasound fusion imaging with real-time navigation for the surveillance after endovascular aortic aneurysm repair.
Ultrasound can be an effective alternative to computed tomography for surveillance following endovascular aneurysm repair (EVAR). Recently, ultrasound fusion imaging with the real-time navigation system was introduced. Here we described 3 patients who underwent post-EVAR surveillance using this novel technique. Complete coregistration was achieved in all patients. The origin of left renal artery was selected for the target of coregistration. Ultrasound fusion imaging was useful to differentiate the confusing lesion and to evaluate the complete resolution of endoleak and newly developed delayed endoleak. Ultrasound fusion image with real-time navigation system can be a feasible imaging tool for post-EVAR surveillance.


Application of histomorphometric methods to the study of bone repair.
Standardized methods for the histomorphometric assessment of bone are essential features of most studies of metabolic bone diseases and their treatments. These methodologies were developed to assess coupled remodeling, focusing primarily on osteoblasts and osteoclasts, the anabolic and catabolic rates of these cells, and structural features of mature bone. Research studies on bone healing and the development of new therapeutic approaches for the enhancement of bone repair also require a comprehensive understanding of the basic cellular and tissue level mechanisms that underlie these processes. However, the histological methods developed for metabolic bone disease studies are not completely suitable for studies of bone repair because they are based on assumptions that there is little variation in tissue composition within a sample of bone and not generally designed to quantify other types of tissues, such as cartilage, that contribute to bone healing. These techniques also do not provide tissue-based structural measurements that are relatable to the specific types of biomechanical and radiographic structural assessments that are used to determine rates of bone healing. These deficiencies in current histological approaches therefore point to the need to establish standardized criteria for the histomorphometric assessments that are specifically adapted for the study of bone repair in models of fracture healing and bone regeneration. In this Perspective, we outline what we believe to be the specific structural, tissue. and cellular aspects that need to be addressed to establish these standardized criteria for the histomorphometric assessment of bone repair. We present the specific technical considerations that need to be addressed to appropriately sample repair tissues to obtain statistically meaningful results and suggest specific procedures and definitions of nomenclatures for the application of this technology to bone repair. Finally, we present how aspects of histomorphometric measurements of bone repair can be related to biomechanical and radiographic imaging properties that functionally define rates of bone healing, and thus, how these tools can be used to provide corroborating data.


A family with complete deficiency of plasma cholesteryl ester transfer protein activities.
A 43-year-old male with a high value of high density lipoprotein cholesterol (151 mg/dl) was found among subjects receiving annual health checks. We investigated the cholesteryl ester transfer protein (CETP) activity and conducted a family study. There was complete deficiency of CETP activities in the proband and his sister, and partial deficiency of CETP activities in his mother and daughter. Genetic analysis revealed a splicing defect (G to A point mutation) in intron 14 of the CETP gene.


Two Cases of Tonic Pupil: Ross and Ross Syndrome Plus.
Ross syndrome is a rare disorder of the peripheral autonomic nervous system characterized by a triad of tonic pupils with light-near dissociation, segmental anhidrosis, and areflexia. Though having a benign course, the disease can cause significant social embarrassment. Both our cases presented with complaints of segmental facial hyperhidrosis. The first case with a one-year history had findings of segmental anhidrosis up to T4 thoracic level, left tonic pupil, and absent right ankle reflex. While the second case with a history of five years had bilateral tonic pupil, absent lower limb reflexes, anhidrosis of left face, neck, and upper trunk up to T4 level, apart from having associated Horner's syndrome. Minor's (starch-iodine) test and dilute pilocarpine test were helpful for diagnosis in both cases, indicating areas of anhidrosis and pupillary cholinergic denervation hypersensitivity respectively. Both cases were provided counseling and managed conservatively.


Length of hospital stay in surgical service patients with nosocomial Hemophilus influenzae infections.
The length of hospitalization was characterized for 24 surgical service patients with nosocomial respiratory tract infections associated with Hemophilus influenzae. The mean number of days were: admission to surgery 8.22; surgery to culture of the nosocomial pathogen 7.00; admission to culture 16.71; admission to discharge 38.75; surgery to discharge 25.48; and culture to discharge 21.42. The length of hospital stay was prolonged for those patients with nosocomial respiratory tract infections related to H influenzae.


Central A1-receptor activation associated with onset of torpor protects the heart against low temperature in the Syrian hamster.
Body temperature drops dramatically during hibernation, but the heart retains the ability to contract and is resistant to induction of arrhythmia. Although adaptive changes in the heart prior to hibernation may be involved in the cold-resistant property, it remains unclear whether these changes are sufficient for maintaining cardiac pulsatility under an extreme hypothermic condition. We forcibly induced hypothermia in Syrian hamsters by pentobarbital anesthesia combined with cooling of the animals. This allows reproduction of a hypothermic condition in the absence of possible hibernation-specific reactions. Unlike hypothermia in natural hibernation, the forced induction of hypothermia caused atrioventricular block. Furthermore, J-waves, which are typically observed during hypothermia in nonhibernators, were recorded on an ECG. The origin of the J-wave seemed to be related to irreversible injury of the myocardium, because J-waves remained after recovery of body temperature. An abnormal ECG was also found when hypothermia was induced in hamsters that were well adapted to a cold and darkened environment or hamsters that had already experienced hibernation. These results suggest that acclimatization prior to hibernation does not have a crucial effect at least on acquisition of cardiac resistance to low temperature. In contrast, an abnormal ECG was not observed in the case of hypothermia induced by central administration of an adenosine A1-receptor agonist and subsequent cooling, confirming the importance of the adenosine system for inducing hibernation. Our results suggest that some specific mechanisms, which may be driven by a central adenosine system, operate for maintaining the proper cardiac pulsatility under extreme hypothermia.


Neuroendocrine neoplasms of the lung.
Neuroendocrine tumors of the lung continue to be difficult nosologic and diagnostic problems, centering on the time-honored terms of "carcinoid," "atypical carcinoid," and "small cell carcinoma." Problems that are encountered in the classification of such neoplasms revolve around the differing criteria that have been advanced for their definition and variable application of such criteria in common practice. This review considers the epithelial and nonepithelial lesions of the lung that may demonstrate neuroendocrine and neuroectodermal differentiation. A proposal is made for a simplified system of classifying the epithelial tumors, dividing them into 3 grades with appended descriptive modifiers.


Optical coherence tomography imaging of telangiectasias during intense pulsed light treatment: a potential tool for rapid outcome assessment.
Vascular malformations commonly occur in the facial region, and can be associated with significant stigma and embarrassment. Studies have shown that even recommended light-based treatments do not always result in complete clearance. This indicates the need for more accurate pre-treatment assessment of vessel morphology to optimize treatment settings and identify possible morphological predictors of the outcome. Fourteen patients (six males, eight females, and aged 37-66 years) with the diagnosis of telangiectasias were enrolled and were all scanned with OCT and digitally photographed before and minutes after IPL treatment. OCT images of the telangiectasias before treatment were displayed as hyporeflective/signal poor bands clearly demarcated from the surrounding tissue. Minutes after treatment, OCT images demonstrated two different reactions. (1) Narrow hyperreflective bands surrounding the vessels, which may indicate edema or insufficient coagulation. (2) Hyperreflective signals within the lumen of the vessels, compatible with the expected irreversible microthrombus formation in the vessels. OCT imaging is capable of real-time assessment of tissue damage during light and laser treatment, including visualization of the perivascular changes. This may offer a more dynamic, more complete understanding of the efficacy and potential outcome of the treatment process. It is hypothesized that these immediate changes may correlate to longer-term treatment outcome.


Structure of a semidilute polymer solution under steady shear.
We use Brownian dynamics computer simulations to investigate the structure of a semidilute polymer solution undergoing a steady, uniform shear flow. We find that the contributions to structure factor from intra- and interchain correlations, which cancel each other almost completely for an equilibrium semidilute solution, are modified in different ways by the shear flow. Incomplete cancellation of these contributions leads to anisotropic patterns that resemble those observed in light scattering experiments on sheared semidilute solutions [Wu et al., Phys. Rev. Lett. 66, 2408 (1991)]. For small wave vectors the structure factor change is dominated by the interchain contribution. We also monitor the distortion of the pair correlation function and show that for small distances it is dominated by the intrachain contribution. Finally, we investigate nonlinear shear viscosity and find that, like the short-distance part of the distortion of the pair correlation function, it is predominantly of intrachain origin.


Self-esteem in adolescence: a comparison of adolescents with diabetes mellitus and leukemia.
This descriptive study examined the self-esteem of adolescents with diabetes mellitus and leukemia. Participants included 22 adolescents with diabetes mellitus and 33 with leukemia. No significant difference was found between the self-esteem of males and females in either group. After the initial diagnosis, over time a decrease in self-esteem in the leukemia group and an increase in self-esteem in the diabetes mellitus group were observed. The adolescents with leukemia who experienced a relapse had moderate self-esteem. Most of the adolescents with leukemia could not attend school because of their illness. The mean scores of self-esteem were high in the diabetes mellitus group and moderate in the leukemia group. In both groups, the mean scores of self-esteem were compared to depressive affect, daydreaming, psychosomatic symptoms, intensity of discussion, and parental interest (subscales in the Rosenberg self-esteem scale). No significant difference was found between the two disease groups. There was a strong and significant correlation between self-esteem and depressive affect in both groups.


Practical management of NSAID gastropathy in rheumatic patients.
Gastropathy is a relatively common complication of treatment with a non-steroidal anti-inflammatory drug (NSAID). Pre-treatment evaluation of patients is therefore of key importance. Contraindications for NSAID treatment may include a history of gastroduodenal ulcer or of NSAID gastropathy, simultaneous administration of anticoagulant therapy and the existence of risk factors for ulcer disease. After NSAID treatment has begun, clinical follow-up is highly desirable. NSAID gastropathy can be asymptomatic despite the presence of a broad spectrum of gastrointestinal mucosal changes on endoscopy, but in the majority of cases there are clinical symptoms. Management of gastropathy may involve withdrawal of NSAID treatment, replacement therapy with a different type of drug or post hoc treatment of the gastropathy with anti-ulcer compounds.


Exploring indications for the Use of direct oral anticoagulants and the associated risks of major bleeding.
Thrombosis is a leading cause of morbidity and mortality in the United States. Arterial and venous thromboses are implicated in the pathogenesis of major disorders, including myocardial infarction, ischemic stroke, and venous thromboembolism. Over the past decade, direct oral anticoagulants (DOACs) (eg, direct thrombin inhibitor and factor Xa [FXa] inhibitors) have been adopted as alternatives to warfarin due to their clinical advantages and efficacy for the treatment of thrombosis. As with all anticoagulants, treatment with DOACs is associated with a risk of major bleeding, including life-threatening gastrointestinal bleeds and intracranial hemorrhages (ICHs). In turn, the burden of bleeding associated with DOAC treatment is itself associated with substantial healthcare costs that are amplified by an increased risk of thromboembolic events and mortality following major bleeding events, especially in patients with ICHs. Given the rapid adoption of the DOACs and projected usage in the large patient population affected by thromboembolic conditions, clinicians are increasingly likely to encounter patients with major bleeding events due to DOAC therapy. Unlike warfarin, effective strategies to manage these bleeds are limited. There is an unmet need for reversal agents for use in the management of patients who receive FXa inhibitors and experience life-threatening bleeding or need emergency surgery. Andexanet alfa and ciraparantag are being evaluated as potential antidotes for both direct and indirect FXa inhibitors.


Characterization of endonuclease G and mitochondria-sarcoplasmic reticulum-related proteins during cardiac hypertrophy.
Endonuclease G (Endo G) is a novel determinant of cardiac hypertrophy. Here, we report the characterization of Endo G and mitochondria-sarcoplasmic reticulum-related proteins during cardiac hypertrophy, and hypothesize that Endo G regulate mitochondrial function partly through Mfn2 and Jp2 during cardiac hypertrophy. Our results show that Endo G levels gradually increased at the beginning of phenylephrine-induced cardiac hypertrophy, accompanied by an abnormal mitochondrial membrane potential. The up-regulation of Mfn2, Jp2, and Endo G appeared at an early stage of cardiac hypertrophy, whereas PGC1α was not up-regulated until a later stage. Abolishing Endo G with siRNA led to the uncoupling of the mitochondrial electron transport chain from ATP production and decreased PGC1α expression, likely by affecting the juxtaposition of the mitochondria and the sarcoplasmic reticulum via Mfn2 and Jp2. Furthermore, abolishing Jp2 altered the expression of Endo G expression and induced mitochondrial dysfunction, suggesting that mitochondrial abnormalities in cardiac hypertrophy are most likely caused by Endo G. Taken together, our study established a link between Endo G and mitochondrial function during cardiac hypertrophy, partly through the effects of Endo G on Mfn2 and Jp2, and revealed a role for Endo G in the crosstalk between the processes controlled by Mfn2 and Jp2 in maladaptive cardiac hypertrophy.


[Experimental radiosurgery in rats using gamma a "gamma knife". Description of a stereotactic device for small laboratory animals].
To allow the experimental use in rats of a Gamma Knife Radiosurgical Unit, a stereotactic device adapted from the conventional Kopf's device was developed. To control the accuracy of the coordinate system based on the De Groot's rat stereotactic atlas, experimental radiosurgical lesions were made on the left striatum. The isodose curve distribution of the 4 mm collimator was calculated with the dose planning software used in Gamma Knife and superimposed on the left striatum target. Doses of 200 Gy were administered to the left striatum in six rats. The results were evaluated 21, 34 and 47 days later. At 34 and 47 days necrotic lesions were exactly as targeted. In a second group of 48 rats receiving a doses of 100 Gy, no lesions were observed after 7, 15, 24, 31, 45 days. However, in all rats sacrificed 59, 72 and 90 days after day radiation, a necrotic lesion was always present and confirmed that at each time the lesions were precisely targeted. This apparatus allows precise and reproducible gamma irradiation lesion in rat brain without expensive and time consuming imaging techniques. This device provides a useful system to observe the experimental effects of radiosurgery on the central nervous system in rats.


[Stress tolerance of different osteosynthetic procedures in pertrochanteric fractures (author's transl)].
To obtain valid information on stress tolerance of pertrochanteric fractures, we exposed three different methods of osteosyntheses to dynamical stress and the results were compared. The methods of osteosyntheses were application of a 130 degrees angle plate, performing an adjoining osteosynthesis with bone cement or using cephalocondylar nails. The measurements in connection with photographical and roentgenological examinations of osteosyntheses under stress gave the following results: Compared with the other methods treatment with condylar nails gave equal results in static behaviour yet was clearly superior in its dynamic reaction.


[Utilization of different sulfur sources by propionic acid bacteria].
The object of this work was to study the ability of propionic bacteria to utilize sulfur compounds having various degrees of oxidation. Propionibacterium shermanii was found to utilize sulfite, thiosulfate, sulfide and elemental sulfur, apart from sulfate, as a sulfur source. When the culture grew in a medium with elemental sulfur, sulfide was produced. The utilization of sulfate by P. shermanii had a peculiar character. In the process of the culture growth, the utilization of sulfate alternated with its release into the medium.


Treatment of refractory generalized status epilepticus with continuous infusion of midazolam.
The optimal therapeutic approach for the patient with refractory generalized status epilepticus remains to be defined. We describe four patients with refractory generalized status epilepticus who were successfully treated with intravenous midazolam. Each patient had prolonged convulsive status epilepticus unresponsive to standard doses of intravenous benzodiazepines, phenytoin, and phenobarbital. The patients subsequently received midazolam administered as an intravenous bolus (200 micrograms/kg) followed by a continuous infusion (0.75 to 11 micrograms/kg/min) lasting 8 hours to 10 days. Clinical examination and scalp electroencephalographic monitoring documented the cessation of seizure activity within minutes of the loading dose in all patients. No significant adverse effects occurred during midazolam treatment. The one patient with prolonged midazolam infusion required fluid boluses and pressors for moderate hypotension, and the remainder of the patients safely tolerated midazolam despite preexistent hemodynamic instability. All patients recovered and maintained good seizure control. Intravenous midazolam appears to be an effective treatment for refractory generalized status epilepticus, and may represent a substantial improvement over current therapeutic approaches such as pentobarbital anesthesia.


Fast switching coil system for sample premagnetization in an unshielded ultra-low-field nuclear magnetic resonance experiment.
We present a system developed to premagnetize liquid samples in an ultra-low-field nuclear magnetic resonance experiment. Liquid samples of a few milliliters are exposed to a magnetic field of about 70 mT, which is abruptly switched off, to leave a transverse microtesla field, where nuclei start precessing. An accurate characterization of the transients and intermediate field level enables a reliable operation of the detection system, which is based on an optical magnetometer.


Lexical profiles of bilingual children with primary language impairment.
This study used lexical tasks to examine associations between languages, tasks, and age in bilingual children with primary language impairment. Participants (n = 41, mean age 8;8 years) lived in the United States, spoke primarily Spanish (L1) at home and English (L2) at school, and were identified with moderate to severe impairments in both languages. A total of eight tasks (four in each language) measured breadth of vocabulary knowledge (receptive and expressive vocabulary) and aspects of lexical processing (rapid automatic naming and nonword repetition). Correlational analyses revealed older children outperformed younger children on lexical tasks in L2 but not L1, as well as relative L2 dominance for most individuals and tasks. Positive associations were found between languages on processing-based tasks but not vocabulary measures. Findings were consistent with literature on typical bilingual learners, albeit with a notable increased risk of plateau in L1 growth. Results are interpreted within a Dynamic Systems framework.


Intussusception in chronic renal failure.
Spontaneous intussusception in adults is rare. Recently, we managed two patients with chronic uremia in whom intussusception developed. Histologic examination of the resected segments showed intestinal intramural hemorrhage. We postulate that the coagulopathy associated with uremia may allow for intestinal intramural hemorrhage, thus forming a nidus from which intussusception may be initiated.


Neighbourhood social capital and obesity: a systematic review of the literature.
Social capital, defined as the resources accessed by individuals and groups through social connections, has been posited to be a social determinant of obesity. However, empirical evidence for this association has been inconsistent - namely, some studies have found a protective association while others have reported no correlation. We sought to conduct a systematic review on the relation between neighbourhood social capital and obesity, considering potential differences on the results based on the measures used and the covariates and mediators included in the studies. PRISMA statement guidelines were followed. Our results indicate that an association between neighbourhood social capital and obesity exists, but that it depends on the measures and covariates used in the study design. Understanding the role of social capital in the development and/or maintenance of obesity will require the use of strong methodological designs and a thorough conceptualization of how this relationship may arise.


Study of chlordecone desorption from activated carbons and subsequent dechlorination by reduced cobalamin.
Since 1972, the French departments of Guadeloupe and Martinique have intensively used organochlorinated pesticides such as chlordecone (CLD) and hexachlorocyclohexane (HCH) isomers to prevent the proliferation of banana weevil (Cosmopolite sordidus). These molecules are stable in the environment, leading to a continuous contamination of soils, water, and food chain in the banana-producing areas. In these polluted areas, water treatment plants are equipped with activated carbon (AC) filters. In order to improve treatment of CLD-contaminated waters by AC, CLD adsorption and desorption kinetic studies are carried out using different ACs produced from sugar cane bagasse as adsorbents and subsequent CLD degradation is performed using reduced vitamin B12 (VB12). A GC-MS method for CLD quantification is as well optimized. This study shows that bagasse ACs are able to capture the pollutant, leading to a CLD concentration decrease from 1 to 73 μg L-1, with an adsorption capacity of 162 μg mg-1. Adsorption capacity increase with the temperature indicates an endothermic process. Polar solvents favor CLD desorption from ACs, suggesting hydrogen bonding between CLD and surface groups of ACs, the best solvent for chemical desorption being ethanol. Subsequent degradation of CLD in ethanol is performed using vitamin B12 reduced by either 1,4-dithiotreitol (DTT) or zerovalent zinc, leading to 90% of CLD removal and to the molecule cage structure opening for formation of a pentachloroindene intermediate product, characterized by GC MS/MS. A pathway for pentachloroindene formation from CLD is proposed.


Surreptitious surgery on Long Island Sound: The oral cancer surgeries of President Grover Cleveland.
Grover Cleveland rose from being the mayor of Buffalo to the governor of New York to the president of the United States. At the start of Cleveland's second term as president, the nation was involved in a severe financial crisis, the extent of which was not known by the general public. President Cleveland was to make a strong appeal to Congress in the coming months to repeal the Sherman Silver Purchase Act of 1890. He thought this would set the nation on the road to fiscal recovery. However, his vice president, Adlai Stevenson, strongly opposed repeal of the Sherman Act. Prior to scheduling his appearance before Congress, President Cleveland noticed a rough spot on his palate. A biopsy confirmed that it was cancer, and it was determined that surgery was needed. Cleveland and his advisors thought the nation would be thrown into a panic if the President's health did not remain a secret. A surgical team, which included a dentist, performed the surgery in secrecy while traveling aboard a yacht. A prosthetic obturator was fabricated by a New York prosthodontist to close the surgical defect. Cleveland recovered well, made a forceful speech before Congress, had the Sherman Act repealed and lived without a recurrence of his oral cancer for the rest of his life. The public remained unaware, for the most part, of the gravity of President Cleveland's health for decades.


Antipyretic and Anti-Inflammatory Effects of Rectal Administration of Reduning Injection in Feverish Rats Induced by Lipopolysaccharide.
This study aimed to explore the antipyretic and anti-inflammatory effects of rectal administration of Reduning injection in feverish rats induced by lipopolysaccharide (LPS), and observe the temperature changes and inflammatory indexes. The selected rats were randomly divided into 6 groups, with 10 rats in each group, named as normal empty group, model group, intravenous group (2 mL/kg), low-dose enema group (1 mL/kg), middle-dose enema group (2 mL/kg), and high-dose enema group (4 mL/kg). The hourly temperature variations in rats injected with LPS in the abdomen were recorded. Five hours later, blood samples from the abdominal aorta were collected to monitor immunoglobulin M (IgM), immunoglobulin A (IgA), interleukin (IL)-6, and tumor necrosis factor (TNF)-α. At 5 hours, the fever peak induced by LPS appeared, and obvious antipyretic effects were observed; the effect was optimal in the medium dose enema group at 4 hours (p < 0.05); the IgM value in the enema groups, the intravenous group, and normal empty group was significantly lower than that in the model group; the IgA value in each group was higher than that in the model group, but there was no statistical significance (p > 0.05); values of IL-6 and TNF-α in each group were lower than those in the model group, and the difference was statistically significant except for the high-dose enema group (p > 0.05). Low-dose and medium-dose rectal administration of Reduning injection have inhibitory effects on IL-6, TNF-α, and IgM in feverish rats induced by LPS, but there is no obvious difference compared to intravenous administration and it could achieve an anti-inflammatory effect. There is a possibility of enhancing IgA immunity with rectal administration, but there is no obvious difference compared to intravenous administration, and rectal administration has no significant effect on mucosal immunity.


The National Health Service Corps: rapid growth and uncertain future.
From inauspicious origins in laudable, but vague, legislation and diffident--even disorganized--implementation, the NHSC has evolved into a major federal program for delivering health care to underserved areas. Tying together the financing of medical education and the delivery of health services for the first time could enable responsiveness to both qualitative and quantitative changes in medical care. But competing demands, within the program and its administration, and in the communities to be served, have made evaluation of the NHSC effort and impact difficult, but all the more necessary for future planning.


Serum cholesterol levels in American (Pima) Indian children and adolescents.
Serum cholesterol levels from birth to adulthood in a population of North American (Pima) Indians are described and compared to those of Caucasian populations. Cholesterol levels at birth (mean +/- SEM, 87 +/- 2.6 mg/100 ml) were similar in Pimas and Caucasians, but levels in Pimas from 5 to 16 years (148 +/- 4.6 mg/100 ml) were 20 to 30 mg/100 ml lower than among most white populations. The levels showed little rise with age from 5 to 16, then rose significantly in both sexes from ages 17 to 25. Cholesterol levels in adult Pimas (190 +/- 1.5 mg/100 ml) were up to 50 to 60 mg/100 ml lower than in American whites, and showed little increase after age 25. Two cohorts of children followed prospectively for six years indicated that the prevalence data reflect sequential changes in the population. Cholesterol levels of those subjects were significantly correlated at the first and last examinations. The Pima, in contrast to Caucasian American populations, have relatively low levels of serum cholesterol and low rates of coronary heart disease, but evidence of a causal relationship with the latter remains to be established.


[End-of-life decision-making process].
Since time immemorial the attitude toward the dying patient has been one of the most difficult issues in medical ethics. The diversity of philosophical, religious, social and legal approaches does not enable one to reach a universal consensus to solve the many problems involved in end-of-life decisions. Within the health care system in Israel there is currently no consensual practice concerning the dying patient. Moreover, there is no published information on the actual decision-making processes within hospitals, hospices and geriatric facilities in Israel concerning the dying patient. A group of investigators in Shaare Zedek Medical Center in Jerusalem recently performed a prospective study to explore the decision-making process concerning DNR orders within this hospital. The results of this study demonstrated that the terminally-ill patients never take part in the decision-making process, they are never consulted about their wishes, and there is no effort to discover their previous wishes concerning their treatment at the terminal stages. Moreover, in many instances even the family was not consulted and did not take part in the decision-making process. In a significant minority, the final decision of a DNR order was undertaken by a single physician. This approach represents an extreme form of unethical paternalism, and it requires an urgent societal intervention to establish an ethically sound decision-making process. Recently, a national committee ("Steinberg Committee") formulated a widely agreed upon legislative proposal organizing all the fundamental and practical issues related to the dying patient. This proposal is based upon a balance between opposing values such as autonomy, life, quality-of-life, beneficence, non-maleficence and "slippery-slope" concerns. It relates to various treatment modalities, such as resuscitation, ventilation, dialysis, medication and sustenance. It establishes a clear position on euthanasia, physician-assisted suicide, withdrawing treatment and withholding treatment. It establishes a hierarchy of decision-making agents, and it validates advanced medical directives. It also promotes a legally-binding requirement of modern palliative care.


Characterization of the GU microbiome in women with self-perceived bladder health over the life course.
A variety of factors influence bladder health, including environmental factors, life experiences, biologic foundations, and coexistent medical conditions. A biologically diverse microbial community exists in the urine that is likely influenced by the microbial inhabitants of the vagina. The relationship between the genitourinary (GU) microbiome and self-perceived bladder health is unknown. To longitudinally define the GU microbiome in women with self-percieved bladder health sampled across multiple time points over a year. Women with no reported lower urinary tract dysfunction or symptoms (LUTS) were recruited from six clinical sites and assessed every 6 weeks for 1 year. Voided urine and vaginal samples were longitudinally collected. Self-perceived bladder health was assessed with select items from the LURN comprehensive assessment of self-reported urinary symptoms (CASUS) tool. We defined four life phases as follows: young (18-34 years, nulliparous), midlife (35-45 years, menstruating), transitional (46-60 years, perimenopausal), mature (>60 years, not using vaginal and/or systemic hormone replacement therapy). DNA was extracted from samples, and the V4 region of the 16S rRNA gene was amplified with region-specific primers. The 16S rRNA sequencing on an Illumina NovaSeq. Microbial beta-diversity was calculated using DEICODE to identify microbial taxa that cluster in the samples. Longitudinal volatility analysis was performed using the gemelli plugin. Log-abundance ratios of microbial features were explored and visualized in Qurro. Fifty-four (N = 16 young, N = 16 midlife, N = 15 transitional, N = 7 mature) women were enrolled and provided baseline data. Most women in each life phase (93%-98%) continued to report self-perceived bladder health throughout the 1-year follow-up as assessed by CASUS items. Temporal-based microbial diversity of urinary and vaginal microbiome remained relatively stable over 1 year in all subjects. The GU microbiomes of mature women were distinct and microbially diverse from that of young, midlife, and transitional women, with genera of Gardnerella, Cupriavidus, and Dialister contributory to the microbial features of the mature microbiome. The mature GU microbiome was statistically different (p < 0.0001) from the midlife, transitional, and young microbiome for the log ratio of Gardnerella and Cupriavidus (in the numerator) and Lactobacillus (in the denominator) for voided samples and Gardnerella and Dialister (in the numerator) and Lactobacillus (in the denominator) for vaginal samples. Differences in the GU microbiome were also demonstrated via longitudinal beta-diversity between women developing urinary frequency as reported by CASUS responses or objectively on bladder diary compared to women without urinary frequency. In women with a self-perceived healthy bladder, the GU microbiome remained stable in all age groups over a 1 year period. Differences were seen with respect to life phase, where mature women were distinct from all other groups, and with respect to self-reported LUTS.


Performance of the Now Malaria rapid diagnostic test with returned travellers: a 2-year retrospective study in a French teaching hospital.
Malaria caused by Plasmodium falciparum remains the major life-threatening parasitic infection in the world. The number of cases in non-endemic countries continues to increase, and it is important that misdiagnosis of malaria should not occur, especially in non-immune travellers, because of the high risk of a fatal outcome. In a retrospective study of 399 sera, the Now Malaria rapid test was compared with the quantitative buffy coat (QBC) test and microbiological examination of thin blood films. Compared with the QBC test and thin blood films, the Now Malaria test had sensitivity and specificity values of 96.4% and 97%, respectively, for the detection of pure P. falciparum infection. A negative predictive value of 99.4% allows this test to be included in diagnostic strategies for patients presenting with clinical suspicion of malaria. Two false-negative results were associated with low levels of parasitaemia in the specimens. Thus, use of the Now Malaria test alone to detect P. falciparum infection in non-endemic countries could lead to misdiagnosis of malaria. This rapid diagnostic test should therefore be performed in association with another prompt traditional method such as examination of thin blood films.


Yohimbine administration prevents over-responsiveness to epinephrine induced by simulated microgravity.
Simulated microgravity produces sustained inhibition of sympathoneural release, turnover, and synthesis of norepinephrine (NE) and hypersensitization of beta-adrenergic pathways. These changes may explain the orthostatic intolerance experienced by astronauts returning from spaceflights. Chronic administration of yohimbine would prevent the increase of beta-adrenergic hypersensitivity to epinephrine (Epi) induced by simulated microgravity. Eight healthy young subjects received 8 mg of yohimbine (an antagonist of alpha2adrenoceptors) orally twice a day during the simulated microgravity achieved through -6 degrees head-down bed rest (HDBR). The catecholamine-induced lipolysis was studied on isolated fat cells from subcutaneous adipose tissue before HDBR and on the fifth day of HDBR. Epi was infused at three graded rates (0.01, 0.02, and 0.03 microg x kg(-1) x min(-1) for 40 min each) before and at the end of the HDBR period. The effects of Epi on the sympathetic nervous system (SNS) activity-assessed by plasma NE levels and spectral analysis of systolic BP and heart rate variability-and on plasma levels of glycerol, non-esterified fatty acids, glucose, and insulin and on energy expenditure were evaluated. Under yohimbine treatment, HDBR failed to modify urinary NE excretion and spectral variability of systolic BP in the mid-frequency range. The beta- and alpha-adrenergic sensitivity of fat cells were not modified by HDBR nor were plasma NE levels and spectral variability of systolic BP induced by Epi infusion. No alteration of Epi-induced changes in heart rate and systolic and diastolic BPs were observed after HDBR. Epi-induced increases in plasma glucose, insulin, glycerol, and non-esterified fatty acid levels as well as energy expenditure were also unmodified by HDBR. Only the Epi-induced plasma lactate level was increased by HDBR. Our data suggest that the increase in the effects of Epi induced during microgravity could be attenuated by chronic administration of yohimbine. An explanation for this effect could be SNS activation brought about by the alpha2-adrenoceptor antagonist properties of yohimbine.