Task-dependent pitch auditory feedback control in cerebellar ataxia.
The purpose of this study was to investigate how ataxia affects the task-dependent role of pitch auditory feedback control in speech. In previous research, individuals with ataxia produced over-corrected, hypermetric compensatory responses to unexpected pitch and formant frequency perturbations in auditory feedback in sustained vowels and single words (Houde et al., 2019; Li et al., 2019; Parrell et al., 2017). In this study, we investigated whether ataxia would also affect the task-dependent role of the auditory feedback control system, measuring whether pitch-shift responses would be mediated by speech task or semantic focus pattern as they are in neurologically healthy speakers. Twenty-two adults with ataxia and 29 age- and sex-matched control participants produced sustained vowels and sentences with and without corrective focus while their auditory feedback was briefly and unexpectedly perturbed in pitch by +/-200 cents. The magnitude and latency of the reflexive pitch-shift responses were measured as a reflection of auditory feedback control. Individuals with ataxia produced larger reflexive pitch-shift responses in both the sustained-vowel and sentence-production tasks than the control participants. Additionally, a differential response magnitude was observed by task and sentence focus pattern for both groups. These findings demonstrate that even though accuracy of auditory feedback control correction is affected by cerebellar damage, as evidenced by the hypermetric responses, the system still retains efficiency in utilizing the task-dependent role of auditory feedback.


An introduction to analytical methods for the postmarketing surveillance of veterinary vaccines.
Any analysis of spontaneous AER data must consider the many biases inherent in the observation and reporting of vaccine adverse events. The absence of a clear probability structure requires statistical procedures to be used in a spirit of exploratory description rather than definitive confirmation. The extent of such descriptions should be temperate, without the implication that they extend to parent populations. It is important to recognize the presence of overdispersion in selecting methods and constructing models. Important stochastic or systematic features of the data may always be unknown. Our attempts to delineate what constitutes an AER have not eliminated all the fuzziness in its definition. Some count every event in a report as a separate AER. Besides confusing the role of event and report, this introduces a complex correlational structure, since multiple event descriptions received in a single report can hardly be considered independent. The many events described by one reporter would then become inordinately weighted. The alternative is to record an AER once, regardless of how many event descriptions it includes. As a practical compromise, many regard the simultaneous submission of several report forms by one reporter as a single AER, and the next submission by that reporter as another AER. This method is reasonable when reporters submit AERs very infrequently. When individual reporters make frequent reports, it becomes difficult to justify the inconsistency of counting multiple events as a single AER when they are submitted together, but as separate AERs when they are reported at different times. While either choice is imperfect, the latter approach is currently used by the USDA and its licensed manufacturers in developing a mandatory postmarketing surveillance system for veterinary immunobiologicals in the United States. Under the proposed system, summaries of an estimated 10,000 AERs received annually by the manufacturers would be submitted to the USDA. In quantitative summaries, AERs received from lay consumers are usually weighted equally with those received from veterinary health professionals, although arguments have been advanced for separate classifications. The emphasis on AER rate estimation differentiates the surveillance of veterinary vaccines by the USDA CVB from the surveillance of veterinary drugs as practiced by the Food and Drug Administration (FDA) Center for Veterinary Medicine (CVM). The FDA CVM does, in fact, perform a retrodictive causality assessment for individual AERs (Parkhie et al., 1995). This distinction reflects the differences between vaccines and drugs, as well as the difference in regulatory philosophy between the FDA and the USDA. The modified Kramer algorithm (Kramer et al., 1979) used by the FDA relies on features more appropriate to drug therapy than vaccination, such as an ongoing treatment regimen which allows evaluation of the response to dechallenge and rechallenge. In tracking AERs, the FDA has emphasized the inclusion of clinical manifestations on labels and inserts, while the USDA has been reluctant to have such information appear in product literature or to use postmarketing data for this purpose. The potential for the misuse of spontaneous AER data is great. Disinformation is likely when the nature of this type of data is misunderstood and inappropriate analytical methods blindly employed. A greater danger lies in the glib transformation of AER data into something else entirely. Since approval before publication is not required, advertisements for veterinary vaccines appear with claims such as "over 3 million doses, 99.9905% satisfaction rating," or "11,500,000 doses, 99.98% reaction free." These claims, presumably based on spontaneous AERs, are almost fraudulent in their deceptiveness. Are we to suppose that 11.5 million vaccinations were observed for reactions? In comparing the two advertisements, we find the second presumed AER rate is double the first. (ABSTRACT TRU


On a Relation Between Packing and Covering Densities of Convex Bodies.
We show that a convex body admits a translative dense packing in R d if and only if it admits a translative economical covering.


Traumatic duodenitis in a dairy cow.
The history, clinical signs, and clinical pathology in a mature Holstein cow were consistent with a diagnosis of intussusception, but the lesion found during exploratory laparotomy consisted of a penetrating wire in the anterior duodenum.


Correction of lower lid retraction combined with entropion using an ear cartilage graft in the anophthalmic socket.
To investigate the surgical results of an ear cartilage graft and supplemental procedures for correcting lower lid retraction combined with entropion in anophthalmic patients. We reviewed retrospectively the medical records of 7 anophthalmic patients with lower lid retraction and entropion, who received a posterior lamellar ear cartilage graft and one or both of lateral tarsal strip or eyelash-everting procedure between March 1998 and March 2003. Preoperative and postoperative lid and socket statuses were also investigated. Ear cartilage grafts were performed in all 7 patients, lateral tarsal strips in 6, and eyelash-everting procedures in 5. Postoperative follow-up durations ranged from 4 to 28 months (average 12.6 months). Retractions were corrected during follow-up in all patients. There were no cases of entropion immediately after surgery. However, the eyelashes of the lower lid returned to an upright position in 4 patients, but not so severe as to touch the ocular prosthesis, and thus did not require surgical correction during follow up. Lower lid retraction combined with entropion in anophthalmic patients can be corrected effectively using an ear cartilage graft with selective, supplemental procedures.


[Evaluation of tobacco smoking addiction and passive exposure to tobacco smoke in students of medicine].
Active and passive tobacco smoking by students of medical universities show that medical studies cannot be an effective barrier against continuing or even commencing smoking. The aim of the study was an evaluation of tobacco smoking and passive exposure to environmental tobacco smoke in the Physiotherapy students of Faculty of Military Medicine Medical University of Lodz. There were 565 Physiotherapy students of between 1-5th year of studies, including 394 women and 171 men (mean age 22.41), who were covered by the questionnaire research. The author's form of the questionnaire consisted of closed questions about demographic data and data on active and passive smoking. Prior to completing the questionnaire each student signed an informed consent as well as had an opportunity to get familiar with the patient's information form and could ask additional questions. The results analysis of the questionnaire indicated that 21.42% of the studied subjects are active smokers whereas 78.58% - non-smokers. No statistically significant differences in tobacco smoking due to sex have been noticed. The highest percentage of smokers was found on the 2nd and 3rd year of studies. In the group of 444 non-smokers there were as many as 67.1% of passive smokers. Tobacco smoking rate among Physiotherapy students amounts to 21.42% and is lower than the rate observed in the Polish population. Among non-smoking physiotherapy students there are 67.1% who are exposed to secondhand tobacco smoke. The obtained result does not largely stray from the result for the adult Polish population.


Morphological and videofluorographic study of the hyoid apparatus and its function in the rabbit (Oryctolagus cuniculus).
The anatomy of the hyoid apparatus and positional changes of the hyoid bone during mastication and deglutition are described in the New Zealand White rabbit (Oryctolagus cuniculus). A testable model is constructed to predict the range of movement during function of the hyoid, a bone entirely suspended by soft tissue. Frame-by-frame analysis of a videofluorographic tape confirms the accuracy of the prediction through observation of hyoid bone excursion during oral behavior. During chewing, translation of the hyoid bone is diminutive and irregular, lacking a clearly discernible path of excursion. However, some movements of the hyoid occur with regularity. During fast opening, anterodorsal movement of the hyoid is interrupted with an abrupt posteroventral depression when the bolus is moved posteriorly toward the cheek teeth by the tongue. This clockwise rotation (when viewed from the right side) of the hyoid accompanies jaw opening and is reversed (posteroventral movement) for the jaw closing sequence. Lateral movements of the hyoid may be slightly coupled to mandibular rotation in the horizontal plane. The findings suggest that the hyoid bone maintains a relatively static position during the dynamics of chewing. The primary function would be to provide a stable base for the movements of the tongue. Another possible function would be to control the position of the larynx within the pharyngeal cavity. Some characteristic features of the rabbit hyoid apparatus may be consequential to relatively erect posture and a saltatory mode of locomotion.


[Sinding-Larsen-Johansson disease: analysis of the associated factors].
To analyse the clinical symptoms, aetiology and biomechanical aspects related to Sinding-Larsen-Johansson (SLJ) disease. A case control study was conducted, analysing the following variables: age, gender, clinical follow up, side of body with the symptoms, sporting activity, radiological stage, presence of patella alta, presence of short hamstring tendons, and increased posterior tibial slope. A total of 15 knees in 14 patients were studied; one case with bilateral disease. The large majority of cases were 85.7% were male, and the mean age was 10.86 (standard deviation 1.61). All of them practised sport and physical activity at a high level with a variable duration of symptoms (1-36 months). There was predominance in side. The majority were radiological grade II (53.3). The data analysis did not show any significant difference (P>.05) between the study groups regarding the Caton and modified Insall indexes. There was a significant difference in the popliteal angle measured in the affected knees (mean: 32.50, SD: 8.9) compared with the control group (mean: 17.67, SD: 8.21). The difference in the posterior slope angle in the affected knees was also statistically significant (mean: 10.47, SD: 2.82) compared with the control (mean: 8.33, SD: 1.4). According to our data, patients have short hamstring tendons and increased posterior tibial slope compared to the control group, and this difference is statistically significant.


Sphingosine 1-phosphate and its type 1 G protein-coupled receptor: trophic support and functional regulation of T lymphocytes.
The lysophospholipid (LPL) growth factors sphingosine 1-phosphate (S1P) and lysophosphatidic acid (LPA) are generated by macrophages, dendritic cells, mast cells, and platelets, which leads to lymph and plasma concentrations of 0.1-1 microM. Distinctive profiles of G protein-coupled receptors (GPCRs) for S1P and LPA are expressed by each type of immune cell and are regulated by cellular activation. At 1-100 nM, S1P signals T cells through their principal S1P(1) GPCRs with consequent protection from apoptosis, enhancement of chemotaxis, and facilitation of optimal regulatory activity of CD4(+)25(+) T cells. At 0.3-3 microM, S1P inhibits T cell chemotaxis and to a lesser extent other functions. These S1P-S1P(1) GPCR signals suppress homing of blood and spleen T cells to secondary lymphoid tissues. S1P(1) GPCR antagonists evoke lymphopenia by permitting blood T cells to enter lymph nodes and blocking S1P(1) GPCR-dependent T cell efflux from lymph nodes. Inversely, there is a decrease in lymphoid tissue traffic of T cells in transgenic mice, which overexpress lymphocyte S1P(1) GPCRs. The immunotherapeutic activity of S1P(1) GPCR antagonists, which limits T cell access to organ grafts and autoimmune antigens, does not reduce other functional capabilities of T cells. LPLs and their GPCRs thus constitute an immunoregulatory system of sufficient prominence for pharmacological targeting in transplantation, autoimmunity, and immunodeficiency.


Fermentation of sugars in orange peel hydrolysates to ethanol by recombinant Escherichia coli KO11.
The conversion of monosaccharides in orange peel hydrolysates to ethanol by recombinant Escherichia coli KO11 has been investigated in pH-controlled batch fermentations at 32 and 37 degrees C. pH values and concentration of peel hydrolysate were varied to determine approximate optimal conditions and limitations of these fermentations. Very high yields of ethanol were achieved by this microorganism at reasonable ethanol concentrations (28-48 g/L). The pH range between 5.8 and 6.2 appears to be optimal. The microorganism can convert all major monosaccharides in orange peel hydrolysates to ethanol and to smaller amounts of acetic and lactic acids. Acetic acid is coproduced in equimolar amounts with ethanol by catabolism of salts of galacturonic acid.


Pathogenesis of nonsteroidal anti-inflammatory drug-related upper gastrointestinal toxicity.
The mechanisms involved in the pathogenesis of ulcers associated with the use of nonsteroidal anti-inflammatory drugs (NSAIDs) have not been fully elucidated. Although studies using acute mucosal injury as a surrogate for clinically relevant outcomes have provided useful information, in practice, acute mucosal injury does not necessarily provide a reliable predictor of clinical ulcers or complications. Several factors that increase the risk of NSAID-associated gastroenteropathy have been identified, and there are data to support or provide speculation for other physiologic factors that might predispose specific subsets of patients to increased mucosal injury. Since clinically significant events occur less frequently than does ulceration, it is the latter determinant, i.e., the identification of the traits that distinguish patients who develop serious NSAID-associated gastrointestinal (GI) toxicity from those who can tolerate these drugs, that may provide the clues necessary to understand the events underlying ulcer pathogenesis.


Sphingosine 1-phosphate regulates regeneration and fibrosis after liver injury via sphingosine 1-phosphate receptor 2.
Sphingosine 1-phosphate (S1P), a bioactive lipid mediator, stimulates proliferation and contractility in hepatic stellate cells, the principal matrix-producing cells in the liver, and inhibits proliferation via S1P receptor 2 (S1P(2)) in hepatocytes in rats in vitro. A potential role of S1P and S1P(2) in liver regeneration and fibrosis was examined in S1P(2)-deficient mice. Nuclear 5-bromo-2'-deoxy-uridine labeling, proliferating cell nuclear antigen (PCNA) staining in hepatocytes, and the ratio of liver weight to body weight were enhanced at 48 h in S1P(2)-deficient mice after a single carbon tetrachloride (CCl(4)) injection. After dimethylnitrosamine (DMN) administration with a lethal dose, PCNA staining in hepatocytes was enhanced at 48 h and survival rate was higher in S1P(2)-deficient mice. Serum aminotransferase level was unaltered in those mice compared with wild-type mice in both CCl(4)- and DMN-induced liver injury, suggesting that S1P(2) inactivation accelerated regeneration not as a response to enhanced liver damage. After chronic CCl(4) administration, fibrosis was less apparent, with reduced expression of smooth-muscle alpha-actin-positive cells in the livers of S1P(2)-deficient mice, suggesting that S1P(2) inactivation ameliorated CCl(4)-induced fibrosis due to the decreased accumulation of hepatic stellate cells. Thus, S1P plays a significant role in regeneration and fibrosis after liver injury via S1P(2).


Batch production and growth kinetics of hybridomas.
Batch cultures of mouse-mouse hybridoma cell lines were carried out and their growth and production kinetics investigated. Three main cell specific production patterns (expressed as pg IgG/cell x hour) were found, which can be used as a classification system for hybridoma cell lines (groups I-III). Cells showing the highest IgG-production at the beginning of the batch culture (during the lag and the onset of the log-phase) were classified as either group I and II. The difference was that cell lines of group II showed a second high cell specific production at the onset of the stationary and death phases. Cell lines of group III had a quite constant production of antibodies during their growth; but IgG secretion completely stopped after the beginning of the stationary phase. The implications of these three production patterns on the design of a production process are discussed.


G-CSF immunotherapy for treatment of acute disseminated murine melioidosis.
Burkholderia pseudomallei, the causative agent of melioidosis, is an important pathogen in tropical regions of Australia and Southeast Asia. Antibiotic therapy can be ineffective in patients with acute septicaemic melioidosis. It has been proposed that adjunctive immunotherapy using granulocyte colony stimulating factor (G-CSF) combined with antibiotics may provide an alternative approach to antibiotics alone. We have developed a murine model for melioidosis that allows novel treatment approaches to be investigated. This study looked at the potential for murine G-CSF therapy both alone and as an adjunct in the treatment of acute disseminated B. pseudomallei infection in BALB/c mice. A number of therapeutic variables involving ceftazidime and recombinant murine G-CSF were studied. Surviving mice were sacrificed and splenic bacterial loads were determined. Combining recombinant murine G-CSF with ceftazidime offered no advantage over ceftazidime alone. Pre-treatment with recombinant murine G-CSF did not demonstrate a significant benefit. This would suggest that adjunct immunotherapy using G-CSF is of limited benefit.


A novel technique for extraction of the appendix in laparoscopic appendectomy.
A new technique for extraction of the appendix by its retrograde introduction into the umbilical port is described. With this technique, appendectomy can be undertaken with a single 10-mm and two 5-mm ports.


Synergistic effects of atomic oxygen and thermal cycling in low earth orbit on polymer-matrixed space material.
Polymer-matrixed materials are widely used in the spacecrafts' structures. However, crafts located in the LEO(Low Earth Orbit) would suffer from hazardous environment factors when orbiting in the space. It has been reported that the space environment factors' integral effect (which represents the factual detriment in space) is not equivalent to the simple summation of each individual. Hence, atomic oxygen and thermal cycling were selected as the starting point for studying the typical LEO synergistic effects on polymer-matrixed space material. In this work, methods such as surface morphology observation, surface components analyzation and inter-laminar-shear strength test were embraced to gather the basic information for the study of degradation. As a result, focusing on the composites selected in this work, synergistic effects do exist between the two factors (AO&TC, representing for atomic oxygen and thermal cycling combined). Besides, a quantified index was proposed to represent synergistic characteristics,so as to lay the foundation for the scientific evolution of material characterization.


Toric intraocular lens combined with a supplementary pinhole implant to treat irregular corneal astigmatism.
A case of refractive lens exchange with the implantation of a supplementary intraocular pinhole (IOPH) to treat a challenging case of keratoconus is presented. Four months after the initial procedure, an intraocular lens (IOL) exchange (under the IOPH) was performed to a toric implant to correct the residual manifest astigmatism. This significantly improved both distance and near acuities and demonstrates how a toric IOL and astigmatism correction in combination with a pinhole device could be used to treat selected cases of keratoconus.


Overcoming the Interobserver Variability in Lung Adenocarcinoma Subtyping: A Clustering Approach to Establish a Ground Truth for Downstream Applications.
The accurate identification of different lung adenocarcinoma histologic subtypes is important for determining prognosis but can be challenging because of overlaps in the diagnostic features, leading to considerable interobserver variability. To provide an overview of the diagnostic agreement for lung adenocarcinoma subtypes among pathologists and to create a ground truth using the clustering approach for downstream computational applications. Three sets of lung adenocarcinoma histologic images with different evaluation levels (small patches, areas with relatively uniform histology, and whole slide images) were reviewed by 17 international expert lung pathologists and 1 pathologist in training. Each image was classified into one or several lung adenocarcinoma subtypes. Among the 4702 patches of the first set, 1742 (37%) had an overall consensus among all pathologists. The overall Fleiss κ score for the agreement of all subtypes was 0.58. Using cluster analysis, pathologists were hierarchically grouped into 2 clusters, with κ scores of 0.588 and 0.563 in clusters 1 and 2, respectively. Similar results were obtained for the second and third sets, with fair-to-moderate agreements. Patches from the first 2 sets that obtained the consensus of the 18 pathologists were retrieved to form consensus patches and were regarded as the ground truth of lung adenocarcinoma subtypes. Our observations highlight discrepancies among experts when assessing lung adenocarcinoma subtypes. However, a subsequent number of consensus patches could be retrieved from each cluster, which can be used as ground truth for the downstream computational pathology applications, with minimal influence from interobserver variability.


In vitro impact of five pesticides alone or in combination on human intestinal cell line Caco-2.
In Burkina Faso, as in most Sahelian countries, the failure to follow good agricultural practices coupled with poor soil and climate conditions in the locust control context lead to high environmental contaminations with pesticide residues. Thus, consumers being orally exposed to a combination of multiple pesticide residues through food and water intake, the digestive tract is a tissue susceptible to be directly exposed to these food contaminants. The aim of our work was to compare in vitro the impact of five desert locust control pesticides (Deltamethrin DTM, Fenitrothion FNT, Fipronil FPN, Lambda-cyalothrine LCT, and Teflubenzuron TBZ) alone and in combination on the human intestinal Caco-2 cells viability and function. Cells were exposed to 0.1-100 μM pesticides for 10 days alone or in mixture (MIX). Our results showed a cytotoxic effect of DTM, FNT, FPN, LCT, and TBZ alone or in combination in human intestinal Caco-2 cells. The most efficient were shown to be FPN and FNT impacting the cell layer integrity and/or barrier function, ALP activity, antioxidant enzyme activity, lipid peroxidation, Akt activation, and apoptosis. The presence of antioxidant reduced lipid peroxidation level and attenuated the pesticides-induced cell toxicity, suggesting that key mechanism of pesticides cytotoxicity may be linked to their pro-oxidative potential. A comparative analysis with the predicted cytotoxic effect of pesticides mixture using mathematical modeling shown that the combination of these pesticides led to synergistic effects rather than to a simple independent or dose addition effect.


Exploring weight initialization, diversity of solutions, and degradation in recurrent neural networks trained for temporal and decision-making tasks.
Recurrent Neural Networks (RNNs) are frequently used to model aspects of brain function and structure. In this work, we trained small fully-connected RNNs to perform temporal and flow control tasks with time-varying stimuli. Our results show that different RNNs can solve the same task by converging to different underlying dynamics and also how the performance gracefully degrades as either network size is decreased, interval duration is increased, or connectivity damage is induced. For the considered tasks, we explored how robust the network obtained after training can be according to task parameterization. In the process, we developed a framework that can be useful to parameterize other tasks of interest in computational neuroscience. Our results are useful to quantify different aspects of the models, which are normally used as black boxes and need to be understood in order to model the biological response of cerebral cortex areas.


Cardiovascular computed tomography in pediatric congenital heart disease: A state of the art review.
Cardiac computed tomography (CCT) has increasingly been used in the assessment of both children and adults with congenital heart disease (CHD), in part due to advances in CCT technology and an increased prevalence of adults with palliated CHD. It serves as a complimentary modality to echocardiography, cardiac magnetic resonance imaging and cardiac catheterization. CCT can provide unique diagnostic information, is less invasive and less likely to require sedation compared to other modalities. Detailed knowledge of individual patient cardiac anatomy, physiology, surgical repair and possible residual lesions are paramount to optimal CCT imaging. This comprehensive review details the use of CCT both pre- and postoperatively for the most common CHD diagnoses. We also aim to highlight some new and innovative technologies that have become available and can further optimize CCT imaging for CHD patients.


Application of biochar to soils may result in plant contamination and human cancer risk due to exposure of polycyclic aromatic hydrocarbons.
Biochars are added to soil to improve agronomic yield. This greenhouse- and field-scale study evaluated polycyclic aromatic hydrocarbon (PAH) contamination in 35 commercial and laboratory-produced biochars, and assessed the effects of biochar amendment of soils on PAH accumulation in vegetables and the risk for cancer. The total and bioavailable PAH concentrations in biochars varied from 638 to 12,347 μg/kg and from below the detection limit (BDL) to 2792 μg/kg, respectively. PAH formation in biochars decreased with increasing production temperature (350-650 °C). Root exudates enhanced PAH release from biochars. The total PAH concentrations in eight edible vegetables growing in biochar-amended soil varied according to biochar and vegetables type from BDL to 565 μg/kg. A health risk assessment framework was integrated with the benzo[a]pyrene toxic equivalency quotient and the incremental lifetime cancer risk (ILCR) to estimate the exposure risk for human beings via ingestion of PAH-contaminated vegetables. The total ILCR for adults was above 10-6, which suggests a risk to human health from direct exposure to PAHs in vegetables grown in biochar-amended soil. These results demonstrate that biochar application may lead to contamination of plants with PAHs, which represents a risk to human health. The PAH levels in biochars produced using different conditions and/or feedstocks need to be evaluated and biochars should be pretreated to remove PAHs before their large-scale agronomic application.


Design, synthesis and biological evaluation of novel pyrazolo-pyrimidinones as DPP-IV inhibitors in diabetes.
We report the design, synthesis, biological activity and docking studies of series of novel pyrazolo[3,4-d]pyrimidinones as DPP-IV inhibitors in diabetes. Molecules were synthesized and evaluated for their DPP-IV inhibition activity. Compounds 5e, 5k, 5o and 6a were found to be potent inhibitors of DPP-IV enzyme. Amongst all the synthesized compounds, 6-methyl-5-(4-methylpyridin-2-yl)-1-phenyl-1H-pyrazolo[3,4-d]pyrimidin-4(5H)-one (5k) was found to be the most active based on in vitro DPP-IV studies and also exhibited promising in vivo blood glucose lowering activity in male Wistar rats.


Validation of IS900- qPCR assay to assess the presence of Mycobacterium avium subs. paratuberculosis in faecal samples according to the OIE procedure.
Paratuberculosis is a chronic enteric progressive disease, caused by Mycobacterium avium subsp. paratuberculosis (MAP). Despite cultural methods being considered the gold standard for the diagnosis of paratuberculosis, quantitative PCR (qPCR) assays have been developed for this purpose. These assays showed sensitivity and specificity comparable to cultural method but provide more rapid analysis results. Aim of our work was the validation of an IS900-qPCR assay for detection of MAP in faeces according to the OIE guidelines relative to the validation of assays for infectious diseases. The analytical and diagnostic characteristics and the reproducibility of the qPCR method were assessed. The robustness of the assay was evaluated using two extraction methods (silica column and magnetic beads DNA capture) and two qPCR systems (STEPONE™ and CFX96™). According to our validation, the analytical specificity, inclusivity and exclusivity were found to be appropriate for the use of this qPCR assay as a diagnostic test. Specifically, the limit of detection was approximately 100 CFU/g or even less if binomial approaches were used for the determination of the 95 % probability of detection (logit and clog-log models) with sufficient repeatability and reproducibility. Estimation of test accuracy was performed using a Bayesian two latent class model, in various scenarios combining different priors for prevalence and accuracy of the two tests used. All models were run considering three different cut-offs for qPCR. Our validation study underlines the good performance of this IS900-qPCR assay for diagnosis of MAP representing a valid and robust alternative to culture. Moreover, coupled with the semiautomatic magnetic beads DNA extraction method, this assay allows the rapid processing of numerous samples.


Enzymatic Reconstitution and Biosynthetic Investigation of the Bacterial Carbazole Neocarazostatin A.
Tricyclic carbazole is an important scaffold in many naturally occurring metabolites, as well as valuable building blocks. Here we report the reconstitution of the ring A formation of the bacterial neocarazostatin A carbazole metabolite. We provide evidence of the involvement of two unusual aromatic polyketide proteins. This finding suggests how new enzymatic activities can be recruited to specific pathways to expand biosynthetic capacities. Finally, we leveraged our bioinformatics survey to identify the untapped capacity of carbazole biosynthesis.


The effect of cytochalasin D and monensin on enveloped vaccinia virus release.
Both monensin and cytochalasin D reduced the production of infectious cell associated virus and infectious extracellular virus with the latter clearly being the more sensitive. The difference in yields were even more clearly seen if the yield of virus particles was monitored instead of yield of infectious virus. Addition of 1 microgram/ml cytochalasin D or 1 microM monensin to the growth medium of vaccinia virus-infected cells inhibited the appearance of extracellular enveloped vaccinia virus (EEV) in the growth medium without affecting the production of intracellular naked vaccinia virus (INV) particles. Although EEV was not released into the medium of cytochalasin D treated cells, EEV was, nevertheless, detectable in CsCl gradients of cell associated virus. Monensin treatment did not affect the synthesis of vaccinia glycoproteins but did significantly reduce the transport of these glycoproteins to the cell surface and also reduced the secretion of proteins. Monensin had the additional effect of causing the appearance of numerous large vacuoles in the cytoplasm. Vaccinia is normally wrapped by cytoplasmic membranes in preparation for release. The monensin induced conversion of cytoplasmic membranes to large vacuoles is presumably the basis for the block in virus wrapping and subsequent release. Cytochalasin D did not alter any of the steps in protein synthesis, transport or secretion. Electronmicroscopic studies confirmed the existence of EEV on the surface of infected cells treated with cytochalasin D. This drug which specifically affects cellular microfilament organisation thus imposes a block on the final release of EEV from the cell surface.


T-CELL LYMPHOCYTIC LEUKEMIA IN A FALLOW DEER (DAMA DAMA).
A 17-yr-old female fallow deer presented with ataxia, inappetence, decreased fecal output, and decreased mentation. A complete blood count demonstrated leukocytosis (24.1×10(3)/μl, n=1.16-7.38×10(3)/μl), characterized by lymphocytosis (22.89×10(3)/μl, n=0.18-3.65×10(3)/μl), anemia (packed cell volume 20%, n=29.0-55.8%), decreased red blood cell count (4.1×10(3)/μl, n=6.86-14.72×10(3)/μl), and decreased hemoglobin (7.5 g/dl, n=9.4-19.2 g/dl). Numerous mature, well-differentiated lymphocytes were noted on the blood film. Despite treatment and clinical improvement, the decision was made to euthanize the deer. Histopathology identified a monomorphic population of CD3 positive, CD79a negative small lymphocytes replacing most of the hematopoietic tissue in the bone marrow without evidence of tissue invasion. Results of viral screening were negative.


Non-MHC-restricted, tissue-specific T cells recognizing autologous oligodendrocytes in the normal SJL/J mouse.
The oligodendrocyte (OD), a glial cell that produces myelin in the central nervous system (CNS), represents a possible target for autoreactive T cells in autoimmune demyelinating processes. To analyze OD/T lymphocyte interactions, we sensitized in vitro SJL/J mouse spleen cells (SC) over Lewis rat OD cultures and maintained them as long-term T-cell lines in interleukin-2 (IL-2)-containing medium. The proliferative response of these lines could be elicited by syngeneic OD as well as by Lewis rat OD, but appeared to be tissue-specific since SC failed to trigger their proliferation. A T-cell clone of the CD3+, CD8+, CD4- phenotype was obtained from these lines. This clone could mount an IL-2-dependent, tissue-specific, non-major histocompatibility complex (MHC)-restricted proliferative response to OD from rat, guinea pig and various strains of mice (including syngeneic OD), but not to SC, whether resting or activated, nor to astrocytes, kidney cells or Langerhans islets. Thus, we showed that SC from normal unimmunized SJL/J mice include a so far undescribed anti-OD autoreactive T-cell population which can be grown in vitro and develop tissue-specific, non-MHC-restricted proliferative responses.


Hemidesmosomes and anchoring fibril collagen appear synchronously during development and wound healing.
Bullous pemphigoid antisera and monoclonal antibodies to type VII collagen were used to localize hemidesmosomes and anchoring fibrils, respectively, in tissues of developing eyes and healing corneal wounds of New Zealand white rabbits. In the 17-day fetal rabbit eye, both antibodies colocalize to the epithelial-stromal junction of the lid and conjunctival region, but neither binds to the cornea, and electron microscopy demonstrates hemidesmosomes only where the antibodies bind. By 20 days of fetal development, the antibodies colocalize in cornea, and, by electron microscopy, hemidesmosomes are shown to be present as well. In healing 7-mm corneal wounds, both antibodies colocalize at the wound periphery within 66 h. By electron microscopy, hemidesmosomes along small segments of basal lamina are also shown to be present at the wound periphery at this time. These demonstrations of the synchronous assembly of hemidesmosomes and anchoring fibrils support the hypothesis of linkage of hemidesmosomes through the basement membrane to anchoring fibrils.


Management of breech presentation with a large pelvic hydatid cyst in late pregnancy in Tibet: a case report.
Hydatid cystic disease (HCD) is primarily a disease of sheep and cattle. Human beings are accidental hosts. It is prevalent in the Tibet Autonomous Region (TAR) of China. In pregnancy, it can cause many complications. We present a multigravida with breech presentation at 37 weeks of pregnancy in whom a large pelvic hydatid cyst and multiple hepatic hydatids were diagnosed by ultrasonography. The large pelvic hydatid cyst was drained through the posterior fornix under the guidance of ultrasound, and an external cephalic version was performed. A healthy baby was delivered vaginally with head presentation at term. HCD during pregnancy presents with management difficulty. It is important to formulate individualized treatment plans according to the actual situation of the patient and the local level of treatment.


Audiovisual spoken word recognition as a clinical criterion for sensory aids efficiency in Persian-language children with hearing loss.
The aim of this study was to examine the role of audiovisual speech recognition as a clinical criterion of cochlear implant or hearing aid efficiency in Persian-language children with severe-to-profound hearing loss. This research was administered as a cross-sectional study. The sample size was 60 Persian 5-7 year old children. The assessment tool was one of subtests of Persian version of the Test of Language Development-Primary 3. The study included two experiments: auditory-only and audiovisual presentation conditions. The test was a closed-set including 30 words which were orally presented by a speech-language pathologist. The scores of audiovisual word perception were significantly higher than auditory-only condition in the children with normal hearing (P<0.01) and cochlear implant (P<0.05); however, in the children with hearing aid, there was no significant difference between word perception score in auditory-only and audiovisual presentation conditions (P>0.05). The audiovisual spoken word recognition can be applied as a clinical criterion to assess the children with severe to profound hearing loss in order to find whether cochlear implant or hearing aid has been efficient for them or not; i.e. if a child with hearing impairment who using CI or HA can obtain higher scores in audiovisual spoken word recognition than auditory-only condition, his/her auditory skills have appropriately developed due to effective CI or HA as one of the main factors of auditory habilitation.


Olva Odlum BDS MSc LDS, the First Female Faculty Member in Manitoba and Dentist to the Indigenous Population.
Olva Odlum qualified as a dentist in England but went on to make a professional life in Canada. She became the first female member of the Manitoba dental faculty and provided care for many dentally-deprived groups, including disabled patients, those with cancer and members of the First Nation tribes.


Cord prolapse: is antenatal diagnosis possible?
Cord presentation was diagnosed antenatally in nine patients at term referred for fetal ultrasound assessment (incidence of 0.61%). Seven patients were delivered by cesarean section; cord position was confirmed in four and suspected in three patients. There were two vaginal deliveries, one following spontaneous version and the other a stillbirth associated with cord prolapse.


Interaction of δ and κ opioid receptors with adenosine A1 receptors mediates cardioprotection by remote ischemic preconditioning.
Multiple initiatives are underway to harness the clinical benefits of remote ischemic preconditioning (rIPC) based on applying non-invasive, brief, intermittent limb ischemia/reperfusion using an external occluder. However, little is known about how rIPC induces protection in cardiomyocytes, particularly through G-protein coupled receptors. In these studies, we determined the role of opioid and adenosine receptors and their functional interactions in rIPC cardioprotection. In freshly isolated cardiomyocytes subjected to 45-min simulated ischemia followed by 60-min simulated reperfusion, we examined the ability of plasma dialysate (derived from blood obtained from rabbits remotely preconditioned by application of brief cycles of hind limb ischemia/reperfusion, rIPC dialysate) to protect cells against necrosis. rIPC dialysate and selective activation of either δ-opioid receptors or κ-opioid receptors significantly reduced the % of dead cells after simulated ischemia and simulated reperfusion. Inhibition of adenosine A1 receptors, but not adenosine A3 receptors, blocked the protection by rIPC dialysate, δ-opioid receptor and κ-opioid receptor activation. In HEK293 cells expressing either hemagglutinin A-tagged δ-opioid receptors or hemagglutinin A-tagged κ-opioid receptors, selective immunoprecipitation of adenosine A1 receptors pulled down both δ-opioid and κ-opioid receptors. This molecular association of adenosine A1 receptors with δ-opioid and κ-opioid receptors was confirmed by reverse pull-down assays. These findings strongly suggest that rIPC cardioprotection requires the activation of δ-opioid and κ-opioid receptors and relies on these receptors functionally interacting with adenosine A1 receptors.


Local gating of information processing through the thalamus.
Inhibitory sculpting of afferent signals in the thalamus is exerted by two types of neurons using gamma-amino butyric acid (GABA) as neurotransmitter. Of them, local-circuit neurons exert their functions via two outputs: axons and presynaptic dendrites. In this issue of Neuron, Govindaiah and Cox reveal that synaptic activation of metabotropic glutamate receptors selectively increases the output of presynaptic dendrites of local interneurons in rat visual thalamus, without affecting the axonal output.


Studies towards neoglycoconjugates from the monosaccharide determinant of Vibrio cholerae O:1, serotype Ogawa using the diethyl squarate reagent.
The effect of reaction time, concentration and molar excess of hapten upon the efficiency of the conjugation of carbohydrates to proteins using the diethyl squarate reagent has been studied using chicken serum albumin (CSA) as the carrier protein and a linker-equipped D-glucose derivative as the hapten. A high degree of incorporation of the latter into CSA was achieved with high efficiency, and the use of a large excess of the ligand was not necessary. Conjugation of the immunodominant monosaccharide determinant of Vibrio cholerae O:1, serotype Ogawa, bearing the same spacer, followed a similar pattern, showing that the nature of the carbohydrate does not substantially affect the outcome of the conjugation and that a predicted degree of antigen-loading onto carrier protein is possible to achieve.


Prevalence of Substance Use in People With First-Episode Psychosis.
People experiencing a first episode of psychosis often have co-occurring substance use, which increases risk of prolonged psychosis and impairs recovery. This article examines the prevalence of substance use in people with first-episode psychosis. The authors searched MEDLINE and other databases for articles published between 1990 and 2009 that described current or lifetime prevalence of substance use, misuse, abuse, or dependence in individuals with first-episode psychosis. Forty-four unique studies provided information. More than 25% of individuals with first-episode psychosis in reviewed studies indicated current or lifetime alcohol use, lifetime alcohol abuse/dependence, current or lifetime cannabis use, or lifetime cannabis abuse or dependence. For all substances, lifetime prevalence of abuse/dependence was higher than current abuse/dependence. Despite variation in assessment methods, findings were generally consistent. Individuals with first-episode psychosis have lower current substance prevalence than lifetime prevalence, suggesting cessation of some substance use prior to seeking treatment for psychosis.


The effects of clipping, nitrogen source and nitrogen concentration on the growth responses and nitrogen uptake of an east african sedge.
Kyllinga nervosa Steud., a sedge from the Serengeti short-grass plains, was subjected to a balanced factorial experiment which included unclipped plants and plants clipped weekly to a 5 cm height, nitrogen supplied as either nitrate or ammonium and three nitrogen concentrations. Tillering rates, green leaf nitrogen, and both green leaf weight and biomass investment in green leaf production increased with nitrogen concentration. Low nitrogen conserved investment in crown production and resulted in adjustments for nitrogen acquisition by increasing biomass allocation to root production. Nitrate nutrition stimulated green leaf weight, tillering rate, nitrogen redistribution and both crown and root nitrogen. Ammonium nutrition increased nitrogen uptake, total plant nitrogen accumulation, reproduction, litter weight and nitrogen loss to decomposers. Clipping increased investment in green leaf production at the expense of stem, root, crown and flower production. Compensatory green leaf production in response to clipping occurred only when plants were grown in ammonium. Clipping stimulated uptake rates of both ammonium and nitrate, and therefore total plant nitrogen accumulation. Results suggest a balanced utilization of both nitrate and ammonium may be necessary for optimal growth in this species.


On the stationarity and normality of the electroencephalographic data during sleep stages.
In this paper, we show techniques to examine the stationarity and the normality of time series as well as results obtained by applying these techniques to EEG data during sleep stages. Many statistical analyses of the EEG data are based on the assumption that the EEG data are stationary and normally distributed. However, the problem is to know the length of data which is most appropriate for the statistical analysis. From the analysis, it is found that a decreased length of data implies an increased degree of stationarity and normality in each stage of sleep. However, for the practical stationary analysis of EEG data, it is shown that it is appropriate to take about 5 s during sleep state. Stationarity is fulfilled to a relatively large extent in REM but more seldom in stage 2. In the case of short data, asymmetric distribution occupies the large parts of non-normal distribution. As the length of data increases the proportion which is symmetric but sharper or flatter than the normal distribution increases.


Factors Affecting Dental Attendance of Children of New Immigrant Parents: A Cross-Sectional Study.
To explore factors affecting children's dental attendance among new immigrants. Participants in this cross-sectional study were 314 new immigrant child-parent pairs. Parents' demographics, oral health knowledge, perceptions, child's caries status, and oral health behaviours were analysed as determinants for dental attendance. Only 43 % of children had a dental visit within the year. Parents believing in the effectiveness of parental checking were four times more likely to seek dental care for their child [adjusted OR (adOR) 4.48, 95 % CI 1.79-11.13]. Parents perceiving dental check-up as a painful experience were 67 % less likely to visit the dentist (adOR 0.33, 95 % CI 0.17-0.63). Lack of insurance and time reduced the odds of a dental visit by 65 and 59 %, respectively (adOR 0.35, 95 % CI 0.16-0.68; adOR 0.41, 95 % CI 0.12-0.99). The care-seeking behaviour of new immigrant children was determined by parents' perceived ability to detect caries, availability of time and dental insurance, and their perceived dental experience.


Effect of Obesity Duration and BMI Trajectories on Cardiovascular Disease: A Narrative Review.
Obesity is an important risk factor for cardiovascular diseases (CVDs). It is crucial to understand the impact of its duration due to the more extended exposure period and the higher frequency of overweight/obesity at younger ages. In the last decade, various studies have discovered that the duration of obesity, in addition to its severity, might have an impact. Therefore, this study aimed to summarize the current literature to investigate the effect of body mass index (BMI) trajectories and overweight/obesity duration on cardiovascular outcomes. To retrieve related articles, we searched PubMed, EMBASE, Google Scholar, Web of Science, Scopus, and Cochrane electronic databases. The duration of overweight/obesity is significantly associated with CVDs, especially heart failure and atrial fibrillation. However, there are contradictory results regarding the association between coronary heart disease and stroke with the duration of obesity. Additionally, no association with peripheral vascular disease has yet to be reported. The absence of this association may be brought on by covariates or various follow-up times. Nevertheless, it seems that both stable overweight and remarkably stable obesity increase the risk of CVDs, as does both stable overweight and notably stable obesity. Metrics showing both the severity and the duration of overweight/obesity are more effective than each alone, and it is recommended to be used for assessing the risk of various CVDs. There are few studies in these areas, and studies with more extended follow-up periods, with a wide age range, while adjusting for some specific covariates, are needed.


Morphological and life-cycle aspects of the parasitic mite Trichosurolaelaps crassipes Womersley, 1956 of trichosurid possums.
All four life-cycle stages of the ubiquitous parasitic possum mite Trichosuroluelups crassipes Womersley, 1956 are illustrated. They comprise the ephemeral, quiescent and nonfeeding larva, an active feeding protonymph, an active feeding deutonymph and the adults. Morphological adaptations with viviparity allow the entire life-cycle of the mite to be completed on the host. The lack of a nidicolous egg or larva and the absence of a quiescent deutonymph implies transmission is via host-host contact.


A special guide for insertion of multiple pins for fracture of the hip: experience with 200 cases.
Two hundred cases of intracapsular femoral neck fractures have been treated with a multiple parallel pin technique similar to that originated by Austin Moore. The technique employs a personally designed guide which facilitates insertion of multiple parallel pins. The fixation obtained satisfactorily resists shear stress and rotational stress and allows for continuous compression at the fracture site. The technique is most effective in treating intracapsular fractures of the femoral neck.


Influence of gastrointestinal activity on the absorption of nilotinib.
Nilotinib has bioavailability (BA) of only about 25% or less. The purpose of this study was to evaluate the influence of gastrointestinal activity on the absorption of nilotinib. In order to change gastrointestinal activity, mosapride was used for enhancement and butylscopolamine was used for suppression. Experiments on oral administration of nilotinib using rats whose gastrointestinal activity was altered by mosapride or butylscopolamine were carried out. The results of oral administration of acetaminophen to rats with peristalsis movement changed showed that the effects of peristalsis and gastric emptying rate (GER) on drug absorption could be evaluated in this experimental system. Similarly, even with nilotinib, no change in Tmax was observed, but Cmax increased and decreased significantly. Due to the change in gastrointestinal activity, Cmax of nilotinib changed greatly. This showed that gastrointestinal activity affected the emulsifying action of bile and that the absorbability changed. As a result of examining the contribution to the emulsifying action, it was found that when the bile does not exist in the gastrointestinal tract, absorption of nilotinib did not change even when gastrointestinal motility was enhanced. Therefore, the results suggested that gastrointestinal activity influenced the emulsifying action of bile and the absorption of nilotinib was changed.


Recommended guidelines for the management of autologous and allogeneic bone marrow transplantation. A report from the Eastern Cooperative Oncology Group (ECOG)
To define the basic state-of-the-art medical care of the patients after bone marrow transplantation as practiced by the Eastern Cooperative Oncology Group. Studies examining the role of bone marrow transplantation in the care of neoplastic disorders identified using computer and bibliographic searches. More than 500 articles, book chapters, and abstracts from meetings, covering all therapeutic and diagnostic aspects of the patient having bone marrow transplantation were critically reviewed; information from over 200 publications is included. Enormous progress has been made in understanding the biology, therapy, and prophylaxis strategies of bone marrow transplantation and in extending the range of potential marrow donors to include unrelated persons. Dramatic advances have occurred in the prevention of serious infection, including cytomegalovirus infection, formerly a cause of a high rate of mortality. The advent of newer-combination, high-dose chemotherapy regimens and advances in cryopreservation and in vitro marrow purging techniques have led to increased use and greater efficacy of autologous transplantation. Recent advances using recombinant hematopoietic growth factors and autologous peripheral stem cells are likely to reduce morbidity and mortality and significantly shorten the length of hospitalization and the cost of bone marrow transplantation. Bone marrow transplantation now is a common procedure done throughout the world. Cooperative groups have assumed a major role in conducting clinical trials. A standardized approach defining basic standards of care will assure uniformity of management, better interpretation of data, and continued progress in the care of the patient who has had a bone marrow transplantation. Basic research and clinical trials are ongoing to define better methods for the treatment and prevention of graft-versus-host disease and hepatic veno-occlusive disease.


Anionic lipids induce a fold-unfold transition in the membrane-translocating Engrailed homeodomain.
Homeoprotein transcription factors have the property of interacting with membranes through their DNA-binding homeodomain, which is involved in unconventional internalization and secretion. Both processes depend on membrane-translocating events but their detailed molecular mechanisms are still poorly understood. We have previously characterized the conformational properties of Engrailed 2 homeodomain (EnHD) in aqueous solution and in micelles as membrane-mimetic environments. In the present study, we used small isotropic lipid bicelles as a more relevant membrane-mimetic model to characterize the membrane-bound state of EnHD. We show that lipid bicelles, in contrast to micelles, adequately reproduce the requirement of anionic lipids in the membrane binding and conformational transition of EnHD. The fold-unfold transition of EnHD induced by anionic lipids was characterized by NMR using 1H, 13C, 15N chemical shifts, nuclear Overhauser effects, residual dipolar couplings, intramolecular and intermolecular paramagnetic relaxation enhancements induced by site-directed spin-label or paramagnetic lipid probe, respectively. A global unpacking of EnHD helices is observed leading to a loss of the native fold. However, near-native propensities of EnHD backbone conformation are maintained in membrane environment, including not only the three helices but also the turn connecting helices H2 and H3. NMR and coarse-grained molecular dynamics simulations reveal that the EnHD adopts a shallow insertion in the membrane, with the three helices oriented parallel to the membrane. EnHD explores extended conformations and closed U-shaped conformations, which are stabilized by anionic lipid recruitment.


The effects of developmental and/or direct lead exposure on FR behavior in the rat.
In the present study, 21-day old female rats were exposed to daily doeses of 750 mg/kg of lead acetate via restricted water intake regimen for 70-80 days prior to mating. Treatment was then continued throughout gesation and nursing. At weaning, litters from half of the treated and control mothers were placed on treatment for the remainder of the experiment. This manipulation yielded four groups for testing: Group Pb/Pb, developmental and direct, postweaning exposure; Group Pb/C, developmental exposure only; group C/Pb, direct exposure only; and Group C/C, no exposure to lead acetate treatment. Beginning at 42--49 days of age, offspring were shaped to bar press on a Fixed Ratio 20 (FR20) schedule of reinforcement and then received 20 sessions each 20 min in length. Analyses revealed that Group Pb/Pb received significantly fewer reinforcements/min across sessions that the other three groups and also took significantly longer to emit each 20 response block. Contrary to previous reports in the literature, it is suggested that rats may not be impervious to postweaning lead exposure, particularly when there is a history of developmental exposure.


Retraction.
Retraction: "Ethanolic extract of leaf of Dillenia pentagynareduces in-vitro cell migration and induces intrinsic pathway of apoptosis via downregulation of NF-κβ in human NSCLC A549 cells", by Debapriya De, Priyanka Chowdhury, Sujogya K. Panda, and Utpal Ghosh, J Cell Biochem. 2019; 19841-19857: The above article, published online on 18 July 2019 in Wiley Online Library (https://onlinelibrary.wiley.com/doi/10.1002/jcb.29289) has been retracted by agreement between the journal's Editor in Chief, Prof. Dr. Christian Behl, and Wiley Periodicals LLC. The authors admit substantial mistakes during figure compilation for this manuscript. Specifically, inconsistencies and duplicated image elements were found in multiple figures. As a result, the data presented in this article are considered insufficient to support the conclusions. The authors disagree with this decision.


Clinical experience and acceptability of the etonogestrel subdermal contraceptive implant.
To evaluate efficacy, adverse effects, and user continuation rate of an etonogestrel subdermal single-rod contraceptive implant. A total of 417 healthy volunteers of childbearing age were included in this multicenter trial. After implant insertion, the women were followed up during the 3 years of contraceptive action. At each visit, clinical findings, side effects, and bleeding patterns were recorded. Efficacy and continuation rates were analyzed using the Pearl Index and Kaplan-Meier life tables, respectively. The observation period totaled 958.5 woman-years (27.5 months per woman). The Pearl Index score was 0. Side effects were reported by 44.4% of users, but the proportion had decreased to 16.5% by the end of the study. The continuation rate was 61.4%. The most common reason for early discontinuation (in 21.1% of the participants) was menstrual disturbances. Etonogestrel subdermal contraceptive implants demonstrated high efficacy and an acceptable continuation rate. Counseling potential users explicitly about the side effects will optimize patient success with this long-acting contraceptive.


Getting with the program.
An Institute of Medicine report recommends that the United States government create a program to provide consistent guidelines for clinical interventions. The reliability of the guidelines will depend on the availability of the clinical data to be assessed.


Post-Acute Care in Inpatient Rehabilitation Facilities Between Traditional Medicare and Medicare Advantage Plans Before and During the COVID-19 Pandemic.
Compare post-acute care (PAC) utilization and outcomes in inpatient rehabilitation facilities (IRF) between beneficiaries covered by Traditional Medicare (TM) and Medicare Advantage (MA) plans during the COVID-19 pandemic relative to the previous year. This multiyear cross-sectional study used Inpatient Rehabilitation Facility-Patient Assessment Instrument (IRF-PAI) data to assess PAC delivery from January 2019 to December 2020. Inpatient rehabilitation for stroke, hip fracture, joint replacement, and cardiac and pulmonary conditions among Medicare beneficiaries 65 years or older. Patient-level multivariate regression models with difference-in-differences approach were used to compare TM and MA plans in length of stay (LOS), payment per episode, functional improvements, and discharge locations. A total of 271,188 patients were analyzed [women (57.1%), mean (SD) age 77.8 (0.06) years], among whom 138,277 were admitted for stroke, 68,488 hip fracture, 19,020 joint replacement, and 35,334 cardiac and 10,069 pulmonary conditions. Before the pandemic, MA beneficiaries had longer LOS (+0.22 days; 95% CI: 0.15-0.29), lower payment per episode (-$361.05; 95% CI: -573.38 to -148.72), more discharges to home with a home health agency (HHA) (48.9% vs 46.6%), and less to a skilled nursing facility (SNF) (15.7% vs 20.2%) than TM beneficiaries. During the pandemic, both plan types had shorter LOS (-0.68 day; 95% CI: 0.54-0.84), higher payment (+$798; 95% CI: 558-1036), increased discharges to home with an HHA (52.8% vs 46.6%), and decreased discharges to an SNF (14.5% vs 20.2%) than before. Differences between TM and MA beneficiaries in these outcomes became smaller and less significant. All results were adjusted for beneficiary and facility characteristics. Although the COVID-19 pandemic affected PAC delivery in IRF in the same directions for both TM and MA plans, the timing, time duration, and magnitude of the impacts were different across measures and admission conditions. Differences between the 2 plan types shrank and performance across all dimensions became more comparable over time.


Role of 5'-TGN-3' motif in the interaction of mycobacterial RNA polymerase with a promoter of 'extended -10' class.
In a systematic approach to understand the transcriptional machinery of mycobacteria, we had previously isolated and characterized mycobacterial promoter regions. In this study, we have investigated molecular interactions between mycobacterial RNA polymerase holoenzyme, reconstituted with different sigma subunits and the promoter element of the Mycobacterium tuberculosis gene pknH (Rv1266c), a representative of promoters belonging to the 'extended -10' class. In vitro transcription assays using the pknH promoter and reconstituted RNA polymerase holoenzyme demonstrated that transcription from the pknH promoter is specifically initiated by sigmaA, the principal sigma factor of mycobacteria. DNase I protection assay and deletion studies with the pknH promoter revealed that the minimal region required for optimal transcription carries the sequence from position -37 to position +6. Moreover, mutation in the TGN motif of the pknH promoter resulted in the loss of >75% of its activity. Binding of RNA polymerase with wild-type promoter as well as its TG- mutant revealed that the TGN motif is required for the transition from a close complex into an open complex. Further, it was observed that the presence of the TGN motif reduces the thermal energy required for the conversion of a close complex into an open complex, necessary for initiation of transcription.


Toxicity study of erucylphosphocholine in a rat model.
To investigate the effect of intraocular erucylphosphocholine (ErPC) on the retina, the retinal pigment epithelium (RPE), and the choroid in an in vivo rat model. Adult male Brown Norway rats were injected intravitreally with ErPC dissolved in balanced salt solution (BSS) at a final concentration of 10 or 100 microM with BSS serving as control. Adverse effects on the anterior and posterior segment were assessed by slit-lamp biomicroscopy and ophthalmoscopy. Retinal toxicity was assessed by electroretinography (ERG), retinal ganglion cell (RGC) quantification, and histology 7 days after intravitreal administration of ErPC. There was neither a statistically significant difference in the clinical examination nor in the ERG waves of treated versus control rats 7 days after intravitreal administration of ErPC. Correspondingly, the number of RGC after BSS injection did not differ significantly from ErPC-injected animals. Histologic sections of the posterior segment of 10 and 100 microM ErPC-injected rats did not show any signs of retinal toxicity. Electron microscopy did not display a difference between the 10 microM and the control group. Only the 100 microM-injected animals showed a discrete irregularity of the Müller cell and the retinal ganglion cell cytoplasm at the ultrastructural level. ErPC can safely be injected into the vitreous of adult rats at a concentration of 10 microM without any retinal toxicity. Even a 10-fold increase in ErPC concentration leads only to a discrete cytoplasmic irregularity of the innermost retinal layers.


How miRs and mRNA deadenylases could post-transcriptionally regulate expression of tumor-promoting protein PLD.
Phospholipase D (PLD) plays a key role in both cell membrane lipid reorganization and architecture, as well as a cell signaling protein via the product of its enzymatic reaction, phosphatidic acid (PA). PLD is involved in promoting breast cancer cell growth, proliferation, and metastasis and both gene and protein expression are upregulated in breast carcinoma human samples. In spite of all this, the ultimate reason as to why PLD expression is high in cancer cells vs. their normal counterparts remains largely unknown. Until we understand this and the associated signaling pathways, it will be difficult to establish PLD as a bona fide target to explore new potential cancer therapeutic approaches. Recently, our lab has identified several molecular mechanisms by which PLD expression is high in breast cancer cells and they all involve post-transcriptional control of its mRNA. First, PA, a mitogen, functions as a protein and mRNA stabilizer that counteracts natural decay and degradation. Second, there is a repertoire of microRNAs (miRs) that keep PLD mRNA translation at low levels in normal cells, but their effects change with starvation and during endothelial-to-mesenchymal transition (EMT) in cancer cells. Third, there is a novel way of post-transcriptional regulation of PLD involving 3'-exonucleases, specifically the deadenylase, Poly(A)-specific Ribonuclease (PARN), which tags mRNA for mRNA for degradation. This would enable PLD accumulation and ultimately breast cancer cell growth. We review in depth the emerging field of post-transcriptional regulation of PLD, which is only recently beginning to be understood. Since, surprisingly, so little is known about post-transcriptional regulation of PLD and related phospholipases (PLC or PLA), this new knowledge could help our understanding of how post-transcriptional deregulation of a lipid enzyme expression impacts tumor growth.


The electron conduction of photosynthetic protein complexes embedded in a membrane.
The conductivity of two photosynthetic protein-pigment complexes, a light harvesting 2 complex and a reaction center, was measured with an atomic force microscope capable of performing electrical measurements. Current-voltage measurements were performed on complexes embedded in their natural environment. Embedding the complexes in lipid bilayers made it possible to discuss the different conduction behaviors of the two complexes in light of their atomic structure.


Validation of tumor-associated macrophage ferritin light chain as a prognostic biomarker in node-negative breast cancer tumors: A multicentric 2004 national PHRC study.
Novel prognostic biomarkers are imperatively needed to help direct treatment decisions by typing subgroups of node-negative breast cancer patients. Large screening of different biological compartments, such as the proteome, by means of high throughput techniques may greatly help scientists to find such markers. The present retrospective multicentric study included 268 node-negative breast cancer patients. We used a proteomic approach of SELDI-TOF-MS screening to identify differentially expressed cytosolic proteins with prognostic impact. The screening cohort was composed of 198 patients. Seventy supplementary patients were included for validation. Immunohistochemistry (IHC) and immunoassay (IA) were run to confirm the prognostic role of the marker identified by SELDI-TOF-MS screening. IHC was also used to explore links between selected marker and epithelial-mesenchymal transition (EMT)-like, proliferation and macrophage markers. Ferritin light chain (FTL) was identified as an independent prognostic marker (HR = 1.30-95% CI: 1.10-1.50, p = 0.001). Validation step by means of IHC and IA confirmed the prognostic value of FTL level. CD68 IHC showed that FTL was stored in tumor-associated macrophages (TAM), which exhibit an M2-like phenotype. We report here, first, the validation of FTL as a breast tumor prognostic biomarker in node-negative patients, and second, the fact that FTL is stored in TAM.


Direct chiral resolution of tartaric acid by ion-pair capillary electrophoresis using an aqueous background electrolyte with (1R,2R)-(-)-1,2-diaminocyclohexane as a chiral counterion.
Chiral resolution of native DL-tartaric acid was achieved by ion-pair capillary electrophoresis (CE) using an aqueous-ethanol background electrolyte with (1R,2R)-(-)-1,2-diaminocyclohexane (R-DACH) as a chiral counterion. Factors affecting chiral resolution and migration time of tartaric acid were studied. By increasing the viscosity of the background electrolyte and the ion-pair formation, using organic solvents with a lower relative dielectric constant, resulted in a longer migration time. The optimum conditions for both high resolution and short migration time of tartaric acid were found to be a mixture of 65% v/v ethanol and 35% v/v aqueous solution containing 30 mM R-DACH and 75 mM phosphoric acid (pH 5.1) with an applied voltage of -30 kV at 25 degrees C, using direct detection at 200 nm. By using this system, the resolution (Rs) of racemic tartaric acid was approximately 1. The electrophoretic patterns of tartaric and malic acids suggest that two carboxyl groups and two hydroxyl groups of tartaric acid are associated with the enantioseparation of tartaric acid by the proposed CE method.


Breaking the binary: Gender versus sex analysis in human brain imaging.
Despite decades of pursuit, human brain imaging has yet to uncover clear neural correlates of male-female behavioral differences. Given that such behavior does not always align with sex categories, we argue that neuroimaging research may find more success by partitioning subjects along nonbinary gender attributes in addition to sex. We review the handful of studies that have done this, several of which find as good or better association between brain measures and "gender" as they do with "sex." Recent advances in operationalizing "gender" as a multidimensional variable should facilitate such studies, along with discovery-based approaches that mine brain imaging data for gender-associated attributes, independent of sex.


Genetic screening protocol for familial hypercholesterolemia which includes splicing defects gives an improved mutation detection rate.
Familial hypercholesterolemia (FH) is a common single gene disorder, which predisposes to coronary artery disease. In a previous study, we have shown that in patients with definite FH around 20% had no identifiable gene defect after screening the entire exon coding area of the low density lipoprotein receptor (LDLR) and testing for the common Apolipoprotein B (ApoB) R3500Q mutation. In this study, we have extended the screen to additional families and have included the non-coding intron splice regions of the gene. In families with definite FH (tendon xanthoma present, n=68) the improved genetic screening protocol increased the detection rate of mutations to 87%. This high detection rate greatly enhances the potential value of this test as part of a clinical screening program for FH. In contrast, the use of a limited screen in patients with possible FH (n=130) resulted in a detection rate of 26%, but this is still of significant benefit in diagnosis of this genetic condition. We have also shown that 14% of LDLR defects are due to splice site mutations and that the most frequent splice mutation in our series (c.1845+11 c>g) is expressed at the RNA level. In addition, DNA samples from the patients in whom no LDLR or ApoB gene mutations were found, were sequenced for the NARC-1 gene. No mutations were identified which suggests that the role of NARC-1 in causing FH is minor. In a small proportion of families (<10%) the genetic cause of the high cholesterol remains unknown, and other genes are still to be identified that could cause the clinical phenotype FH.


Antiarrhythmic effects of ginsenoside Rg2 on calcium chloride-induced arrhythmias without oral toxicity.
Malignant arrhythmias require drug therapy. However, most of the currently available antiarrhythmic drugs have significant side effects. Ginsenoside Rg2 exhibits excellent cardioprotective effects and appears to be a promising candidate for cardiovascular drug development. So far, the oral toxicity and antiarrhythmic effects of Rg2 have not been evaluated. Acute oral toxicity of Rg2 was assessed by the Limit Test method in mice. Subchronic oral toxicity was determined by repeated dose 28-day toxicity study in rats. Antiarrhythmic activities of Rg2 were evaluated in calcium chloride-induced arrhythmic rats. Antiarrhythmic mechanism of Rg2 was investigated in arrhythmic rats and H9c2 cardiomyocytes. The results of toxicity studies indicated that Rg2 exhibited no single-dose (10 g/kg) acute oral toxicity. And 28-day repeated dose treatment with Rg2 (1.75, 3.5 and 5 g/kg/d) demonstrated minimal, if any, subchronic toxicity. Serum biochemical examination showed that total cholesterol in the high-dose cohort was dramatically decreased, whereas prothrombin time was increased at Day 28, suggesting that Rg2 might regulate lipid metabolism and have a potential anticoagulant effect. Moreover, pretreatment with Rg2 showed antiarrhythmic effects on the rat model of calcium chloride induced arrhythmia, in terms of the reduced duration time, mortality, and incidence of malignant arrhythmias. The antiarrhythmic mechanism of Rg2 might be the inhibition of calcium influx through L-type calcium channels by suppressing the phosphorylation of Ca2+/calmodulin-dependent protein kinase II. Our findings support the development of Rg2 as a promising antiarrhythmic drug with fewer side effects for clinical use.


The AIDS pandemic: a catalyst for women's rights.
The rates of HIV infection among women are rising at a higher rate than among men. After citing the reasons for women's vulnerability, this report argues the potential role of gynecologists and obstetricians through integration of HIV/AIDS into sexual and reproductive health services to strengthen the response to the epidemic.


Incidence and morphological study of zygomaticofacial and zygomatico-orbital foramina in dry adult human skulls: the non-metrical variants.
Morphological diversity in the form of multiple zygomaticofacial (ZF) foramina was studied in 165 dry, unsexed adult human skulls. Zygomatic bones revealed variation in the number of foramina on their orbital and facial aspects. These were absent in 72 (21.8%) sides. A single ZF foramen was seen in 148 (44.9%) sides. Two ZF foramina were found in 92 (27.9%) sides, out of which 29 (8.8%) sides had one zygomatico-orbital (ZO) foramen, while 63 (19.1%) sides had two ZO foramina. Three ZF foramina, a relatively uncommon occurrence, were found in 17 (5.1%) sides, which included eight (2.4%) sides with one less and nine (2.7%) sides with the same number of ZO foramina. Four ZF foramina were seen in one (0.3%) side with three on the orbital aspect, a feature not reported before. The zygomatic nerve may be disrupted on elevating periorbita from the lateral wall during orbital surgery, and care should be taken to prevent injury to the nerve during the lateral orbitotomy approach to access intraorbital soft tissue tumors.


Medicaid: out-of-pocket expenses and access to care.
A 27-page report by the Kaiser Family Foundation examined 13 different studies showing how increased out-of-pocket costs keep people out of care.


Pertussis toxin inhibits endothelium-dependent relaxations evoked by fluoride.
Fluoride is a direct activator of G-proteins. In isolated rings of canine coronary artery, fluoride caused relaxation of rings with endothelium, but only slight contraction of rings denuded of endothelium. The endothelium-dependent relaxations to fluoride were inhibited by pertussis toxin, an inhibitor of G-proteins, or by methylene blue, an inhibitor of soluble guanylate cyclase. Therefore, fluoride causes endothelium-dependent relaxations in part by activating a pertussis toxin-sensitive G-protein in the endothelial cells.


Oxygen-deficient bismuth molybdate nanocatalysts: Synergistic effects in boosting photocatalytic oxidative coupling of benzylamine and mechanistic insight.
Herein, bismuth molybdate (Bi2MoO6) nanocatalysts containing oxygen vacancies (OVs) are found to considerably promote the photocatalytic performance toward oxidative coupling of benzylamine to N-benzylidenebenzylamine under visible light irradiation. The structure-activity relationship for this interesting catalyst is revealed for the first time. The oxygen-deficient Bi2MoO6 nanoplatelets (BMO-NPs) are synthesized using ethylene glycol-ethanol solvent mixture as a reaction medium in solvothermal method. A comparison with hydrothermally prepared Bi2MoO6 square-like sheets (BMO-SHs) suggests that the nanoplatelets are much smaller in size and contain higher amount of OVs. Benzylamine conversion over the BMO-NPs is ca. 4.0 times higher than that over the BMO-SHs and ca. 3.8 and ca. 34.6 times higher than that over the commercial benchmark TiO2 P25 and BiVO4 catalysts, respectively. The BMO-NPs achieve more than 80% product yield within 2 h of irradiation regardless of substituents of benzylamine derivatives. The enhanced activity of BMO-NPs is due to synergistic roles of high surface-to-volume ratio and OVs, providing enlarged active area, extended light absorption range and improved charge separation and transfer efficiency as evidenced from UV-vis DRS, BET surface area, photocurrent response, electrochemical impedance spectroscopy, and time-resolved fluorescence decay measurements. EPR-trapping and radical scavenging experiments indicate O2- as a main active species rather than 1O2 and a plausible imine formation mechanism via O2--assisted charge transfer is proposed accordingly. The work offers an alternative facile preparation method to design efficient semiconductor photocatalysts and for the first time reveals a possible benzylamine coupling mechanism over the oxygen-deficient Bi2MoO6 nanocatalyst.


Regulation of noise in the expression of a single gene.
Stochastic mechanisms are ubiquitous in biological systems. Biochemical reactions that involve small numbers of molecules are intrinsically noisy, being dominated by large concentration fluctuations. This intrinsic noise has been implicated in the random lysis/lysogeny decision of bacteriophage-lambda, in the loss of synchrony of circadian clocks and in the decrease of precision of cell signals. We sought to quantitatively investigate the extent to which the occurrence of molecular fluctuations within single cells (biochemical noise) could explain the variation of gene expression levels between cells in a genetically identical population (phenotypic noise). We have isolated the biochemical contribution to phenotypic noise from that of other noise sources by carrying out a series of differential measurements. We varied independently the rates of transcription and translation of a single fluorescent reporter gene in the chromosome of Bacillus subtilis, and we quantitatively measured the resulting changes in the phenotypic noise characteristics. We report that of these two parameters, increased translational efficiency is the predominant source of increased phenotypic noise. This effect is consistent with a stochastic model of gene expression in which proteins are produced in random and sharp bursts. Our results thus provide the first direct experimental evidence of the biochemical origin of phenotypic noise, demonstrating that the level of phenotypic variation in an isogenic population can be regulated by genetic parameters.


Correction to: Dental core training: the trainee perspective.
Author's correction note: Education article Br Dent J 2020; 228: 782-790.When initially published the wording on the y-axis of figures 4-15 was ordered incorrectly.The original article has been corrected.


Evaluation of a combined lysate/recombinant antigen anti-HTLV-I/II ELISA in high and low endemic areas of HTLV-I/II infection.
The Wellcozyme HTLV-I/II ELISA (Murex Diagnostics) was evaluated in 7800 samples of various serum panels. Repeat activity was found by Wellcozyme in (A) 1/2181 (0.05%) Dutch blood donors, (B) 44/3036 (1.4%) Curaçao (Caribbean area) blood donors, (C) 46/2533 (1.8%) individuals of different Ethiopian population subsets, (D) 30/30 (100%) confirmed anti-HTLV-I positive samples and (E) 20/20 (100%) HTLV-II PCR-positive samples. All 91 Wellcozyme-positive samples were tested for confirmation by Western blot (WB, Diagnostic Biotechnology). Among Wellcozyme HTLV-I/II ELISA-positive individuals, HTLV-I/II WB positivity was found in 0/1 Dutch blood donors, 40/44 (88.9%) Curaçao blood donors and 20/46 (43.5%) Ethiopian individuals. HTLV-I positivity was found in 40 (1.3%) WB-positive Curaçao blood donors and in 9 (0.35%) Ethiopian individuals. HTLV-II positivity was found in 11 (0.43%) WB-positive Ethiopian individuals. The Wellcozyme HTLV-I/II ELISA had a specificity of 99.95% in Dutch blood donors and a sensitivity of 100% on confirmed HTLV-I- and HTLV-II-positive samples. In Ethiopia 55% of the HTLV-I/II WB-positive individuals were exclusively HTLV-II positive, whereas in Curaçao no HTLV-II infections were found.


Early Renal Replacement Therapy Versus Standard Care in the ICU: A Systematic Review, Meta-Analysis, and Cost Analysis.
Renal replacement therapy (RRT) is the treatment of choice for severe acute kidney injury, but there are no firm guidelines as to the time of initiation of RRT in the critically ill. The primary objective of this study is to determine 1-month mortality rates of early versus late dialysis in critical care. As secondary end points, we provide a cost analysis of early versus late RRT initiation, intensive care unit (ICU) length of stay (LOS), hospital LOS, and number of patients on dialysis at day 60 postrandomization. We identified all randomized controlled trials (RCTs) through EMLINE and MEDBASE that examined adult patients admitted to critical care who were randomized to receiving early dialysis versus standard of care. Inclusion criteria: (1) RCTs conducted after the year 2000, (2) the population evaluated had to be adults admitted to ICU, (3) the intervention had to be early RRT versus standard care, and (4) outcomes had to measure patient mortality. Two independent investigators reviewed search results and identified appropriate studies. Information was extracted using standardized case report forms. Overall, 7 RCTs were included with a total of 1400 patients. Early RRT showed no survival benefit when compared to standard treatment (odds ratio [OR], 0.90 95% confidence interval [95% CI] 0.70-1.15, P = .39). There was no significant difference in length of hospital stay in patients with early RRT (-1.55 days [95% CI -4.75 to 1.65, P = .34]), in length of ICU stay (-0.79 days [95% CI -2.09 to 0.52], P = .24), or proportion of patients on dialysis at day 60 (OR 0.93 [95% CI 0.62 to 1.43], P = .79). Per patient, there is likely a small increase in costs (<US$1000) owing to increased total dialysis. Across all measured domains, there is no clear benefit to early RRT. Moreover, this intervention may result in increased costs and exposes patients to an invasive therapy with potential harm.


Insulin resistance phenotypes and coronary artery disease in a native Pakistani cohort.
To determine the relationship between insulin resistance (IR) and atheroma burden in Pakistanis. A prospective case-control study of 400 patients selected for the presence/absence of angiographic disease. Coronary atheroma burden was quantified and IR and cardiovascular risk factors were measured. The patients were divided into two groups by QuickI score. Waist circumference (90 +/- 10 vs. 90 +/- 9 cm; p = 0.7) was similar but the groups differed in body mass index (26.5 +/- 3.7 vs. 24.2 +/- 3.5 kg/m(2); p < 0.001) and waist:hip ratio (0.94 +/- 0.09 vs. 0.90 +/- 0.06; p < 0.001). Lipid parameters showed similar high-density lipoprotein cholesterol (HDL-C) (0.77 +/- 0.23 vs. 0.82 +/- 0.22 mmol/l; p = 0.1) differences in triglycerides [1.32 (0.08-3.98) vs. 1.12 (0.37-3.61) mmol/l; p = 0.01], but no difference in low-density lipoprotein cholesterol (LDL-C) (2.75 +/- 1.00 vs. 2.90 +/- 0.94 mmol/l; p = 0.14). In insulin-resistant patients C-reactive protein (CRP) [6.8 (0.3-175.1) vs. 3.9 (0.2-57.9) mg/l: p < 0.001], sialic acid (82 +/- 14 vs. 77 +/- 15 mg/l; p < 0.001) aspartate transaminase [24 (7-171) vs. 21 (7-83) IU/l; p < 0.001] and gamma-glutamyl transferase [27 (8-482) vs. 21 (7-168) IU/l; p = 0.005] levels were increased. In insulin-resistant patients (n = 187), coronary artery disease (CAD) burden correlated (r = 0.55) with age (beta = 1.62; p < 0.001), HDL-C (beta = -53.2; p < 0.001), lipoprotein (a) (beta = 11.4; p = 0.007), smoking (beta = 7.98; p = 0.004), CRP (beta = 6.06; p = 0.03) and QuickI index (beta = -146; p = 0.04). In contrast in insulin-sensitive patients (n = 178) CAD burden (r = 0.46) correlated with LDL-C (beta = 10.0; p = 0.02), CRP (beta = 7.13; p = 0.03), HDL-C (beta = -38.1; p = 0.03), and weakly with age (beta = 0.73; p = 0.07) and smoking (beta = 5.52; p = 0.09). Indian Asians show a dichotomous insulin-resistance phenotype. Atheroma is associated with low HDL-C and inflammation associated in all but LDL-C is a factor in the insulin sensitive in contrast to age and extent of IR in the insulin resistant.


Isolation and characterization of SSR and EST-SSR loci in Chamaecyparis formosensis (Cupressaceae).
Simple sequence repeat (SSR) and expressed sequence tag (EST)-SSR markers were developed as tools for marker-assisted selection of Chamaecyparis formosensis and for the molecular differentiation of cypress species. Based on the SSR-enriched genomic libraries and transcriptome data of C. formosensis, 300 primer pairs were selected for initial confirmation, of which 19 polymorphic SSR and eight polymorphic EST-SSR loci were chosen after testing in 92 individuals. The number of alleles observed for these 27 loci ranged from one to 17. The levels of observed and expected heterozygosity ranged from 0.000 to 1.000 and from 0.000 to 0.903, respectively. Most markers also amplified in C. obtusa var. formosana. The developed SSR and EST-SSR sequences are the first reported markers specific to C. formosensis. These markers will be useful for individual identification of C. formosensis and to distinguish cypress species such as C. obtusa var. formosana.


Incidence and risk factors of neurological deficits of surgical correction for scoliosis: analysis of 1373 cases at one Chinese institution.
A retrospective study. To investigate the incidence of neurologic deficits after scoliosis correction at 1 institution and identify the risk factors for such deficits in scoliosis correction. Neurologic deficit is one of the risks of surgical correction of scoliosis. Reports of the incidence of the neurologic deficits involving a large number of cases at 1 institution are rare. Statistical analysis of the neurologic deficits was performed in 1373 scoliosis cases treated at 1 institution by etiologies in light of the patients' sex, age, sagittal profile, surgical approach, Cobb's angle, and the type of surgery. The total incidence of neurologic deficits was 1.89% and that of serious and mild ones was 0.51% and 1.38%, respectively. The total incidence of neurologic deficits were 1.06% in adolescent idiopathic scoliosis patients and 2.89% in congenital scoliosis (CS) patients, 3.32% in scoliosis patients with hyperkyphosis (>40 degrees ) and 1.38% in those without hyperkyphosis, 3.43% for combined procedures and 1.24% for single posterior procedures, 3.69% in patients with Cobb's angle more than 90 degrees and 1.45% in those with an angle less than 90 degrees , 1.68% with primary surgery and 5.97% with revision surgery, the difference between them was significant (P < 0.05). In adolescent idiopathic scoliosis patients, the incidence were 3.85% for combined procedures and 0.64% for posterior procedure, 0.82% with primary surgery and 8.33% with revision surgery, 4.17% with hyperkyphosis and 0.61% without hyperkyphosis, the difference between them was significant (P < 0.05). The incidence of CS patients with Cobb's angle more or less than 90 degrees were 7.23% and 1.68% and the difference was significant (P < 0.05). In surgical correction of scoliosis, the risk factors for neurologic deficits include CS, scoliosis with hyperkyphosis, scoliosis correction by combined procedures, scoliosis with a Cobb's angle more than 90 degrees , and a revision surgery.


Quality of life and its predictive factors among women with obstetric fistula in Ethiopia: A cross-sectional study.
Living with obstetric fistulas is detrimental to the quality of life of women with fistulas. This study aimed to assess the quality of life and predictive factors among women with obstetric fistula in Ethiopia. A cross-sectional study was employed on consecutively selected 478 women. Linear regressions were used for data analysis. The mean quality of life in physical, psychological, social, and environmental health domains and the overall quality of life were 40.59 ± 1.58, 38.10 ± 1.78, 29.59 ± 1.97, 34.21 ± 1.65, and 44.61 ± 3.99 respectively. Repair outcome without urinary inconsistence (β = 5.2; 95% CI = 0.72, 9.64), self-esteem (β = 1.3; 95% CI = 0.96, 1.57), negative attitude (β = 5.1; 95% CI = 1.86, 8.33), waiting treatment (β = -8.4; 95% CI = -15.54, -1.10), and low intention (β = 4.7; 95% CI = 1.52, 7.93) were predictors of the quality of life in physical domain. Repair outcome without urinary inconsistence (β = 5.9; 95% CI = 1.73, 9.99), self-esteem (β = 1.8; 95% CI = 1.47, 2.11), negative attitude (β = -6.4; 95% CI = -9.60, -3.25), fathers at primary school (β = 12.5; 95% CI = 0.08, 24.82), living only with parents (β = 4.9; 95% CI = 0.99, 8.90), time of care-seeking (β = -0.01, 95% CI = -0.02, -0.002), and duration lived with fistula (β = -5.4; 95% CI = -9.12, -1.68) were predictors of psychological domain. Dead birth (β = -5.2; 95% CI = -9.86, -0.51), self-esteem (β = 1.1; 95% CI = 0.72, 1.43), and living only with parents (β = 5.5; 95% CI = 0.30, 10.69), and living only with husband (β = 7.8; 95% CI = 2.01, 13.55) were predictors of social domain. Living in rural (β = -6; 95% CI = -9.22, -2.79), women at secondary school (β = 14.1; 95% CI = 3.67, 24.48), self-esteem (β = 1.3; 95% CI = 0.99, 1.55), negative attitude (β = -5.1; 95% CI = -7.97, -2.29) were predictors of quality of life in environmental domain. Repair outcome without urinary inconsistence (β = 8.3; 95% CI = 0.62, 16.02), self-esteem (β = 2.1; 95% CI = 1.34, 2.79), and living only with parents (β = 2.9; 95% CI = 1.06, 4.76) were significant predictors of the overall quality of life. The quality of life of women with obstetric fistula was low. Repair outcomes, self-esteem, negative attitudes, rural residence, living with parents, and time of care-seeking were significant predictors of quality of life. Urgent measures should be taken to address these factors to improve the quality of life of women with fistula.


The influence of calcium and pH on the uptake and toxicity of copper in Folsomia candida exposed to simplified soil solutions.
The aim of the present study was to investigate the influence of Ca and pH on the uptake and effects of Cu in Folsomia candida (Collembola). Assuming that soil pore water is the main route of exposure, F. candida were exposed for seven days to Cu in simplified soil solutions at different Ca concentrations and different pH levels. A hormetic-type effect was seen for the effect of Cu on F. candida survival. Toxicity of Cu was slightly decreased and Cu uptake increased at the highest Ca concentrations. Cu toxicity and uptake were not significantly affected by pH. Conditional binding constants for Cu(2+), Ca(2+) and H(+), calculated with a Langmuir isotherm, were used to relate Cu toxicity to the fraction of occupied binding sites (BL). The estimated 50% effect level (fCu-BL50) was 0.14 when all data were combined. To verify the Langmuir parameters, binding constants calculated based on internal Cu concentrations were used to estimate predicted effects and compared with the measured values. A good correlation between predicted and measured survival indicated that the principles of a biotic ligand model may be applicable to explain Cu toxicity to F. candida in simplified soil solutions.


Theoretical investigation of the enzymatic phosphoryl transfer of β-phosphoglucomutase: revisiting both steps of the catalytic cycle.
Enzyme catalyzed phosphate transfer is a part of almost all metabolic processes. Such reactions are of central importance for the energy balance in all organisms and play important roles in cellular control at all levels. Mutases transfer a phosphoryl group while nucleases cleave the phosphodiester linkages between two nucleotides. The subject of our present study is the Lactococcus lactis β-phosphoglucomutase (β-PGM), which effectively catalyzes the interconversion of β-D-glucose-1-phosphate (β-G1P) to β-D-glucose-6-phosphate (β-G6P) and vice versa via stabile intermediate β-D-glucose-1,6-(bis)phosphate (β-G1,6diP) in the presence of Mg(2+). In this paper we revisited the reaction mechanism of the phosphoryl transfer starting from the bisphosphate β-G1,6diP in both directions (toward β-G1P and β-G6P) combining docking techniques and QM/MM theoretical method at the DFT/PBE0 level of theory. In addition we performed NEB (nudged elastic band) and free energy calculations to optimize the path and to identify the transition states and the energies involved in the catalytic cycle. Our calculations reveal that both steps proceed via dissociative pentacoordinated phosphorane, which is not a stabile intermediate but rather a transition state. In addition to the Mg(2+) ion, Ser114 and Lys145 also play important roles in stabilizing the large negative charge on the phosphate through strong coordination with the phosphate oxygens and guiding the phosphate group throughout the catalytic process. The calculated energy barrier of the reaction for the β-G1P to β-G1,6diP step is only slightly higher than for the β-G1,6diP to β-G6P step (16.10 kcal mol(-1) versus 15.10 kcal mol(-1)) and is in excellent agreement with experimental findings (14.65 kcal mol(-1)).


Molecular distribution, sources and potential health risks of fine particulate-bound polycyclic aromatic hydrocarbons during high pollution episodes in a subtropical urban city.
Abundance of fine particulate-bound 16 priority polycyclic aromatic hydrocarbons (PAHs) was investigated to ascertain its sources and potential carcinogenic health risks in Varanasi, India. The city represents a typical urban settlement of South Asia having particulate exposure manyfold higher than standard with reports of pollution induced mortalities and morbidities. Fine particulates (PM2.5) were monitored from October 2019 to May 2020, with 32% of monitoring days accounting ≥100 μgm-3 of PM2.5 concentration, frequently from November to January (99% of monitoring days). The concentration of 16 priority PAHs varied from 24.1 to 44.6 ngm-3 (mean: 33.1 ± 3.2 ngm-3) without much seasonal deviations. Both low (LMW, 56%) and high molecular weight (HMW, 44%) PAHs were abundant, with Fluoranthene (3.9 ± 0.4ngm-3) and Fluorene (3.5 ± 0.3ngm-3) emerged as most dominating PAHs. Concentration of Benzo(a)pyrene (B(a)P, 0.5 ± 0.1ngm-3) was lower than the national standard as it contributed 13% of total PAHs mass. Diagnostic ratios of PAH isomers indicate predominance of pyrogenic sources including emissions from biomass burning, and both from diesel and petrol-driven vehicles. Source apportionment using receptor model revealed similar observation of major PAHs contribution from biomass burning and fuel combustion (54% of source contribution) followed by coal combustion for residential heating and cooking purposes (44%). Potential toxicity of B[a]P equivalence ranged from 0.003 to 1.365 with cumulative toxicity of 2.13ngm-3. Among the PAH species, dibenzo[h]anthracene contributed maximum toxicity followed by B[a]P, together accounting 86% of PAH induced carcinogenicity. Incremental risk of developing cancer through lifetime exposure (ILCR) of PAHs was higher in children (3.3 × 10-4) with 56% contribution from LMW PAHs, primarily through ingestion and dermal contact. Adults in contrast, were more exposed to inhale airborne PAHs with cumulative ILCR of 2.2 × 10-4. However, ILCR to PM2.5 exposure is probably underestimated considering unaccounted metal abundance thus, require source-specific control measures.