The Application of Adaptive Minimum Match k-Nearest Neighbors to Identify At-Risk Students in Health Professions Education.
When learners fail to reach milestones, educators often wonder if any warning signs could have allowed them to intervene sooner. Machine learning can predict which students are at risk for failing a high-stakes certification examination. If predictions can be made well before the examination, educators can meaningfully intervene before students take the examination to reduce their chances of failing. The authors used already-collected, first-year student assessment data from 5 cohorts in a single Master of Physician Assistant Studies program to implement an "adaptive minimum match" version of the k-nearest neighbors algorithm using changing numbers of neighbors to predict each student's future examination scores on the Physician Assistant National Certifying Exam (PANCE). Validation occurred in 2 ways by using leave-one-out cross-validation (LOOCV) and by evaluating predictions in a new cohort. "Adaptive minimum match" version of the k-nearest neighbors algorithm achieved an accuracy of 93% in LOOCV. "Adaptive minimum match" version of the k-nearest neighbors algorithm generates a predicted PANCE score for each student one year before they take the examination. Students are classified into extra support, optional extra support, or no extra support categories. Then, one year remains to provide appropriate support to each category of student. Predictive analytics can identify at-risk students who might need additional support or remediation before high-stakes certification examinations. Educators can use the included methods and code to generate predicted test outcomes for students. The authors recommend that educators use predictive modeling responsibly and transparently, as one of many tools used to support students. More research is needed to test alternative machine learning methods across a variety of educational programs.


Marketing and primary health care: an approach to planning in a Tongan village.
Expensive health facilities have failed to meet most health needs. This perceived failure has been interpreted as failure of scientific medicine with subsequent alienation of health facilities. It is therefore necessary to develop primary health care (PHC) programmes shaped around the life patterns of the population it serves. This paper discusses the similarity between marketing concept and providing PHC, and as it was applied to development planning in a village of the Kingdom of Tonga. The issues, circumstances, and approaches discussed, reflect what is desirable from the villagers' viewpoint. In the absence of epidemiological data, there was greater reliance on the villagers' observations to rank priorities. The process of arriving at the final objectives used the marketing approach to provide direction and guideline for discussions for the problems. Marketing can be of assistance in the search for more acceptable and accessible health service and will help to focus on the communities' perspective of their total needs.


Oxygen gradients in small and big sparged insect-cell bioreactors.
It should be clear from the above that the calculations described here are at best rough estimations yielding order-of-magnitude values. Even though, the following general conclusions can be drawn. The gradients in stagnant layers surrounding the particles which are characteristic for animal-cell bioreactors are relatively small as compared to the gradients which can be expected in the bulk-liquid phases of the three bioreactors considered, in particular to the gradients in the stagnant layer surrounding the air bubbles. It can be concluded that under almost all circumstances gradients are likely to exist and can be very steep in larger vessels and in particular at high cell densities. The effects of gradients, however, are largely unknown; therefore research on the effects of gradients on specific and volumetric productivities and product quality seems to be an interesting area.


Evaluation of the gap in delivery of opioid agonist therapy among individuals with opioid-related health problems: a population-based retrospective cohort study.
Although opioid-related harms have reached new heights across North America, the size of the gap in opioid agonist therapy (OAT) delivery for opioid-related health problems is unknown in most jurisdictions. This study sought to characterize the gap in OAT treatment using a cascade of care framework, and determine factors associated with engagement and retention in treatment. A population-based retrospective cohort study. Ontario, Canada. Individuals who sought medical care for opioid-related health problems or died from an opioid-related cause between 2005 and 2019. Monthly treatment status for buprenorphine/naloxone or methadone OAT between 2013 and 2019 (i.e. 'off OAT', 'retained on OAT < 6 months', 'retained on OAT ≥ 6 months'). Of 122 811 individuals in the cohort, 97 516 (79.4%) received OAT at least once during the study period. There was decreasing 6-month treatment retention over time. Model results indicated that males had higher odds of being on OAT each month [odds ratio (OR) = 1.26, 95% confidence interval (CI) = 1.23-1.28] but lower odds of OAT retention (OR = 0.90, 95% CI = 0.88-0.92), while the reverse was observed for older individuals (monthly: OR = 0.76 per 10-year increase, 95% CI = 0.76-0.77; retention: OR = 1.36 per 10-year increase, 95% CI = 1.34-1.38) and individuals with higher neighbourhood income (e.g. highest income quintile, monthly: OR = 0.79, 95% CI = 0.77-0.82; highest income quintile, retention: OR = 1.15, 95% CI = 1.11-1.20). Individuals residing in rural areas and with a history of mental health diagnoses had poorer outcomes overall, including lower odds of being on OAT each month (rural: OR = 0.75, 95% CI = 0.73-0.78; mental health: OR = 0.89, 95% CI = 0.87-0.92) and OAT retention (rural: OR = 0.79, 95% CI = 0.77-0.82; mental health: OR = 0.81, 95% CI = 0.78-0.83), as well as higher risk of starting/stopping OAT [rural, starting OAT: hazard ratio (HR) = 1.07, 95% CI = 1.05-1.10; mental health, starting OAT: HR = 1.20, 95% CI: 1.18-1.23; rural, stopping OAT: HR = 1.24, 95% CI: = 1.22-1.26; mental health, stopping OAT: HR = 1.11, 95% CI = 1.09-1.13]. Individuals with a history of mental health diagnoses also had a higher risk of death, regardless of OAT status (off OAT death: HR = 1.49, 95% CI = 1.33-1.66; on OAT death: HR = 1.20, 95% CI = 1.09-1.31). Factors influencing engagement and declining retention in treatment with opioid agonist therapy in Ontario's health system include age, sex and neighbourhood income, as well as mental health diagnoses or residing in rural regions.


Separation of mouse spleen haematopoietic cell fractions for transplantation purposes in lethally irradiated mice.
Mouse cells were fractioned by adherence chromatography on glass beads. The fractionation of spleen cells yielded 5 different fractions, whose transplantation effect was tested by the method of Till and McCulloch. The results of these transplantation tests showed that the chosen form of adherence chromatography was not sufficiently successful, since none of the resultant fractions contained solely colony-forming units (CFU) and in none of them were CFU present in an exeptionally high concentration.


Transcriptional regulation of the mucosal immune system mediated by T-bet.
The immune system faces the arduous task of defending the mucosal surfaces from invading pathogens, but must simultaneously repress responses against commensal organisms and other inert antigens that are abundant in the external environment, as inappropriate immune activation might expose the host to increased risk of autoimmunity. The behavior of individual immune cells is governed by the expression of transcription factors that are responsible for switching immune response genes on and off. T-bet (T-box expressed in T cells) has emerged as one of the key transcription factors responsible for controlling the fate of both innate and adaptive immune cells, and its expression in different immune cells found at mucosal surfaces is capable of dictating the critical balance between permitting robust host immunity and limiting susceptibility to autoimmunity and allergy.


Plasma membrane lipids of human diploid fibroblasts from normal individuals and patients with cystic fibrosis.
The lipid composition of isolated plasma membranes of human skin fibroblasts is described for the first time. Plasma membranes from a number of strains of fibroblasts from patients with cystic fibrosis and matched normals were isolated by a recently described procedure and analysed for major phospholipid classes, cholesterol and fatty acids. No differences in the quantities of these compounds were detected between cells of the two different origins. The fetal calf serum used to supplement the growth medium contained relatively more palmitoleate and oleate but less stearate than the membranes. There were also no consistent differences between cystic fibrosis and normal membranes in terms of the fatty acid compositions of their individual phospholipid classes. Consistent with this lack of chemical change in the lipids of membranes of cystic fibrosis cells, the degree of fluorescence polarization of diphenylhexatriene, an index of fluidity, was also unchanged.


Genitourinary paraganglioma: Demographic, pathologic, and clinical characteristics in the surveillance, epidemiology, and end results database (2000-2012).
Extra-adrenal paragangliomas (PGLs) are infrequent, benign, and neuroendocrine tumors arising from chromaffin cells of the autonomic nervous system. Most PGLs are sporadic, but up to 32% are associated with inherited syndromes such as neurofibromatosis type 1, von Hippel-Lindau disease, and familial PGL. Although most PGLs develop above the umbilicus, they have been reported in the genitourinary (GU) tract. Owing to the paucity of literature on the rates of GU PGL, the objective of our study is to describe the demographic, pathologic, and clinical characteristics of GU PGL, and compare them to non-GU sites of PGL using the surveillance, epidemiology, and end results (SEER) database. The SEER 18 database was used to identify all cases of PGL from 2000 to 2012. Demographic, pathologic, and clinical characteristics were described using chi-square and t-test for categorical and continuous variables, respectively. The Kaplan-Meier method was used to compare overall survival (OS) between GU and non-GU PGL. Statistical significance was defined as P<0.05. All analyses were performed using excel and SAS/Stat version 9.4. A total of 299 cases of PGL were retrieved from SEER, and 20 (6.7%) of the total PGL arose from the GU tract. The mean age at diagnosis was higher in non-GU than GU PGL (50.4±17.2 vs. 40.8±15.6, P = 0.026). Furthermore, 75% of GU PGLs developed in the bladder, followed by the kidneys/renal pelvis, and spermatic cord (20%). Non-GU PGL developed most frequently within the endocrine system (43%). PGL, overall, was more common in men than in women, and it was more common in whites than all other races. Although 55.5% of GU PGLs were organ confined, only 22.2% of non-GU PGLs were localized at diagnosis. All cases of PGL were treated with surgery. There were 2 cause-specific deaths in the GU PGL groups between 2000 and 2012. The 5-year OS was 93.3% for GU PGL vs. 65.5% in non-GU PGL (P = 0.062). GU PGL remains rare with low incidence (6.7% of all PGL cases) in the US population between 2000 and 2012. Bladder PGL represents just 5% of all PGL. Moreover, GU PGL had better OS compared to PGL developing outside of the GU tract although the P-value only approached statistical significance. The bladder represents the most common site of involvement, and surgery is the mainstay of treatment for GU PGL. Clearer prognostic factors, including tumor grade and stage, are needed to better elucidate PGL management in the future; thus, pooled studies from various institutions with detailed clinical information are needed to delineate these prognostic factors.


Investigation of the Gender Distribution of National Institutes of Health Grants Across Six Surgical Specialties From 2015 to 2020: Toward Promoting Equity in Academic Surgery.
There is a paucity of literature evaluating research-funding differences between male and female surgeons. Our study aims to evaluate possible disparities in the National Institutes of Health (NIH) grant awards by surgeon gender, type of medical degree (MD/DO), and advanced degrees among six surgery specialties: general surgeons, neurosurgeons, urologists, obstetricians/gynecologists, plastic, and orthopedic surgeons, from 2015 to 2020. A retrospective cohort study was performed investigating the number of NIH grants received by male and female surgeon-scientists overall and within each listed specialty, 2015-2020. As a surrogate for grants submitted, the proportion of active surgeon-scientists per specialty was used. A priori level of significance was defined as P < 0.05. After adjusting for confounders, male surgeons had a higher mean number of NIH grants and higher grant funding than female surgeons (P < 0.001 for both). Type of medical degree (MD/DO) was not significantly associated with NIH funding. An advanced degree was associated with NIH funding among neurosurgeons only (P < 0.05). Differences in the proportion of active surgeon-scientists and proportion of NIH grants received by male and female surgeon-scientists were found only in the fields of orthopedic surgery (5.8% female surgeons and received 20.7% of grants, P = 0.003) and plastic surgery (17.2% female surgeons and received 33.3% of grants, P = 0.01). Male surgeons received most of the total surgical NIH grants. However, funding for female surgeons in orthopedic and plastic surgery outpaces that of their male counterparts when compared to gender proportions in their respective field. Future studies should further investigate the effects of additional applicant demographics on securing NIH grant funding.


The selective detection of cell surface determinants by means of antibodies and acetylated avidin attached to highly fluorescent polymer microspheres.
Procedures are described for the synthesis of 500 A-diameter polymer microspheres containing a novel fluorescent cross-linking agent. These microspheres have very high fluorophore concentration without quenching of the fluorescence and show very low nonspecific interaction with cells. When monoclonal anti-Thy-1.2 is attached to the fluorescent microspheres, specific binding results in 10(4) spheres being attached per thymocyte while non-specific binding is less than 1%. Similar values are obtained for an indirect staining procedure. The high non-specific binding of cationic avidin to negative cell surfaces is shown to be decreased to negligible levels by acetylation of the amine groups of the protein without decreasing its high-affinity binding to biotin. The use of acetyl-avidin (pI = 6.7) directly, or when attached to fluorescent microspheres, resulted in a highly selective detection of biotinyl groups on the erythrocyte or lymphocyte cell surface. Attachment of biotinyl groups to the hinge carbohydrates of antibodies did not affect their specificity. It allowed their detection by means of microspheres-acetyl-avidin conjugates.


Supervision of redeployed surgical trainees during the COVID-19 pandemic: what have we learnt and how can we improve?
When cases of patients presenting with Coronavirus Disease 2019 (COVID-19) overwhelmed existing services in the United Kingdom (UK), surgical trainees were redeployed to assist frontline efforts. This project characterises the effects of redeployment on the supervision of these trainees. The resulting generation of practical recommendations could be implemented for future crises. A qualitative approach was utilised, comprised of seven phenomenological interviews with surgical and intensive care consultants, as well as redeployed surgical trainees. Interview recordings were transcribed and subsequently analysed using Thematic Analysis. The project utilised participants currently in surgical training within the London deanery across a variety of surgical specialties representing several UK National Health Service (NHS) Trusts. Three types of participants were interviewed. Four interviews were conducted with redeployed surgical trainees, across all stages of training, in full time employment who were redeployed for two weeks or more. One interview was conducted with an educational supervisor of multiple redeployed trainees. The third group comprised two consultant intensivists who supervised redeployed trainees within their respective departments. Four themes were developed: 'Responding to an unforeseen crisis', 'Maintaining surgical identity and culture; A fish out of water?', 'Trainee supervision and support' and 'Preparation and sequelae'. Participants described a sense of obligation to the pandemic effort. Many described a significant interruption to training, however communication of this to surgical supervisors was suboptimal with minimal mitigation. Supervisors on the frontline were challenged by the assessment of trainee competence and acceptance into a new community of practice. Both trainees and supervisors described the management of uncertainty, advocating for the use of reflective practice to ensure preparation for the future. This project presents an insight into several potentially long-lasting effects on surgical training. The recommendations generated may be applicable to trainees returning to work from time out of training, increasing the utility of this work.


Improving compliance with Occupational Safety and Health Administration standards.
Health care facilities can be dangerous places. The mission of the Occupational Safety and Health Administration (OSHA) is to improve the safety of the American workplace by developing and implementing standards that prevent occupational injury, illness, and death. Perioperative services are performed in environments where exposure to bloodborne pathogens is a daily occurrence, making implementation and compliance with OSHA standards very important. Employees and employers must remain current with workplace safety requirements, including use of personal protective equipment. This article presents implications of the OSHA standards for employers, educators, and employees.


[Human Caring at Different Stages After Disasters].
Disasters, including natural disasters and man-made ones, occur rather frequently in recent years. Disasters bring destruction to the resources and expectations of local residents and cause varying degrees of physical and mental damage to the victims, some of whom suffered persistent post-traumatic stress disorder (PTSD). Human caring is the essence and core of nursing. Giving consideration to the needs and wishes of patients, the nursing staff respect the personal values of patients, give patients sincere attention and care, satisfy the reasonable needs of patients, protect the dignity of patients, stimulate patients' positive inner drive, alleviate their physical and mental pain, and facilitate the smooth recovery of patients. However, very few studies have been done to look into the human caring at various stages after a disaster hit and there is little relevant guidance available. Herein, we examined the characteristics and needs of victims of disasters in the post-disaster response stage and recovery stage and elaborated on specific and feasible measures for the implementation of human caring after disasters, including prehospital emergency care, which includes on-the-site emergency care and transportation of patients, in-hospital treatment, which includes an environment of human caring and human caring for patients and their families, especially for the special-needs populations, and post-hospital recovery stage, which includes post-hospital continuing and community-based human caring. We intend to provide guidance and reference for the practice of human caring when major disasters occur.


Jonathan Kipnis.
In an interview with Neuron, Jony Kipnis discusses his formative academic years and subsequent discoveries in meningeal lymphatics. He is enthusiastic about the prospect of therapeutic developments in neuroimmunology arising from focusing on the brain's borders.


Dopamine clearance in critically ill patients.
To examine the validity of the low-dose "renal" dopamine regimen in critically ill patients by investigating the steady-state clearance of dopamine. A prospective clinical study. Teaching hospital intensive care unit. 48 haemodynamically stable patients receiving a dopamine infusion. Sampling of arterial blood and dopamine infusates. Plasma and infusate dopamine levels were measured by liquid chromatography with electrochemical detection. Steady-state clearance was determined by dividing the actual infusion rate by the steady-state plasma concentration. Dopamine clearance for the whole group was 46.4 +/- 35.9 ml/kg per min (mean +/- SD), which is significantly lower than 70 +/- 15.2 ml/kg per min reported for elective surgical patients (p = 0.01). Twelve patients with renal dysfunction had significantly lower dopamine clearances (36 +/- 16.6 ml/kg per min) than the remaining 36 patients (61 +/- 38.5 ml/kg per min, p = 0.022). There was a very poor correlation between plasma dopamine level and infusion rate for the group as a whole (r = 0.47), and this worsened (r = 0.31) when only those patients on a "renal" dose of 2-5 microg/kg per min were considered (n = 30). Plasma dopamine clearance is lower in critically ill patients and there is a large interindividual variation. It is therefore impossible to predict the plasma level from the infusion rate. Consequently, the concept of a selective renovascular low-dose dopamine infusion is invalid in critically ill patients.


Enhancing biomass and ethanol production by increasing NADPH production in Synechocystis sp. PCC 6803.
This study demonstrates that increased NADPH production can improve biomass and ethanol production in cyanobacteria. We over-expressed the endogenous zwf gene, which encodes glucose-6-phosphate dehydrogenase of pentose phosphate pathway, in the model cyanobacterium Synechocystis sp. PCC 6803. zwf over-expression resulted in increased NADPH production, and promoted biomass production compared to the wild type in both autotrophic and mixotrophic conditions. Ethanol production pathway including NADPH-dependent alcohol dehydrogenase was also integrated with and without zwf over-expression. Excessive NADPH production by zwf over-expression could improve both biomass and ethanol production in the autotrophic conditions.


Investigation of novel rapidly synergistic cloud point extraction pattern for bismuth in water and geological samples coupling with flame atomic absorption spectrometry determination.
Rapidly synergistic cloud point extraction (RS-CPE) greatly simplified and accelerated the procedure of traditional cloud point extraction (CPE). In order to expand the application of RS-CPE, this work was carried out after the establishment of the improved extraction technique. The new established extraction method was firstly applied for bismuth extraction and determination coupled with flame atomic absorption spectrometry (FAAS) in this work. The improved RS-CPE was accomplished in the room temperature in 1 min. Non-ionic surfactant Triton X-100 (TX-100) was used as extractant. Octanol worked as cloud point revulsant and synergic reagent. TX-100 has a relatively high cloud point temperature (CPT), which limited its application in CPE. In this work, TX-100 accomplished the RS-CPE procedure in room temperature successfully. The factors influencing RS-CPE, such as concentrations of reagents, pH, conditions of phase separation, effect of environmental temperatures, salt effect and instrumental conditions, were studied systematically. Under the optimal conditions, the limit of detection (LOD) for bismuth was 4.0 μg L(-1), with sensitivity enhancement factor (EF) of 43. The proposed method greatly improved the sensitivity of FAAS for the determination of bismuth and was applied to the determination of trace bismuth in real and certified samples with satisfactory analytical results. The proposed method was rapid, simple, and sensitive.


Biodegradation and decolourization of textile dye wastewater using Ganoderma lucidum.
In this work, treatment of textile dye wastewater was carried in a batch reactor using Ganoderma lucidum. The characteristics of textile dye wastewater were studied. The effect of process parameters like pH, temperature, agitation speed and dye wastewater concentration on dye decolourization and degradation were studied. These parameters were optimized using response surface methodology (RSM). From the results, the optimized conditions were: pH 6.6, temperature 26.5 °C, agitation speed 200 rpm and dye wastewater concentration 1:2. At these optimized conditions, the maximum decolourization and COD reduction were found to be 81.4 and 90.3 %. Kinetic studies were carried out using different models like first-order, diffusional and Singh model. From the results, it was found that the degradation follows the first-order reaction model.


Evaluation of the physico-chemical properties of hydrocarbons-exposed bacterial biomass.
In the efforts for the removal of hazardous materials from the environment biological processes are a valuable tool. Although much attention has been paid to the changes in bacteria at the omics level, another, physical-chemical perspective on the issue is essential, as little is known of microbial response to continuous exposition on harmful substances. This study provides in-depth characterization of the physical-chemical parameters of bacterial biomass after hydrocarbons exposure. To provide comparability of the harmful effects of chlorotoluenes and xylenes non-exposed and 12-months hydrocarbons exposed cells were analyzed, using the advanced spectrometric methods, inverse gas chromatography and low-temperature N2 sorption to evaluate acid-base as well as dispersive properties of the studied biomass. Presented results indicate P. fluorescens B01 cells strategy aimed at protecting the cell, thus lowering its' biodegradation efficiency as a result of metabolic stress. The outcome of the study was that prolonged exposure to pollutants might reduce the bioavailability of hydrocarbons to bacteria cells, and consequently decrease the effectiveness of decontamination of polluted sites by indigenous microorganisms.


LncRNA polymorphisms and breast cancer risk.
Breast cancer (BC) is the most prevalent cancer in females and the second reason of cancer-related mortality in females in the world. It is thought to be a complex interaction of variables like personal lifestyle, climate, genetics, and reproductive factors. Many polymorphisms have been linked to cancer in genome-wide association experiments, and they are linked to long non-coding RNAs (lncRNAs). LncRNAs, which have > 200 nucleotides in their transcripts, affect many biological processes, including differentiation, migration, apoptosis, cell cycle, and cell proliferation. Different lncRNAs with tumor suppressor and oncogenic roles have been shown to have elevated expression levels in the development of BC. Single-nucleotide polymorphisms (SNPs) in lncRNAs can affect the expression level, structure, and function of lncRNAs. LncRNA polymorphisms are predictive of cancer incidence, making them useful for early detection and customized therapy control. SNPs may affect genetic susceptibility to BC. This study was set to see whether there was a link between lncRNA polymorphisms and the risk of BC. Accordingly, the individual and combined genotypes of lncRNA-related variants could predict BC and clinical and care outcomes. However, further large-scale trials of diverse ethnic groups and comprehensive health records should be performed to validate the results. Furthermore, adequate functional assessments should be carried out to shed light on the etiology of BC. DATA AVAILABILITY: Not applicable.


Colorimetric As (V) detection based on S-layer functionalized gold nanoparticles.
Herein, we present simple and rapid colorimetric and UV/VIS spectroscopic methods for detecting anionic arsenic (V) complexes in aqueous media. The methods exploit the aggregation of S-layer-functionalized spherical gold nanoparticles of sizes between 20 and 50 nm in the presence of arsenic species. The gold nanoparticles were functionalized with oligomers of the S-layer protein of Lysinibacillus sphaericus JG-A12. The aggregation of the nanoparticles results in a color change from burgundy-red for widely dispersed nanoparticles to blue for aggregated nanoparticles. A detailed signal analysis was achieved by measuring the shift of the particle plasmon resonance signal with UV/VIS spectroscopy. To further improve signal sensitivity, the influence of larger nanoparticles was tested. In the case of 50 nm gold nanoparticles, a concentration of the anionic arsenic (V) complex lower than 24 ppb was detectable.


CSF neopterin and quinolinic acid are biomarkers of neuroinflammation and neurotoxicity in FIRES and other infection-triggered encephalopathy syndromes.
Infection-triggered encephalopathy syndromes (ITES) are potentially devastating neuroinflammatory conditions. Although some ITES syndromes have recognisable MRI neuroimaging phenotypes, there are otherwise few biomarkers of disease. Early detection to enable immune modulatory treatments could improve outcomes. We measured CSF neopterin, quinolinic acid, kynurenine and kynurenine/tryptophan ratio using a liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) system. The CSF of 18 children with ITES were compared with acute encephalitis (n = 20), and three control groups, namely epilepsy (n = 20), status epilepticus (n = 18) and neurogenetic controls (n = 20). The main ITES phenotypes in 18 patients were acute encephalopathy with biphasic seizures and late restricted diffusion (AESD, n = 4), febrile infection-related epilepsy syndrome (FIRES n = 4) and other ITES phenotypes. Influenza A was the most common infectious trigger (n = 5), and 50% of patients had a preceding notable neurodevelopmental or family history. CSF neopterin, quinolinic acid and kynurenine were elevated in ITES group compared to the three control groups (all p < 0.0002). The ROC (area under curve) for CSF neopterin (99.3%, CI 98.1-100) was significantly better than CSF pleocytosis (87.3% CI 76.4-98.2) (p = 0.028). Elevated CSF neopterin could discriminate ITES from other causes of seizures, status epilepticus and febrile status epilepticus (all p < 0.0002). The elevated CSF metabolites normalised during longitudinal testing in two patients with FIRES. CSF neopterin and quinolinic acid are neuroinflammatory and excitotoxic metabolites. This CSF metabolomic inflammatory panel can discriminate ITES from other causes of new onset seizures or status epilepticus, and rapid results (4 h) may facilitate early immune modulatory therapy.


Outcome of trimodality protocol for invasive bladder cancer patients at Karachi, Pakistan.
To evaluate the outcomes of patients with muscle invasive bladder cancer managed by trimodality protocol with assessment of factors that may predict treatment response, risk of recurrences and survival of such patients in our population. A prospective, single arm study was conducted between July 2006 and December 2009 at the Sindh Institute of Urology and Transplantion, Karachi. One hundred and sixteen patients with muscle invasive bladder cancer T2-T3N0M0 were treated with concurrent chemoradiation (total dose 6500 cGy) after maximal transurethral resection. Complete response was defined as no tumour seen on check cystoscopy and biopsy. The disease control and overall survival were determined by Kaplan and Meier method and statistical inferences with the log-rank test. Cox regression analysis was used to find different prognostic factors. At the median follow up of 36 months (14-43), out of total 116, 62(51.6%) surviving patients the bladder was functioning well, while 18(15%) had local recurrence; in 9 patients superficial tumour recurred and required further transurethral resection and intravesical drug therapy and nine patients who had muscle invasive recurrence; underwent radical cystectomies. Concurrent chemoradiation was well tolerated. The overall survival at 3 years was 54%. Initial complete response, primary tumour stage and absence of hydronephrosis were the most important prognostic factors for survival (all p=<0.0001). Trimodality treatment was found to be an effective therapy in patients with invasive bladder cancer and complete TURBT, tumour stage and no hydronephrosis at time of presentation were found important prognostic factors for treatment response, disease free and over all survival rates.


Modulation of multidrug resistance by taxuspine C and other taxoids from Japanese yew.
Taxuspine C (1), a new taxoid from the Japanese yew Taxus cuspidata, increasing the cellular accumulation of vincristine (VCR) in multidrug-resistant tumor cells as potent as verapamil enhanced the chemotherapeutic effect of VCR in P388/VCR-bearing mice. When taxuspine C (1) was given i.p. daily at 200 mg/kg with 0.2 mg/kg VCR for 5 days, a treated/control (T/C) value of 138% was obtained. The other new taxoids, taxezopidines G (8) and H (9), from the yew also increased the VCR accumulation as potent as verapamil. These results suggest that some taxoids may be useful for overcoming multidrug resistance in tumor cells.


Hymenolepis muris-sylvaticae and H. microstoma: immunological cross-reaction in mice.
A primary infection with Hymenolepis microstoma strongly protects against cross-infection with H. muris-sylvaticae and also against secondary infection with H. microstoma in NMRI mice, resulting in an accelerated loss of worms and a weight reduction of the remaining worms. A primary infection with H. muris-sylvaticae causes an accelerated rejection of secondary infection with H. muris-sylvaticae but it has no effect on cross-infection with H. microstoma, neither with regard to worm recovery nor with regard to worm biomass. Determinations by enzyme-linked immunosorbent assay of antibody concentrations in the mouse sera revealed that: (1) the antibody response evoked by H. microstoma infection is much greater than by H. muris-sylvaticae infection; (2) a cross-infection with H. muris-sylvaticae boosts the antibody response evoked by H. microstoma infection; (3) H. microstoma antigen can be used to measure antibody concentration against both H. microstoma and H. muris-sylvaticae; and (4) although H. muris-sylvaticae is rejected faster in a cross-infection (i.e., after a primary H. microstoma infection) than in a secondary infection (i.e., after a primary H. muris-sylvaticae infection), antibodies evoked by the primary H. microstoma infection show little cross-reaction with H. muris-sylvaticae antigen. This suggests that it is doubtful whether serum antibodies are the direct effectors in worm rejection.


Esophageal plexiform fibromyxoma: A case report with molecular analysis for MALAT1-GLI1 fusion.
Plexiform fibromyxoma (PFM) is a rare gastrointestinal tract tumor that develops in the stomach in most cases. Here, we report an extremely rare case of esophageal PFM. A female in her mid-30 s presented with difficulty in swallowing and breathing. Endoscopic examination revealed a submucosal tumor measuring approximately 45 × 50 mm in the upper thoracic esophagus. The biopsied specimen did not show definite histological evidence of gastrointestinal stromal tumors (GISTs). Since imatinib administration based on a clinical diagnosis of GIST did not show a therapeutic effect for tumor reduction, tumor resection was performed. The resected tumor exhibited proliferation of spindle tumor cells with abundant myxoid and vascular stroma separated by a muscular layer, indicating a plexiform arrangement. Immunohistochemical analysis demonstrated that the tumor cells diffusely expressed vimentin and alpha-smooth muscle actin, but not desmin, c-kit, DOG1, and CD34. MALAT1-GLI1 fusion was detected in formalin-fixed paraffin-embedded tissue using RT-PCR and Sanger sequencing. The results suggested that a fibromyxoid tumor can develop in the esophagus, showing an identical histology and MALAT1-GLI1 fusion to gastric PFM.


Limitations of secondary data sets for road traffic injury epidemiology: a study from Karachi, Pakistan.
To evaluate the completeness of descriptive information and accuracy of ambulance data when compared with two other secondary sources of data, namely, medical examiners' reports and medical records, in the city of Karachi, Pakistan. A retrospective chart review was carried out of all road traffic injury (RTI) victims transported by an ambulance service in Karachi from January 1 to December 31, 2003. Information on the name, age, and gender of the victim, date, time, and place of injury, and type of injury and its outcome was obtained, and then compared with medical examiners' data and inpatient medical records. The variables missing in the ambulance service database were identified. The ambulance service transported 1,245 RTI patients during 2003, 81% of whom were taken to one of three trauma centers. Twenty percent died before reaching hospital. There were missing variables in 340 (27%) cases, the majority of which were the unique identifiers such as name and age (67%). Data on other variables were reported in 95% or more cases. None of the medical examiners' reports or inpatient medical records contained a description of event of injury. The agreement rate for the variables available in the three data sets ranged from 61% to 100%. Secondary data on injuries, such as ambulance reports, medical examiners' reports, and medical records, have significant limitations. In Karachi, except with regard to unique identifiers, ambulance data seem to provide information just as accurate as that in medical examiners' and inpatient medical records.


Whole-body protein metabolism assessed by leucine and glutamine kinetics in adult patients with active celiac disease.
To assess the effect of increased renewal of intestinal epithelial cells on leucine and glutamine (Gln) turnover, 4-hour intravenous infusions of L-[1-(13)C]leucine and L-[2-(15)N]Gln were administered to five adult patients with active celiac disease in the postabsorptive state. There was a 35% increase in leucine flux (micromoles per kilogram per hour) in patients (117 +/- 17) compared with healthy controls (96 +/- 11, P < .03). Gln flux was increased by 13% in patients (377 +/- 35) versus controls (335 +/- 16, P < .04). These results suggest that active celiac disease, characterized by villous atrophy and crypt cell hyperplasia, is associated with a dramatic increase in whole-body protein breakdown as assessed by 13C-leucine, which may contribute per se to the protein malnutrition status of the patients. The increase in Gln utilization as assessed by L-[2-(15)N]Gln was moderate, but may have been offset due to the villose atrophy and ensuing reduced intestinal epithelial cell mass. The results are consistent with the concept that increased renewal of intestinal epithelial cells represents a sizable fraction of whole-body protein turnover and that Gln is an important fuel for epithelial intestinal cells in vivo.


Search for familial clustering of multiple myeloma with any cancer.
Multiple myeloma (MM) is a disease of immunoglobulin-producing plasma cells, which reside mainly in the bone marrow. Family members of MM patients are at a risk of MM, but whether other malignancies are in excess in family members is not established and is the aim of this study. MM patients (24 137) were identified from the Swedish Cancer Registry from years 1958 to 2012. Relative risks (RRs) were calculated for MM defined by any cancer diagnosed in first-degree relatives and compared with individuals whose relatives had no cancer. MM was reliably associated with relative's colorectal, breast and prostate cancers, non-thyroid endocrine tumors, leukemia and cancer of unknown primary; in addition, MM was associated with subsites of bone and connective tissue tumors and of non-Hodgkin lymphoma, including lymphoplasmacytic lymphoma/Waldenström macroglobulinema (RR 3.47). MM showed a strong association (RR 1.91) in colorectal cancer families, possibly as part of an unidentified syndrome. All the associations of MM with discordant cancers are novel suggesting that MM shares genetic susceptibility with many cancers. The associations of MM bone and connective tissue tumors were supported by at least two independent results. Whether the results signal bone-related biology shared by MM and these tumors deserves further study.


Attitudes to viability of preterm infants and their effect on figures for perinatal mortality.
To examine how local attitudes to management of extreme preterm labour can influence data on perinatal mortality. One year prospective study in a geographically defined population. The 17 perinatal units of Trent region. All preterm infants of less than or equal to 32 weeks' gestation in the Trent region. Infants who had been considered viable at birth were referred for intensive care; those who had been considered non-viable received terminal care. Whether each infant was born alive, dead, or alive but considered non-viable. Large differences were observed among units in the rates of delivery of infants of less than or equal to 27 weeks' gestation (rates varied from 7.2 to 0 per 1000 births). These differences were not present in the data relating to infants of between 28 and 32 weeks' gestation. The variation seemed to result from different approaches to the management of extreme preterm labour--that is, whether management took place in a labour ward or a gynaecology ward. Place of delivery of premature babies (less than or equal to 27 weeks' gestation) may influence classification and hence figures for perinatal mortality. In addition, the fact that the onus of judgment regarding viability and classification is often placed on relatively junior staff might also affect the figures for perinatal mortality. The introduction of a standard recording system for all infants greater than 500 g would be advantageous.


Numerical analysis of water and solute transport in variably-saturated fractured clayey till.
This study numerically investigates the influence of initial water content and rain intensities on the preferential migration of two fluorescent tracers, Acid Yellow 7 (AY7) and Sulforhodamine B (SB), through variably-saturated fractured clayey till. The simulations are based on the numerical model HydroGeoSphere, which solves 3D variably-saturated flow and solute transport in discretely-fractured porous media. Using detailed knowledge of the matrix, fracture, and biopore properties, the numerical model is calibrated and validated against experimental high-resolution tracer images/data collected under dry and wet soil conditions and for three different rain events. The model could reproduce reasonably well the observed preferential migration of AY7 and SB through the fractured till, although it did not capture the exact depth of migration and the negligible impact of the dead-end biopores in a near-saturated matrix. A sensitivity analysis suggests fast flow mechanisms and dynamic surface coating in the biopores, and the presence of a plough pan in the till.


Early treadmill running delays rotator cuff healing via Neuropeptide Y mediated inactivation of the Wnt/β-catenin signaling.
Defining the optimal rehabilitation programs for rotator cuff healing remains a challenge. Early treadmill running may have negative effects on tendon-bone interface (TBI) healing with increased expression of Neuropeptide Y (NPY). However, the underlying mechanism is still unknown. The mice were randomly assigned to four groups: control group, treadmill group, treadmill + BIBO3304 group and BIBO3304 group alone. Specifically, the control group was allowed free cage activity without any treatment after surgery. The treadmill group received early treadmill running initiated from postoperative day 2. The treadmill + BIBO3304 group received treadmill running combined with intra-articular injection of BIBO3304 postoperatively. The BIBO3304 group only received type 1 NPY receptor (Y1 receptor, Y1R) antagonist BIBO3304 postoperatively. Healing outcomes of the rotator cuff were evaluated by histological analysis, synchrotron radiation micro-computed tomography (SR-μCT) scanning, and biomechanical testing at 4 and 8 weeks after surgery. The expression of NPY and its Y1 receptor during the treadmill running were tested by immunofluorescence. In addition, the related signaling pathway of Neuropeptide Y among all groups was detected by immunohistochemistry and western-blot. Immunofluorescence results show that early treadmill training could lead to a significant increase in the expression of NPY at the healing site, and Y1R was widely expressed in both normal or injured rotator cuff without statistical difference. At the same time, early treadmill running delayed the healing of rotator cuff, as indicated with unsatisfactory outcomes, including a significantly lower histological score, decreased bone formation and inferior biomechanical properties at postoperative week 4 and 8. Moreover, the use of BIBO3304 could partly alleviate the negative effects of early treadmill running on the healing of rotator cuff and promote the natural healing process of rotator cuff, as evidenced by significant differences observed between the treadmill and treadmill + BIBO3304 groups, as well as observed between the control and BIBO3304 groups. On the other hand, the expressions of Wnt3a and β-catenin in the treadmill group were significantly lower compared with the other groups, while the expression in the BIBO3304 group was the highest, as evaluated by immunohistochemistry and western-blot. Early treadmill running increased the expression of NPY at the RC healing site, which might burden the expression of Wnt3a/β-catenin and delay the healing process, inhibition of Y1 receptor with BIBO3304 could promote bone-tendon healing through the Wnt/β-catenin signaling.The translational potential of this article: This is the first study to evaluate the specific role of the NPY-Y1R axis and its underlying mechanism by which early treadmill running delays bone-tendon healing. Further, our study may provide references of precise and individualized exercise-based rehabilitation strategies for TBI healing in clinic. This is the first study to evaluate the specific role of the NPY-Y1R axis and its underlying mechanism by which early treadmill running delays bone-tendon healing. Further, our study may provide references of precise and individualized exercise-based rehabilitation strategies for TBI healing in clinic.


Research diversity from the perspective of the Science of Unitary Human Beings.
This column presents an overview of Rogerian nursing science-based research. In Rogers' own words, she fostered the development of nursing science with a diversity of methods. She encouraged both qualitative and quantitative approaches and the development of new research methods and tools. Her only caveat was that theoretical conceptualization and research method must match.


Production of biosurfactant from a new and promising strain of Pseudomonas aeruginosa PA1.
The Pseudomonas aeruginosa PA1 strain, isolated from the water of oil production in Sergipe, Northeast Brazil, was evaluated as a potential rhamnolipid type of biosurfactant producer. The production of biosurfactants was investigated using different carbon sources (n-hexadecane, paraffin oil, glycerol, and babassu oil) and inoculum concentrations (0.0016-0.008 g/L). The best results were obtained with glycerol as the substrate and an initial cell concentration of 0.004 g/L. A C:N ratio of 22.8 led to the greatest production of rhamnolipids (1700 mg/L) and efficiency (1.18 g of rhamnolipid/g of dry wt).


Efficient itaconic acid production by Aspergillus terreus: Overcoming the strong inhibitory effect of manganese.
Itaconic acid (IA), a building block platform chemical, is produced industrially by Aspergillus terreus utilizing glucose. Lignocellulosic biomass can serve as a low cost source of sugars for IA production. However, the fungus could not produce IA from dilute acid pretreated and enzymatically saccharified wheat straw hydrolyzate even at 100-fold dilution. Furfural, hydroxymethyl furfural and acetic acid were inhibitory, as is typical, but Mn2+ was particularly problematic for IA production. It was present in the hydrolyzate at a level that was 230 times over the inhibitory limit (50 ppb). Recently, it was found that PO43- limitation decreased the inhibitory effect of Mn2+ on IA production. In the present study, a novel medium was developed for production of IA by varying PO43- , Fe3+ and Cu2+ concentrations using response surface methodology, which alleviated the strong inhibitory effect of Mn2+ . The new medium contained 0.08 g KH2 PO4 , 3 g NH4 NO3 , 1 g MgSO4 ·7H2 O, 5 g CaCl2 ·2 H2 O, 0.83 mg FeCl3 ·6H2 O, 8 mg ZnSO4 ·7H2 O, and 45 mg CuSO4 ·5H2 O per liter. The fungus was able to produce IA very well in the presence of Mn2+ up to 100 ppm in the medium. This medium will be extremely useful for IA production in the presence of Mn2+ . This is the first report on the development of Mn2+ tolerant medium for IA production by A. terreus.


Nisin production utilizing skimmed milk aiming to reduce process cost.
Nisin is a natural additive for conservation of food, pharmaceutical, and dental products and can be used as a therapeutic agent. Nisin inhibits the outgrowth of spores, the growth of a variety of gram-positive and gram-negative bacteria. This study was performed to optimize large-scale nisin production in skimmed milk and subproducts aiming at low-costs process and stimulating its utilization. Lactococcus lactis American Type Culture Collection (ATCC) 11454 was developed in a rotary shaker (30 degrees C/36 h/100 rpm) in diluted skimmed milk and nisin activity, growth parameters, and media components were also studied. Nisin activity in growth media was expressed in arbitrary units (AU/mL) and converted to standard nisin concentration (Nisaplin, 25 mg of pure nisin is 1.0x10(6) AU/mL). Nisin activity in skimmed milk 2.27 g(total solids) was up to threefold higher than transfers in skimmed milk 4.54 g(total solids) and was up to 85-fold higher than transfers in skimmed milk 1.14 g(total solids). L. lactis was assayed in a New Brunswick fermentor with 1.5 L of diluted skimmed milk (2.27 g(total solids)) and airflow of 1.5 mL/min (30 degrees C/36/200 rpm), without pH control. In this condition nisin activity was observed after 4 h (45.07 AU/mL) and in the end of 36 h process (3312.07 AU/mL). This work shows the utilization of a low-cost growth medium (diluted skimmed milk) to nisin production with wide applications. Furthermore, milk subproducts (milk whey) can be exploited in nisin production, because in Brazil 50% of milk whey is disposed with no treatment in rivers and because of high organic matter concentrations it is considered an important pollutant. In this particular case an optimized production of an antimicrobial would be lined up with industrial disposal recycling.


Current communication practices for biomarker testing in non-small cell lung cancer: Exploring patient and clinician perspectives.
We qualitatively explored patient and clinician experiences with biomarker testing in one academic health system to identify current communication practices and unmet testing information needs. We conducted 1:1 in-depth interviews with 15 clinicians (i.e., nurses, oncologists, pathologists) and 12 patients diagnosed with non-small cell lung cancer between January and May 2022. Participants described experiences with biomarker testing as well as associated communication practices and needs. Interviews were audio-recorded and transcribed. Analysis was informed by the Framework Method. Patients described challenges retaining information early in their patient journey. While patients were generally aware of biomarkers and their effect on treatment options, they expressed limited knowledge of expected time delays between testing and receiving results. Additionally, many did not know their testing results. Clinicians and patients both noted no standard education material on biomarker testing is currently available. They suggested such materials could support patient knowledge and decision-making. Communication between patients and clinicians about biomarker testing is largely delivered through verbal counseling at a time when patients may be cognitively compromised. All participants supported the idea of delivering standard, tangible education materials on biomarker testing to patients. Education materials may enhance counseling efforts and patient knowledge.


Phytoplankton communities in the littoral zone of lakes: Observations on structure and dynamics in oligotrophic and eutrophic systems.
The structure and seasonal dynamics of phytoplankton communities in the littoral zone were compared between oligotrophic and eutrophic lakes in the southeastern United States. Differences in diversity and species composition between lakes could be ascribed to long-term variation in nutrients corresponding to trophic status. However, significant within-lake variation could not be accounted for by microstratification of nutrients or other abiotic variables. Local biotic factors, perhaps dominated by the spawning activities of centrarchid fishes, resuspend periphyton and generate tychoplankton which becomes a persistent and integral part of the phytoplankton community in eutrophic systems. The patchy distribution of these biotic factors and resultant tychoplankton may lead to the observed variation. Grazing by herbivorous zooplankton was considered to be the major factor affecting the relative abundance of phytoplankton in the littoral zone, completely overriding the effects of nutrient concentration and biotic interactions between phytoplankton species during spring and summer.The relative importance of tychoplankton and grazing as regulatory factors operates independently of the trophic status or geographical location of a lake, making comparisons of different studies difficult and perhaps meaningless if traditional analyses based only on nutrients and interactions between species of phytoplankton are used. Limnetic as well as littoral components must be considered in future studies of phytoplankton communities in the littoral zone.


ORIENTATE: automated machine learning classifiers for oral health prediction and research.
The application of data-driven methods is expected to play an increasingly important role in healthcare. However, a lack of personnel with the necessary skills to develop these models and interpret its output is preventing a wider adoption of these methods. To address this gap, we introduce and describe ORIENTATE, a software for automated application of machine learning classification algorithms by clinical practitioners lacking specific technical skills. ORIENTATE allows the selection of features and the target variable, then automatically generates a number of classification models and cross-validates them, finding the best model and evaluating it. It also implements a custom feature selection algorithm for systematic searches of the best combination of predictors for a given target variable. Finally, it outputs a comprehensive report with graphs that facilitates the explanation of the classification model results, using global interpretation methods, and an interface for the prediction of new input samples. Feature relevance and interaction plots provided by ORIENTATE allow to use it for statistical inference, which can replace and/or complement classical statistical studies. Its application to a dataset with healthy and special health care needs (SHCN) children, treated under deep sedation, was discussed as case study. On the example dataset, despite its small size, the feature selection algorithm found a set of features able to predict the need for a second sedation with a f1 score of 0.83 and a ROC (AUC) of 0.92. Eight predictive factors for both populations were found and ordered by the relevance assigned to them by the model. A discussion of how to derive inferences from the relevance and interaction plots and a comparison with a classical study is also provided. ORIENTATE automatically finds suitable features and generates accurate classifiers which can be used in preventive tasks. In addition, researchers without specific skills on data methods can use it for the application of machine learning classification and as a complement to classical studies for inferential analysis of features. In the case study, a high prediction accuracy for a second sedation in SHCN children was achieved. The analysis of the relevance of the features showed that the number of teeth with pulpar treatments at the first sedation is a predictive factor for a second sedation.


[Chemical Constituents from Drypetes hainanensis Stems and Leaves].
To study the chemical constituents from the stems and leaves in Drypetes hainanensis. The constituents were isolated and purified by various chromatography, and the structures were identified by extensive spectral analysis. Eleven compounds were isolated and identified as syringaresinol-4-O-glycoside (1), koaburaside (2), abietin (3) syringin (4), kelampayoside A (5), 7,7'-bis-(4-hydroxy-3,5-dimethoxyphenyl)-8,8'-dihydroxymethyl-tetrahydrofuran-4-O-β-glucopyranoside (6), amentoflavone (7), 3,4,5-trimethoxyphenyl-β-D-glucopyranoside (8),1,4-di-O-methyl-myo-inositol (9), glycerol (10) and succinic acid (11). All the compounds are isolated from this plant for the first time.


Prophylaxis of intracranial hemorrhage due to vitamin K deficiency in infants.
To determine the most effective way of preventing intracranial hemorrhage due to vitamin K deficiency in infants, we first performed a comparative study using Normotest on the effects of several regimens for the oral administration of vitamin K2. Based on the results, we gave vitamin K2 orally, 2 mg, at birth, and then, 4 mg, at 1 week of age (on discharge from the newborn nursery) to all infants except premature and low-birth-weight infants born in Nagasaki Prefecture, Japan. Since then, as the number of infants with vitamin K2 prophylaxis increased, patients with intracranial hemorrhage due to vitamin K deficiency decreased in number, and no patient was found in 1984. The incidence of this disease in infants with vitamin K prophylaxis was 1/68,500, which was one-twentieth of that (1/3,500 live births) before the period when most neonates received vitamin K prophylaxis. From the results, we concluded that the oral administration of vitamin K2 at birth and 1 week of age prevents this disease.


Characterization of a colchicine receptor protein in rat epididymal adipose tissue.
Colchincine was found to be taken up by adipose tissue and therein to bind to a soluble macromolecule not sedimented by centrifugation for 2 h at 100 000 x g. A similar binding occurred when soluble extracts of adipose tissue were incubated with colchicine. The binding reaction reaction is temperature dependent and shows a pH optimum between 6.8 and 7.0. Double reciprocal plots of colchicine concentration versus amounts of colchicine bound to protein in the steady state disclosed an apparent Km of 0.250 to 1.5 muM. The colchicine binding activity of soluble tissue extracts decreased when the extracts were incubated at 37 degree C. Addition of guanosine triphosphate and Mg-2+ retarded the loss of colchicine binding activity. The molecular weight of the colchicine complex was estimated to be 115 000 and its sedimentation coefficient 5.8 S. All of these characteristics are remarkably similar to those of the protein tubulin which has been isolated from other tissues. Since it is now well known that tubulin is a protein subunit of cytoplasmic microtubules, it is suggested that the previously reported metabolic effects of colchicine on adipose tissue result from the dissolution of microtubules by colchicine.


Determining optimal clothing ensembles based on weather forecasts, with particular reference to outdoor winter military activities.
Military and civil defense personnel are often involved in complex activities in a variety of outdoor environments. The choice of appropriate clothing ensembles represents an important strategy to establish the success of a military mission. The main aim of this study was to compare the known clothing insulation of the garment ensembles worn by soldiers during two winter outdoor field trials (hike and guard duty) with the estimated optimal clothing thermal insulations recommended to maintain thermoneutrality, assessed by using two different biometeorological procedures. The overall aim was to assess the applicability of such biometeorological procedures to weather forecast systems, thereby developing a comprehensive biometeorological tool for military operational forecast purposes. Military trials were carried out during winter 2006 in Pokljuka (Slovenia) by Slovene Armed Forces personnel. Gastrointestinal temperature, heart rate and environmental parameters were measured with portable data acquisition systems. The thermal characteristics of the clothing ensembles worn by the soldiers, namely thermal resistance, were determined with a sweating thermal manikin. Results showed that the clothing ensemble worn by the military was appropriate during guard duty but generally inappropriate during the hike. A general under-estimation of the biometeorological forecast model in predicting the optimal clothing insulation value was observed and an additional post-processing calibration might further improve forecast accuracy. This study represents the first step in the development of a comprehensive personalized biometeorological forecast system aimed at improving recommendations regarding the optimal thermal insulation of military garment ensembles for winter activities.


The utilisation of radiology for the teaching of anatomy in Canadian medical schools.
To determine the utilisation of diagnostic imaging (radiology) as a department and/or imaging medium in the teaching of anatomy at the Canadian undergraduate medical education level. The study objectives were achieved through the use of a questionnaire and a literature review. The anatomy department head at each English-based Canadian Medical School was contacted, and the individual most responsible for anatomy teaching in the medical school curriculum was identified. This individual was subsequently asked to complete a questionnaire that evaluated the involvement of radiology for anatomy teaching in their curriculum. The use and integration of radiology is a common practice in the teaching of anatomy in Canadian undergraduate medicine. Although the methods and extent of its use varied among institutions, every English-based Canadian medical school, except one, was using diagnostic imaging material in their instruction of anatomy. Furthermore, half of the institutions had a radiologist as a faculty member of their anatomy department to help teach and to use imaging to its full potential. This audit of anatomy departments suggests that diagnostic imaging has an important role to play in anatomy teaching in Canadian English-speaking medical schools.


How SNARE molecules mediate membrane fusion: recent insights from molecular simulations.
SNARE molecules are the core constituents of the protein machinery that facilitate fusion of synaptic vesicles with the presynaptic plasma membrane, resulting in the release of neurotransmitter. On a molecular level, SNARE complexes seem to play a quite versatile and involved role during all stages of fusion. In addition to merely triggering fusion by forcing the opposing membranes into close proximity, SNARE complexes are now seen to also overcome subsequent fusion barriers and to actively guide the fusion reaction up to the expansion of the fusion pore. Here, we review recent advances in the understanding of SNARE-mediated membrane fusion by molecular simulations.


Labeling Acidic Compartments of Neutrophils with Cresyl Violet.
We introduce the acidotropic marker cresyl violet to stain acidic granules in live neutrophils. Cresyl violet is less phototoxic, more photostable, and more cost-effective than other commercially available acidotropic markers. Additionally, it does not photoconvert to fluorescent species of a different color, a limitation of other commonly used acidotropic markers. Staining can be readily detected by fluorescence microscopy or by flow cytometry, and can be used as a readout of degranulation in activated neutrophils.


Management of thyroid disease in pregnancy.
Thyroid disease occurs in pregnant at the rate of 0.2-0.6%. Graves' disease is the most common thyroid disorder in our pregnant patients. Thyroid disease is suspected if a significant goitre is detected during pregnancy. Confirmation by thyroid function tests is necessary but their interpretation requires an understanding of changes in the maternal thyroid physiology. Pharmacotherapy with the thionamides is the treatment of choice. Changes in dosage are influenced by the effect of pregnancy on the disease and the optimal level of control for the mother and the foetus. The dose is best titrated against the free thyroxine index or free thyroxine level. The outcome of pregnancy depends on early diagnosis and skillful manipulation of antithyroid drugs.


[Studies on HPLC-ELSD fingerprint of the Coleus forskohlii introduced in Tongcheng].
To establish HPLC-ELSD fingerprint of Coleus forskohlii. Chromatographic fingerprint of Coleus forskohlii was investigated by HPLC-ELSD and gradient elution mode was applied to chromatographic separation. The HPLC-ELSD fingerprint of Coleus forskohlii was established preliminarily. HPLC-ELSD fingerprint method is repeated and can be used in quality control of Coleus forskohlii. The active constituent in Coleus forskohlii is probably at equal pace between introduced in Tongcheng and provenance.


In vitro assay for ABA 8'-hydroxylase: implications for improved assays for cytochrome P450 enzymes.
In vitro assays for cytochrome P450 enzymes developed from plant-derived microsomal extracts have not been used extensively for the characterization and quantification of enzyme activities in plant tissues. We describe here an in vitro assay for abscisic acid (ABA) 8'-hydroxylase that was developed using microsomes extracted from (+)-ABA-induced corn suspension cultures. This assay may be useful for further characterization and monitoring of ABA 8'-hydroxylase activities in germinating seeds, seedlings, and other tissues. Additionally, the optimization protocols provided here may be adapted towards improving in vitro enzyme assays for other cytochrome P450 enzymes expressed in plants.


A tongue resection in macroglossia due to primary amyloidosis.
A lady of 62 years with longstanding macroglossia from primary amyloidosis had the anterior two-thirds of her tongue successfully resected. She had Factor X deficiency. The surgical, anaesthetic and haematological problems encountered are described.


In vitro neurite extension by granule neurons is dependent upon astroglial-derived fibroblast growth factor.
When grown in the absence of astroglial cells, purified mouse cerebellar granule neurons survive less than 36 hr and do not extend neurites. Here we report that low concentrations of basic fibroblast growth factor (bFGF, 1-25 ng/ml) maintained the viability and promoted the differentiation of purified granule neurons. The effect of bFGF on granule cell neurite outgrowth was dose dependent. Neurite outgrowth was stimulated markedly in the presence of 1-25 ng/ml bFGF, but effects were not seen below 1 ng/ml or above 50 ng/ml. When affinity-purified antibodies against bFGF (1-5 micrograms/ml) were added either to purified granule cells or to co-cultures of neurons and astroglial cells, process extension by granule neurons was severely impaired. The inhibition of neurite outgrowth in the presence of anti-bFGF antibodies was reversed by the addition of 25 ng/ml of exogenous bFGF. In addition to neuronotrophic effects, bFGF influenced the rate of growth of the astroglial cells. This result depended on whether the astroglia were grown in isolation from neurons, where low doses of bFGF (10-25 ng) stimulated glial growth, or in coculture with neurons, where much higher doses of bFGF (100-250 ng/ml) were needed for glial mitogenesis. Immunoprecipitation of lysates from 35S-labeled cerebellar astroglial cells with anti-bFGF antibodies revealed a single band after SDS-PAGE at 18,000 Da, the molecular weight of bFGF. These results indicate that glial cells synthesize bFGF and are possibly an endogenous source of bFGF in cerebellar cultures. Thus, astroglial cells synthesize soluble factors needed for neuronal differentiation.


Halo lipids. 10. Synthesis and cytostatic activity of O-alkylglycerophospho-L-serine analogs.
The synthesis of O-alkylglycerophospho-L-serine analogs is described, which represent a new class of cytostatically active agents based on phospholipids. The new compounds were obtained by conversion of O-alkylglycerophosphoric ester analogs by means of phospholipase D and by condensing O-alkylglycerophosphoric acid analogs with protected L-serine followed by the removal of protective groups of the resulting intermediates. The structure of the O-alkylglycerophospho-L-serines was confirmed by fast atom bombardment mass spectrometry. The compounds were found to inhibit the growth of Ehrlich ascites tumor cells in vitro. Half maximum inhibition was observed at concentrations between 7 and 15 microM. For the 1-O-alkyl-2-methoxy glycerophosphoserine only a higher value (30 microM) was found. With most of the substances tested growth was completely inhibited at a concentration of 40 microM.


The "French Paradox" and beyond: neuroprotective effects of polyphenols.
Chronic ethanol ingestion is known to cause oxidative damage to a number of organs including the brain. This is partly due to the ability of ethanol to enhance oxygen free radical production and lipid peroxidation. Increase in oxidative stress has been regarded as an important underlying factor for a number of human health problems including cardiovascular diseases, aging, as well as many age-related neurodegenerative diseases. The strikingly low incidences of coronary heart diseases (CHD) in France, despite intake of a high-fat diet, have been attributed to the consumption of red wine containing high levels of polyphenolic compounds. In recent years, understanding the "French Paradox" has stimulated new research interest to investigate whether polyphenolic antioxidants may offer protective effects beyond the cardiovascular system, and whether polyphenols from other botanical sources may similarly offer beneficial effects to human health. Our studies with animal models have provided information clearly indicating the ability of grape polyphenols to ameliorate neuronal damages due to chronic ethanol consumption. Studies with resveratrol, an important component of grape polyphenols, also show protective effects on neuron cell death induced by ethanol and other oxidative agents. These studies demonstrate an urgent need to extend research beyond the "French Paradox" towards better understanding molecular mechanisms of action of polyphenolic compounds and their application to human health.


Status of lipid peroxidation, some antioxidant enzymes and erythrocytic fragility of crossbred cattle naturally infected with Theileria annulata.
Erythrocytic lipid peroxidation, activities of some antioxidant enzymes and osmotic fragility of red blood cells was studied in adult (>1 year) crossbred cattle naturally infected with Theileria annulata. Twenty clinically healthy animals (group I) and 15 clinical cases (group II) of tropical theileriosis were selected. Cattle suffering from theileriosis had significantly higher (p<0.01) erythrocytic lipid peroxidation and osmotic fragility. Activities of antioxidant enzymes, viz. glucose-6-phosphate dehydrogenase (G6PD) and glutamate peroxidase (GPx) were also significantly increased (p<0.01) in group II. However, superoxide dismutase and catalase did not show significant changes. The results indicated that infection with theileria led to increased oxidative stress to the animals, and even a significant rise in the activities of antioxidant enzymes. G6PD and GPx could not lower this oxidative stress. However, the increase in the activities of antioxidant enzymes pointed towards the body's defence mechanism against lipid peroxidation during oxidative stress in theileriosis.


Pain Assessment Disparities by Race, Ethnicity, and Language in Adult Hospitalized Patients.
Nurses assess patients' pain using several validated tools. It is not known what disparities exist in pain assessment for medicine inpatients. Our purpose was to measure differences in pain assessment across patient characteristics, including race, ethnicity, and language status. Retrospective cohort study of adult general medicine inpatients from 2013 to 2021. The primary exposures were race/ethnicity and limited English proficiency (LEP) status. The primary outcomes were 1) the type and odds of which pain assessment tool nursing used and 2) the relationship between pain assessments and daily opioid administration. Of 51,602 patient hospitalizations, 46.1% were white, 17.4% Black, 16.5% Asian, and 13.2% Latino. 13.2% of patients had LEP. The most common pain assessment tool was the Numeric Rating Scale (68.1%), followed by the Verbal Descriptor Scale (23.7%). Asian patients and patients with LEP were less likely to have their pain documented numerically. In multivariable logistic regression, patients with LEP (OR 0.61, 95% CI 0.58-0.65) and Asian patients (OR 0.74, 95% CI 0.70-0.78) had the lowest odds of numeric ratings. Latino, Multi-Racial, and patients classified as Other also had lower odds than white patients of numeric ratings. Asian patients and patients with LEP received the fewest daily opioids across all pain assessment categories. Asian patients and patients with LEP were less likely than other patient groups to have a numeric pain assessment and received the fewest opioids. These inequities may serve as the basis for the development of equitable pain assessment protocols.


Otx2-Genetically Modified Retinal Pigment Epithelial Cells Rescue Photoreceptors after Transplantation.
Inherited retinal degenerations are blinding diseases characterized by the loss of photoreceptors. Their extreme genetic heterogeneity complicates treatment by gene therapy. This has motivated broader strategies for transplantation of healthy retinal pigmented epithelium to protect photoreceptors independently of the gene causing the disease. The limited clinical benefit for visual function reported up to now is mainly due to dedifferentiation of the transplanted cells that undergo an epithelial-mesenchymal transition. We have studied this mechanism in vitro and revealed the role of the homeogene OTX2 in preventing dedifferentiation through the regulation of target genes. We have overexpressed OTX2 in retinal pigmented epithelial cells before their transplantation in the eye of a model of retinitis pigmentosa carrying a mutation in Mertk, a gene specifically expressed by retinal pigmented epithelial cells. OTX2 increases significantly the protection of photoreceptors as seen by histological and functional analyses. We observed that the beneficial effect of OTX2 is non-cell autonomous, and it is at least partly mediated by unidentified trophic factors. Transplantation of OTX2-genetically modified cells may be medically effective for other retinal diseases involving the retinal pigmented epithelium as age-related macular degeneration.


Development of a protocol to quantify local bone adaptation over space and time: Quantification of reproducibility.
In vivo micro-computed tomography (µCT) scanning of small rodents is a powerful method for longitudinal monitoring of bone adaptation. However, the life-time bone growth in small rodents makes it a challenge to quantify local bone adaptation. Therefore, the aim of this study was to develop a protocol, which can take into account large bone growth, to quantify local bone adaptations over space and time. The entire right tibiae of eight 14-week-old C57BL/6J female mice were consecutively scanned four times in an in vivo µCT scanner using a nominal isotropic image voxel size of 10.4µm. The repeated scan image datasets were aligned to the corresponding baseline (first) scan image dataset using rigid registration. 80% of tibia length (starting from the endpoint of the proximal growth plate) was selected as the volume of interest and partitioned into 40 regions along the tibial long axis (10 divisions) and in the cross-section (4 sectors). The bone mineral content (BMC) was used to quantify bone adaptation and was calculated in each region. All local BMCs have precision errors (PE%CV) of less than 3.5% (24 out of 40 regions have PE%CV of less than 2%), least significant changes (LSCs) of less than 3.8%, and 38 out of 40 regions have intraclass correlation coefficients (ICCs) of over 0.8. The proposed protocol allows to quantify local bone adaptations over an entire tibia in longitudinal studies, with a high reproducibility, an essential requirement to reduce the number of animals to achieve the necessary statistical power.


Ultrastructural scoring of skin biopsies for diagnosis of vascular Ehlers-Danlos syndrome.
Vascular Ehlers-Danlos syndrome (vEDS) results from a mutation in the gene encoding alpha-1, type III pro-collagen (COL3A1) and confers fragility to skin, ligament and vascular tissue. We tested the value of skin biopsy for diagnosis of vEDS through an ultrastructure scoring procedure. Study design was a multicentric, case-control, blinded trial consisting of two phases: phase 1 was to identify an ultra-structure score providing the best discriminative value for vEDS and phase 2 was to replicate this result in a different population. We enrolled 103 patients, 66 cases defined through the revised nosology for Ehlers-Danlos syndromes and 37 control subjects selected from patients referred for other pathologies. Ultrastructure of extracellular matrix was read by three to five experienced pathologists blinded for diagnosis. We used the receiver operating curves and logistic regression analysis for ranking ultrastructure scores. We created a detailed description of lesions observed in vEDS patients with 27 items (coded 0 or 1). In the phase 1 (17 cases and 20 controls), abnormal fibroblast shape, presence of lysosomes in the fibroblast and abnormal basal lamina were found to be independent discriminative items. Addition of these three items (defining an ultrastructure score) had the best diagnosis value (area under the curve (AUC) = 0.96). In the phase 2 (49 cases, 17 controls), ultrastructure score provided odds ratio of 9.76 (95 % CI 2.91-32.78), and AUC of 0.90. The ultrastructure score of skin biopsy has predictive value for the diagnosis of vEDS. Presence of two or more signs (either abnormal fibroblast, presence of lysosomes in the fibroblast or abnormal basal lamina) is very evocative of vEDS.


Modulating properties of polysaccharides nanocomplexes from enzymatic hydrolysis of chitosan.
Synthesis of nanocomplexes is a simple and low-cost technique for the production of encapsulation systems aiming industrial applications, based on the interaction of at least two oppositely charged molecules. Gellan gum (anionic) is a water-soluble biopolymer resistant to stomach pH conditions, therefore an interesting alternative as an encapsulating matrix. Chitosan (cationic) is also widely used due to its biocompatibility and mucoadhesive properties, although its low water solubility is an important step to be overcome for the production of the complexes. To improve this property, many techniques have been employed, but most of them use unsustainable techniques and chemical agents. The enzymatic hydrolysis of chitosan using proteases emerges as an alternative to these drawbacks and, therefore, this study aimed to evaluate the electrostatic nanocomplexation of native (C) or hydrolyzed (HC) chitosan (by porcine pepsin protease) with gellan gum (G). Polysaccharides and nanocomplexes formed with different G:C or G:HC ratio were evaluated by zeta potential measurements, particle size distribution, X-ray diffraction (XRD), Fourier Transform Infrared Spectroscopy (FTIR), Scanning Transmission Electron Microscopy (STEM), intrinsic viscosity and turbidity analyses. Chitosan hydrolysis allowed the formation of a smaller (445.3 nm in pH 4.5) and more soluble structure (3 kDa), which positively influenced the formation of the complexes. The ratios G:HC of 7:3 and 8:2 formed complexes with lower values of zeta potential (13.9 mV and -5.0 mV, respectively), particle size (635.8 nm and 533.6 nm, respectively) and polydispersity (0.28 and 0.23) compared to complexes formed with native chitosan. Overall, our results show that enzymatic hydrolysis of chitosan favored the formation of electrostatic complexes with reduced size and low polydispersity, which can be used as efficient encapsulating matrices for improved targeted delivery and controlled release of bioactive compounds.


Selection criteria and match results for postgraduate residency programs: A cross-sectional model from a major academic center in Jordan.
With the continuous uptrend in the number of medical student graduates and the limited availability of postgraduate residency positions, the process of selecting the most appropriately qualified candidates to fill these positions remains challenging. This necessitates implementing objectively measured, distinguishing, and transparent selection process. The purpose of this study is to share our model of single-center resident selection for postgraduate residency programs to serve as a guide for other institutions. We reviewed the process of residency program selection at our institution. Data were collected about the application process, demographic characteristics, medical school location and GPA, entry exam score, requested specialties, and match results. The proposed selection criteria and their association with the match results were reported. Factors associated with matching with the first two selections were analyzed. 785 physicians applied to fill 96 positions at nineteen residency programs. 443 (56%) were males, 686 (87%) graduated from Jordanian medical schools. Half failed the entry exam and were excluded from competition. Seventy-two out of 96 (75%) matched with either of their first two requested specialties. The highest-in-demand programs were ophthalmology, otolaryngology and dermatology. Although a GPA of more than 80% increased the likelihood of matching with the top two requested specialties, an entry exam score of more than 70% was the main determining factor (AOR 8.7, 95% C.I. 2.4, 31.9). The selection process for postgraduate residency programs is highly competitive. To avoid selection bias, transparent and objectively measured criteria are applied in the selection model. Clinical performance and medical knowledge reflected by the cumulative GPA and entry exam score are the most significant determinants for acceptance.


Repetitive transcranial magnetic stimulation combined with olanzapine and amisulpride for treatment-refractory schizophrenia.
Treatment-refractory schizophrenia (TRS), accounting for approximately 30% of all schizophrenia cases, has poor treatment response and prognosis despite treatment with antipsychotic drugs. To analyze the therapeutic effectiveness of repetitive transcranial magnetic stimulation (rTMS) combined with olanzapine (OLZ) and amisulpride (AMI) for TRS and its influence on the patient's cognitive function. This study enrolled 114 TRS patients who received treatment at the First Affiliated Hospital of Zhengzhou University between July 2019 and July 2022. In addition to the basic OLZ + AMI therapy, 54 cases of the control group (Con group) received modified electroconvulsive therapy, while 60 cases of the research group (Res group) received rTMS. Data on therapeutic effectiveness, safety (incidence of drowsiness, headache, nausea, vomiting, or memory impairment), Positive and Negative Symptom Scale, Montreal Cognitive Assessment Scale, and Schizophrenia Quality of Life Scale were collected from both cohorts for comparative analyses. The Res group elicited a higher overall response rate and better safety profile when compared with the Con group. Additionally, a significant reduction was observed in the post-treatment Positive and Negative Symptom Scale and Schizophrenia Quality of Life Scale scores of the Res group, presenting lower scores than those of the Con group. Furthermore, a significant increase in the Montreal Cognitive Assessment Scale score was reported in the Res group, with higher scores than those of the Con group. The treatment of TRS with rTMS and OLZ + AMI is effective and safe. Moreover, it can alleviate the patients' mental symptoms, improve their cognitive function and quality of life, and has a high clinical application value.


[A multicenter evaluation of efficacy and tolerance in the local treatment with dolgit cream of patients with osteoarthrosis].
A 2-week clinical trial of cream dolgit efficacy and tolerance in 76 patients with primary osteoarthritis in 8 rheumatological centers showed this preparation to reduce local pain syndrome in 70.1%, palpation pain sensitivity in 79.2%, joint swelling in 75.5% of patients. Unwanted reactions in the form of skin itch at the site of application were reported in 7.8% of cases.


Hitting them where it hurts: Pantothenate kinase targeting in pathogenic fungi.
In this issue of Structure, Gihaz and colleagues develop antifungal compounds targeting fungal pantothenate kinase (PanK). Through utilization of high-throughput chemical screening, along with biochemical and functional analyses and newly solved fungal PanK crystal structures, high-affinity inhibitors are produced, and invaluable insights into the mechanisms of fungal PanK are gained.


Exogenous AdoMet and its analogue sinefungin differentially influence DNA cleavage by R.EcoP15I--usefulness in SAGE.
While it has been demonstrated that AdoMet is required for DNA cleavage by Type III restriction enzymes, here we show that in the presence of exogenous AdoMet, the head-to-head oriented recognition sites are cleaved only on a supercoiled DNA. On a linear DNA, exogenous AdoMet strongly drives methylation while inhibiting cleavage reaction. Strikingly, AdoMet analogue sinefungin results in cleavage at all recognition sites irrespective of the topology of DNA. The cleavage reaction in the presence of sinefungin is ATP dependent. The site of cleavage is comparable with that in the presence of AdoMet. The use of EcoP15I restriction in presence of sinefungin as an improved tool for serial analysis of gene expression is discussed.


Penicillium oxalicum putative methyltransferase Mtr23B has similarities and differences with LaeA in regulating conidium development and glycoside hydrolase gene expression.
Putative methyltranferase LaeA and LaeA-like proteins, which are conserved in many filamentous fungi, regulate the sporogenesis and biosynthesis of secondary metabolites. In this study, we reported the biological function of a LaeA-like methyltransferase, Penicillium oxalicum Mtr23B, which contains a methyltransf_23 domain and an S-adenosylmethionine binding domain, in controlling spore pigment formation and in the expression of secondary metabolic gene cluster and glycoside hydrolase genes. Additionally, we compared Mtr23B and LaeA, and determined their similarities and differences in terms of their roles in regulating the above biological processes. mtr23B had the highest transcriptional level among the 12 members of the methyltransf_23 family in P. oxalicum. The colony color of Δmtr23B (deletion of mtr23B) was lighter than that of ΔlaeA, although Δmtr23B produced ~ 19.2-fold more conidia than ΔlaeA. The transcriptional levels of abrA, abrB/yA, albA/wA, arpA, arpB, and aygA, which are involved in the dihydroxynaphtalene-melanin pathway, decreased in Δmtr23B. However, Mtr23B had a little effect on brush-like structures and conidium formation, and had a different function from LaeA. Mtr23B extensively regulated glycoside hydrolase gene expression. The absence of Mtr23B remarkably repressed prominent cellulase- and amylase-encoding genes in the whole culture period, while the effect of LaeA mainly occurred in the later phases of prolonged batch cultures. Similar to LaeA, Mtr23B was involved in the expression of 10 physically linked regions containing secondary metabolic gene clusters; the highest regulatory activities of Mtr23B and LaeA were observed in BrlA-dependent cascades. Although LaeA interacted with VeA, Mtr23B did not interact with VeA directly. We assumed that Mtr23B regulates cellulase and amylase gene transcription by interacting with the CCAAT-binding transcription factor HAP5 and chromatin remodeling complex.


The discovery of IgE.
In 1919, the search began for the factor, later called reagin, that could mediate an allergy, such as allergic asthma, in sera of allergic subjects. In 1967, the fifth class of immunoglobulins, IgE, was discovered and found to be able to carry reagin activity. This discovery has had immense importance for the understanding, diagnosis, and treatment of allergic diseases.


Visualization of chromatin distribution in living PTO cells by Hoechst 33342 fluorescent staining.
Chromatin distribution was visualized in living cells with the selective DNA fluorochrome Hoechst 33342. This dye was shown to be non-toxic on the rat kangaroo PTO cell line by measuring the labelled cell growth rate. The aim of this work was firstly to visualize chromatin distribution without fixation or dehydration and secondly to demonstrate that quantitative determination of DNA content was possible under these non-toxic labelling conditions. During interphase, condensed, decondensed and thin network chromatin configurations were visualized. In nucleolar regions the fluorochrome revealed well-defined chromocentres. During mitosis, fluorescent chromosome banding was observed in vital conditions and chromocentres on fixed chromosomes. Chromatin segregation was visualized after micronucleation, which induced chromosomal set distribution in individual micronuclei. By this means, we demonstrated that the chromocentres observed in interphase nuclei were part of nuclear organizer region (NOR)-bearing chromosomes. This vital staining of chromatin was shown to be compatible with the quantitative determination of DNA content, both in living PTO cells and in isolated nuclei.


Odontoblast markers and dentine reactions in carious primary molars with and without hypomineralised enamel defects.
Wnt/β-Catenin signalling and DMP1 have key roles in tertiary dentinogenesis. To compare the relationship between remaining dentine thickness (RDT), tertiary dentine thickness (TDT), β-catenin and dentine matrix protein 1 (DMP1) in carious second primary molar teeth with normal (SPM) and hypomineralised enamel (HSPM). Extracted carious SPM and HSPM were fixed, sectioned (5 μm) and stained with haematoxylin and eosin or with indirect immunofluorescence for β-catenin and DMP1. Image analysis was performed to determine RDT, TDT, β-catenin and DMP1 intensity in the odontoblast layer and dentine-pulp complex. Carious SPM (n = 11; mean RDT = 1536.1 μm) and HSPM (n = 12; mean RDT = 1179.9 μm) had mean TDT 248.6 μm and 518.1 μm, respectively (P = .02). There were no significant differences in intensity values in the odontoblast layer and dentine-pulp complex for β-catenin and DMP1 for both groups. There was no observable variation in Wnt/β-catenin and DMP1 expression between HSPM and SPM despite a statistically significant twofold increased TDT in HSPM compared with SPM that had similar RDT. Thus, the observed increased TDT in HSPM is more likely due to an earlier onset of repair processes rather than an amplified response to caries.


The spectral color of natural and anthropogenic time series and its impact on the statistical significance of cross correlation.
The cross-correlation between time series is a common tool to study and quantify the impact of climatic and anthropogenic changes on ecosystems. The traditional method for estimating the statistical significance of correlation relies on the assumption that the data are independent, but time series found in nature are often strongly auto-correlated because of low-frequency environmental variability and ecosystem inertia. Previous authors have used Monte Carlo simulations to study the impact of serial auto-correlation on the significance of cross-correlations. Most studies have used random time series that are often a poor representation of those found in nature, e.g., low-order auto-regressive models with normally distributed noise. Moreover, we are not aware of any tests of the applicability of those methods to anthropogenic time series. Here, we study the effect of serial auto-correlation on the performance of two methods for estimating the significance of cross-correlations determined from Monte Carlo simulations with time series that are generated synthetically based on power-law specification of spectral characteristics. Such time series have an auto-correlation structure defined by a single parameter, their spectral "color", and are generally more convenient representations of natural time series than the autoregressive models. Our results show that one of the two methods considered here accurately reproduces prescribed error rates for the wide range of spectral colors representative of climatic, ecological and anthropogenic time series. For this, we characterized roughly 1800 observational records in different categories of spectral colors, including climate variability, abundance of vertebrate species, and pollution. We specifically focus on time series with annual sampling over data records of at least 40 years, which are particularly relevant for climate studies. The methodology advocated in this study provides a simple and realistic assessment of the significance of sample estimates of cross correlation for time series with any sample interval and record length.


Immuno-stimulatory activity of Astragalus polysaccharides in cyclophosphamide-induced immunosuppressed mice by regulating gut microbiota.
Evidence has indicated the immune-stimulatory effect of Astragalus polysaccharides (APS), yet it remains unknown whether the potential mechanism is associated with gut microbiota. In this study, we aimed to investigate the role of gut microbiota in APS-initiated immune-enhancing activity in mice. BALB/c mice were injected with cyclophosphamide to establish a mouse immunosuppression model. We found that APS significantly ameliorated the immunosuppression in mice, indicative of the increased immune organ indices, the promoted proliferation of immune cells, and the up-regulated intestinal inflammation. Western blot analysis demonstrated that APS treatment significantly activated Toll-like receptor 4 (TLR4) and mitogen-activated protein kinase (MAPK) pathways in the intestine. By 16S rDNA sequencing, APS treatment reversed the gut microbiota dysbiosis in immunocompromised mice. At the genus level, APS increased the abundance of bacteria (like Lactobacillus, Bifidobacteria, Roseburia, and Desulfovibrio) and decreased the content of several bacteria (like Oscillibacter, Tyzzerella, and Lachnoclostridium). However, APS had no immune-enhancing effect on immunocompromised mice with gut microbiota depletion. In conclusion, APS can enhance immune responses in immunocompromised mice by modulating gut microbiota dysbiosis.


Comparison of Anorectic Potencies of the Trichothecenes T-2 Toxin, HT-2 Toxin and Satratoxin G to the Ipecac Alkaloid Emetine.
Trichothecene mycotoxins, potent translational inhibitors that are associated with human food poisonings and damp-building illnesses, are of considerable concern to animal and human health. Food refusal is a hallmark of exposure of experimental animals to deoxynivalenol (DON) and other Type B trichothecenes but less is known about the anorectic effects of foodborne Type A trichothecenes (e.g., T-2 toxin, HT-2 toxin), airborne Type D trichothecenes (e.g. satratoxin G [SG]) or functionally analogous metabolites that impair protein synthesis. Here, we utilized a well-described mouse model of food intake to compare the anorectic potencies of T-2 toxin, HT-2 toxin, and SG to that of emetine, a medicinal alkaloid derived from ipecac that inhibits translation. Intraperitoneal (IP) administration with T-2 toxin, HT-2 toxin, emetine and SG evoked anorectic responses that occurred within 0.5 h that lasted up to 96, 96, 3 and 96 h, respectively, with lowest observed adverse effect levels (LOAELs) being 0.1, 0.1, 2.5 and 0.25 mg/kg BW, respectively. When delivered via natural routes of exposure, T-2 toxin, HT-2 toxin, emetine (oral) and SG (intranasal) induced anorectic responses that lasted up to 48, 48, 3 and 6 h, respectively with LOAELs being 0.1, 0.1, 0.25, and 0.5 mg/kg BW, respectively. All four compounds were generally much more potent than DON which was previously observed to have LOAELs of 1 and 2.5 mg/kg BW after IP and oral dosing, respectively. Taken together, these anorectic potency data will be valuable in discerning the relative risks from trichothecenes and other translational inhibitors of natural origin.