Size-, mass-, and density-controlled preparation of TiO2 nanoparticles in a spherical coordination template.
TiO2 nanoparticles were prepared with strict control of the size, molecular mass (PDI = 1.02), and mass density by hydrolyzing a precursor, Ti(acac)2(biphen), in the presence of a well-defined hollow Pd12L24 spherical endotemplate. The formation of highly monodisperse TiO2 nanoparticles was confirmed by (1)H NMR spectroscopy, MALDI-TOF mass spectrometry, transmission electron microscopy, and atomic force microscopy. The template was easily removed by treating the sphere with a metal scavenger followed by calcination to give crystalline TiO2 nanoparticles (2.0 ± 0.2 nm in diameter), which showed photocatalytic activity.


Cervical Transdural Discectomy with Laminoplasty for the Treatment of Multi-segment Cervical Spinal Stenosis Accompanied with Cervical Disc Herniation: Technical Note and Clinical Outcome.
To describe the surgical technique of cervical transdural discectomy with laminoplasty (CTDL) for the treatment of multi-segment cervical spinal stenosis (CSS) accompanied with cervical disc herniation (CDH) and investigate its surgical outcomes and complications. This was a clinical study. Between 2012 and 2018, 31 patients (13 males and 18 females) with multi-segment CSS (over two cervical segments) accompanied with huge CDH and underwent CTDL were enrolled in this study. The details of CTDL technique with general anesthesia was described by the authors. The average follow-up period of patients was 65.03 months (range from 24 to 126 months). Perioperative parameters such as age, sex, operative level, operative time, estimated blood loss, ambulation time, and operative complications were recorded. The results of clinical metrics such as the visual analog scale (VAS) and Japanese Orthopaedic Association (JOA) scores in the preoperative and during the follow-up period were obtained and used to evaluate clinical outcomes. Radiographic improvement was evaluated by the compression ratio, sagittal maximum spinal cord compression (SMSCC), and cervical range of motion (ROM). The preoperative and postoperative follow-up parameters (VAS, JOA, Compression ratio, SMSCC, and ROM) were assessed with paired t test. A P-value <0.05 was considered statistically significant. In the study, the mean age of the 31 patients was 55.23 ± 10.97 years. The mean operative time was 192.45 ± 24.17 min (ranging from 150 to 245 min), and intraoperative blood loss was 322.58 ± 129.00 mL (ranging from 150 to 600 mL). The VAS neck pain was improved significantly over the follow-up period (P < 0.05, respectively). The VAS arm pain improved significantly from 6.26 ± 0.93 preoperatively to 1.74 ± 0.63 at 24 months postoperatively (P < 0.001). There was no significant difference in improvement of VAS arm pain between 24 months postoperatively and final follow-up (P = 0.180). Compared with preoperative JOA score, JOA score was significantly improved at 24 months postoperatively (14.79 ± 1.84 vs 9.66 ± 2.81, P < 0.001). Meanwhile, there were no statistically significant differences between the final follow-up and the postoperative JOA scores (15.08 ± 1.71 vs 14.79 ± 1.84, P = 0.051). Postoperative patients showed significantly higher index of compression ratio (58.30 ± 8.51 vs 27.17 ± 3.89, P < 0.001) and lower SMSCC (25.12 ± 5.67 vs 33.66 ± 5.38, P < 0.001). In addition, there was no significant difference between preoperative and postoperative cervical ROM (P = 0.740). One patient observed postoperative symptom of C6 nerve root injury, which was resolved within 24 months after the surgery; meanwhile, the neurological monitoring also reflected the intraoperative stretching of the C6 nerve root. Two cases involved postoperative cerebrospinal fluid (CSF) leakage which may have been related to laceration of dura mater. This study suggested that CTDL technique could acquire satisfactory surgical outcomes for patients with multi-segment CSS accompanied with CDH, but the surgical indications of the patients need to be selected strictly.


A small trypsin inhibitor from the frog of Odorrana grahami.
A novel peptide inhibitor (OGTI) of serine protease with a molecular weight of 1949.8, was purified from the skin secretion of the frog, Odorrana grahami. Of the tested serine proteases, OGTI only inhibited the hydrolysis activity of trypsin on synthetic chromogenic substrate. This precursor deduced from the cDNA sequence is composed of 70 amino acid residues. The mature OGTI contains 17 amino acid residues including a six-residue loop disulfided by two half-cysteines (AVNIPFKVHFRCKAAFC). In addition to its unique six-residue loop, the overall structure and precursor of OGTI are different from those of other serine protease inhibitors. It is also one of the smallest serine protease inhibitors ever found.


Nature and course of cognitive function in late-life schizophrenia: a systematic review.
To systematically review the literature on the nature and course of cognition in late-life schizophrenia (LLS). We conducted a literature search using Medline. Search terms included schizophrenia, cognition, memory, and other cognitive search terms. We limited our search to age 45 and above. All titles or abstracts were read, and relevant papers were reviewed. Only cross-sectional studies with healthy control groups or longitudinal studies of cognition in LLS are presented in this review. We identified 23 publications reporting on cross-sectional studies comparing cognition in subjects with LLS and healthy controls, and 19 publications reporting on cognitive changes during longitudinal follow-up. The cross-sectional reports suggest that patients with LLS are most consistently impaired in executive function, visuospatial ability, and verbal fluency. Impairment has less consistently been observed in memory, attention, and working memory. Longitudinal studies suggest that patients with LLS start to decline cognitively around the age of 65, and that this decline may first affect visuospatial abilities. However, most of these studies have been conducted in institutionalized patients, rather than the typical ambulatory population. Other limitations include small sample sizes, short follow-up periods, and lack of comprehensive neuropsychological assessments. The existing literature suggests that the nature and course of cognition in LLS is heterogeneous. Larger and longer studies using both comprehensive and specific cognitive assessments are needed to understand the causes and consequences of this heterogeneity.


Fixation strength of meniscal repair devices.
The aim of this study was to measure and compare the ultimate failure strengths and cyclic fatigue strengths of currently available meniscal suturing devices. No statistically significant difference in failure load was found between a vertical loop suture (mean 46.3 N), a horizontal mattress suture (52.5 N), the T-fix Device (47.5 N), and the 16-mm (39.2 N) and 13-mm (32.8 N) Bionix Arrow. Statistically inferior results were seen with the 10-mm Bionix Arrow (18.8 N), the S.D. Sorb Stapler (4.3 N), and the 12-mm Arthrex Meniscal Dart (10.5 N) (P<0.01). The Mitek Meniscal Repair System (28.1 N) performed intermediate, with significantly better results than the S.D. Sorb Stapler and the 12-mm Arthrex Dart, but significantly worse than the vertical and horizontal sutures, the T-fix and Bionix 16-mm Device (P<0.01). Cyclic fatigue strength was significantly less for the 10-mm Bionix Arrow, the S.D. Sorb Stapler, and the 12-mm Arthrex Meniscal Dart (P<0.01) compared to all other devices. We conclude that the 13- and 16-mm Bionix Arrow and the T-fix Device have comparable ultimate failure strengths and cyclic fatigue strengths to conventional meniscal suturing techniques. The 10-mm Bionix Arrow, S.D. Sorb Stapler, and the 12-mm Arthrex Meniscal Dart, however, have far inferior failure and cyclic fatigue strengths and their clinical application should be questioned.


Perceived barriers and facilitators of day-case surgery for major foot and ankle procedures? A cross-sectional survey of United Kingdom surgeons.
Advances in minimally invasive surgery and improved post-operative pain management make it possible to consider performing even major foot/ankle operations as day-case. This could have significant benefits for patients and the health service. However there are theoretical concerns about post-operative complications and patient satisfaction due to pain. To scope the current practice of foot and ankle surgeons on day-case surgery for major foot and ankle procedures in the United Kingdom (UK). An online survey (19 questions) was sent to UK foot and ankle surgeons via the British Orthopaedic Foot & Ankle Society membership list in August 2021. Major foot and ankle procedures were defined as surgery that is usually performed as an inpatient in majority of centres and day-case as same day discharge, with day surgery as the intended treatment pathway. 132 people responded to the survey invitation with 80% working in Acute NHS Trusts. Currently 45% of respondents perform less than 100 day-case surgeries per year for these procedures. 78% felt that there was scope to perform more procedures as day-case at their centre. Post-operative pain (34%) and patient satisfaction (10%) was not highly measured within their centres. Lack of adequate physiotherapy input pre/post-operatively (23%) and lack of out of hours support (21%) were the top perceived barriers to performing more major foot and ankle procedures as day-case. There is consensus among UK surgeons to do more major foot/ankle procedures as day-case. Out of hours support and physiotherapy input pre/ post-op were perceived as the main barriers. Despite theoretical concerns about post-operative pain and satisfaction this was only measured by a third of those surveyed. There is a need for nationally agreed protocols to optimise the delivery of and measurement of outcomes in this type of surgery. At a local level, the provision of physiotherapy and out of hours support should be explored at sites where this is a perceived barrier.


Systematic Review of Mindfulness Practice for Reducing Job Burnout.
A systematic search and critical appraisal of interdisciplinary literature was conducted to evaluate the evidence for practicing mindfulness to treat job burnout and to explore implications for occupational therapy practitioners. Eight articles met inclusion criteria. Each study was assessed for quality using the Physiotherapy Evidence Database scale. We used the U.S. Agency for Health Care Policy and Research guidelines to determine strength of evidence. Of the studies reviewed, participants included health care professionals and teachers; no studies included occupational therapy practitioners. Six of the 8 studies demonstrated statistically significant decreases in job burnout after mindfulness training. Seven of the studies were of fair to good quality. There is strong evidence for the use of mindfulness practice to reduce job burnout among health care professionals and teachers. Research is needed to fill the gap on whether mindfulness is effective for treating burnout in occupational therapy practitioners.


Development and Verification of a Digital Twin Patient Model to Predict Specific Treatment Response During the First 24 Hours of Sepsis.
To develop and verify a digital twin model of critically ill patient using the causal artificial intelligence approach to predict the response to specific treatment during the first 24 hours of sepsis. Directed acyclic graphs were used to define explicitly the causal relationship among organ systems and specific treatments used. A hybrid approach of agent-based modeling, discrete-event simulation, and Bayesian network was used to simulate treatment effect across multiple stages and interactions of major organ systems (cardiovascular, neurologic, renal, respiratory, gastrointestinal, inflammatory, and hematology). Organ systems were visualized using relevant clinical markers. The application was iteratively revised and debugged by clinical experts and engineers. Agreement statistics was used to test the performance of the model by comparing the observed patient response versus the expected response (primary and secondary) predicted by digital twin. Medical ICU of a large quaternary- care academic medical center in the United States. Adult (> 18 year yr old), medical ICU patients were included in the study. No additional interventions were made beyond the standard of care for this study. During the verification phase, model performance was prospectively tested on 145 observations in a convenience sample of 29 patients. Median age was 60 years (54-66 d) with a median Sequential Organ Failure Assessment score of 9.5 (interquartile range, 5.0-14.0). The most common source of sepsis was pneumonia, followed by hepatobiliary. The observations were made during the first 24 hours of the ICU admission with one-step interventions, comparing the output in the digital twin with the real patient response. The agreement between the observed versus and the expected response ranged from fair (kappa coefficient of 0.41) for primary response to good (kappa coefficient of 0.65) for secondary response to the intervention. The most common error detected was coding error in 50 observations (35%), followed by expert rule error in 29 observations (20%) and timing error in seven observations (5%). We confirmed the feasibility of development and prospective testing of causal artificial intelligence model to predict the response to treatment in early stages of critical illness. The availability of qualitative and quantitative data and a relatively short turnaround time makes the ICU an ideal environment for development and testing of digital twin patient models. An accurate digital twin model will allow the effect of an intervention to be tested in a virtual environment prior to use on real patients.


Case finding for chronic obstructive pulmonary disease in primary care: a pilot randomised controlled trial.
Chronic obstructive pulmonary disease (COPD) is a major cause of morbidity and mortality. However, much of the disease burden remains undiagnosed. To compare the yield and cost effectiveness of two COPD case-finding approaches in primary care. Pilot randomised controlled trial in two general practices in the West Midlands, UK. A total of 1634 ever-smokers aged 35-79 years with no history of COPD or asthma were randomised into either a 'targeted' or 'opportunistic' case-finding arm. Respiratory questionnaires were posted to patients in the 'targeted' arm and provided to patients in the 'opportunistic' arm at routine GP appointments. Those reporting at least one chronic respiratory symptom were invited for spirometry. COPD was defined as pre-bronchodilator forced expiratory volume in 1 second/forced vital capacity (FEV1/FVC)<0.7 and FEV1<80% of predicted. Primary outcomes were the difference in the proportion of patients diagnosed with COPD and the cost per case detected. Twenty-six per cent (212/815) in the 'targeted' and 13.6% (111/819) in the 'opportunistic' arm responded to the questionnaire and 78.3% (166/212) and 73.0% (81/111), respectively, reported symptoms; 1.2% (10/815) and 0.7% (6/819) of patients in the 'targeted' and 'opportunistic' arms were diagnosed with COPD (difference in proportions = 0.5% [95% confidence interval {CI} = -0.5% to 3.08%]). Over a 12-month period, the 'opportunistic' case-finding yield could be improved to 1.95% (95% CI = 1.0% to 2.9%). The cost-per case detected was £424.56 in the 'targeted' and £242.20 in the 'opportunistic' arm. Opportunistic case finding may be more effective and cost effective than targeting patients with a postal questionnaire alone. A larger randomised controlled trial with adequate sample size is required to test this.


Two autopsy cases of sudden unexpected death from Dravet syndrome with novel de novo SCN1A variants.
Dravet syndrome (DS) is characterized by high epilepsy-related premature mortality with a markedly young age at death, however, autopsy report of sudden unexpected death with DS has been fewer than expected. We report two autopsy cases with sudden unexpected death from DS. Case 1 was a 13-year-old male who drowned in a bathtub, and Case 2 was a 3-year-old female who died while sleeping. In Case 1, the blood concentration of the anticonvulsant, valproic acid, was below the recommended therapeutic range. Neuropathological investigation and genetic analysis of 402 cardiovascular disease-related and 146 epilepsy-related genes by next generation sequencing were applied. No significant neuronal loss with gliosis was observed in the brain of either patient. Although possible mild malformations of cortical development were found in both, the degree thereof was similar to that of age-matched controls. Genetic analysis identified a novel variant in SCN1A intron 23 (c.4477-3T > C) in Case 1 that falls outside of the minor splicing consensus sequence. In vitro splicing functional assays with minigene constructs revealed that this intronic variant leads to a 2-bp insertion immediately before exon 24 that results in protein truncation. Similarly, a novel de novo missense mutation of unknown significance, SCN1A_Arg187Pro, was identified in Case 2. In both cases, we also identified cardiomyopathy-related variants classified as likely pathogenic; however, the effect of these variants at death was minimal because there was an absence of pathological change indicating inherited cardiomyopathy. The present cases emphasize the need for multifaceted examination of DS cases so as to obtain a definitive autopsy diagnosis and to explore the mechanism of sudden unexpected death.


Physical exercise prevents amyloid β1-40-induced disturbances in NLRP3 inflammasome pathway in the hippocampus of mice.
Amyloid beta (Aβ), one of the main hallmarks of Alzheimer's Disease (AD), may stimulate pattern recognition receptors (PRR) such as the NLRP3 inflammasome, inducing a pro-inflammatory state in the brain that contributes to disease development. Physical exercise can have multiple beneficial effects on brain function, including anti-inflammatory and neuroprotective roles. The objective of this study was to investigate the prophylactic effect of moderate treadmill exercise for 4 weeks on inflammatory events related to NLRP3 signaling in the hippocampus of mice after intracerebroventricular Aβ1-40 administration. Our results show that Aβ1-40 administration (400 pmol/mouse, i.c.v.) significantly increased the immunocontent Iba-1 (a microglial reactivity marker), NLRP3, TXNIP, and caspase-1 in the hippocampus of mice. However, physical exercise prevented the hippocampal increase in Iba-1, TXNIP, and activation of the NLRP3 inflammasome pathway caused by Aβ1-40. Moreover, physical exercise per se reduced the TXNIP and caspase-1 immunocontent in the hippocampus. No alterations were observed on the immunocontent of GFAP, ASC, and IL-1β in the hippocampus after Aβ1-40 and/or physical exercise. These results reinforce the role of NLRP3 inflammasome pathway in AD and point to physical exercise as a possible non-pharmacological strategy to prevent inflammatory events triggered by Aβ1-40 in mice.


Comparative studies of radiotracer citrates in oncological models--2. 153Sm-citrate and 67Ga-citrate.
153Sm-citrate solutions were prepared from enriched 152Sm2O3 which had been irradiated at 10(12)n cm-2 s-1 in the University of Alberta Slowpoke reactor. 153Sm was rapidly bound (93% in 2 h) by Melanoma 2AB cells in tissue culture upon incubation in the presence of 153Sm-citrate (1.9 nmol 10(6) cells). In vitro ultracentrifugation studies of 153Sm-citrate solutions showed that colloid formation under incubation conditions could have been responsible in part for the uptake by cultured cells. Low uptakes (less than 1% in 2h) of 67Ga-citrate were seen under similar conditions. 153Sm-citrate injected into BDF1 mice (Lewis lung carcinoma) and Copenhagen x Fisher rats (Dunning R3327-H prostatic tumors) was concentrated mainly in the liver, with some tumor and bone uptake. The percent of injected dose per organ for 153Sm and 67Ga in the murine and rat models respectively, 24 h after i.v. dosing, was 17.2 +/- 4.7 and 2.0 +/- 0.6 (tumor), 63.9 +/- 7.9 and 14.4 +/- 1.4 (liver) and 0.6 +/- 0.1 and 1.3 +/- 0.1 (blood); % injected dose g-1 femur was 6.2 +/- 2.7 and 12.9 +/- 2.7, respectively. Scintigrams of rats showed qualitative biodistributions similar to the quantitative mouse data obtained by dissection studies. The high hepatic uptake detracted from the otherwise superior tumor localization of 153Sm-citrate when compared to 67Ga-citrate in these models. The murine Lewis lung tumor index (% injected dose g-1 tumor x tumor: blood) was 303.6 for 153Sm-citrate and 48.9 for 67Ga-citrate, 24 h after injection.


Particle disease: a unique cause of hypercalcemia.
Hypercalcemia as a result of ectopic 1, 25-dihydroxyvitamin D (1,25-(OH)2 D) production has been well-described in sarcoidosis and other granulomatous diseases. The 1-alpha-hydroxylase enzyme in activated macrophages is not subject to physiologic regulations, resulting in hypercalcemia with inappropriately normal or elevated 1,25-(OH) 2 D. Particle disease is the local inflammatory response provoked by an overwhelming production of wear debris from a failed joint prosthesis. Enhanced focal bone resorption in particle disease has been described due to local production of inflammatory cytokines. However, this process previously has not been reported to cause hypercalcemia. We describe a patient with hypercalcemia, low parathyroid hormone levels, and elevated 1,25-(OH)2 D whose failed prosthetic joint generated a large amount of inflammatory debris, forming a soft tissue mass with lymphadenopathy. Biopsy of the mass demonstrated activated macrophages and foreign body granuloma, resulting in unregulated production of 1,25-(OH)2 D and hypercalcemia. We present the first case of hypercalcemia associated with elevated 1,25-(OH)2 D in particle disease due to a failed prosthetic hip.


Clinical problems in atrial synchronous ventricular inhibited pacing: a long-term follow-up of 54 patients.
Fifty-four VDD-paced patients were followed for more than 12 months; they were studied retrospectively in order to assess possible clinical problems and their management. The patients were between 19 and 84 years of age (mean, 66 +/- 11). Twenty-four of the 54 received a VDD pacemaker as a primary implant and 30 had had VVI pacemakers which were changed to VDD mainly because of limited exercise tolerance or symptoms of AV asynchrony. The follow-up time was between 12 and 69 months (mean, 39 +/- 17). Pacemaker and lead problems were neither frequent nor serious. Six patients had spontaneous paroxysmal supraventricular tachyarrhythmias, four had pacemaker-mediated supraventricular tachycardias and six had ventricular tachyarrhythmias. Treatment of tachyarrhythmias included drugs, DC conversion, reprogramming, or combinations of these measures. Frequency of hospitalization was not greater than expected. In conclusion, VDD pacing appears safe and reliable, with problems mainly associated with the underlying cardiac disease rather than to the pacing mode itself.


Quantifying the forces guiding microbial cell adhesion using single-cell force spectroscopy.
During the past decades, several methods (e.g., electron microscopy, flow chamber experiments, surface chemical analysis, surface charge and surface hydrophobicity measurements) have been developed to investigate the mechanisms controlling the adhesion of microbial cells to other cells and to various other substrates. However, none of the traditional approaches are capable of looking at adhesion forces at the single-cell level. In recent years, atomic force microscopy (AFM) has been instrumental in measuring the forces driving microbial adhesion on a single-cell basis. The method, known as single-cell force spectroscopy (SCFS), consists of immobilizing a single living cell on an AFM cantilever and measuring the interaction forces between the cellular probe and a solid substrate or another cell. Here we present SCFS protocols that we have developed for quantifying the cell adhesion forces of medically important microbes. Although we focus mainly on the probiotic bacterium Lactobacillus plantarum, we also show that our procedures are applicable to pathogens, such as the bacterium Staphylococcus epidermidis and the yeast Candida albicans. For well-trained microscopists, the entire protocol can be mastered in 1 week.


Prophylactic vasectomy.
A prospective study of the value of vasectomy with prostatectomy is presented. Vasectomy significantly reduced the incidence of epididymitis after prostatectomy in the presence of pre-operative urinary tract infection and prolonged indwelling urethral catheterisation.


Transfer of drug-resistance plasmids by conjugation from nosocomial strains of Serratia marcescens to Escherichia coli in biological fluids of human origin.
Six independent isolates of multi-resistant Serratia marcescens associated with nosocomial infections were examined for their ability to transfer drug-resistance plasmids by conjugation to Escherichia coli in biological fluids of human origin, such as normal and pathological urine, faeces, blood plasma and ascitic fluid. Luria broth was used as a control. Positive transfer was found in all media assayed. The different patterns of linked transferable resistance found in the transconjugants corresponded to the phenotypic expression of five plasmids. The frequencies of transfer varied with plasmid types and media employed. The culture media did not affect the phenotypic expression of the plasmids.


Sp1 and Sp3 regulate the basal transcription of receptor activator of nuclear factor kappa B ligand gene in osteoblasts and bone marrow stromal cells.
Receptor activator of nuclear factor kappa B ligand (RANKL), a potent regulator of osteoclast formation and function, is expressed by osteoblasts and bone marrow stromal cells. However, the molecular mechanism underlying RANKL expression in osteoblast/stromal cells remains largely unclear. Here, we characterized the molecular mechanism controlling RANKL basal transcription in osteoblast/stromal cells. We cloned a 1,103-bp murine RANKL promoter (from -953 to +150, relative to the transcription start site). Using a series of deletion mutants of the 1,103-bp promoter, we identified a 100-bp region (-154 to -54) mediating RANKL basal transcription in both osteoblasts and bone marrow stromal cells. Electrophoretic mobility shift assay (EMSA) using five overlapping oligonucleotides (Probes 1-5) spanning the 100-bp region showed that Probes 1 and 2 specifically bound nuclear proteins with high affinity from both cell types. Computer analysis revealed that Probes 1 and 2 contain a putative Sp1-binding site. Supershift assays with Sp1 and Sp3 antibodies confirmed that the nuclear proteins binding to Probes 1 and 2 are Sp1 and Sp3. Functionally, the mutation of the Sp1/Sp3 site in Probe 1 profoundly reduced the basal promoter activity while the mutation of the one in Probe 2 resulted in moderate reduction in the basal promoter activity. Moreover, the mutation of both sites abrogated the RANKL basal promoter activity, indicating that Sp1 and Sp3 play a key role in the RANKL basal transcription in osteoblasts and bone marrow stromal cells.


Use of integrative medicine approaches for treating adults with sleep disturbances.
This study determined the prevalence, perceived efficacy, and influencing factors and evaluated the sources of information as well as the barriers and facilitators for the use of integrative medicine approaches (IMA) within the previous 12 months to treat adults with sleep disturbances. No studies have assessed the use of integrative medicine approaches in adults with sleep disturbances. A cross-sectional postal survey was conducted. Adults with sleep disturbances were purposively selected from the sleep center of a hospital in Taiwan. Binary logistic regression was used to analyze and compare the differences between groups of use and non-use IMA. The response rate was 94.5% (n = 515). The prevalence for the use of integrative medicine approaches was 53.4% (n = 275). The most prevalent IMA was music. However, the most perceivably efficacious integrative medicine approaches was acupressure, followed by music and meditation. Independent explanatory variables were educational attainment, the number of chronic diseases/symptoms, and healthy lifestyles for the use of IMA. The most common source of information was television/radio. The top facilitator and barrier for the use of integrative medicine approaches were side effects of sleeping pills and limited accessibility. IMA are used by a considerable proportion of adults for treating sleep disturbances in Taiwan. The findings of this study identified the significant explanatory variables of IMA use and provided a fundamental understanding the aspects of IMA use in the adults with sleep disturbances is particularly important for health care providers in practice associated with their patients.


Bayesian network enables interpretable and state-of-the-art prediction of immunotherapy responses in cancer patients.
Immune checkpoint inhibitors, especially PD-1/PD-L1 blockade, have revolutionized cancer treatment and brought tremendous benefits to patients who otherwise would have had a limited prognosis. Nonetheless, only a small fraction of patients respond to immunotherapy, and the costs and side effects of immune checkpoint inhibitors cannot be ignored. With the advent of machine and deep learning, clinical and genetic data have been used to stratify patient responses to immunotherapy. Unfortunately, these approaches have typically been "black-box" methods that are unable to explain their predictions, thereby hindering their responsible clinical application. Herein, we developed a "white-box" Bayesian network model that achieves accurate and interpretable predictions of immunotherapy responses against nonsmall cell lung cancer (NSCLC). This tree-augmented naïve Bayes (TAN) model accurately predicted durable clinical benefits and distinguished two clinically significant subgroups with distinct prognoses. Furthermore, our state-of-the-art white-box TAN approach achieved greater accuracy than previous methods. We hope that our model will guide clinicians in selecting NSCLC patients who truly require immunotherapy and expect our approach to be easily applied to other types of cancer.


Morphological affinities of a fossil ulna (KNM-WS 65401) from Buluk, Kenya.
The morphological affinities of a primate proximal ulna (KNM-WS 65401) recovered from the late Early Miocene site Buluk, Kenya, are appraised. Nineteen three-dimensional landmarks on ulnae from 36 extant anthropoid species (n = 152 individuals) and KNM-WS 65401, as well as a subset of 14 landmarks on six ulnae belonging to other East African Miocene catarrhine taxa, were collected. To quantify ulnar shape, three-dimensional geometric morphometric techniques were used and linear dimensions commonly cited in the literature were derived from the landmark data. KNM-WS 65401 is situated between monkeys and hominoids in the principal components morphospace. KNM-WS 65401 shares features such as a short olecranon process, broad trochlear notch, and laterally oriented radial notch with extant hominoids, whereas features such as an anteriorly directed trochlear notch and flat, proximodistally elongated, and anteroposteriorly narrow radial notch are shared with extant monkeys. Principal component scores and linear metrics generally align KNM-WS 65401 with both suspensors and arboreal quadrupeds, but quadratic and linear discriminant analyses of principal component score data provide posterior probabilities of 80% and 83%, respectively, for assignment of KNM-WS 65401 to the suspensory group. Compared with fossil ulnae from other Miocene primates, KNM-WS 65401 is morphologically most distinct from KNM-LG 6, attributed to Dendropithecus macinnesi, and morphologically most similar to KNM-WK 16950R, attributed to Turkanapithecus kalakolensis. The KNM-WS 65401 individual likely possessed more enhanced capabilities for elbow joint extension, perhaps during suspensory behaviors, compared with other Miocene primates in the sample.


Dextran sulphate 500 delays and prevents mouse scrapie by impairment of agent replication in spleen.
Treatment of scrapie-infected mice with dextran sulphate (DS) 500 resulted in considerably reduced spleen titres over a long period of time. Subsequently, the central nervous system disease was delayed or even prevented during the 350-day period of observation. Both effects increased after multiple injections of the compound. The potency of DS 500 to protect against scrapie was greatest when treatment and infection were carried out simultaneously. Under these conditions the lethality of 500 to 1000 LD50 was reduced to almost zero. Treatment as early as 10 weeks before infection still prolonged the incubation periods. Of several other polyions tested, dextran sulphate 5 and pentosan polysulphate also impaired scrapie pathogenesis.


The clinical significance of increased large cells in marginal zone lymphoma.
Marginal zone lymphoma (MZL) is a primary, indolent small B-cell lymphoma. Subtypes include nodal, splenic, and those of extranodal mucosa-associated lymphoid tissue (MALT). These are slow growing and generally exhibit low rates of transformation to diffuse large B-cell lymphoma (DLBCL). At initial diagnosis, there can be an increase in large cells (LCs) that does not meet criteria for DLBCL. Prior studies have noted this finding, but the clinical significance of these LCs has not been well established. A total of 161 cases of MZL from 1994 to 2021 were evaluated, including all subtypes. There were 33 cases with increased LCs (>10 LCs per high-power field [hpf]), with the majority containing >15 LCs/hpf (28/33) and 128 cases without increased LCs. Cases with increased LCs were significantly more likely to have a Ki-67 proliferation index of ≥30% (P < .0001). Overall survival was not significantly different between the groups but progression-free survival was significantly worse in the LC group (P < .0001). MZL with increased LCs was also associated with a higher stage at diagnosis (P = .0035), was more likely to transform to DLBCL (P = .0016), and had a greater frequency of relapse (P < .0001). Subgroup analysis showed that both nodal and MALT LC groups had a worse progression-free survival and a higher rate of relapse than their standard nodal and MALT lymphoma counterparts, but only within the MALT subgroup did the LC cases present at a higher stage and have a higher rate of transformation to DLBCL than the standard cases. Although larger studies are needed for validation, these results suggest that the presence of LCs in MZL may serve as a useful prognostic indicator and potentially help guide clinical decision-making.


Discovery and validation of hsa_circ_0001953 as a potential biomarker for proliferative diabetic retinopathy in human blood.
This study aimed to determine whether circular RNAs (circRNAs) in whole blood could be served as novel non-invasive biomarkers for proliferative diabetic retinopathy (PDR). This retrospective cross-sectional study comprised 34 healthy participants, 34 PDR patients and 34 non-proliferative DR (NPDR) patients. High-throughput whole transcriptome sequencing was performed to explore the expression profile of circRNAs in the whole blood, and the candidate circRNAs were validated by quantitative real-time polymerase chain reaction (qRT-PCR). Receiver operating characteristic (ROC) analysis evaluated the ability of these candidate circRNAs in discriminating PDR patients from NPDR patients and healthy subjects. Finally, the networks of circRNA-miRNA-mRNA based on the candidate circRNAs were constructed. Using sequencing and qRT-PCR, hsa_circ_0001953 was found to be elevated in PDR patients in contrast with the other two groups. Statistical analysis showed that the expression levels of hsa_circ_0001953 in PDR patients were positively related to the duration of diabetes and HbAc1. Receiver operating characteristic (ROC) curve analysis revealed that hsa_circ_0001953 was associated with a high diagnostic accuracy in discriminating PDR patients from NPDR patients and healthy controls, resulting in an area under the curve (AUC) of 0.87 and 0.92, respectively. The circRNA-miRNA-target gene networks for hsa_circ_0001953 showed that hsa_circ_0001953 could interact with dozens of miRNAs and some targeted mRNAs have been potentially involved in the pathogenesis of diabetes. The present findings indicate that hsa_circ_0001953 in the whole blood may serve as a novel diagnostic biomarker and potential therapeutic target for PDR.


Ultra-low-dose emergency chest computed tomography protocols in three vendors: A technical note.
In suspected community-acquired pneumonia (CAP), chest CT is superior to the routinely obtained radiographs (CXR), but administers higher radiation doses. However, ultra-low-dose CT (ULDCT) has shown promising results. To compare radiation dose and image quality using standard and ULDCT protocols designed for a multicenter study encompassing three CT scanner models from GE, Canon, and Siemens. Patients with suspected CAP were referred for non-contrast standard dose chest CT (NCCT) and ULDCT. Effective radiation dose and Contrast-to-Noise Ratio (CNR) was calculated. Mean effective doses were GE (n = 10) 6.93 mSv in NCCT and 0.27 mSv in ULDCT; Canon (n = 9) 3.48 in mSv NCCT and 1.11 mSv in ULDCT; Siemens (n = 10) 2.85 mSv in NCCT and 0.45 mSv in ULDCT. CNR was reduced by 29-39% in ULDCT. The proposed CT protocols yielded dose reductions of 96%, 68%, and 84% using a GE, Canon, and Siemens scanner, respectively.


Vegetation analysis of Wadi Al-Jufair, a hyper-arid region in Najd, Saudi Arabia.
Wadi Al-Jufair, a tributary of Wadi Nisah, is one of the important wadis of Najd region (Saudi Arabia) sheltering a rich diversity of higher plants. The study area is extended into approximately 15 km(2) encompassing the commonest geomorphological features encountered in desert wadis. The wadi supports several rare plants, including Maerua crassifolia Forssk., a regionally endangered tree, and Acacia oerfota (Forssk.) Schweinf., a rare shrub with restricted distribution. The present study aims to analyze the vegetation of wadi Al-Jufair and propose its designation as an important plant reserve. The vegetation type is fundamentally of chamaephytic nature with some phanerophytes, and distinguished into associations where the dominant perennial species give the permanent character of plant cover in each habitat. Four vegetation groups were identified with the application of TWINSPAN, DCA and CCA programs and named after the characteristic species as follows: Lycium shawii; A. oerfota; Acacia raddiana-Rhazya stricta and Artemisia monosperma. These plant associations demonstrate significant variation in soil texture, moisture, organic matter, pH, EC, and minerals of Wadi Al-Jufair.


Tissue engineered skin: Apligraf, a bi-layered living skin equivalent.
Driven by the need for a readily available, non-immunological tissue that possesses many of the characteristics of normal human skin, tissue-engineered skin has been developed. For over a decade, laboratory grown or processed skin has been under investigation and, in some cases, available as an alternative to autologous grafts. Apligraf, derived from neonatal foreskin and bovine type I collagen, is the first bi-layered living skin equivalent approved in the US and other countries for use in venous ulcers. Apligraf is effective both in the treatment of refractory venous ulcers and for acute wounds such as surgical excision sites and split thickness donor sites. Apligraf is safe and is not clinically rejected. Its ultimate fate is not known, so it may well work to aid healing in a variety of ways including graft 'take' and as a stimulus for healing.


Ten years (2004-2014) of Chagas disease surveillance and vector control in Ecuador: successes and challenges.
To describe the current situation of Chagas disease in Ecuador and to evaluate the impact of vector control for the period 2004-2014. Since 2004, the Ministry of Public Health has formalized activities for the surveillance and control of Chagas disease and we analyzed here available records. More than 200 000 houses were surveyed, and 2.6% were found to be infested (95% CI: 2.6-2.7), and more than 51 000 houses were sprayed with residual insecticide, with important yearly variations. A total of 915 cases of T. cruzi infection were registered. The Amazon region is emerging as a high priority area, where nearly half of T. cruzi infection cases originate. The costal region and the southern highland valleys remain important high-risk area. Vector control efforts over the past 10 years have been effective in the coastal region, where T. dimidiata predominates, and resulted in important reductions in house infestation indices in many areas, even reaching negligible levels in some parishes. Vector efforts need to be sustained and expanded for the elimination of T. dimidiata to be feasible. Novel vector control interventions need to be designed to reduce intrusion by several triatomine species present in the Amazon region and southern Ecuador. Strong political commitment is needed to sustain current achievements and improve the national coverage of these programmes.


Three-dimensional evaluation of effectiveness of hand and rotary instrumentation for retreatment of canals filled with different materials.
The aim of this study was to measure the remaining filling volume of different obturation materials from root-filled extracted teeth by using 2 removal techniques. Eighty single-rooted teeth were collected and decoronated, and the root canal was prepared by using the ProTaper nickel-titanium rotary files. The teeth were randomly allocated into 4 groups, and each group was obturated by using a different material. Group 1 was filled with gutta-percha and TubliSeal sealer, group 2 was filled with EndoRez points and EndoRez sealer, group 3 was filled with RealSeal points and RealSeal sealer, and Group 4 was filled with a gutta-percha point and GuttaFlow sealer. Teeth were scanned with a micro-computed tomography scan, and then root fillings were removed by using ProTaper retreatment files or hand K-files. Teeth were scanned again, and volume measurements were carried out with micro-computed tomography software. Statistical analysis showed significant differences between the 2 removal techniques for gutta-percha and for both techniques between gutta-percha and the other groups. The present study showed that all tested filling materials were not completely removed during retreatment by using hand or rotary files. Gutta-percha was more efficiently removed by using hand K-files.


[Comparison of the protective effect of garlic extract and cell defense during adaptive response].
The resistance of human cell DNA to damaging doses of CdCl2 or gamma radiation has been investigated after pretreatment with garlic extract (GE) or with adaptive doses of the same mutagens. The adaptive response (AR) and pretreatment with GE stabilize the DNA structure in a similar way. In experiments with 4-nitroquinoline-1-oxide (4-NQO), GE does not stabilize DNA structure but increases the rate and volume of repair of induced breaks. 3-Aminobenzamide (3-AB) increases the number of DNA breaks induced in experiments with CdCl2, gamma radiation, and 4-NQO. This suggests that poly(ADP-ribose)polymerase participates defense of cells from mutagens. Thus, it has been demonstrated that cell defense from CdCl2 or gamma radiation in experiments with GE and AO is mediated by stabilization of DNA structure and in experiments with 4-NQO, by activation of repair of DNA breaks induced.


Chemical Methods for N- and O-Sulfation of Small Molecules, Amino Acids and Peptides.
Sulfation of the amino acid residues of proteins is a significant post-translational modification, the functions of which are yet to be fully understood. Current sulfation methods are limited mainly to O-tyrosine (sY), which requires negatively charged species around the desired amino acid residue and a specific sulfotransferase enzyme. Alternatively, for solid-phase peptide synthesis, a de novo protected sY is required. Therefore, synthetic routes that go beyond O-sulfation are required. We have developed a novel route to N-sulfamation and can dial-in/out O-sulfation (without S-sulfurothiolation), mimicking the initiation step of the ping-pong sulfation mechanism identified in structural biology. This rapid, low-temperature and non-racemising method is applicable to a range of amines, amides, amino acids, and peptide sequences.


Hybrid constructed wetlands for the treatment of wastewater from a fertilizer manufacturing plant: Microcosms and field scale experiments.
Wastewater from a fertilizer manufacturing plant requires improvement prior to its environmental disposal. Ammonium is the critical contaminant to be removed. The aim of this study was to evaluate the feasibility of using free water surface wetlands (FWSWs), horizontal subsurface flow wetlands (HSSFWs), and their combination in hybrid wetlands (HWs) for the final treatment of wastewater with high ammonium concentration from a fertilizer manufacturing plant. Substrates and macrophytes were evaluated in microcosm experiments during three months. There were no significant differences in contaminant removal among HSSFWs with LECA or FWSWs planted with Typha domingensis or Canna indica. In a second stage, two configurations of pilot-scale HWs were constructed at the manufacturing facilities. Configuration A: HSSFW(A1)-FWSW(A2) and Configuration B: FWSW(B1)-HSSFW(B2) were evaluated during 12 months. There were no significant differences in contaminant removal (%) between the two configurations of HWs for COD (A: 74.5 ± 12.2/B: 81.5 ± 9.4), ammonium (A: 59.5 ± 17.5/B: 57.9 ± 21.4), nitrite (A: 79.8 ± 24.2/B: 80.6 ± 16.8) and dissolved inorganic nitrogen (DIN) (A: 59.4 ± 17.3/B: 50.3 ± 24.4). However, nitrate concentration (9.83 ± 3.11 mg N L-1) was significantly lower after Configuration A than after Configuration B (18.8 ± 5.2 mg N L-1). Comparing FWSWs and HSSFWs, they did not present significant differences in ammonium removal, while FWSWs presented the highest DIN removal. T. domingensis and C. indica in HSSFWs and T. domingensis in FWSWs tolerated wastewater conditions. T. domingensis presented the highest productivity. In further research, FWSWs in series planted with T. domingensis should be studied.


Enhanced Thermoelectric Properties of Te Doped Polycrystalline Sn0.94Pb0.01Se.
Thermoelectric materials can directly convert heat and electricity, which is a kind of promising energy material. In view of cost and mechanical properties, polycrystalline SnSe material with high zT value is greatly desired. In this study, polycrystalline Sn0.94Pb0.01Se1-xTex samples were prepared by the vacuum melting-hot pressing sintering method. Sn vacancies, Pb and Te atoms were simultaneously introduced into the polycrystalline SnSe. The power factor of Sn0.94Pb0.01Se1-xTex samples was decreased, which could be attributed to the generation of n-type semiconductor SnSe2. In addition, the phonons were strongly scattered by point defects and dislocations, which led to the decrease of thermal conductivity-from 0.43 Wm-1K-1 to 0.29 Wm-1K-1 at 750 K. Finally, the polycrystalline Sn0.94Pb0.01Se0.96Te0.04 sample achieved the maximum zT value of 0.60 at 750 K.


Evidence supporting performance measures of laparoscopic appendectomy through a novel surgical proficiency assessment tool and low-cost laparoscopic training system.
Laparoscopic training remains inaccessible for surgeons in low- and middle-income countries, limiting its widespread adoption. We developed a novel tool for assessment of laparoscopic appendectomy skills through ALL-SAFE, a low-cost laparoscopy training system. This pilot study in Ethiopia, Cameroon, and the USA assessed appendectomy skills using the ALL-SAFE training system. Performance measures were captured using the ALL-SAFE verification of proficiency tool (APPY-VOP), consisting of a checklist, modified Objective Structured Assessment of Technical Skills (m-OSATS), and final rating. Twenty participants, including novice (n = 11), intermediate (n = 8), and expert (n = 1), completed an online module covering appendicitis management and psychomotor skills in laparoscopic appendectomy. After viewing an expert skills demonstration video, participants recorded their performance within ALL-SAFE. Using the APPY-VOP, participants rated their own and three peer videos. We used the Kruskal-Wallis test and a Many-Facet Rasch Model to evaluate (i) capacity of APPY-VOP to differentiate performance levels, (ii) correlation among three APPY-VOP components, and (iii) rating differences across groups. Checklist scores increased from novice (M = 21.02) to intermediate (M = 23.64) and expert (M = 28.25), with differentiation between experts and novices, P = 0.005. All five m-OSATS domains and global summed, total summed, and final rating discriminated across all performance levels (P < 0.001). APPY-VOP final ratings adequately discriminated Competent (M = 2.0), Borderline (N = 1.8), and Not Competent (M = 1.4) performances, Χ2 (2,85) = 32.3, P = 0.001. There was a positive correlation between ALL-SAFE checklist and m-OSATS summed scores, r(83) = 0.63, P < 0.001. Comparison of ratings suggested no differences across expertise levels (P = 0.69) or location (P = 0.66). APPY-VOP effectively discriminated between novice and expert performance in laparoscopic appendectomy skills in a simulated setting. Scoring alignment across raters suggests consistent evaluation, independent of expertise. These results support the use of APPY-VOP among all skill levels inside a peer rating system. Future studies will focus on correlating proficiency to clinical practice and scaling ALL-SAFE to other settings.


Impacts of crab bioturbation and local pollution on sulfate reduction, Hg distribution and methylation in mangrove sediments, Rio de Janeiro, Brazil.
Mercury (Hg) and methylmercury (MeHg) are highly toxic and poorly studied in mangroves. Burrowing Uca crabs change sediment topography and biogeochemistry and thus may affect Hg distribution and MeHg formation. We studied added (203)Hg distribution, Me(203)Hg formation and sulfate reduction rates (SRR) in sediment aquariums containing Uca leptodactyla; and analyzed profiles of Me(203)Hg formation and SRR in sediment cores from two mangroves with distinct environmental impacts. MeHg formation and SRR were higher in the top (≤6cm) sediment and there was no significant difference in Hg methylation in more or less impacted mangroves. In aquariums, crab bioturbation favored Hg retention in the sediment. In the treatment without crabs, Hg volatilization and water Hg concentrations were higher. Hg methylation was higher in bioturbated aquariums but SRR were similar in both treatments. These findings suggest that bioturbating activity favors Hg retention in sediment but also promotes MeHg formation near the surface.


Characterization of Cerebral Edema in Acute-on-Chronic Liver Failure.
The nature of cerebral edema in acute-on-chronic liver failure (ACLF) is not well studied. We aimed to characterize cerebral edema in ACLF using magnetization transfer ratio (MTR) and diffusion tensor imaging (DTI). Forty-six patients with cirrhosis and acute decompensation were included. Patients were divided into groups A (no cerebral failure, n = 39) and B (cerebral failure, n = 7). Group A was subdivided into no-ACLF (n = 11), grade 1 (n = 10), grade 2 (n = 9) and grade 3 (n = 9) ACLF as per CANONIC study. MRI brain and plasma TNF-alpha, IL-1beta and IL-6 were measured at baseline and 7-10 days after admission. Ten age- and sex-matched healthy controls were also included. Mean diffusivity (MD) values, an MRI marker of water content, progressively increased from controls to no-ACLF to ACLF grade 1, 2 and 3 in group A in frontal white matter (FWM) and basal ganglia (P < 0.0001). MD values improved only in survivors on follow-up. MD values correlated with IL-6 levels at baseline. On multivariate analysis MELD score ≥28 and MD values (>8 × 10-9 M2/s) in FWM were independent predictors of 90-day mortality. There was no significant difference in clinical and MRI parameters between group A and B. Cerebral edema increases with severity of ACLF. Correlation between MD values and IL-6 levels suggests pathogenic role of inflammation in cerebral edema. Patients with grade 3 ACLF have cerebral edema irrespective of presence of clinically evident cerebral failure. MELD score and cerebral edema have prognostic significance in ACLF.


The Endocannabinoid Analgesic Entourage Effect: Investigations in Cultured DRG Neurons.
The endocannabinoid 2-Arachidonyl glycerol (2-AG) exerts dose-related anti-nociceptive effects, which are potentiated by the related but inactive 2-palmitoyl glycerol (2-PG) and 2-linoleoyl glycerol (2-LG). This potentiation of analgesia and other in vivo measures was described as the "entourage effect". We investigated this effect on TRPV1 signalling in cultured dorsal root ganglion (DRG) nociceptors. Adult rat DRG neurons were cultured in medium containing NGF and GDNF at 37°C. 48 h later cultures were loaded with 2 µM Fura2AM for calcium imaging, and treated with 2-AG, 2-PG and 2-LG, individually or combined, for 5 min, followed by 1 µMol capsaicin. The amplitude and latency of capsaicin responses were measured (N=3-7 rats, controls N=16), and analysed. In controls, 1 µMol capsaicin elicited immediate calcium influx in a subset of neurons, with average latency of 1.27 ± 0.2 s and amplitude of 0.15 ± 0.01 Units. 2-AG (10-100 µMol) elicited calcium influx in some neurons. In the presence of 2-AG (0.001-100 µMol), capsaicin responses were markedly delayed in 64% neurons by up to 320 s (P<0.001). 2-PG increased capsaicin response latency at 0.1 nMol-100 µMol (P<0.001), in 60% neurons, as did 2-LG at 0.1-100 µMol (P<0.001), in 76% neurons. Increased capsaicin response latency due to 2-AG and 2-PG was sensitive to the CB2 but not to the CB1 receptor antagonist. Combined application of 1 µMol 2-AG, 5 µMol 2-PG and 10 µMol 2-LG, also resulted in significantly increased capsaicin response latency up to 281.5 ± 41.5 s (P<0.001), in 96% neurons, that was partially restored by the CB2, but not the CB1 antagonist. 2-AG, 2-LG and 2-PG significantly delayed TRPV1 signalling in the majority of capsaicin-sensitive DRG neurons, that was markedly increased following combined application. Further studies of these endocannabinoids are required to identify the underlying mechanisms.


Keratins in oral cancer: necessity of mass spectrometry for validation of antibody based identifications.
Keratins are intermediate filament family proteins which are predominantly expressed in the epithelial cells. Most of the studies which evaluate the status of keratins in clinical samples of the oral cavity are based on the identification of their presence and localization by immunohistochemistry using monoclonal antibodies. It is very well known that many monoclonal/polyclonal antibodies show cross-reactivity with the other closely related or non-related proteins. This cross-reactivity might be the result of epitope similarity, but it is not always necessary. Therefore studies done with only antibody based techniques can mislead interpretation unless they are validated with additional techniques like mass-spectrometry. In this investigation we have evaluated the status of keratin 18 in cancer of buccal mucosa using 1DE, 2DE and western blotting with monoclonal antibody to keratin 18. The patterns emerging showed aberrant as well as differential expression of K18 in adjacent normal versus tumor tissue samples of buccal mucosa. Mass spectrometry analysis of the immunodetected spots however revealed that it is keratin 13. Thus this study emphasizes the necessity of validation of antibody based findings when dealing with proteins of a large family having similarity/homology in amino acid sequence.


Topographical distribution in the adult rat brain of neurotrophic activities directed to central nervous system targets.
Cortex, hippocampus, septum and striatum of day 18 rat embryos were grafted to several brain regions of young adult rats which had been lesioned in the chosen area 4 days earlier. Thirty days after transplantation, the grafts were fixed and morphometrically analysed under light microscope. The volumes, neuronal densities and total number of neurons of the transplants were compared. Each graft survived best when transplanted to its original region. Good survival was also achieved by heterotopic grafts between regions that are anatomically related. Striatal grafts showed reasonable survival only when transplanted to their original site. In a second series of experiences, the neurons from the same embryonic brain regions were cultured in a defined medium, to which was added tissue extracts from the lesioned regions of the adult brain. The neuronal survival was estimated. The in vitro results are closely related to those obtained in vivo. This experimental evidence agrees with the theory of the existence of a retrograde transport of NGF from the hippocampus to the septum, sustaining the survival of the latter. On the other hand, our results demonstrate the existence of other unidentified neurotrophic factors in the central nervous system which differ from one region to another.


Efficacy, safety, and molecular response predictors of oral ixazomib and short-course rituximab in untreated iNHL.
Patients with indolent B-cell non-Hodgkin lymphoma (iNHL) generally require treatment but experience normal survival, emphasizing the need for simpler, safer therapies. Proteasome inhibitors target aberrant signaling pathways within iNHL and have manageable toxicities. We evaluated the oral proteasome inhibitor ixazomib as initial monotherapy, and combined with rituximab, for first-line treatment of iNHL. Treatment-naïve patients with iNHL needing therapy received oral ixazomib 4 mg weekly until progressive disease or unacceptable adverse events. A 4-week course of rituximab was added during month 7. The primary end point was overall response rate (ORR) during the ixazomib monotherapy window. Correlations included gene expression profiling and response to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccination. Thirty-three patients with follicular lymphoma (FL) (n = 20), marginal zone lymphoma (n = 7), and other iNHL were treated with a median follow-up of 30.3 months. During the 6-month ixazomib window, the ORR was 24%, including 35% in FL. The best ORR over the entire study period was 52% overall and 65% in FL; complete response was achieved in 33% and 45%, respectively. The median duration of response was 25.8 months (range, 0-49.7), and the 24-month progression-free and overall survival rates were 51% (95% confidence interval [CI], 32-67) and 91% (95% CI, 74-97), respectively. Ixazomib was well tolerated. Baseline downregulation of proteasome genes, PSMB9 (P = .03) and PSMB8 (P = .007), were associated with response. All evaluated patients generated anti-S antibodies to SARS-CoV-2 vaccination, with a median of 254.9 binding arbitrary unit per mL. Ixazomib demonstrated efficacy alone and with short-course rituximab in untreated iNHL while exhibiting favorable toxicity, convenience, and retention of the B-cell immune response. This trial is registered at www.clinicaltrials.gov as NCT02339922.


Molecular causes of an evolutionary shift along the parasitism-mutualism continuum in a bacterial symbiont.
Symbiosis with microbes is a ubiquitous phenomenon with a massive impact on all living organisms, shaping the world around us today. Theoretical and experimental studies show that vertical transmission of symbionts leads to the evolution of mutualistic traits, whereas horizontal transmission facilitates the emergence of parasitic features. However, these studies focused on phenotypic data, and we know little about underlying molecular changes at the genomic level. Here, we combined an experimental evolution approach with infection assays, genome resequencing, and global gene expression analysis to study the effect of transmission mode on an obligate intracellular bacterial symbiont. We show that a dramatic shift in the frequency of genetic variants, coupled with major changes in gene expression, allow the symbiont to alter its position in the parasitism-mutualism continuum depending on the mode of between-host transmission. We found that increased parasitism in horizontally transmitted chlamydiae residing in amoebae was a result of processes occurring at the infectious stage of the symbiont's developmental cycle. Specifically, genes involved in energy production required for extracellular survival and the type III secretion system-the symbiont's primary virulence mechanism-were significantly up-regulated. Our results identify the genomic and transcriptional dynamics sufficient to favor parasitic or mutualistic strategies.


Leaf-twig carbon isotope ratio differences in photosynthetic-twig desert shrubs.
Carbon isotope ratios were determined for bulk tissues of both leaves and current season twigs of 29 species of Mohave Desert shrubs. Leaf and twig tissues were found to differ in their carbon isotope ratios, only in those species which had photosynthetic twigs. These data suggest that the twigs of these species operate at lower intercellular CO2 values than leaves, an interpretation which is consistent with available gas-exchange data. An effect of microhabitat was also evident between the mean isotope ratios of leaves from wash versus slope habitats.


Genetic polymorphisms of the sortase A gene and early childhood caries in two-year-old children.
To explore and compare the genetic polymorphisms of the sortase A (srtA) gene found in Streptococcus mutans (S. mutans) infecting two-year-old children suffering early childhood caries to those found in caries-free children through molecular identification methods. Clinical S. mutans strains were isolated from the dental plaques of two-year-old children. Fifteen strains of S. mutans from the caries-active group and 15 strains of S. mutans from the caries-free group were collected. Genomic DNA was extracted from the S. mutans isolates. DNA fragments, including the srtA gene, were amplified by PCR. The PCR products were purified, sequenced and analyzed. A chi-square test and BioEdit software were used to analyze the sequencing results. All 30 clinically isolated S. mutans strains had a 741 base pair (bp) srtA gene. There were no nucleotide sequence insertions or deletions observed in the srtA genes. Twenty mutations were identified in the srtA genes that taken from the 30 clinical strains. There were 10 silent point mutations at the 78, 99, 150, 165, 186, 222, 249, 261, 312, and 636bp positions. The other 10 mutations were point mutations resulting in a missense mutation at the 23, 34, 36, 47, 112, 114, 168, 176, 470, and 671bp positions. None of the positions were enzyme-activity sites of srt A. The missense mutation rates of the two groups did not exhibit statistically significant differences. There were no genetic polymorphisms of the sortase A gene associated with early childhood caries in two-year-old children.


Delays in starting morning operating lists: an analysis of more than 20,000 cases in 22 German hospitals.
Delays in the start of the first operation of the day often lead to conflicts among the involved physicians and nurses. Data on such delays have already been published for individual hospitals, but robust comparative data from a large number of institutions have not been available till now. The study is based on the operating room (OR) documentation of four surgical services (general surgery, trauma/orthopedic surgery, gynecology, and ear nose throat [ENT] surgery) in 22 German hospitals over a nine-month period. Three process points ("patient arrival in OR suite," "anesthesia ready," and "incision") were analyzed for the first operation of the day in each OR. 21,357 operations in the first position were analyzed. The percentage of delays differed markedly for the three process points. The incision was delayed in more than 70% of the general surgical and trauma/orthopedic cases, but less often in gynecological (61 ± 24%) and ENT cases (42 ± 29%). The frequency of delays longer than 10 minutes was between 20% and 40%. The mean delay in delayed cases ranged from 14.1 ± 5.4 to 21.6 ± 8.2 minutes depending on the type of service and process point. The processes for the first operation of the day are not optimally structured in the hospitals whose cases were analyzed in this study. Delayed starts were common.


FTIR detection of water reactions in the oxygen-evolving centre of photosystem II.
Flash-induced Fourier transform infrared (FTIR) difference spectroscopy has been used to study the water-oxidizing reactions in the oxygen-evolving centre of photosystem II. Reactions of water molecules were directly monitored by detecting the OH stretching bands of weakly H-bonded OH of water in the 3700-3500 cm(-1) region in FTIR difference spectra during S-state cycling. In the S1-->S2 transition, a band shift from 3588 to 3617 cm(-1) was observed, indicative of a weakened H-bond. Decoupling experiments using D2O:H2O (1:1) showed that this OH arose from a water molecule with an asymmetric H-bonding structure and this asymmetry became more significant upon S2 formation. In the S2-->S3, S3-->S0 and S0-->S1 transitions, negative bands were observed at 3634, 3621 and 3612 cm(-1), respectively, representing formation of a strong H-bond or a proton release reaction. In addition, using complex spectral features in the carboxylate stretching region (1600-1300 cm-(1)) as 'fingerprints' of individual S-state transitions, pH dependency of the transition efficiencies and the effect of dehydration were examined to obtain the information of proton release and water insertion steps in the S-state cycle. Low-pH inhibition of the S2-->S3, S3-->S0 and S0-->S1 transitions was consistent with a view that protons are released in the three transitions other than S1-->S2, while relatively high susceptibility to dehydration in the S2-->S3 and S3-->S0 transitions suggested the insertion of substrate water into the system during these transitions. Thus, a possible mechanism of water oxidation to explain the FTIR data is proposed.


Interleukin 6 SNP rs1800797 associates with the risk of adult-onset asthma.
Interleukin 6 (IL6) is an inflammatory cytokine that has been suggested to have an important role in the pathogenesis of asthma. IL6 single-nucleotide polymorphisms (SNPs) have been associated with levels of IL6, and with childhood and prevalent adult asthma. A recent study also suggested that IL6 SNPs associate especially with atopic asthma. However, association of IL6 SNPs with adult-onset asthma has not been studied. In a population-based study of 467 incident adult-onset asthma cases and 613 disease-free controls from South Finland, we analyzed association of 6 tagging SNPs of the IL6 locus with the risk of adult-onset asthma and with atopy. Asthma was clinically diagnosed, and atopy was defined based on Phadiatop test. IL6 SNP rs1800797 associated with the risk of adult-onset asthma in a log additive model, with adjusted odds ratio (aOR) 1.31 (95% confidence interval 1.09-1.57), and especially with the risk of atopic adult-onset asthma when compared with non-atopic controls, aOR 1.46 (95% CI 1.12-1.90). This is the first study to show an association of IL6 with adult-onset asthma, and especially with atopic adult-onset asthma.


Intracellular antibody immunity.
Antibodies allow the immune system to target pathogens despite their tremendous diversity and rapid evolution. Once bound to a pathogen, antibodies induce a broad range of effector mechanisms, including phagocytosis and complement. However, these mechanisms are all initiated in the extracellular space, meaning that pathogens like viruses evade them upon infection of their target cells. Recently, it has been shown that, in addition to mediating extracellular immune responses, antibodies also activate immunity inside infected cells. Antibodies that are bound to the surface of non-enveloped viruses or bacteria are carried into the cell during pathogen entry. Once inside the cell, these pathogen-attached antibodies are recognised by a highly conserved, high affinity cytosolic antibody receptor called TRIM21. TRIM21 initiates both sensor and effector responses that reduce viral replication and induce an antiviral state. These responses are an important part of antiviral immunity and the removal of TRIM21 results in uncontrolled viraemia and death in a mouse model of infection.


Phytase from antarctic yeast strain Cryptococcus laurentii AL27.
Cryptococcus laurentii strain AL27 demonstrating significant potential for intracellular phytase production was selected by 2-step screening of Antarctic yeasts. The strain showed increased phytase activity in a culture medium with 40 g/L sucrose, KH2PO4 providing 5 mg/L phosphorus, and cultivation temperature of 24 degrees C, which relates it to psychrotrophic microorganisms. The enzyme kinetic characteristics according to sodium phytate were Km = 0.98 mmol/L, vlim = 33.3 micromol g(-1) min(-1). The enzyme had maximum activity at 40 degrees C and acted within a wide pH range: from 2.0 to 5.5, which is of positive significance for its direct inclusion into the feed of monogastric animals.


Hemodynamic effects of different preparations of stroma free hemolysates in the isolated perfused rat kidney.
We have examined the effects of Stroma Free Hemolysate (SFH) solutions in the isolated perfused rat kidney. Three types of SFH, stored for 6 to 8 months at 4 degrees C, were tested: 1) unmodified, 2) glyoxalated and lightly cross linked and 3) pyridoxalated and polymerized. All three SFH solutions, added to the perfusate at a concentration of approximately 420 mg/100ml, increased renal vascular resistance (RVR) and reduced glomerular filtration rate (GFR). Unmodified, glyoxalated and lightly cross linked and pyridoxalated polymerized SFH resulted in a rise in RVR of 55%, 38% and 33% respectively and a fall in GFR of 42%, 57% and 83% respectively. In order to determine whether storage had altered the effect of SFH on renal function, one of the forms of SFH (glyoxalated and lightly cross linked) was studied only 4-6 weeks after preparation. While this preparation caused an increase in RVR of 41% it did not alter GFR; filtration fraction (FF) rose. However, after further storage of this preparation for 6-7 months, the solution resulted in a marked decrease in GFR of 47% as well as a rise in RVR of 23%. We conclude that three different SFH preparations resulted in marked vasoconstriction and reductions in GFR. These deleterious effects on renal hemodynamics were noted at a concentration of hemoglobin well below that necessary to effectively improve oxygen content. Storage of the SFH solutions may cause or contribute to their effects on renal function. SFH solutions intended for use as blood substitutes should be tested for vasoconstrictor activity.


Differences in the diagnostic value between fiberoptic and high definition laryngoscopy for the characterisation of pharyngeal and laryngeal lesions: A multi-observer paired analysis of videos.
High definition laryngoscopy (HDL) could lead to better interpretation of the pharyngeal and laryngeal mucosa than regularly used fiberoptic laryngoscopy (FOL). The primary aim of this study is to quantify the diagnostic advantage of HDL over FOL in detecting mucosal anomalies in general, in differentiating malignant from benign lesions and in predicting specific histological entities. The secondary aim is to analyse image quality of both laryngoscopes. Retrospective paired analysis with multiple observers evaluating endoscopic videos simulating daily clinical practice. A tertiary referral hospital. In 36 patients, both FOL and HDL videos were obtained. Six observers were provided with additional clinical information, and 36 FOL and HDL videos were evaluated in a randomised order. Sensitivity, specificity, positive and negative predictive value and diagnostic accuracy of observers using both flexible laryngoscopes were calculated for detection of mucosal lesions in general and uncovering malignant lesions. Sensitivities were calculated for prediction of specific histological entities. Image quality (scale 1-10) was assessed for both flexible laryngoscopes. HDL reached higher sensitivity compared to FOL for detection of mucosal abnormalities in general (96.0% vs 90.4%; P = .03), differentiating malignant from benign lesions (91.7% vs 79.8%; P = .03) and prediction of specific histological entities (59.7% vs 47.2%; P < .01). Image quality was judged better with HDL in comparison with FOL (mean: 8.4 vs 5.4, P < .01). HDL is superior to FOL in detecting mucosal anomalies in general, malignancies and specific histological entities. Image quality is considered as superior using HDL compared to FOL.


The meaning of the Tp-Te interval and its diagnostic value.
The interval between T peak (Tp) and T end (Te) has been proposed as a measure of transmural dispersion of repolarization, but experimental and clinical studies to validate Tp-Te have given conflicting results. We have investigated the meaning of Tp-Te and its diagnostic potential. We used a digital model of the left ventricular wall to simulate the effect of varying action potential durations on the timing of Tp and Te. Furthermore, we used the vectorcardiogram to explain the relationships between Tp locations in the precordial electrocardiogram leads. Prolongation or ischemic shortening of action potentials in our model did not result in substantial Tp shifts. The phase relationships revealed by the vectorcardiogram showed that Tp-Te in the precordial leads is a derivative of T loop morphology. Tp-Te is the resultant of the global distribution of the repolarization process and is a surrogate diagnostic parameter.


Blood viscosity and its relationship to iron deficiency, symptoms, and exercise capacity in adults with cyanotic congenital heart disease.
This study sought to determine the relationship between blood viscosity and iron deficiency and their impact on symptoms and exercise function in adults with cyanotic congenital heart disease. Iron deficiency is believed to raise whole blood viscosity in cyanotic congenital heart disease, although available data are inconsistent. Thirty-nine cyanotic adults were prospectively assessed for iron deficiency (transferrin saturation < or =5%), hyperviscosity symptoms, and exercise capacity. Same-day measurement of whole blood viscosity and hematocrit (Hct) adjusted viscosity (cells resuspended in autologous plasma to Hct of 45%) was performed at shear rates ranging from 0.277 s(-1) to 128.5 s(-1). Viscosity did not differ between patients with iron deficiency (n = 14) and those without (n = 25). Whole blood viscosity correlated with Hct (r = 0.63, p < 0.001 at low shear and r = 0.84, p < 0.001 at high shear) but not with red blood cell size or iron indices. Hyperviscosity symptoms were independent of iron indices but directly correlated with increased Hct-adjusted viscosity (r = 0.41, p = 0.01). Exercise capacity did not differ in iron-deficient patients. However, peak oxygen consumption was higher in those with Hct > or = 65% (12.6 +/- 3.4 ml/kg/m2 vs. 9.8 +/- 2.6 ml/kg/m2, mean +/- SD, p = 0.036) despite higher whole blood viscosity in these same individuals (p < 0.01 for all shear rates). Iron deficiency is common in cyanotic adults but does not alter viscosity. Hyperviscosity symptoms are associated with a higher Hct-adjusted viscosity independent of cell size or iron stores. Higher Hct is associated with better exercise capacity. Further work to understand the origin of hyperviscosity symptoms is warranted.


Endophytic fungi isolated from medicinal plants: future prospects of bioactive natural products from Tabebuia/Handroanthus endophytes.
Medicinal plants are a rich source of natural products used to treat many diseases; therefore, they are the basis for a new drug discovery. Plants are capable of generating different bioactive secondary metabolites, but a large amount of botanical material is often necessary to obtain small amounts of the target substance. Nowadays, many medicinal plants are becoming rather scarce. For this reason, it is important to point out the interactions between endophytic microorganisms and the host plant, because endophytes are able to produce highly diverse compounds, including those from host plants that have important biological activities. Thence, this review aims at presenting the richness in bioactive compounds of the medicinal plants from Tabebuia and Handroanthus genera, as well as important aspects about endophyte-plant interactions, with emphasis on the production of bioactive compounds by endophytic fungi, which has been isolated from various medicinal plants for such a purpose. Furthermore, bio-prospection of natural products synthesized by endophytes isolated from the aforementioned genera used in traditional medicine could be used to treat illnesses.


New insights in endogenous modulation of ligand-gated ion channels: histamine is an inverse agonist at strychnine sensitive glycine receptors.
Histamine is involved in many physiological functions in the periphery and is an important neurotransmitter in the brain. It acts on metabotropic H1-H4 receptors mediating vasodilatation, bronchoconstriction and stimulation of gastric acid secretion. In the brain histamine is produced by neurons in the tuberomamillary nucleus (TMN), which controls arousal. Histamine is also a positive modulator of the inhibitory Cys-loop ligand-gated ion channel GABAA. We investigated now its effect on the second member of inhibitory Cys-loop ligand-gated ion channels, the strychnine sensitive glycine receptor. We expressed different human and rat glycine receptor subunits in Xenopus laevis oocytes and characterized the effect of histamine using the two electrode voltage clamp technique. Furthermore we investigated native glycine receptors in hypothalamic neurons using the patch-clamp technique. Histamine inhibited α1β glycine receptors with an IC50 of 5.2±0.3 mM. In presence of 10 mM histamine the glycine dose-response curve was shifted, increasing the EC50 for glycine from 25.5±1.4 μM to 42.4±2.3 μM. In addition, histamine blocked the spontaneous activity of RNA-edited α3β glycine receptors. Histamine inhibited glycine receptors expressed in hypothalamic TMN neurons with an IC50 of 4.6±0.3 mM. Our results give strong evidence that histamine is acting on the same binding site as glycine, being an inverse agonist that competitively antagonizes glycine receptors. Thus, we revealed histamine as an endogenous modulator of glycine receptors.


Pilot Study: Next-generation Sequencing of the Semen Microbiome in Vasectomized Versus Nonvasectomized Men.
Approximately half a million vasectomies are performed every year in the USA. There is a paucity of literature on the impact of male sterilization on the semen microbiome and whether it prompts microbiota dysbiosis. To investigate if vasectomy induces changes in the seminal microbiome via comparison of semen samples from men before and after vasectomy, and if the seminal microbiome profiles for vasectomized men follow a particular pattern with respect to diversity and abundance. From July 2021 to February 2022, we prospectively collected and analyzed semen samples from 58 men at one outpatient clinic. Eighteen men provided a semen sample before and 3 mo after vasectomy. We also collected semen samples from 22 fertile nonvasectomized men and from a further 18 vasectomized men at 3 mo after vasectomy. Semen microbiome α-diversity, beta-diversity, and relative abundance were compared initially between paired and then between unpaired vasectomized and nonvasectomized samples. Analysis of variance (ANOVA), permutational multivariate ANOVA, and analysis of the composition of microbiomes with bias correction were used to assess differences. In both paired and unpaired sets of samples, a decreasing trend for α-diversity in semen after vasectomy was observed. Shannon diversity, the relative abundance of species with an abundance >2%, and composition were not significantly changed. Sphingomonas, Brevundimonas, and Paracoccus abundance decreased after vasectomy, while Corynebacterium abundance increased. The results may be limited by the sample size and lack of demographic heterogeneity. Vasectomy is followed by a decrease in α-diversity and changes in the relative abundance of bacterial species in the semen microbiome. Further investigation is necessary to understand the clinical significance of these changes after vasectomy. We evaluated changes in the bacteria species in semen after vasectomy. We found that vasectomy decreased the richness and evenness of bacteria species in semen, but the overall bacterial community remained similar. Further studies are needed to assess the implications of changes in semen bacteria after vasectomy.


Molsidomine enhances the protective activity of valproate against pentylenetetrazole-induced seizures in mice.
Molsidomine (25 mg kg(-1)), a donor of nitric oxide, commonly used in the treatment of coronary artery disease, enhanced the protective activity of valproate against the clonic phase of pentylenetetrazole-induced seizures in mice, significantly reducing the ED(50) of valproate from 123.5 to 78 mg kg(-1). Molsidomine was found to be ineffective with respect to the protective action of clonazepam, ethosuximide and phenobarbital. Alone, molsidomine in a dose of 25 mg kg(-1) was ineffective in this model of seizures. Since N(G)-nitro-L-arginine, an inhibitor of nitric oxide synthase, failed to reverse the effect of molsidomine on valproate, an involvement of nitric oxide in the mechanism of the anticonvulsive efficacy of valproate does not seem to be probable. Molsidomine (25 mg kg(-1)) significantly elevated the free plasma level of valproate from 33.8 to 46.2 microg ml(-1). Therefore, we conclude that the interaction of molsidomine with valproate is at the pharmacokinetic level. The combination of valproate with molsidomine appears beneficial because is free from adverse effects, in terms of motor impairment and long-term memory deficit. Our results suggest that the dosage of valproate in patients with coronary artery disease treated with molsidomine should be decreased. It would allow us to reduce adverse effects of valproate.


Secondary stroke prevention with apixaban in nonvalvular atrial fibrillation: A subgroup analysis of the STANDARD study.
Data on independent risk factors for stroke recurrence in Japanese patients with nonvalvular atrial fibrillation are limited. We performed a subgroup analysis of a postmarketing surveillance study of apixaban (STroke prevention ANticoagulant Drug Apixaban Real-world Data study) in Japanese patients with nonvalvular atrial fibrillation receiving oral apixaban (5 mg/2.5 mg twice daily) in routine clinical practice. Patients were categorized into primary and secondary prevention groups based on the absence or presence of a history of ischemic stroke/transient ischemic attack, respectively. Patients in the secondary prevention group (1101 of 6306 patients [17.5%] analyzed; mean observation period, 15.7 months) had a higher risk of ischemic stroke or hemorrhage than those in the primary prevention group. The incidence rates of major (3.92%/year vs 2.06%/year), intracranial (1.87%/year vs 0.55%/year), and cerebral (1.14%/year vs 0.37%/year) hemorrhage and effectiveness outcomes (ischemic stroke/systemic embolism/transient ischemic attack, 3.25%/year vs 0.57%/year) were significantly higher (all P < 0.001) in the secondary prevention group than in the primary prevention group. Multivariate analysis identified no independent risk factors in the secondary prevention group, while prior major bleeding, alcohol abuse, advanced age, male sex, lower body weight, higher serum creatinine, and antiplatelet drug use were identified as risk factors for major hemorrhage, and advanced age and antiplatelet drug use for effectiveness outcomes in the primary prevention group. Among Japanese patients with nonvalvular atrial fibrillation who received apixaban, presence of a history of ischemic stroke/transient ischemic attack was associated with higher incidence rates of hemorrhage and thromboembolic events.


Different approaches for dealing with rare variants in family-based genetic studies: application of a Genetic Analysis Workshop 17 problem.
Rare variants are becoming the new candidates in the search for genetic variants that predispose individuals to a phenotype of interest. Their low prevalence in a population requires the development of dedicated detection and analytical methods. A family-based approach could greatly enhance their detection and interpretation because rare variants are nearly family specific. In this report, we test several distinct approaches for analyzing the information provided by rare and common variants and how they can be effectively used to pinpoint putative candidate genes for follow-up studies. The analyses were performed on the mini-exome data set provided by Genetic Analysis Workshop 17. Eight approaches were tested, four using the trait's heritability estimates and four using QTDT models. These methods had their sensitivity, specificity, and positive and negative predictive values compared in light of the simulation parameters. Our results highlight important limitations of current methods to deal with rare and common variants, all methods presented a reduced specificity and, consequently, prone to false positive associations. Methods analyzing common variants information showed an enhanced sensibility when compared to rare variants methods. Furthermore, our limited knowledge of the use of biological databases for gene annotations, possibly for use as covariates in regression models, imposes a barrier to further research.


Bioconcentration ratio of diazinon by freshwater fish and snail.
The bioconcentration ratios of diazinon from water by freshwater fishes were generally larger than that of crayfish and snails. Among fishes, the bioconcentration ratio of diazinon by topmouth gudgeon was the highest value, 152 being average. However, elimination of diazinon from fish body was linearly rapid. The influence of test concentration centration ratio of diazinon in whole body of topmouth gudgeon was increased proportional to the body weight.


Complex form of instrumental behavior of dogs.
A complex form of higher nervous activity, conditional reflex transswitching or switching, was elaborated in four dogs under conditions of unrestrained movements, freedom of reinforcement choice (food or water), place of reinforcement determined by situation factors as well as independent switching on of conditional stimuli. It was shown that motivated goal-directed behavior of the animals was determined by activation of forward and backward connections. The chains of instrumental conditioned reflexes forming in the final analysis the complex behavior are developed according to the "trial-and-error" principle. In the instrumental conditioned reflexes there are two constantly coexisting types of mutually complementing conditioned connections of the signal with reinforcement--direct and indirect. In experiments with unrestrained conditions of animals and independent regulation of the experiment, one of the frequently encountered physiological mechanisms of generalization appears to be efferent and afferent generalization.


Cytogenetical and hematological analysis of chronic myelogenous leukemia patients with a novel case 52XX, t (1;9;22) (q23.3; q34; q11.2), +6, +8, i(9) (q10;q10), +18,+19,+21+der22 t(9;22)(q34;q11).
(9;22) (q34; q11) translocation is appear in above ninety percent of chronic myelogenous leukemia patients while variant/complex translocations were observed in almost 5% to 8% chronic myelogenous leukemia (CML) positive cases. Gleevec (Imatinib Mesylate) is the first choice breakpoint cluster region (BCR)/ABL targeted oral therapy that produced a complete response almost in 71% to 80% of patients affected with CML. A complete blood count (CBC) of 37 patients was done during diagnosis, however only 21 showed abnormal CBC values which were selected for the study. Karyotyping study using bone marrow samples was performed on 21 CML patients for the conformation of 9;22, however, fluorescence in situ hybridisation was performed for the detection of the BCR-ABL fusion gene of 15 patients. Out of 21, 17 patients showed Ph-positive (9;22) (q34; q11) translocation. Sixteen CML patients showed standard translocation however only CML patients showed a three-way variant/complex translocation with six additional chromosomes, 52XX, t(1;9;22) (q23.3;q34;q11),+6,+8, i(9)(q10;q10), +18,+19,+21 + der22 t(9;22)(q34;q11)). Here we report we report a novel case of six additional chromosomes with the three-way translocation of 52XX, t(1;9;22) (q23.3;q34;q11),+6,+8, i(9)(q10;q10), +18,+19,+21 + der22 t(9;22)(q34;q11) in blast phase.


Variations in white blood count, thromboxane B2 levels and hematocrit in chronic venous hypertension.
To analyze variations in leukocyte count and thromboxane B2 production in the femoral vein of patients with chronic venous hypertension (CVH). Prospective clinical study, controlled, non randomized and open. Hospital das Clínicas, Faculdade de Medicina da Universidade de São Paulo, referral center, university hospital. 15 patients with recurring stasis ulcer were analyzed, selected randomly from the venous diseases outpatient center, and 4 without lower limb venous alterations were also analyzed. Blood samples from the femoral and brachial veins were drawn following supine and 45 degrees reverse Trendelenburg. Direct leukocyte count and analysis of the thromboxane B2 with enzyme linked immunosorbent assay test. After 30 minutes in reverse Trendelenburg, patients with CVH showed a leukocyte count reduced by +/- 27% (p = 0.02) and thromboxane B2 levels increased by +/- 158% (p = 0.02). We suggest that future studies of medications for stasis ulcers include their effects on leukocyte entrapment and thromboxane B2 production in the lower limb venous system.


Magnetic Resonance Imaging of Fistula-In-Ano.
This article explains the pathogenesis of fistula-in-ano and details the different classifications of fistula encountered, describe their features on MR imaging, and explains how imaging influences subsequent surgical treatment and ultimate clinical outcome. Precise preoperative characterization of the anatomic course of the fistula and all associated infection via MR imaging is critical for surgery to be most effective. MR imaging is the preeminent imaging modality used to answer pertinent surgical questions.


SMG6 regulates DNA damage and cell survival in Hippo pathway kinase LATS2-inactivated malignant mesothelioma.
Many genes responsible for Malignant mesothelioma (MM) have been identified as tumor suppressor genes and it is difficult to target these genes directly at a molecular level. We searched for the gene which showed synthetic lethal phenotype with LATS2, one of the MM causative genes and one of the kinases in the Hippo pathway. Here we showed that knockdown of SMG6 results in synthetic lethality in LATS2-inactivated cells. We found that this synthetic lethality required the nuclear translocation of YAP1 and TAZ. Both are downstream factors of the Hippo pathway. We also demonstrated that this synthetic lethality did not require SMG6 in nonsense-mediated mRNA decay (NMD) but in regulating telomerase reverse transcriptase (TERT) activity. In addition, the RNA-dependent DNA polymerase (RdDP) activity of TERT was required for this synthetic lethal phenotype. We confirmed the inhibitory effects of LATS2 and SMG6 on cell proliferation in vivo. The result suggests an interaction between the Hippo and TERT signaling pathways. We also propose that SMG6 and TERT are novel molecular target candidates for LATS2-inactivated cancers such as MM.


Morphology, physiology, and biochemistry of zinc-stressed caraway plants.
A greenhouse pot experiment was conducted to evaluate the impact of zinc supply (0, 1, and 2 mM Zn as ZnSO4) on morpho-physiological and biochemical parameters of caraway (Carum carvi L.). Exposure to different Zn concentrations for 12 weeks compromised severely all growth parameters (plant height, number of secondary branches, diameter of primary and secondary branches, fresh and dry weight of aerial parts and roots) yield and its components (number of umbels per primary branches and secondary branches; number of umbel per plant; number of seeds per plant; and the weight of 1000 seeds). These manifestations were intimately linked with excessive accumulation of Zn in roots and leaves, alteration of the content of photosynthetic pigments, and extended lipid peroxidation. A manifest increment of proline and soluble sugar content was also observed in response to Zn application. Lipid content in seeds was dropped in Zn-treated plants and the fatty acid profiles were profoundly affected as they were enriched with saturated fatty acids at the expense of unsaturated ones. While improving their oxidative stability as revealed by the reduced values calculated oxidizability and oxidative susceptibility, Zn treatment reduced the lipid nutritional quality of caraway seeds. Moreover, Zn treatment reduced the essential oil yield and its main component carvone while it enhanced the content of its precursor limonene. It also induced alteration of terpene metabolism as revealed in the redirection of the carbon flux to the shikimate/phenylpropanoid pathway resulting in the stimulation of the production of phenolic compounds and their subsequent antioxidant activities.


Comparison of Outcomes of Two Femoral Fixation Devices in Hamstring Anterior Cruciate Ligament Reconstruction.
Tear of the anterior cruciate ligament (ACL) is a common ligamentous injury of the knee. Reconstruction of this ligament is often required to restore functional stability of the knee. Outcome of ACL reconstruction is significantly affected by how the graft is fixed to the bone. This study is to determine if there is a different clinical outcome after cortical versus cortical-cancellous suspension femoral fixation in hamstring based anterior cruciate ligament (ACL) reconstruction. This is a retrospective comparative study conducted between 2006 and 2010. We enrolled patients who underwent arthroscopic ACL reconstruction. Sixty two patients met inclusion criteria and 41 agreed to come for followup assessment. Median age was of 28 years (range 18-39 years). Demographic baseline profile of both groups was similar. The femoral fixation devices were cortical (n = 16) and cortical-cancellous suspension techniques (n = 25). The average period of evolution at the time of assessment was 40 months (range 12-72 months). The patients were examined according to Lachman test (using Rolimeter knee tester), anterior drawer test, pivot shift test, International Knee Documentation Committee questionnaire, and Tegner-Lysholm knee scoring scale. The objective evaluation of the patients (Lachman test) showed better results in terms of stability in the group of patients who underwent the cortical-cancellous suspension method. These differences were not reflected in the assessment of activity level (Tegner-Lysholm), where both groups showed the same results. ACL reconstruction with both cortical and cortical-cancellous suspension femoral fixation techniques show the same clinical results at medium long followup. However, cortical-cancellous fixations seem to provide greater stability to the reconstruction.


Parathyroid hormone - a bone anabolic and catabolic agent.
A key factor in the control of bone remodelling is parathyroid hormone (PTH), the principal regulator of calcium homeostasis. Elevated levels of PTH increase bone turnover, leading to either anabolic or catabolic effects on the skeleton depending upon the pattern and duration of elevation. New evidence indicating that downregulation of an osteocyte signal (sclerostin, the SOST gene product) occurs in response to intermittent PTH has rekindled interest in the key role played by osteocytes and bone-lining cells in co-ordinating surface anabolic activity. Microarray analysis has also delineated many genes and pathways regulated by intermittent and continuous PTH in osteoblasts and whole bones.


The role of platelets in peripheral arterial disease: therapeutic implications.
Peripheral arterial disease (PAD) is associated with platelet hyperaggregability as well as an increase in morbidity and mortality from myocardial infarction and stroke. Enhanced platelet activation in PAD may substantially contribute to these adverse outcomes. A relative resistance to aspirin therapy has been reported in patients with PAD. Therefore, clopidogrel may be superior to aspirin in treatment of PAD. Furthermore, the aspirin + clopidogrel combination could be more effective than monotherapy but its risk-benefit ratio has yet to be evaluated. Clopidogrel is preferable to ticlopidine because of its safer profile and the convenience of once-daily administration. The glycoprotein (Gp) IIb/IIIa inhibitors may also find a place as short-term therapy after peripheral angioplasty. There is a need to consider the use of clopidogrel in patients who cannot tolerate aspirin. Patients who have an event while taking aspirin also present a problem. One possibility here is to substitute aspirin with clopidogrel or to add clopidogrel to the aspirin. Although these options are currently not evidence based in patients with PAD, there is emerging evidence showing that they are realistic choices.


Modifications of laccase activities of copper efflux oxidase, CueO by synergistic mutations in the first and second coordination spheres of the type I copper center.
The redox potential of type I copper in the Escherichia coli multicopper oxidase CueO was shifted in the positive or negative direction as a result of the single, double, and triple mutations in the first and second coordination spheres: the formation of the NH···S(-)(Cys500 ligand) hydrogen bond, the breakdown of the NH(His443 ligand)···O(-)(Asp439) hydrogen bond, and the substitution of the Met510 ligand for the non-coordinating Leu or coordinating Gln. Laccase activities of CueO were maximally enhanced 140-fold by virtue of the synergistic effect of mild mutations at and at around the ligand groups to type I copper.