Prolonged exposure to particulate Cr(VI) is cytotoxic and genotoxic to fin whale cells.
Hexavalent chromium [Cr(VI)] is a human lung carcinogen and global marine pollutant. High Cr concentrations, resembling the ones observed in occupationally exposed workers, have been observed in fin whales (Balaenoptera physalus) in the Gulf of Maine. This outcome suggests Cr might be disrupting the health of fin whale populations. Indeed, Cr in acute (24 h) exposure does cause toxicity in fin whale cells. However, human cell culture data indicate prolonged exposures (120 h) induce a higher amount of toxicity compared to 24 h exposure due to an inhibition of homologous recombination repair. However, whether prolonged exposure causes similar outcomes in fin whale cells is unknown. Due to the importance of assessing prolonged exposure toxicity, this study focuses on characterizing acute and prolonged exposure of Cr(VI) in male and female fin whale cells. Cytotoxicity was measured by the clonogenic assay, also known as colony forming assay, which measures the ability of cells to proliferate and form colonies after the treatment. DNA double strand breaks were analyzed by neutral comet assay. Clastogenicity was measured using the chromosome aberration assay. Intracellular Cr levels were measured with Graphite Furnace Atomic Absorption Spectrometry (GFAAS) with Syngistix Software. In this study, we demonstrate that particulate Cr(VI) induces cytotoxicity and genotoxicity in a treatment-dependent manner after 24 h and 120 h exposures. Cytotoxicity levels were generally low with relative survival above 64 %. DNA double strand break data and chromosome aberration data were elevated after a 24 h exposure, but decreased after a 120 h exposure. While cytotoxicity was similar after 24 h and 120 h exposures, less DNA double strand breaks and chromosomal instability occurred with prolonged exposure. Particulate Cr(VI) is cytotoxic and genotoxic to fin whale cells after acute and prolonged exposures. The reduction of genotoxicity we have observed after 120 h exposure may be partly explained by lower intracellular Cr levels after 120 h. However, the decrease in intracellular levels is not reflected by a similar decrease in chromosome aberrations suggesting other mechanisms may be at play. Male fin whale cells appear to be more susceptible to the genotoxic effects of particulate Cr(VI) while female cells are less susceptible possibly due to increased cell death of damaged cells, but more work is needed to clarify if this outcome reflects a sex difference or interindividual variability. Overall, the study shows particulate Cr(VI) does induce toxicity at both acute and prolonged exposures in fin whales cells indicating Cr(VI) exposure is a health risk for this species.


Effects of MTNR1B genetic variants on the risk of type 2 diabetes mellitus: A meta-analysis.
Whether melatonin receptor 1B (MTNR1B) variants are associated with type 2 diabetes mellitus (T2DM) remains unclear. Therefore, we performed this meta-analysis to better explore correlations between MTNR1B variants and T2DM. Literature research was performed in PubMed, Medline, and Embase. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated. Totally 21 studies were enrolled to analyses. Pooled overall analyses showed that MTNR1B rs10830963 variant was significantly correlated with the susceptibility to T2DM (allele model: p = 0.02, OR = 0.97, 95% CI 0.95-1.00). Further subgroup analyses by ethnicity of participants revealed that rs10830963 variant was significantly correlated with the susceptibility to T2DM in South Asians, but not in Caucasians or East Asians. No any other positive results were found in overall and subgroup analyses. Our findings indicated that MTNR1B rs10830963 variant might serve as a genetic biomarker of T2DM, especially in South Asians.


Membrane currents of rat satellite cells attached to intact skeletal muscle fibers.
Muscle satellite cells play an important role in the postnatal growth of skeletal muscle and in the regeneration of damaged muscle during adult life. Little is known about the physiological properties of satellite cells in their dormant state as they lie adjacent to the intact muscle fibers, underneath the basement membrane. Our recent experiments, using patch clamp techniques, indicate that no tight electrical coupling is present between satellite cells and the muscle fiber dissociated from rat flexor digitorum brevis. Satellite cells possess sodium channels with low sensitivity to tetrodotoxin and at a much lower density than muscle. In addition, satellite cells are insensitive to acetylcholine (ACh) for at least 24 hr after having been removed from the animal, even when detached from their muscle fiber. However, we could measure ACh-evoked currents from satellite cells 48-72 hr in culture, indicating that ACh sensitivity develops with time.


Different therapies of Chinese herbal medicine for diarrhea-predominant irritable bowel syndrome: A network meta-analysis of double-blinded, placebo-controlled trials.
Shuganjianpi Therapy (SGJP), Jianpi Therapy (JP), Shugan Therapy (SG), Jianpiwenshen Therapy (JPWS), and Shuganjianpiwenshen Therapy (SGJPWS), consisting of formulas from Chinese herbal medicine (CHM), have been tremendously applied to irritable bowel syndrome (IBS). However, it remains uncertain when exploring the preferable option among different CHM therapies for diarrhea-predominant irritable bowel syndrome (IBS-D). To compare and rank the efficacy and safety of different CHM therapies for IBS-D. We searched randomized, double-blinded, placebo-controlled trials through mainstream databases from their inception to October 31, 2022. Eligible randomized controlled trials (RCTs) applied one of the CHM therapies as the experimental group and placebo as the control group. Two authors independently extracted data into a form and evaluated the quality of the retrieved articles by the Cochrane Risk of Bias Tool. At least one of the following outcomes was assessed: Serotonin, Neuropeptide Y (NPY), Incidence of Adverse Events (AE), and Irritable Bowel Syndrome-Severity Scoring System (IBS-SSS) with its subscales of Severity of Abdominal Pain (SAP), Frequency of Abdominal Pain (FAP), Severity of Abdominal Distension (SAD), Dissatisfaction with Bowel Habits (DBH), and Interference with Quality of Life (IQOL). A Bayesian network meta-analysis on a random-effect model was conducted using R 4.2.2 software. 1367 records were retrieved from databases in an initial search. Fourteen studies involving six interventions with 2248 participants were identified. Provided pairwise comparisons, the surface under the cumulative ranking curve (SUCRA) ranking, and cluster analysis, JPWS was the best option for ameliorating clinical symptoms simultaneously, which included IBS-SSS, SAP, FAP, SAD, DBH, and IQOL. As for AE, JPWS contributed to fewer adverse events than others as well. In respect of serum indicators, we noticed the dominance of SGJP in regulating both serotonin and NPY. JPWS and SGJP were the most prominent CHM therapies for IBS-D in terms of clinical symptoms, including abdominal pain, distension, bowel habits, and improvement of quality of life. The effect of JP and SG for IBS-D required further investigation. As a potential candidate, SGJP may well treat IBS-D by mediating dysmotility, visceral hypersensitivity, and the gut-brain axis with an increase of NPY and a reduction of serotonin. For safety, JPWS was ideal for the fewest adverse events in the treatment of IBS-D. On account of a small sample size and possible geographical publication bias, more double-blinded and placebo-controlled trials with larger samples worldwide would be necessary for strengthening current evidence.


Construction of recombinant vaccinia virus strains using single-stranded DNA insertion vectors.
The ability of single-stranded (ss) DNA, isolated from recombinant M13 bacteriophage, to direct the insertion of foreign genetic elements into the vaccinia virus (VV) genome was examined. An identical chimeric transcriptional unit [VV promoter/chloramphenicol acetyl transferase (CAT) gene embedded in DNA sequences encoding vaccinia virus thymidine kinase (TK)] was inserted into either the previously characterized plasmid insertion vector, pGS20, or into M13mp18. It was found that the ss vector (M13mp18:TK/CAT) was four times more efficient than the plasmid vector (pGS20:CAT) in catalyzing homologous recombination of the cat gene by marker transfer into the VV genome. Furthermore, Southern blot analyses and CAT enzymatic activity assays confirmed that the structure of the M13-derived recombinant genomes were as expected and that the chimeric genes were fully active. Although the precise mechanism responsible for the ss DNA-catalyzed insertion event is not known, these results are discussed with respect to the advantages of using M13-based vectors with which to manipulate and insert genetic information into infectious VV recombinants.


A Study of Individualized Diagnosis and Treatment for Depression with Atypical Features (iDoT-AFD): study protocol for a randomized clinical trial and prognosis study.
Major depressive disorder (MDD) with atypical features, namely depression with atypical features (AFD), is one of the most common clinical specifiers of MDD, closely associated with bipolar disorder (BD). However, there is still a lack of clinical guidelines for the diagnosis, treatment, and prognosis of AFD. Our study mainly focuses on three issues about how to identify AFD, what is the appropriate individualized treatment for AFD, and what are the predictive biomarkers of conversion to BD. The Study of Individualized Diagnosis and Treatment for Depression with Atypical Features (iDoT-AFD) is a multicenter, prospective, open-label study consisting of a 12-week randomized controlled trial (RCT) and a continued follow-up until 4 years or reaching the study endpoint. It is enrolling 480 patients with AFD (120 per treatment arm), 100 patients with BD, and 100 healthy controls (HC). Multivariate dimension information is collected including clinical features, cognitive function, kynurenine pathway metabolomics, and multimodal magnetic resonance imaging (MRI) data. Firstly, multivariate informatics analyses are performed to recognize patients with AFD from participants including the first-episode and recurrent atypical depression, patients with BD, and patients with HC. Secondly, patients with atypical depression are randomly allocated to one of the four treatment groups including "single application of selective serotonin reuptake inhibitor (SSRI) or serotonin-noradrenaline reuptake inhibitor (SNRI)", "SSRI/SNRI combined with mood stabilizer," "SSRI/SNRI combined with quetiapine (≥ 150 mg/day)," or "treatment as usual (TAU)" and then followed up 12 weeks to find out the optimized treatment strategies. Thirdly, patients with atypical depression are followed up until 4 years or switching to BD, to explore the risk factors of conversion from atypical depression to BD and eventually build the risk warning model of conversion to BD. The first enrolment was in August 2019. The iDoT-AFD study explores the clinical and biological markers for the diagnosis, treatment, and prognosis of AFD and further provides evidence for clinical guidelines of AFD. ClinicalTrials.gov NCT04209166. Registered on December 19, 2019.


Characterization and functional performance of a commercial human conjunctival epithelial cell line.
The conjunctiva is a complex tissue that covers the eye beginning at the corneal limbus and extending over the inner surfaces of the eyelids. Due to its important functions in maintaining the health of the ocular surface, adequate in vitro models of conjunctival structure and function are essential to understand its roll in different pathologies. Because there is scarcity of human conjunctival tissue that can be used in research, cell lines are often the only option for initial studies. An immortalized human conjunctival epithelial cell (IM-HConEpiC) line is now commercially available; however, it is not very well characterized. In this study, we have developed a new protocol to culture these cells without the use of collagen-coated culture surfaces, but with a defined cell culture medium. We characterized IM-HConEpiCs cultured under these conditions and corroborated that the cells maintained a conjunctival epithelial phenotype, including acidic and neutral mucins, junctional proteins E-cadherin and zonula occludens 1, and expression of CK8 and CK19, among others. In addition, we analyzed the response to oxidative stress and inflammatory stimuli and found that IM-HConEpiCs respond as expected for conjunctival epithelial tissue. For instance, cells exposed to oxidative stress increased the production of reactive oxygen species, and that increase was blocked in the presence of an antioxidant agent. In addition, after stimulation with TNF-α, IM-HConEpiCs significantly increased the production of IL-1β, IL-6, IL-8, and IP-10. Therefore, with this study we conclude that IM-HConEpiCs can be a useful tool in functional studies to determine the response of the conjunctiva to pathological conditions and/or to test new therapeutic strategies.


Antioxidants: the good, the bad and the ugly.
Inflammation has a fundamental role in mediating all stages of atherosclerotic disease. The key role of oxidation in linking lipids and inflammation to atherosclerosis is compelling and is supported by experimental evidence. However, the relevance of the antioxidant hypothesis for the treatment of patients with atherosclerosis has not been definitively proven. Results of randomized trials with 'antioxidant' vitamins have been disappointing, and there are potentially important problems associated with their use, including their potential pro-oxidant effects. Probucol has reduced postpercutaneous coronary intervention (PCI)-restenosis and progression of carotid atherosclerosis in clinical trials. The antioxidant vascular protectant AGI-1067 has also been effective at preventing atherosclerosis in all tested animal models. The nonintervened reference coronary segments of the PCI vessel demonstrated improvements with AGI-1067 in the Canadian Antioxidant Restenosis Trial-1 (CART-1), evidence supportive of a clinical effect on slowing atherosclerosis progression. Two trials test the antioxidant/anti-inflammatory hypothesis with AGI-1067; CART-2 assesses its value for the reduction of both atherosclerosis progression and post-PCI restenosis, and Aggressive Reduction of Inflammation Stops Events (ARISE), which is evaluating its effects on hard cardiovascular outcomes.


Enhancing magnetoresistance in tetrathiafulvalene carboxylate modified iron oxide nanoparticle assemblies.
We report a facile approach to stabilize Fe3O4 nanoparticles (NPs) by using tetrathiafulvalene carboxylate (TTF-COO(-)) and to control electron transport with an enhanced magnetoresistance (MR) effect in TTF-COO-Fe3O4 NP assemblies. This TTF-COO-coating is advantageous over other conventional organic coatings, making it possible to develop stable Fe3O4 NP arrays for sensitive spintronics applications.


Network Load Balancing Using Modular Arithmetic Computations.
Load-balanced routing has attracted considerable attention, especially in the recent years, where huge data volumes are carried over the computer networks. It is particularly important for non-all-to-all networks, where there is no direct communication between all the nodes of the network.Telecommunication and network systems constitute complex dynamic systems with an ever-increasing number of users and network services. It has become apparent that, new routing demands can not be easily satisfied by conventional routing methods. Thus, intelligent optimization methods (e.g., nature-inspired methodologies) have arisen to improve network efficiency.This paper presents a computational method that is based on modular arithmetic for achieving dynamic load balancing on data networks. The proposed algorithm organizes the overall communication into equal-sized packets, it divides the communication into a series of communication steps between the network nodes, and performs packet transfer. The last section includes discussion on the main costs each network routing operation inures: the data movement cost, the load information cost and the data reordering cost.


Gossypetin- based therapeutics for cognitive dysfunction in chronic unpredictable stress- exposed mice.
Chronic unpredictable stress (CUS) is a promising model for induction of cognition impairment. Stress induced memory dysfunction is linked to the activation of kynurenine (KYN) pathway. This pathway indicates that, chronic stress primarily promotes the release of excessive cortisol from the adrenal gland, which tends to activate microglia and further increases kynurenine and its downstream pathway, resulting in excessive quinolinic acid (QA), which further impairs brain derived neurotrophic factor (BDNF) levels and leads to neurodegeneration. Prior studies already established anti-oxidant and anti-depressant activity of gossypetin. This research study was mainly conducted to elaborate neuroprotective activity of gossypetin against CUS-induced cognition impairment via acting on kynurenine pathway. In this study, Swiss albino mice were exposed to various stressors for five weeks and then administered with gossypetin (5, 10 and 20 mg/kg, i.p.) from the 4th to the 7th week (from day 22 to 49). Several behavioral tests were carried out between days 36 to 49 (6th and 7th week) and further corticosterone, neurotransmitters, oxidative stress, and brain-derived neurotrophic factor (BDNF) levels were measured. Results state that CUS exposed mice showed significant improvement in the behavioral pattern after gossypetin treatment. Corticosterone levels and oxidative stress was also found to be significantly decreased in gossypetin (10 and 20 mg/kg, i.p.) treated mice when compared with CUS exposed mice. Whereas, serotonin, norepinephrine and BDNF levels were also found to be increased after gossypetin treatment. Hence, gossypetin can be considered as a neuroprotective agent against cognition impairment caused by chronic unpredictable stress.


Selective inhibition of collagen biosynthesis in the dental pulps of glucocorticoid-treated rabbits.
Prednisolone (2 mg/kg) was injected daily for 14 days. Collagen and protein biosynthesis were measured in isolated molar and incisor pulps by the incorporation of [14C]-proline into protein and collagen in vitro. Collagen solubility, free proline content, prolyl hydroxylase activity, collagenolytic activity and DNA and RNA contents were also assayed. Rabbits injected with saline or starved served as controls. Collagen synthesis was inhibited selectively in both prednisolone and starvation groups. No other aspect of collagen and protein metabolism was affected by either prednisolone treatment or starvation. Thus glucocorticoid administration reduces collagen formation in the pulp, resembling the anti-anabolic effect of starvation. Glucocorticoid treatment at high daily dosages, therefore, may disturb normal development and metabolism of teeth.


Association between periodontitis and metabolic syndrome in the Hamburg City Health Study.
Previous studies demonstrated an association between severe chronic periodontitis (CP) and metabolic syndrome (MetS). However, these studies mostly used the outdated National Cholesterol Education Program-Adult Treatment Panel (NCEP-ATP) III case definition of MetS. Additionally, CP was rarely diagnosed based on a full-mouth examination. Thus, the aim of the current study was to re-evaluate the potential association between CP and MetS in the Hamburg City Health Study (HCHS), a large population-based survey of middle-aged and elderly men and women in Germany, in view of more current definitions of MetS and CP. A cross-sectional study was performed with baseline-data from participants of the HCHS. Periodontitis severity grades were determined in a random sample of 6,209 participants of which 5,456 had sufficient data to call absence or presence of MetS. Variables defining MetS according to the currently valid harmonized definition were determined and a full-mouth examination was performed, including determination of the clinical attachment loss, bleeding on probing, and dental plaque index. CP was classified in three grades of severity (none/mild, moderate, and severe). The Kruskal-Wallis test or the Chi-squared test were used for descriptive statistics and multivariate logistic regression models with and without adjustments for potential confounders (age, sex, smoking, high-sensitivity C-reactive protein [hsCRP], energy intake, and physical activity) were used to test for associations. The prevalence of MetS (39.0%) increased according to the severity grades of periodontitis (none/mild: 33.6%; moderate: 38.7%, and severe: 46.8%). Multivariate logistic regression analyses demonstrated that severe but not moderate CP was associated with MetS after adjusting for age and sex (odds ratio [OR], 1.24; 95% confidence interval [CI], 1.03 to 1.48; P = 0.02). However, the association was attenuated after additional adjustment for smoking (OR, 1.19; 95% CI, 0.99 to 1.43; P = 0.058) and hsCRP, energy intake, and physical activity (OR, 1.11; 95% CI, 0.91 to 1.36; P = 0.294). The use of the more current definitions for MetS and CP confirmed previous observations of an age- and sex-adjusted association between severe CP and MetS. Smoking, high-energy intake, and low physical activity were identified as important lifestyle-related confounders. Abdominal obesity, as indicated by elevated waist circumference, was determined as the most important component of MetS in relationship to CP.


How important is the CO2 chemoreflex for the control of breathing? Environmental and evolutionary considerations.
Haldane and Priestley (1905) discovered that the ventilatory control system is highly sensitive to CO2. This "CO2 chemoreflex" has been interpreted to dominate control of resting arterial PCO2/pH (PaCO2/pHa) by monitoring PaCO2/pHa and altering ventilation through negative feedback. However, PaCO2/pHa varies little in mammals as ventilation tightly couples to metabolic demands, which may minimize chemoreflex control of PaCO2. The purpose of this synthesis is to (1) interpret data from experimental models with meager CO2 chemoreflexes to infer their role in ventilatory control of steady-state PaCO2, and (2) identify physiological causes of respiratory acidosis occurring normally across vertebrate classes. Interestingly, multiple rodent and amphibian models with minimal/absent CO2 chemoreflexes exhibit normal ventilation, gas exchange, and PaCO2/pHa. The chemoreflex, therefore, plays at most a minor role in ventilatory control at rest; however, the chemoreflex may be critical for recovering PaCO2 following acute respiratory acidosis induced by breath-holding and activity in many ectothermic vertebrates. An apparently small role for CO2 feedback in the genesis of normal breathing contradicts the prevailing view that central CO2/pH chemoreceptors increased in importance throughout vertebrate evolution. Since the CO2 chemoreflex contributes minimally to resting ventilation, these CO2 chemoreceptors may have instead decreased importance throughout tetrapod evolution, particularly with the onset and refinement of neural innovations that improved the matching of ventilation to tissue metabolic demands. This distinct and elusive "metabolic ventilatory drive" likely underlies steady-state PaCO2 in air-breathers. Uncovering the mechanisms and evolution of the metabolic ventilatory drive presents a challenge to clinically-oriented and comparative respiratory physiologists alike.


Clinical features and outcome of eight patients with Chlamydia psittaci pneumonia diagnosed by targeted next generation sequencing.
The clinical symptoms of Chlamydia psittaci pneumonia are still poorly understood. This study was designed to summarize the clinical features and outcome of eight C. psittaci pneumonia patients diagnosed by targeted next generation sequencing (tNGS). We included eight C. psittaci pneumonia patients admitted to our hospital from January 2021 to July 2022. The tNGS was performed to the samples collected from bronchoalveolar lavage fluid of each patient. Their clinical data were analysed, including baseline features, clinical symptoms, chest radiographic findings and laboratory examinations. The tNGS sequence number for C. psittaci was in a range of 10 to 1722. The radiographic characteristics were mainly featured by patchy consolidation shadows, ground-glass density shadows, air bronchogram and slight pleural effusion. Within 1-3 days after hospitalization, most patients showed increased neutrophil ratio, C-reactive protein and erythrocyte sedimentation rate, and decreased lymphocyte count, total protein, albumin and prealbumin. Some patients showed increased glutamic-pyruvic transaminase, glutamic-oxaloacetic transaminase and lactate dehydrogenase levels. Three critically ill patients showed increased creatine kinase, creatine kinase isoenzyme and high-sensitivity troponin T (hs-TnT) levels. A poultry or bird contact history, typical flu-like symptoms, patchy consolidation, ground-glass density shadow and air bronchogram may contribute to the diagnosis and treatment of C. psittaci pneumonia. Increase in creatine kinase, creatine kinase isoenzyme and hs-TnT may indicate a severe condition. Moxifloxacin and minocycline were effective in the management of C. psittaci pneumonia.


Mycobacterial infection in the ferret.
Mycobacteriosis is an important disease in the feral ferret (Mustela putorius furo) of New Zealand; elsewhere, reports of tuberculosis in the ferret are sporadic. Genus Mycobacterium consists of aerobic, non-spore-forming, gram-positive, nonmotile bacteria that characteristically feature a cell wall rich in mycolic acids and esters. The epidemiology of mycobacteriosis in the ferrets of New Zealand involves complex interactions between ferrets, possums, and livestock. Investigators have shown that the ferret is highly susceptible only to Mycobacterium bovis infection and is more resistant to infection by other Mycobacterium spp. The principal site of all mycobacterial infection in the ferret is the gastrointestinal tract.


First trimester surgical abortion practices: a survey of National Abortion Federation members.
The objective of this study was to survey first trimester surgical abortion practices of North American providers. A survey was mailed to the 310 active member clinics of the National Abortion Federation, the professional organization of abortion providers in North America. The 236 respondents (76%) comprising the study group provided approximately 30% and 33% of all first trimester abortions in the US and Canada, respectively. Of the 529 practitioners who perform most abortions at the clinics, 68% are obstetrician-gynecologists, and 18% are family physicians; 51% are at least 50 years old. The majority of respondents (156 of 236, 66%) confirm gestational age with ultrasonography. Of the 202 clinics that favor one method of anesthesia, 58% use local cervical block with or without oral pre-medication, 32% combine local anesthesia with intravenous sedation, and 10% use general anesthesia. Most clinics open the cervix with tapered dilators (207 of 233, 89%), evacuate the uterus by using curved rigid plastic cannulas (157 of 229, 69%) and an electric vacuum source (191 of 233, 82%), and explore the uterus with a metal curette following aspiration (133 of 232, 57%). A large majority of respondents examine the uterine aspirate on-site (91%), prescribe antibiotics (91%), and offer oral contraceptives (95%) when requested. Although variations exist, surgical techniques and postoperative practices are quite uniform among clinics. Local anesthesia is presently the most frequent method of pain control. Ultrasonographic dating before abortion is common practice. The "graying" of skilled practitioners raises concerns about the future availability of abortion.


The New Zealand cataract and refractive surgery survey 1997/1998.
This study documents the current practice for cataract and refractive surgery in New Zealand. A postal questionnaire was distributed in late 1997 to all consultant members of the Ophthalmological Society of New Zealand that were resident in the country at that time. Most questions were identical to the 1997 survey of the American Society of Cataract and Refraction Surgeons (ASCRS) to enable a comparison. There were 98 returns from 101 surveys distributed. Of the returns, 72 performed cataract surgery, 23 performed PRK and 11 performed LASIK. ASCRS members did more refractive surgery than did New Zealanders: 28 versus 1% of 1-5 RK per month, 7 versus 1% of 1-2 clear lens extractions per month and 85 versus 51% had access to an excimer laser. For cataract surgery, ASCRS members used more topical anaesthesia (30 vs 5.5%), used no sutures more often (73 vs 51%), used more preoperative antibiotics (76 vs 26%) and used fewer injections of antibiotic/steroids (38 vs 61%). Otherwise the two groups were broadly similar.


Environmental determinants of Kaposi's sarcoma-associated herpesvirus (KSHV) transmission in rural Uganda (ENDKU study): Contributions to research on KSHV infection and reactivation in African children; A longitudinal cohort study.
The Environmental Determinants of KSHV transmission in rural Uganda (ENDKU) study began enrollment in February 2020 with the purpose of defining the relationship between malaria, primarily caused by Plasmodium falciparum in sub-Saharan Africa, and KSHV susceptibility and reactivation. Uganda is an ideal study site, because both malaria and KSHV are endemic and widespread, even among young children. ENDKU is a longitudinal cohort study of infants enrolled at six months of age and followed until three years of age. The main study, and one smaller sub-study, is nested within the General Population Cohort (GPC), a long-standing population cohort in rural Uganda. The ENDKU study was created to test the hypothesis that P. falciparum malaria increases an infant's susceptibility to KSHV infection. A sub-study to evaluate the effects of P. falciparum on KSHV reactivation involves an additional cohort of 5-10-year-old children with and without acute malaria who presented to the GPC study clinic. For each study, participants provided demographic and behavioral data through administered questionnaires and blood and saliva samples. Despite barriers presented by the COVID-19 pandemic, the study team was able to leverage the long-standing relationship of the UK Medical Research Council and the Uganda Virus Research Institute (MRC/UVRI) with the community, a strong commitment to research, and a multi-disciplinary team of experts to successfully implement the ENDKU study. The results of this multi-pronged approach will answer important questions about the etiology and transmission of KSHV in sub-Saharan Africa and the data and samples collected will be an important future resource for scientific research in the region.


Do hypothalamic regulatory factors other than luteinizing hormone releasing factor exert a priming effect?
Extracts of hypothalamic tissue were applied to pituitary tissue in vitro to see whether they could exert a priming effect on gonadotrophin, adrenocorticotrophin, thyrotrophin or prolactin secretion. A clear-cut priming effect was seen only for gonadotrophin secretion.


RBBP6, a RING finger-domain E3 ubiquitin ligase, induces epithelial-mesenchymal transition and promotes metastasis of colorectal cancer.
RBBP6 has been implicated in tumorigenesis but its role in tumor metastasis and progression has not been evaluated. Interestingly, here we show that RBBP6 is upregulated in colorectal cancer (CRC) where its expression level is positively correlated with distant metastasis. In this study, we identified RBBP6, a RING Finger-domain E3 ubiquitin ligase, served as an independent prognostic factor and predicted poor outcome for CRC patients. RBBP6 promoted cell proliferation, migration, and invasion in CRC cells and promoted tumor growth, lung metastasis, and liver metastasis in mouse models. Mechanistically, we revealed that RBBP6 bound and ubiquitylated IκBα, an inhibitor of the NF-κB-signaling pathway. RBBP6-mediated ubiquitination and degradation of IκBα significantly enhanced p65 nuclear translocation, which triggered the activation of NF-κB pathway and then induced the epithelial-mesenchymal transition (EMT) process and cell metastasis. Furthermore, by DNA methylation results and ChIP analysis, we demonstrated that the promoter of RBBP6 was hypomethylated, and was activated by multi-oncogenic transcription factors. In conclusion, our findings suggest that RBBP6 may be a potential prognostic biomarker and therapeutic target for CRC invasion and metastasis.


Laparoscopic gastric resection for gastrointestinal stromal tumors.
This study aimed to review clinical outcomes for patients selected to undergo laparoscopic resection for gastrointestinal stromal tumor (GIST) of the stomach. All 112 laparoscopic gastric resections performed from February 1995 to March 2007 were reviewed. Pre- and postoperative variables were analyzed, and data are given as mean +/- standard deviation. Laparoscopic gastric resection was attempted for 63 GIST in 61 patients (31 men and 30 women) with a mean age was 59.1 +/- 19 years. The tumors were located at the fundus (n = 19), antrum (n = 18), body (n = 17), gastroesophageal junction/cardia (n = 7), and pylorus (n = 2). Common presentations were upper gastrointestinal bleed (n = 29) and incidental finding on esophagogastroduodenoscopy (n = 17). The laparoscopic procedures performed were partial gastrectomy (n = 52), antrectomy (n = 4), esophagogastrectomy (n = 3), and endoscopically assisted and/or transgastric resection (n = 3). There was one conversion to open procedure for control of bleeding from the spleen. The mean tumor size was 3.8 +/- 1.8 cm. Negative surgical margins were achieved in all but one case. The mean operative time was 151.9 +/- 67.3 min, and the mean estimated blood loss was 97.4 +/- 200.7 ml. A regular diet was resumed at a mean of 2.9 +/- 1.6 days, and the mean length of hospital stay was 3.9 +/- 2.2 days. The perioperative complication rate was 16.4% including deep vein thrombosis postoperative bleed, anastomotic stricture, and incisional hernia. One mortality occurred, due to respiratory failure. The GISTs included 48 rated as low risk, six rated as intermediate risk, and nine rated as high malignant potential. At a mean follow-up period of 15 +/- 21.8 months (range, 0-103 months), three of nine patients with high malignant potential GIST experienced, respectively, metastatic disease to the liver, liver and lung, and peritoneum. At this writing, all the other patients are disease free. Laparoscopic gastric resection for GIST is a feasible option. Adequate oncologic resection was achieved with 98.4% of patients chosen for laparoscopic resection. Resection margin positivity and recurrence rates are low after laparoscopic approaches for appropriately selected patients with GIST, demonstrating favorable characteristics.


Resistance to HSV-1 infection in the epithelium resides with the novel innate sensor, IFI-16.
Toll-like receptors (TLRs) are innate sentinels required for clearance of bacterial and fungal infections of the cornea, but their role in viral immunity is currently unknown. We report that TLR signaling is expendable in herpes simplex virus (HSV)-1 containment as depicted by plaque assays of knockout mice (MyD88(-/-), Trif(-/-) and MyD88(-/-) Trif(-/-) double knockout) resembling wild-type controls. To identify the key sentinel in viral recognition of the cornea, in vivo knockdown of the DNA sensor IFI-16/p204 in the corneal epithelium was performed and resulted in a loss of IFN-regulatory factor-3 (IRF-3) nuclear translocation, interferon-α production, and viral containment. The sensor seems to have a similar function in other HSV clinically relevant sites such as the vaginal mucosa in which a loss of p204/IFI-16 results in significantly more HSV-2 shedding. Thus, we have identified an IRF-3-dependent, IRF-7- and TLR-independent innate sensor responsible for HSV containment at the site of acute infection.


Modifications in avoidance reactions of mice, on a second exposure to the hot plate, resist to various amnesia-inducing treatments.
The avoidance responses of mice exposed to the hot plate (55 degrees C) were found to be modified when tested a second time. In fact, when forepaws licking was no longer observed, the rearing was clearly anticipated (7 s instead of 15 s) as well as jumping (24 s instead of 55 s). These modifications of avoidance strategies as well as their latencies were still observed even 24 days after the first exposure. Avoidance responses were prevented by morphine or haloperidol injected prior to the first exposure, but not with scopolamine or diazepam. These modifications were not affected in mice injected with morphine or submitted to either a supramaximal electroshock or to ether anesthesia delivered immediately after the first hot plate exposure. Among the various known types of memory, these modifications could be linked to procedural memory.


Spatial patterns of tumour growth impact clonal diversification in a computational model and the TRACERx Renal study.
Genetic intra-tumour heterogeneity fuels clonal evolution, but our understanding of clinically relevant clonal dynamics remain limited. We investigated spatial and temporal features of clonal diversification in clear cell renal cell carcinoma through a combination of modelling and real tumour analysis. We observe that the mode of tumour growth, surface or volume, impacts the extent of subclonal diversification, enabling interpretation of clonal diversity in patient tumours. Specific patterns of proliferation and necrosis explain clonal expansion and emergence of parallel evolution and microdiversity in tumours. In silico time-course studies reveal the appearance of budding structures before detectable subclonal diversification. Intriguingly, we observe radiological evidence of budding structures in early-stage clear cell renal cell carcinoma, indicating that future clonal evolution may be predictable from imaging. Our findings offer a window into the temporal and spatial features of clinically relevant clonal evolution.


Use of recombinant factor VIIa to treat persistent bleeding following dental extractions in two cirrhotic patients.
A single dose of recombinant factor VIIa (rFVIIa) has been shown to be effective and safe in correcting the prothrombin time (PT) in cirrhotic patients, but no clinical data exists demonstrating its efficacy in arresting active bleeding. rFVIIa was used in two cirrhotic patients for persistent bleeding following dental extractions despite repeated treatment at the wound site and, in one case, repeated administrations of fresh-frozen plasma (FFP). Bleeding stopped promptly in both patients after administration of rFVIIa. However, bleeding recurred in the patient who had not received concomitant treatment at the extraction sites. No recurrence of bleeding was observed in the second patient, who underwent local treatment 15 min after rFVIIa. Recombinant factor VIIa arrested bleeding after dental extractions in two cirrhotic patients who had been unsuccessfully treated with FFP. However, additional local treatment is needed to limit the risk of recurrence as a result of the short half-life of rFVIIa.


Native American Children and Their Reports of Hope: Construct Validation of the Children's Hope Scale.
Child reports of hope continue to be utilized as predictors of positive adjustment; however, the utilization of the hope construct has not been assessed within the culturally diverse Native American child group. The present study investigated the applicability of the Hope theory among 96 Native American children in the Midwest. Measures included the Children's Hope Scale and a Hope Interview. Native American children in the current sample appear to conceptualize hope as a way to reach goals as did the children in the normative sample. Results from the factor analysis demonstrate that the factor structure found in the current study was similar to the factor structure found in the standardization sample. Because of the similar Hope theory conceptualization and factor structure, interventions focused on the positive psychology construct of hope may be applicable within a Native American child population.


Facial nerve grafting: a thirty year retrospective review.
It is axiomatic that nerve grafting be considered as an integral part of every ablative or traumatic injury involving the extratemporal main trunk and branches of the facial nerve. Our experience with 170 autogenous grafts has established the efficacy of the technique and has proved the high regenerative and adaptive capacities of the facial nerve system.


Changes in ventricular fibrillation threshold during repeated short-term coronary occlusion and release.
The time course of changes in VFT was determined during the 1st phase of arrhythmia following coronary occlusion and during consecutive reperfusion in five repeated periods of occlusion and reperfusion in 10 mongrel dogs (17--24 kg). VFT was determined using a square wave pulse series of 140 ms duration which was triggered by the R-wave of the ECG and placed into the vulnerable period of the cardiac cycle. After acute occlusion VFT decreased to a minimum level within a few minutes and then increased again slowly up to the control value which was reached about 20 min after the ligation. When the occlusions were repeated several times the extent of the decrease in VFT became increasingly less and its duration increasingly shorter until finally there was no significant decrease in VFT. Reperfusion after coronary occlusion led to an abrupt decrease in VFT within 1 min, followed by a rapid increase to the control value. This time course did not depend upon the number of prior occlusions. The results show that in the case of repeating short-term coronary occlusions one cannot expect comparable VFT time courses for the consecutive periods of occlusion except for the 1st and 2nd ones. Differing mechanisms leading to the occurrence of VF after coronary ligation and during reperfusion are discussed.


Raman studies of the conformation of the basic pancreatic trypsin inhibitor.
The vibrational Raman spectra of the basic pancreatic trypsin inhibitor in aqueous solution, as lyophilized powder and in a single crystal and presented. The thermal stability of this protein is demonstrated by the fact that minor alterations in the spectrum, mainly in the amide III band near 1260 cm-1, occur in the solution spectrum only at temperatures above 75 degrees C. No significant spectral changes appear when the pH value of the solution is varied in the range from 1.5 to 8.7. The distinct differences of the powder spectrum compared to that of the solution, show that lyophilization causes appreciable conformational changes both in the main-chain and in the side-chains. A difference in main chain conformation of the basic pancreatic trypsin inhibitor in single crystal and in solution is suggested by different amide III frequencies.


Hydrolysis of dolichyl esters by rat liver lysosomes.
Dolichyl ester hydrolase activity is broadly distributed among the organs of the rat. The highest activity was found in spleen, brain, lung, and thyroid tissues, whereas this activity is very low in stomach and intestine. The esterase involved is localized to the lumen of lysosomes and, to some extent, in the plasma membranes. Hydrolysis occurs with both alpha-saturated and alpha-unsaturated polyisoprenes esterified with different fatty acids, but the rate of hydrolysis is strongly dependent on the nature of the substrate. The enzyme involved is inhibited by divalent cations, EDTA and EGTA and also by one of the products, dolichol. The esterase is activated by 3-[(3-cholamidopropyl) dimethylammonio]-1-propranesulfonic acid and taurodeoxycholate and inhibited by Triton X-100. Dolichyl esterase activity is completely inhibited by alpha- and beta-naphthyl acetate, phenylmethylsulfonyl fluoride, and beta-chloromethylmercurisulfate. These inhibitors, as well as the pH optimum for dolichyl ester hydrolysis, clearly differentiate the enzyme involved from cholesteryl esterase and triglyceride lipase. Microsomal phospholipase A hydrolyzes dolichyl esters at a slow rate only. In vivo labeling experiments with [3H]mevalonate demonstrated that newly synthesized dolichol is transported in esterified form to the lysosomes, where this lipid is slowly hydrolyzed by the esterase. The possibility is raised that the role of the fatty acyl moiety may be to target dolichol to its final location in the cell.


Unexpected difficult intubation. Asymptomatic epiglottic cysts as a cause of upper airway obstruction during anaesthesia.
Two cases of asymptomatic epiglottic cysts which presented as partial upper airway obstruction following induction of anaesthesia are described. The incidence, pathology and anaesthetic management are discussed.


Coronary Artery Axial Plaque Stress and its Relationship With Lesion Geometry: Application of Computational Fluid Dynamics to Coronary CT Angiography.
The purpose of this study was to characterize the hemodynamic force acting on plaque and to investigate its relationship with lesion geometry. Coronary plaque rupture occurs when plaque stress exceeds plaque strength. Computational fluid dynamics was applied to 114 lesions (81 patients) from coronary computed tomography angiography. The axial plaque stress (APS) was computed by extracting the axial component of hemodynamic stress acting on stenotic lesions, and the axial lesion asymmetry was assessed by the luminal radius change over length (radius gradient [RG]). Lesions were divided into upstream-dominant (upstream RG > downstream RG) and downstream-dominant lesions (upstream RG < downstream RG) according to the RG. Thirty-three lesions (28.9%) showed net retrograde axial plaque force. Upstream APS linearly increased as lesion severity increased, whereas downstream APS exhibited a concave function for lesion severity. There was a negative correlation (r = -0.274, p = 0.003) between APS and lesion length. The pressure gradient, computed tomography-derived fractional flow reserve (FFRCT), and wall shear stress were consistently higher in upstream segments, regardless of the lesion asymmetry. However, APS was higher in the upstream segment of upstream-dominant lesions (11,371.96 ± 5,575.14 dyne/cm(2) vs. 6,878.14 ± 4,319.51 dyne/cm(2), p < 0.001), and in the downstream segment of downstream-dominant lesions (7,681.12 ± 4,556.99 dyne/cm(2) vs. 11,990.55 ± 5,556.64 dyne/cm(2), p < 0.001). Although there were no differences in FFRCT, % diameter stenosis, and wall shear stress pattern, the distribution of APS was different between upstream- and downstream-dominant lesions. APS uniquely characterizes the stenotic segment and has a strong relationship with lesion geometry. Clinical application of these hemodynamic and geometric indices may be helpful to assess the future risk of plaque rupture and to determine treatment strategy for patients with coronary artery disease. (Evaluation of FFR, WSS, and TPF Using CCTA; NCT01857687).


Novel microsatellite markers for evaluation of genetic diversity in the tetraploid flame azalea, Rhododendron calendulaceum (Ericaceae).
Microsatellite markers have become a popular and useful tool for investigating evolutionary processes at shallow taxonomic scales such as within a species or between extremely closely related species. Rhododendron sect. Pentanthera is a closely related group of deciduous azaleas that demonstrate both naturally occurring and horticulturally derived hybridization. Two species, flame azalea and Cumberland azalea, represent a particularly recalcitrant evolutionary problem, which will benefit from the development of rapidly evolving molecular markers. Microsatellite markers were specifically developed for Rhododendron calendulaceum, the flame azalea, for use in studies of genetic structure and potential hybridization with its close relative Rhododendron cumberlandense, the Cumberland azalea. Forty-eight primer pairs designed from paired-end Illumina MiSeq data were screened for robust amplification. Sixteen of these pairs were PCR-amplified in the presence of fluorescently labeled primers and genotyped in 66 flame azalea individuals from three geographically dispersed populations. Fifteen primer pairs were both reliable and polymorphic and exhibit ample variability for use in downstream population-level investigations. Cross-amplification in all other members of Rhododendron sect. Pentanthera was highly successful, suggesting broad utility across the entire clade. The novel microsatellite markers presented here functioned well within the target species and amplified with high success in the remaining members of the clade. They represent a significant improvement to the genetic toolkit available for Rhododendron sect. Pentanthera, and particularly for the flame/Cumberland azalea evolutionary problem.


Assessment of variation in microbial community amplicon sequencing by the Microbiome Quality Control (MBQC) project consortium.
In order for human microbiome studies to translate into actionable outcomes for health, meta-analysis of reproducible data from population-scale cohorts is needed. Achieving sufficient reproducibility in microbiome research has proven challenging. We report a baseline investigation of variability in taxonomic profiling for the Microbiome Quality Control (MBQC) project baseline study (MBQC-base). Blinded specimen sets from human stool, chemostats, and artificial microbial communities were sequenced by 15 laboratories and analyzed using nine bioinformatics protocols. Variability depended most on biospecimen type and origin, followed by DNA extraction, sample handling environment, and bioinformatics. Analysis of artificial community specimens revealed differences in extraction efficiency and bioinformatic classification. These results may guide researchers in experimental design choices for gut microbiome studies.


The effect of indomethacin on the psychomotor function of patients with rheumatic diseases.
We have compared the effect of single and multiple doses of indomethacin and placebo on objective measurements of psychomotor impairment in patients. Following a single 50 mg dose (n = 8), indomethacin produced psychomotor disturbance in only those patients who had no recent history of NSAID exposure. After multiple doses of indomethacin (25 and 50 mg tid for 5 days), significant psychomotor impairment was observed. We conclude that other NSAIDs may induce cross-tolerance to the psychomotor effects of indomethacin. Tachyphylaxis may develop to the psychomotor disturbance caused by indomethacin.


Osteomesopycnosis. A new case.
A 10-year old, mentally and physically normally developed girl complained of back pain following an accident. Radiological findings revealed an inhomogeneous, coarse osteosclerosis along the endplates and posterior parts of the ovoid shaped vertebral bodies, in the proximal and distal femora-, proximal humeri and tibia epiphyses, and along the lateral aspects of the femoral necks. The pelvis presented a mottled appearance of the trabecular pattern at the symphysis pubis and the lateral parts of ischii and sclerosis in the acetabular region. The only laboratory abnormality was a high serum parathormone level. The family history revealed an autosomal dominant inheritance. The name proposed by Maroteaux for this skeletal disorder is osteomesopycnosis, as the lesions were localized to the spine, pelvis and sometimes proximal femur. The skull, ribs, long bones, hands and feet were not involved in the reported 12 cases. Radiographs of this patient showed in addition changes in the hand. This disease must be distinguished from atypical axial osteomalacia.


Production of H₂O₂ in the endoplasmic reticulum promotes in vivo disulfide bond formation.
Oxidative protein folding in the luminal compartment of endoplasmic reticulum (ER) is thought to be accompanied by the generation of H₂O₂, as side-product of disulfide bond formation. We aimed to examine the role of H₂O₂ produced in the lumen, which on one hand can lead to redox imbalance and hence can contribute to ER stress caused by overproduction of secretory proteins; on the other hand, as an excellent electron acceptor, H₂O₂ might serve as an additional pro-oxidant in physiological oxidative folding. Stimulation of H₂O₂ production in the hepatic ER resulted in a decrease in microsomal GSH and protein-thiol contents and in a redox shift of certain luminal oxidoreductases in mice. The oxidative effect, accompanied by moderate signs of ER stress and reversible dilation of ER cisternae, was prevented by concomitant reducing treatment. The imbalance also affected the redox state of pyridine nucleotides in the ER. Antibody producing cells artificially engineered with powerful luminal H₂O₂ eliminating system showed diminished secretion of mature antibody polymers, while incomplete antibody monomers/dimers were accumulated and/or secreted. Evidence are provided by using in vivo models that hydrogen peroxide can promote disulfide bond formation in the ER. The results indicate that local H₂O₂ production promotes, while quenching of H₂O₂ impairs disulfide formation. The contribution of H₂O₂ to disulfide bond formation previously observed in vitro can be also shown in cellular and in vivo systems.


Influenza virus-specific T cells fail to reduce lung virus titres in cyclosporin-treated, infected mice.
Cyclosporin A (CsA) inhibited the function(s) of transferred influenza-specific K,D-restricted cytotoxic T cells, which led to clearance of virus in the lungs of influenza virus-infected mice. CsA had no effect on the migration of the transferred cells to the lungs. The pattern of migration and the number of cells recovered from the lungs were similar when cells were transferred into normal, untreated, infected or CsA-treated, infected mice. CsA had no effect on the in vitro expression of cytotoxic activity by the K,D-restricted cytotoxic T cells. These findings strongly suggest that the in vivo clearance of influenza virus by K,D-restricted cytotoxic T cells involves a lymphokine mechanism.


Construction of a selective cleavage system for a protein displayed on the cell surface of yeast.
We constructed a novel protein-purification system in which Saccharomyces cerevisiae with a protein displayed on the cell surface is harvested and the displayed protein is then cleaved from the cell surface. GFPuv was used as a model protein in this cell surface engineering experiment. In this system, the C-terminal 320 amino acids of alpha-agglutinin were bound to the C-terminal of GFPuv for display on the cell surface. In this novel system, the insertion of the recognition sequence-encoding gene of protease factor Xa between GFPuv and alpha-agglutinin was successfully carried out. The GFPuv, displayed by the insertion, was successfully cleaved from yeast cell surface by treatment with factor Xa, and could be easily recovered. By removing such a protease with well-known properties, the displayed protein could be isolated and purified with relative ease.


Temperature-jump studies on benzeneboronic acid-serine protease interactions. Subtilisin and alpha-chymotrypsin.
The interactions of benzeneboronic acid (BBA) as a transition state analog with subtilisin (EC 184.108.40.206) and with alpha-chymotrypsin (EC 18.104.22.168) were investigated kinetically by the temperature-jump method using pH indicators. For both enzymes, the concentration dependence of the relaxation time was consistent with a two-step mechanism involving a fast bimolecular association followed by a slow, unimolecular process. The possibility of a trigonal-tetrahedral interconversion of BBA at the active site of the enzyme is discussed.


Sequence of the argF gene encoding ornithine transcarbamoylase from Neisseria gonorrhoeae.
The gonococcal argF gene encoding ornithine transcarbamoylase (OTCase) contains an open reading frame of 993 nucleotides which starts with a GUG codon and encodes a peptide with a deduced Mr of 36,731. We compared the predicted amino acid (aa) sequence to OTCase sequences previously determined for Escherichia coli and Pseudomonas aeruginosa and found that highly conserved regions in the genes from these organisms were also conserved in Neisseria gonorrhoeae, including those aa known to be important for carbamoyl phosphate and ornithine binding. In the flanking regions of the gene were found 15-bp inverted repeats that may serve as transcriptional termination signals, and which contain the neisserial DNA-uptake sequence.


Roles of the Werner syndrome protein in pathways required for maintenance of genome stability.
Werners syndrome is a disease of premature aging where the patients appear much older than their chronological age. The gene codes for a protein that is a helicase and an exonuclease, and recently we have learned about some of its protein interactions. These interactions are being discussed as they shed light on the molecular pathways in which Werner protein participates. Insight into these pathways brings insight into the aging process.


Recombinant adeno-associated virus-mediated delivery of antisense angiotensin II receptor 1 gene attenuates hypertension development.
The renin-angiotensin system plays a crucial role in the development and establishment of hypertension, and the pharmacological blockade of the system results in a reduction in blood pressure. In the present study, we investigated whether the effects of a novel, double-stranded, recombinant adeno-associated virus vector (rAAV)-mediated antisense angiotensin II receptor 1 (AT1R) gene efficiently prevents the development of hypertension induced by a high-salt diet in adult, male Sprague-Dawley (SD) rats. A rAAV was prepared with a cassette containing a cytomegalovirus promoter and partial cDNA (660 base pairs) for the AT1R inserted in the antisense direction (rAAV-AT1-AS). A single tail vein injection of the rAAV-AT1-AS or rAAV-GFP (green fluorescent protein, a reporter gene) was performed in adult, male SD rats. Two weeks after injection, the animals were fed a diet containing 8% NaCl, and the systolic blood pressure was measured weekly using the tail-cuff method for 12 weeks. The high-salt diet induced a significant rise in systolic blood pressure in the rAAV-GFP-treated animals; however, the rAAV-AT1-AS treatment attenuated the rise in blood pressure (142.7+/-4.5 mmHg vs 117+/-3.8 mmHg, P<0.01), and the hypotensive effect was maintained until the experiments ended at 12 weeks. In the rAAV-GFP-treated animals AT1 was overexpressed in various tissues, especially in the aorta and kidney at mRNA levels; in contrast, rAAV-AT1-AS treatment markedly attenuated AT1 expression. Furthermore, rAAV-AT1-AS treatment prevented target organ damages from hypertension, including cardiac dysfunction and renal injury compared to the rAAV-GFP group. These results suggest that rAAVmediated anti-AT1 delivery attenuates the development of hypertension and protects against renal injury and cardiac remodeling.


Loss of inhibitor of apoptosis proteins as a determinant of polyamine analog-induced apoptosis in human melanoma cells.
We have previously shown that the clinically relevant polyamine analog N1,N11-diethylnorspermine (DENSPM) causes rapid apoptosis in human melanoma SK-MEL-28 cells via a series of events that include mitochondrial release of cytochrome c and activation of the caspase cascade. Upstream to these events, DENSPM downregulates polyamine biosynthesis and potently upregulates polyamine catabolism at the level of spermidine/spermine N1-acetyltransferase (SSAT). In searching for downstream effectors that either contribute to or abrogate the apoptotic response, we observed that DENSPM treatment of SK-MEL-28 cells for 30 h led to cytosolic release of Smac/Diablo, a mitochondrial protein known to bind and inhibit the function of inhibitor of apoptosis proteins (IAPs). Subsequently, we found that DENSPM markedly lowered survivin and ML-IAP protein (but not XIAP) levels by 18 h via an apparently Smac/Diablo-independent pathway. Proteasome inhibitors fully prevented survivin and ML-IAP protein loss as well as apoptosis, suggesting that the proteasome-mediated degradation of survivin and ML-IAP is causally linked to the cellular outcome. We also observed that structural analogs of DENSPM which differentially induced SSAT and apoptosis lowered survivin and ML-IAP levels in a manner that correlated with enzyme activity. The linkage between IAPs and SSAT was more directly established by the finding that selective prevention of SSAT induction by small interfering RNA prevented survivin and ML-IAP loss as well as apoptosis during DENSPM treatment. Among the melanoma cell lines (SK-MEL-28, MALME-3M, A375 and LOX), survivin degradation correlated temporally with the onset of DENSPM induced apoptosis or growth inhibition. By contrast, ML-IAP degradation occurred only during rapid apoptosis seen in SK-MEL-28 cells. These data suggest a sequence of events whereby DENSPM induction of SSAT leads to loss of IAP proteins and a more fulminate apoptotic response. The findings implicate survivin and ML-IAP as important determinants of polyamine analog drug action in melanoma cells.


Microstructural changes precede depression in patients with relapsing-remitting Multiple Sclerosis.
Multiple Sclerosis lesions in the brain and spinal cord can lead to different symptoms, including cognitive and mood changes. In this study we explore the temporal relationship between early microstructural changes in subcortical volumes and cognitive and emotional function in a longitudinal cohort study of patients with relapsing-remitting Multiple Sclerosis. In vivo imaging in forty-six patients with relapsing-remitting Multiple Sclerosis was performed annually over 3 years magnetic resonance imaging. Microstructural changes were estimated in subcortical structures using the free water fraction, a diffusion-based MRI metric. In parallel, patients were assessed with the Hospital Anxiety and Depression Scale amongst other tests. Predictive structural equation modeling was set up to further explore the relationship between imaging and the assessment scores. In a general linear model analysis, the cohort was split into patients with higher and lower depression scores. Nearly all subcortical diffusion microstructure estimates at the baseline visit correlate with the depression score at the 2 years follow-up. The predictive nature of baseline free water estimates and depression subscores after 2 years are confirmed in the predictive structural equation modeling analysis with the thalamus showing the greatest effect size. The general linear model analysis shows patterns of MRI free water differences in the thalamus and amygdala/hippocampus area between participants with high and low depression score. Our data suggests a relationship between higher levels of free-water in the subcortical structures in an early stage of Multiple Sclerosis and depression symptoms at a later stage of the disease.


Reliable control of spike rate and spike timing by rapid input transients in cerebellar stellate cells.
Granule cell activity in cerebellar cortex directly excites Purkinje cells via parallel fibers, but it also inhibits Purkinje cells via cerebellar cortical interneurons. This contribution of inhibitory interneurons to cerebellar cortical processing remains poorly understood. In the present study we examined the response properties of stellate cells in vitro to input patterns that may result from granule cell activity in vivo. We constructed input waveforms that represented the sum of inputs from all individual synapses and applied these waveforms to the soma of stellate cells during whole cell recordings in acute brain slices. The stimulus waveforms contained fluctuations in a broad range of frequencies and were applied at different amplitudes. To determine the contribution of synaptic shunting to stellate cell spike responses we applied the same input waveforms either as a simulated synaptic conductance using dynamic clamping or as a direct current injection stimulus. Only the dynamic clamp stimulus has the shunting properties of real synapses, i.e. leads to different-sized synaptic current as a function of membrane potential. We found that stellate cells spike with millisecond precision in response to fast temporal fluctuations in the total synaptic input. Transient increases in excitatory input frequency led to pronounced stellate cell spike responses, indicating that this pathway may be very responsive to even small assemblies of co-activated granule cells. This was observed regardless of whether the input waveform was applied as a conductance with dynamic clamping, or as a direct current injection. Thus the shunting properties of a conductance input did not play a major role in determining the control of precisely timed spiking. In contrast, a more tonic increase in excitatory conductance did not lead to a sustained spike response as obtained with prolonged positive current injection. However, even with tonic current injection the precision of spiking was lost, as previously observed. Overall, the synaptic response function of stellate cells suggests that this cell type may pick out transients in granule cell activity, and may generate precisely timed inhibition of Purkinje cells during behavior.


Lactoferrin and the development of salivary stones: a pilot study.
Salivary stones (sialoliths) are calcified structures located in the ductal system of the major salivary glands. Their exact cause is not clear but in general they are characterized by concentric inorganic (hydroxyapatite) layers. The formation is a slow intermittent process which may result in enlargement of the sialolith causing obstruction of saliva secretion resulting in mealtime related pain and swelling of the affected salivary gland. Various studies reported the presence of organic material such as proteins and lipids in the core of sialoliths. In the present study the protein composition of twenty submandibular sialoliths was analyzed. It was found that proteins contributed on average 5% to the dry weight of submandibular stones whereby small salivary stones contained more extractable proteins than large salivary stones. Using a combination of SDS-PAGE gel electrophoresis and Western blotting, we identified α-amylase (in all stones; 100%), lysozyme (95%), lactoferrin (85%), secretory-IgA (75%), MUC7 (60%), complement C4 (60%) and C-reactive protein (35%). The presence, and the combinations, of lactoferrin, lysozyme, s-IgA and α-amylase in sialoliths was confirmed by ELISA. The gradually increasing size of a sialolith might provoke a local inflammatory response in the duct of the submandibular gland whereby the relatively low concentrations of lactoferrin and lysozyme may originate from neutrophils. The interaction of lactoferrin with s-IgA could contribute to the accumulation of lactoferrin in sialoliths. In summary, these results suggest a new pathophysiological role for lactoferrin, in the formation of sialoliths.


COVID-19 Outbreak: Neurological Manifestations Beyond Cough and Fever.
COVID-19 is one of the most disastrous respiratory diseases (after the 1918 influenza outbreak) spreading in the community. So far, it has killed 7,37,417 individuals. High variability in the viral genome and its greater ability to spread in the human community are badly affecting the comorbid individuals. Although infected individuals mainly possess respiratory issues, neurological manifestations in these individuals cannot be overlooked. The literature search is based on the recent development in the concerned field. We searched databases like PubMed, Google Scholar, and ScienceDirect using the keywords "COVID-19", "neurological manifestations", "CNS", and "PNS". The major neurological complications observed in these patients are encephalitis, necrotising haemorrhagic encephalopathy, Guillain-Barré Syndrome, smell/taste impairment, epileptic seizures, and abnormal states of consciousness. COVID-19 infection is just more than a cough, fever, and respiratory illness; it can cause indirect neurological complications in infected patients. It is therefore advised to treat and have a careful observation of the COVID-19 patients for neurological manifestations.


Capsaicin-induced depolymerization of axonal microtubules mediates analgesia for trigeminal neuropathic pain.
Capsaicin is a specific agonist of transient receptor potential vanilloid 1 (TRPV1), which is enriched in nociceptors. Capsaicin not only produces acute pain but also leads to long-lasting analgesia in patients with chronic pain. Although capsaicin-induced TRPV1 and Ca 2+ /calpain-dependent ablation of axonal terminals is necessary for long-lasting analgesia, the mechanisms underlying capsaicin-induced ablation of axonal terminals and its association with analgesia are not fully understood. Microtubules are composed of tubulin polymers and serve as a main axonal cytoskeleton maintaining axonal integrity. In this study, we hypothesized that capsaicin would increase the depolymerization of microtubules and lead to axonal ablation and analgesia for trigeminal neuropathic pain. Paclitaxel, a microtubule stabilizer, decreased capsaicin-induced ablation of axonal terminals in time-lapsed imaging in vitro. Capsaicin increases free tubulin in dissociated sensory neurons, which was inhibited by paclitaxel. Consistently, subcutaneous injection of paclitaxel prevented capsaicin-induced axonal ablation in the hind paw skin. Capsaicin administration to the facial skin produced analgesia for mechanical hyperalgesia in mice with chronic constriction injury of the infraorbital nerve, which was prevented by the coadministration of paclitaxel and capsaicin. Whole-mount staining of facial skin showed that paclitaxel reduced capsaicin-induced ablation of peptidergic afferent terminals. Despite the suggested involvement of TRPV1 Ser801 phosphorylation on microtubule integrity, capsaicin-induced analgesia was not affected in TRPV1 S801A knock-in mice. In conclusion, capsaicin-induced depolymerization of axonal microtubules determined capsaicin-induced ablation of nociceptive terminals and the extent of analgesia. Further understanding of TRPV1/Ca 2+ -dependent mechanisms of capsaicin-induced ablation and analgesia may help to improve the management of chronic pain.


Presurgical thalamocortical connectivity is associated with response to vagus nerve stimulation in children with intractable epilepsy.
Although chronic vagus nerve stimulation (VNS) is an established treatment for medically-intractable childhood epilepsy, there is considerable heterogeneity in seizure response and little data are available to pre-operatively identify patients who may benefit from treatment. Since the therapeutic effect of VNS may be mediated by afferent projections to the thalamus, we tested the hypothesis that intrinsic thalamocortical connectivity is associated with seizure response following chronic VNS in children with epilepsy. Twenty-one children (ages 5-21 years) with medically-intractable epilepsy underwent resting-state fMRI prior to implantation of VNS. Ten received sedation, while 11 did not. Whole brain connectivity to thalamic regions of interest was performed. Multivariate generalized linear models were used to correlate resting-state data with seizure outcomes, while adjusting for age and sedation status. A supervised support vector machine (SVM) algorithm was used to classify response to chronic VNS on the basis of intrinsic connectivity. Of the 21 subjects, 11 (52%) had 50% or greater improvement in seizure control after VNS. Enhanced connectivity of the thalami to the anterior cingulate cortex (ACC) and left insula was associated with greater VNS efficacy. Within our test cohort, SVM correctly classified response to chronic VNS with 86% accuracy. In an external cohort of 8 children, the predictive model correctly classified the seizure response with 88% accuracy. We find that enhanced intrinsic connectivity within thalamocortical circuitry is associated with seizure response following VNS. These results encourage the study of intrinsic connectivity to inform neural network-based, personalized treatment decisions for children with intractable epilepsy.


Comparison of heater probe and YAG laser in endoscopic treatment of major bleeding from peptic ulcers.
In our experience using the YAG laser and the heater probe in endoscopic treatment of major hemorrhage from peptic ulcers, ultimate hemostatic success was achieved in 19 of 20 (95%) ulcers treated with the heater probe, versus 24 of 35 (69%) YAG laser-treated ulcers (p less than 0.05). The heater probe was considerably faster, more convenient to use, and safer than YAG laser. We presently consider the heater probe to be the preferred endoscopic hemostatic device.


Continent diversion: five decades of developments and evolution.
To provide a chronological overview of the evolution of continent urinary diversion (CUD) over the last 50 years and to highlight important milestones. We performed an extensive literature review and analysed different forms of urinary diversion worldwide. After the evaluation of surgical techniques, we assessed the advantages and disadvantages of assorted CUD approaches based on published long-term follow-up data. A wide variety of surgical options for CUD is available and feasible to date, although consensus among urologists regarding the 'gold standard' is still lacking. Several forms of orthotopic bladder substitutes and continent cutaneous urinary reservoirs have been shown to provide excellent long-term results. The last 50 years of CUD have seen constant evolution and refinement of techniques, but the best surgical approach remains unclear and there is no 'one-size-fits-all' option, but rather tailor-made approaches are necessary to ensure patient satisfaction.


Potent antagonism of calmodulin activity in vitro, but lack of antiproliferative effects on keratinocytes by the novel leukotriene biosynthesis inhibitor MK-886.
MK-886, a leukotriene biosynthesis inhibitor, which prevents the translocation and activation of 5-lipoxygenase, has been proposed as an effective drug for the treatment of inflammatory disorders, including psoriasis. In the present study, we investigated the effects of MK-886 on calmodulin as a potential target protein of anti-inflammatory drug activity, and on the proliferation of cultured human keratinocytes, a calmodulin-dependent cellular response with indicative value for antipsoriatic drug activity. Despite potent calmodulin-antagonistic activity in vitro, MK-886 failed to block cell proliferation in a human keratinocyte line, whereas trifluoperazine, a well characterized calmodulin antagonist with similar effects on calmodulin activity in our in vitro assays, inhibited cell proliferation in a dose-dependent manner. Further investigations on the mechanism of action revealed that, in contrast with trifluoperazine, calmodulin antagonism by MK-886 in vitro is likely to be mediated at the level of the allosteric calmodulin-recognition site of phosphodiesterase, rather than by binding to calmodulin itself. Therefore, our data do not conflict with the proposed role of calmodulin in the regulation of cell proliferation, but demonstrate that drug-induced antagonism of calmodulin, detected by inhibition of calmodulin-dependent enzymes in vitro, is not necessarily linked to antiproliferative activity in human keratinocytes.


A review on post-COVID-19 impacts and opportunities of agri-food supply chain in Malaysia.
Malaysia is strongly supported by the agriculture sector as the backbone to drive the economy. However, COVID-19 has significantly affected agriculture across the production, supply, and marketing chains. It also disturbs the balance of food supply and demand in Malaysia. COVID-19 was an unexpected pandemic that resulted in shock and panic and caused a huge global impact. However, the impacts of this pandemic on the agriculture sector in Malaysia, particularly in the production and supply chains, are still unclear and scarce. This review offers insights into the challenges, particularly in sustaining agri-food production and supply chains. It also highlights the opportunity and relevant measures towards sustainability in agriculture to avoid agri-food disasters in the future. This study was carried out through a desk review of the secondary source of information covering the impact of COVID-19 in Malaysia particularly in the agri-food aspect, and a wide range of strategies and initiatives as the effective measures to overcome the crisis of this pandemic. Online desk research of the government published data and customer desk research were utilized to complete this study. Search engines such as Google Scholar and the statistical data from the official websites including the Department of Statistics Malaysia (DOSM) and the Food and Fertilizer Technology Center for the Asian and Pacific Region (FFTC-AP), were utilized. Keywords such as impact of COVID-19, pandemic, and agri-food supply chain were used to conduct the searches. The articles identified to be related to the study's objective were then downloaded and included in the study. Descriptive methods were used as the primary analysis technique following the descriptive analysis and visual data analysis in performing the sources obtained. This devastating impact damages the lives by causing 4.3 million confirmed infections and more than 290,000 deaths. This disease presents an unprecedented challenge to the public health. The lockdown restriction under the movement control order (MCO), for more than of the world's population in the year 2020 to control the virus from spreading, has disrupted most of the economic sectors. The agriculture industry was seen as one of the essential industries and allowed to operate under strict standard operating procedures (SOP). Working under strict regulations came with a huge price paid for almost all industries. This pandemic has affected the national agri-food availability and accessibility in Malaysia. This outbreak created a reflection of opportunity for sharing a more flexible approaches in handling emergencies on agricultural food production and supply chains. Therefore, the government should be ready with the roadmap and enforce the measures to control the pandemic without disrupting the agri-food supply chain in the near future.


Pembrolizumab for patients with melanoma or non-small-cell lung cancer and untreated brain metastases: early analysis of a non-randomised, open-label, phase 2 trial.
Immunotherapy targeting the PD-1 axis has activity in several tumour types. We aimed to establish the activity and safety of the PD-1 inhibitor pembrolizumab in patients with untreated brain metastases from melanoma or non-small-cell lung cancer (NSCLC). In this non-randomised, open-label, phase 2 trial, we enrolled patients aged 18 years or older with melanoma or NSCLC with untreated brain metastases from the Yale Cancer Center. Patients had at least one untreated or progressive brain metastasis between 5 and 20 mm in diameter without associated neurological symptoms or the need for corticosteroids. Patients with NSCLC had tumour tissue positive for PD-L1 expression; this was not required for patients with melanoma. Patients were given 10 mg/kg pembrolizumab every 2 weeks until progression. The primary endpoint was brain metastasis response assessed in all treated patients. The trial is ongoing and here we present an early analysis. The study is registered with ClinicalTrials.gov, number NCT02085070. Between March 31, 2014, and May 31, 2015, we screened 52 patients with untreated or progressive brain metastases (18 with melanoma, 34 with NSCLC), and enrolled 36 (18 with melanoma, 18 with NSCLC). A brain metastasis response was achieved in four (22%; 95% CI 7-48) of 18 patients with melanoma and six (33%; 14-59) of 18 patients with NSCLC. Responses were durable, with all but one patient with NSCLC who responded showing an ongoing response at the time of data analysis on June 30, 2015. Treatment-related serious and grade 3-4 adverse events were grade 3 elevated aminotransferases (n=1 [6%]) in the melanoma cohort, and grade 3 colitis (n=1 [6%]), grade 3 pneumonitis (n=1 [6%]), grade 3 fatigue (n=1 [6%]), grade 4 hyperkalemia (n=1 [6%]), and grade 2 acute kidney injury (n=1 [6%]) in the NSCLC cohort. Clinically significant neurological adverse events included transient grade 3 cognitive dysfunction and grade 1-2 seizures (n=3 [17%]) in the melanoma cohort. Pembrolizumab shows activity in brain metastases in patients with melanoma or NSCLC with an acceptable safety profile, which suggests that there might be a role for systemic immunotherapy in patients with untreated or progressive brain metastases. Merck and the Yale Cancer Center.


Continuous in-vivo monitoring of arterial oxygenation in chronic obstructive lung disease.
We monitored arterial oxygen saturation (SaO2) in 10 patients with severe chronic obstructive lung disease during 24 h of breathing room air followed by 24 h of breathing two litres of oxygen per minute. Three subjects without chronic obstructive lung disease were monitored while breathing room air. Greatest declines in SaO2 occurred during sleep, with intermittent decreases as great as 44% saturation (range, 12% to 44% saturation. Baseline SaO2 was significantly higher while patients breathed low-flow oxygen (94% versus 86% saturated), and declines in SaO2 during sleep were less noteworthy (1% to 27% saturation). Subjects without chronic obstructive lung disease showed declines in SaO2 of much lesser magnitude (3% to 11% saturation) with sleep. These declines were not primarily due to alveolar hypoventilation. Our data indicate that there are patients with chronic airways obstruction who suffer profound intermittent desaturation at night that can readily be relieved with low-flow oxygen administration.


Culture Negative Endocarditis Masquerading as Recurrent Supraventricular Tachycardia.
Supraventricular tachycardia are common dysrhythmias seen in hospitalized patients. Electrolyte derangements and cardiomyopathy are among the most common causes. Rarely, blood culture negative endocarditis can lead to unexplained recurrentsupraventricular tachycardia. Herein, we present a case of recurrent atrioventricular nodal reentrant tachycardia in a patient with no previous history of cardiovascular disease.


Skin sparing mastectomy and robotic latissimus dorsi-flap reconstruction through a single incision.
Robotic latissimus dorsi-flap reconstruction (RLDFR) after skin-sparing mastectomy (SSM) for breast cancer (BC) has been performed through a single nipple incision. We report results of SSM with RLDFR, mainly with analysis of feasibility, morbidity, indications, and technique standardization. We determined characteristics of patients, previous treatment of BC, and type of reconstruction. Surgical technique, duration of surgery, and complication rate were reported according to three successive periods: P1-3. Forty RLDFR, with breast implant for 16 patients, with previous breast radiotherapy in 30% had been performed. In logistic regression, factors significantly associated with duration of surgery ≥ 300 min were P2 (OR 0.024, p = 0.004) and P3 (OR 0.012, p = 0.004) versus P1. The median mastectomy weight was 330 g and 460 g for BMI < and ≥ 23.5 (p = 0.025). Length of hospitalization was 4 days. Total complication rate was 20% (8/40): seven breast complications (four re-operations) and one RLDF complication with re-operation. Periods were significantly predictive of complications (p = 0.045). SSM with RLDFR is feasible, safe, and reproducible. We reported a decrease of duration of surgery, length of post-operative hospitalization, and complication rate.


Treatment of multi-resistant urinary tract infections in patients with multiple sclerosis.
Multi-resistant nosocomial urinary tract infections occurring in thirty patients with neurogenic bladder dysfunction were treated with ciprofloxacin 100 mg twice daily Early bacteriological eradication was obtained in 48.3%, persistence in 34.5% and superinfection in 17.2%. Recurrence, 30 days after initial eradication occurred in 50% while late eradication was maintained in 35% and reinfection with another germ was noted in 15%. Most recurring and reinfecting germs were still susceptible to ciprofloxacin. The drug was well tolerated. The overall outcome was mainly determined by the vesico-urethral dysfunction itself and by the presence of indwelling bladder catheters.


Acute and chronic changes of myocardial energetics in the mammalian and human heart.
In earlier studies using papillary muscles of the rat left ventricle and highly sensitive thermopiles we demonstrated that the heat liberated per gram of myocardium per unit of developed tension-time integral is decreased when the rats suffered from hypothyroidism or renal hypertension. This increase in economy of force production was shown to be associated with a decrease in myosin-ATPase activity and a change in isomyosin composition. In a recent study we showed an increase in heat per gram of mammalian myocardium per tension-time integral of 70% after application of isoproterenol. In order to study the relationship between energy costs and developed tension-time integral in the human heart, haemodynamics and myocardial oxygen consumption were measured. The data were obtained using a Millar microtip catheter pressure transducer and the argon method. Haemodynamics and myocardial energetics were analysed in 8 patients without significant heart disease before and after application of isoproterenol and in 10 patients with dilative cardiomyopathy (NYHA II-III). During one cardiac cycle, myocardial oxygen consumption per gram of LV myocardium per beat (MVO2/g x beat) is related to LV stress-time integral (integral of sigma xt). The economy of myocardial contraction (EC) was calculated by (formula; see text) EC was 11.3 +/- 3.2 in normal and 14.3 +/- 4.7 dyn x s x g/cm2 x mu cal in dilative cardiomyopathic hearts (NS). Isoproterenol decreased EC from 11.3 +/- 3.2 to 5.5 +/- 1.6 dyn x s x g/cm2 x mu cal in the normal hearts (p less than 0.01). In the rat myocardium, changes in economy of force generation were found due to catecholamines, pressure overload and hypothyroidism. In the human heart, similar energetic changes were observed due to catecholamines. No significant differences in energy of force production were seen between normal and dilative cardiomyopathic hearts. The effect of catecholamines in the mammalian and human myocardium is explained by changes in activation processes and in chemomechanical energy transduction at the level of the contractile proteins.


Determination of the fate and biological responses of food additive silica particles in commercial foods.
Synthetic amorphous silica (SAS) is widely added to commercial foods as an anticaking agent. Concern about the potential application of nanosized silica in foods has increased as nanomaterials are not intended for use as food additives. This study evaluated the particle size distributions and biological responses of food additive SAS. An accurate, sensitive, and cost-effective analytical method for probing SAS was established, and quantitative analysis of its presence in commercial foods was performed. The results demonstrate that food additive SAS is an aggregated material composed of nanosized particles with nanosized aggregates of silica particles identified in commercial foods. Food additive SAS did not exhibit acute cytotoxicity compared to both general-grade nano (G-nano) and bulk (G-bulk) silica. Moreover, intestinal transport amounts of food additive SAS were significantly lower than for G-nano. Taken together, we find that food additive SAS does not exhibit acute toxicity resulting from nanosized materials.


CINcere Modelling: What Have Mouse Models for Chromosome Instability Taught Us?
Chromosomal instability (CIN) is a process leading to errors in chromosome segregation and results in aneuploidy, a state in which cells have an abnormal number of chromosomes. CIN is a hallmark of cancer, and furthermore linked to ageing and age-related diseases such as Alzheimer's. Various mouse models have been developed to explore the role of CIN in ageing and cancer. While these models reveal only a modest contribution of CIN to the initiation of cancer, they also clearly show that CIN is a powerful accelerator of cancer in a predisposed background. Other than cancer, CIN also appears to provoke premature ageing in some of the CIN models. In this review, we discuss the phenotypes of the various available mouse models, what we have learnt so far, and importantly, also which questions still need to be addressed.


Giant hepatic angiomyolipoma presenting with severe anemia: A surgical case report and review of literature.
Angiomyolipoma (AML) is a solid benign neoplasm with mesenchymal features. The clinical signs and symptoms of hepatic angiomyolipoma are nonspecific, and treatment strategy is variable. A 35-years-old male patient has admitted to the hospital with symptoms of severe anemia. Abdominal multi-slice computed tomography (MSCT) and Gadoxetic acid (GA)-enhanced magnetic resonance imaging (MRI) revealed a heterogeneous 23 × 17 cm-in-sized tumor with heterogeneous enhancement and increased angiogenesis. Percutaneous hepatic biopsy was proceeded and in immunohistochemistry, tumor cells responded positively to the HMB-45, SMA, and Glutamine stains, the CD-34 stain was positive for blood vessels as well as Ki-67 sporadically positive, but the Heppar1, S-100, CK stains reacted negative. The final pathologic result was consistent with the primary hepatic angiomyolipoma. The final surgical management was extended right hepatectomy with ligation of the right hepatic artery (RHA) and vein (RHV), as well as the middle hepatic vein (MHV), one month after portal embolization as well as to enlarge the remnant liver volume. A hepatic angiomyolipoma (HAML) primary origin is relatively rare, the clinical manifestations are variable and non-specific. Histological examination and immunohistochemistry staining are considered as the gold standard for HAML diagnosis. HAML are commonly expressed benign behaviors, but HAML cases with malignant behaviors were reported in a cumulative incidence. Radical surgery must be still the most effective and major treatment approach. The present case being the first case with initial syndrome of severe anemia accounted in the English literature. Giant angiomyolipoma is composed of blood vessels and could lead to extensive internal tumoral hemorrhage. We here present a report of this case with had primary hepatic angiomyolipoma with clinical picture of severe anemia.


Photo-induced coupling reactions of tetrazoles with carboxylic acids in aqueous solution: application in protein labelling.
The photo-induced reactions of diaryltetrazoles with carboxylic acids in aqueous solution were investigated. Besides measuring the apparent second-order rate constant and evaluating the functional group compatibility of these reactions, we further incorporated the tetrazoles into SAHA, leading to a new active-site-directed probe for labelling HDACs in both cell lysates and living cells.


Response of coastal marine eco-environment to river fluxes into the sea: a case study of the Huanghe (Yellow) River mouth and adjacent waters.
The impact of the Huanghe (Yellow) River outflows on its estuary was investigated with river gauging and shipboard hydrographic observations. The river flux has been decreasing dramatically; the discharges of water and sediment in the 1990s dropped to 27.4% and 31.9% of those in the 1950s, respectively, resulting in frequent and lengthy events of downstream channel dry-up since the 1970s. There were accumulatively 897 zero-flow days during the 1990s in the river course below the Lijin Hydrological Station, 100 km upstream from the river mouth, which is 82.4% of that in 1972. As freshwater input decreases, river-borne nutrients to the estuarine increased significantly. Concentration of dissolved inorganic nitrogen (DIN) in the 1990s was four times of that in 1950s. Changes in amount and content of the riverine inputs have greatly affected the estuarine ecosystem. Over the past several decades, sea surface temperature and salinity in the estuary and its adjacent waters increased and their distribution pattern altered in response to the reduction of freshwater inflow. The distribution of and seasonal succession in nutrient concentrations in the surface layer have also changed with a shift of river outlet and the decrease in riverine nutrient loads. Furthermore, deterioration of estuarine ecosystem by less river input has decreased primary productivity in the deltaic region waters, and in turn depressed the fishery.


Longitudinal quantification of aqueous flare and cells in Vogt-Koyanagi-Harada disease.
To quantitatively evaluate the changes of aqueous flare and cells in eyes with Vogt-Koyanagi-Harada (VKH) disease. This prospective study included 35 initial-onset VKH patients (70 eyes) and 46 recurrent VKH patients (92 eyes) following immunotherapy. Aqueous flare and cells were quantified using the laser flare-cell meter before treatment, 2 weeks, 1, 3, 6 and 9 months after treatment. Before treatment, mean aqueous flare (ph/ms) in initial-onset and recurrent VKH eyes were 8.1 (SD 4.1) vs 43.6 (20.7) (p = 0.000). Following treatment, recurrent VKH eyes showed a significantly higher flare value than initial-onset VKH eyes at 2 weeks, 1, 3 and 6 months. Prior to treatment, mean cell counts (cells/0.5 mm3) in initial-onset and recurrent VKH eyes were 2.0 (1.9) vs 39.4 (23.1) (p = 0.000). Following treatment, recurrent VKH eyes showed significantly higher cell counts than initial-onset VKH eyes at 2 weeks, 1 and 3 months. Our study shows that recurrent VKH patients displayed a more striking and long-lasting breakdown of the BAB and more severe inflammation than initial-onset VKH patients. Our study also indicates that the disruption of BAB lasted longer than aqueous cells either in initial-onset or in recurrent VKH patients.


H2 production from CO, formate or starch using the hyperthermophilic archaeon, Thermococcus onnurineus.
The hyperthermophilic archaeon, Thermococcus onnurineus, was grown in media supplemented with either CO, formate, or starch. H(2) was produced with each substrate with respective maximum rates of 1.55, 3.83 and 2.66 mmol H(2)/l h. The yields (mol H(2)/mol substrate) were 0.98, 1 and 3.13, respectively. This microbe is the first example where a single microorganism can grow and produce H(2) using CO, formate or starch as substrate.


Ambivalence in modelling oblique saccades.
Human oblique saccades might be made by synchronized but independent vertical and horizontal pulse generators; we call this the Cartesian theory. Another hypothesis is that the oblique amplitude and angle are coded centrally and trigonometrically-derived signals are sent to the horizontal and vertical muscles (the polar coordinate theory). We took a Cartesian model and cross-coupled the two generators to produce saccades identical to those of a polar coordinate model. This is disproof by counter-example: the experimental evidence claimed to support the polar coordinate model does not necessarily do so. Moreover, the behavior reported for oblique saccades is so variable, contradictory, idiosyncratic, and species-dependent that any model of the central organization of oblique saccades is probably premature.


Small-molecule-driven hepatocyte differentiation of human pluripotent stem cells.
The differentiation of pluripotent stem cells to hepatocytes is well established, yet current methods suffer from several drawbacks. These include a lack of definition and reproducibility, which in part stems from continued reliance on recombinant growth factors. This has remained a stumbling block for the translation of the technology into industry and the clinic for reasons associated with cost and quality. We have devised a growth-factor-free protocol that relies on small molecules to differentiate human pluripotent stem cells toward a hepatic phenotype. The procedure can efficiently direct both human embryonic stem cells and induced pluripotent stem cells to hepatocyte-like cells. The final population of cells demonstrates marker expression at the transcriptional and protein levels, as well as key hepatic functions such as serum protein production, glycogen storage, and cytochrome P450 activity.


Activation of Solid-State Emission and Photostability through Molecular Confinement: The Case of Triptycene-Fused Quinacridone Dyes.
We report the facile, metal-free convergent synthesis and the characterization of novel quinacridone dyes in which two triptycene units end-cap and sterically confine the quinacridone chromophore. A precise comparison of the confined dyes with their known homologues reveals that the reduction of π-π interactions in triptycene-fused quinacridone dyes compared to classical quinacridone results not only in an increase of solubility and processability but also in an enhancement of fluorescence quantum yield and photostability in the solid state.