Effect of target cell availability on HIV-1 production in vitro.
The recovery of CD4 target cells following antiretroviral therapy may facilitate virus production and escape from antiretroviral suppression. To address this hypothesis, we directly examined whether the CD4 target cell number increases viral production in the presence of suboptimal therapy. The effect of the CD4 T cell number on HIV-1 replication with a suboptimal dose of zidovudine was studied in vitro. Varying numbers of CD4 T cells were infected with HIV-1 and treated with 1 nM zidovudine. Virus production was measured by p24 antigen capture enzyme-linked immunosorbent assay. Partial sequencing of HIV-1 pol was performed to assess zidovudine-resistant mutations. Wild type virus production was found to increase eightfold in cultures with 100 x 10(4) cells compared with cultures with 10 x 10(4) cells. The IC90 of zidovudine was 4 logs higher in cultures with 16 x 10(4) cells compared with cultures with 1 x 10(4) cells. No zidovudine-resistant mutations were found. Target cell availability may play a direct role in wild type HIV-1 resurgence following therapy.


Laparoscopic Oblique Rectus Abdominis Myocutaneous Flap Harvest for Perineal Reconstruction After Abdominoperineal Resection.
Treatment of perineal defects after abdominoperineal resection or salvage surgery for either locally advanced rectal cancer or anal carcinoma can be challenging. Myocutaneous flap reconstruction has proven to reduce perineal morbidity and abscess formation in the pelvis; however, it is associated with significant donor-site morbidity. To our knowledge, this is the first report of a laparoscopic oblique rectus abdominis myocutaneous flap harvesting for perineal reconstruction. This technical note aimed to demonstrate the feasibility of the technique. Introduction of a laparoscopic technique in harvesting of this flap can potentially further reduce morbidity associated with this flap creation by minimizing abdominal wall trauma and obviating the need for laparotomy for tunneling of the flap intra-abdominally. This report describes a technique using a 6-port laparoscopy, in which the harvesting of the myocutaneous flap was performed after a standardized abdominoperineal resection. The flap itself is passed through the rectus sheath toward the pelvis with the help of a retractor. Two patients successfully underwent a laparoscopic oblique rectus abdominis flap reconstruction after abdominoperineal resection. This report describes our initial experience with laparoscopic harvesting of an oblique rectus abdominis flap for perineal reconstruction after abdominoperineal resection. We believe the technique is easy and reproducible for laparoscopic surgeons and can reduce donor-site morbidity. However, further studies will be needed to confirm this observation.


Modulation of Aptamer-Ligand-Binding by Complementary Oligonucleotides: A G-Quadruplex Anti-Ochratoxin A Aptamer Case Study.
Short oligonucleotides are widely used for the construction of aptamer-based sensors and logical bioelements to modulate aptamer-ligand binding. However, relationships between the parameters (length, location of the complementary region) of oligonucleotides and their influence on aptamer-ligand interactions remain unclear. Here, we addressed this task by comparing the effects of short complementary oligonucleotides (ssDNAs) on the structure and ligand-binding ability of an aptamer and identifying ssDNAs' features that determine these effects. Within this, the interactions between the OTA-specific G-quadruplex aptamer 1.12.2 (5'-GATCGGGTGTGGGTGGCGTAAAGGGA GCATCGGACA-3') and 21 single-stranded DNA (ssDNA) oligonucleotides complementary to different regions of the aptamer were studied. Two sets of aptamer-ssDNA dissociation constants were obtained in the absence and in the presence of OTA by isothermal calorimetry and fluorescence anisotropy, respectively. In both sets, the binding constants depend on the number of hydrogen bonds formed in the aptamer-ssDNA complex. The ssDNAs' having more than 23 hydrogen bonds with the aptamer have a lower aptamer dissociation constant than for aptamer-OTA interactions. The ssDNAs' having less than 18 hydrogen bonds did not affect the aptamer-OTA affinity. The location of ssDNA's complementary site in the aptamer affeced the kinetics of the interaction and retention of OTA-binding in aptamer-ssDNA complexes. The location of the ssDNA site in the aptamer G-quadruplex led to its unfolding. In the presence of OTA, the unfolding process was longer and takes from 20 to 70 min. The refolding in the presence of OTA was possible and depends on the length and location of the ssDNA's complementary site. The location of the ssDNA site in the tail region led to its rapid displacement and wasn't affecting the G-qaudruplex's integrity. It makes the tail region more perspective for the development of ssDNA-based tools using this aptamer.


Carbon flux through photosynthesis and central carbon metabolism show distinct patterns between algae, C3 and C4 plants.
Photosynthesis-related pathways are regarded as a promising avenue for crop improvement. Whilst empirical studies have shown that photosynthetic efficiency is higher in microalgae than in C3 or C4 crops, the underlying reasons remain unclear. Using a tailor-made microfluidics labelling system to supply 13CO2 at steady state, we investigated in vivo labelling kinetics in intermediates of the Calvin Benson cycle and sugar, starch, organic acid and amino acid synthesis pathways, and in protein and lipids, in Chlamydomonas reinhardtii, Chlorella sorokiniana and Chlorella ohadii, which is the fastest growing green alga on record. We estimated flux patterns in these algae and compared them with published and new data from C3 and C4 plants. Our analyses identify distinct flux patterns supporting faster growth in photosynthetic cells, with some of the algae exhibiting faster ribulose 1,5-bisphosphate regeneration and increased fluxes through the lower glycolysis and anaplerotic pathways towards the tricarboxylic acid cycle, amino acid synthesis and lipid synthesis than in higher plants.


Hypofractionated versus conventionally fractionated radiotherapy for ductal carcinoma in situ (DCIS) of the breast.
Hypofractionated radiotherapy (RT) in the setting of early invasive breast cancer has been shown to have similar local control rates and cosmetic outcomes as conventionally fractionated RT. This study compares ipsilateral recurrence rates between hypofractionated and conventional RT, with and without a boost. The effect of hypofractionated RT and chest wall separation (CWS) on cosmetic outcome was also assessed. All patients with ductal carcinoma in situ (DCIS) treated between 1998 and 2012 across two sites of a single cancer institution were retrospectively studied. Patients were analysed according to those receiving conventional RT (≤2 Gy per fraction) and those receiving hypofractionated RT (>2 Gy per fraction), as well as the presence or absence of a tumour bed boost. Data were collected through electronic medical records and local cancer registry. Cosmetic outcome was scored by physicians on a four-point scale during clinical follow-up appointments. One hundred and ninety-seven patients were treated for DCIS during the study period. One hundred and forty-one were treated with conventional RT, and 56 with hypofractionated RT. After a median follow up of 4.4 years, there were 12 ipsilateral recurrences, of which seven were invasive disease and five DCIS. Ten recurrences occurred in patients who received conventional RT (7.1% recurrence rate) and two in those who received hypofractionated RT (3.6% recurrence rate) (P = 0.48). Cosmetic outcomes were not significantly different between conventional and hypofractionated RT (P = 0.06). Hypofractionation represents a suitable alternative for treating DCIS in the absence of randomised data.


A study of ventricular electrogenesis and contractility in acute pressure overload of the right heart ventricle.
A model of fat embolism induced acute pressure overload of rabbit right ventricle of heart is presented. A decrease in right ventricular contractility is registered as early as the first minutes after loading. Changes in spatial magnitude and polar coordinates of vectors of ventricular depolarization, and myocardial ischemia manifestations are recorded. The changes in electrogenesis described are attributed basically to ensuing development of hemodynamic disorders.


Recovery and repurposing of waste isopropanol with CO2-switchable deep eutectic solvents.
Large amounts of waste isopropanol (IPA) are generated in industry, rendering the recovery of IPA highly desirable due to the economic and environmental benefits. Because it forms an azeotropic mixture with water, IPA is difficult to separate from the waste stream. In the present work, a novel CO2-switchable monoethanolamine-butanol deep eutectic solvent (DES) ([MEA][BuOH]) was identified as a superior medium for separating IPA and water at ambient temperature by forming butanol-IPA mixtures. The switchable solvent system combines the advantages of homogeneous and heterogeneous systems, i.e., rapid mixing due to the low mass transfer limitations and facile product separation, respectively. The low viscosity of [MEA][BuOH], the similar physical features (polarity, dipole moment, and dielectric constant) of butanol and IPA, and the H-bonding interactions of [BuOH] with IPA are thought to enable effective IPA capture from water by the butanol. Recovery of the IPA and formation of a butanol-IPA mixture is appealing because the resultant mixture could serve as an additive or substitute for alternative fuels. The results suggest that the developed process will provide a low-cost, energy-saving, effective, and environmentally benign route to recycling and repurposing waste IPA, an environmental hazard, as a potential alternative fuel.


Encapsulation of ω-3 fatty acids in nanoemulsion-based delivery systems fabricated from natural emulsifiers: Sunflower phospholipids.
Nanoemulsions have considerable potential for encapsulating and delivering ω-3 fatty acids, but they are typically fabricated from synthetic surfactants. This study shows that fish oil-in-water nanoemulsions can be formed from sunflower phospholipids, which have advantages for food applications because they have low allergenicity and do not come from genetically modified organisms. Nanoemulsions containing small droplets (d<150 nm) could be produced using microfluidization, by optimizing phospholipid type and concentration, with the smallest droplets being formed at high phosphatidylcholine levels and at surfactant-to-oil ratios exceeding unity. The physical stability of the nanoemulsions was mainly attributed to electrostatic repulsion, with droplet aggregation occurring at low pH values (low charge magnitude) and at high ionic strengths (electrostatic screening). These results suggest that sunflower phospholipids may be a viable natural emulsifier to deliver ω-3 fatty acids into food and beverage products.


Hepatitis C Screening in Community-Based Voluntary Counselling and Testing Services in Europe: An Observational Study from the COBATEST Network 2014-2018.
The COBATEST Network links community-based voluntary counselling and testing (CBVCT) services in the European region and collects testing data using standardised data collection tools. This study aims to describe the population being screened for anti-HCV antibodies in the COBATEST Network and identify risk factors associated with a reactive HCV screening test result in the period 2014-2018. Clients aged > 16 screened for HCV in the period 2014-2018 at one of the Network's CBVCT services were included in the study. In the 5 year period, 7426 clients were screened for HCV in 22 centres in 10 countries and anti-HCV antibodies were detected in 113 (1.5%). The majority of people screened were aged 25-44, men who have sex with men (MSM), not HIV+ , not reporting a history of injecting drug use or sex work. Detection of anti-HCV antibodies was associated with being HIV + MSM (aOR 9.1, 95% CI 3.8; 21.8 compared to HIV-clients) and being a person who injects drugs (PWID, aOR 28.1, 95% CI 17.6; 45.0, compared to people with no history of injecting drug use). This study demonstrates that HIV-MSM with no history of injection drug use are using CBVCT services for HCV screening, but reactive screening test is associated with being HIV+ or PWID. The integration of HCV screening into the CBVCT service model may widen access to testing for populations that may otherwise not be tested.


Ovarian development in mice requires the GATA4-FOG2 transcription complex.
We have demonstrated previously that mammalian sexual differentiation requires both the GATA4 and FOG2 transcriptional regulators to assemble the functioning testis. Here we have determined that the sexual development of female mice is profoundly affected by the loss of GATA4-FOG2 interaction. We have also identified the Dkk1 gene, which encodes a secreted inhibitor of canonical beta-catenin signaling, as a target of GATA4-FOG2 repression in the developing ovary. The tissue-specific ablation of the beta-catenin gene in the gonads disrupts female development. In Gata4(ki/ki); Dkk1(-/-) or Fog2(-/-); Dkk1(-/-) embryos, the normal ovarian gene expression pattern is partially restored. Control of ovarian development by the GATA4-FOG2 complex presents a novel insight into the cross-talk between transcriptional regulation and extracellular signaling that occurs in ovarian development.


[Combination chemotherapy of Hodgkin's disease with vincristine, cyclophosphamide, procarbazine, and prednisolone].
Nineteen patients with stages II B-IV of Hodgkin's disease were treated with a combination of vincristine, cyclophosphamide, procarbazine, and prednisolone (CVPP regimen), given in a 3 week intervals for 3 months. Eighteen patients were available for responses. A complete remission (CR) was obtained in 9 (50%) out of 18 patients. The median duration of CR and relapse-free survival were 7 months and 26+ months, respectively. Hematologic toxicity was mild, although occasionally cumulative for aged patients and for previously treated patients. Other toxicities such as alopecia and neurotoxicity were troublesome but reversible. Thus, a further study by modification of the regimen warranted to improve the response rate and relapse-free survival.


ERICA: prevalence of fish consumption and its association with cardiovascular risk factors and healthy behavior in Brazilian adolescents.
To describe the prevalence of fish consumption and its association with cardiovascular risk factors and healthy behavior in Brazilian adolescents. The authors investigated data from 71,533 participants of the Study of Cardiovascular Risk in Adolescents (Estudo de Riscos Cardiovasculares em Adolescentes - ERICA), a nationwide, cross-sectional, school-based study. Of these, 37,815 adolescents were included for blood analyses. All prevalence estimates were presented proportionally with their 95% confidence intervals. Bivariate relationships were evaluated with Pearson's Chi-square test, and a multinomial logistic regression model was applied, considering p < 0.05. Prevalence of fish consumption in the 7 days prior to the interview was 28.6% (95%CI 26.9-30.3), significantly higher among male adolescents (p = 0.0049), Asian descendants (p = 0.0270), private and rural school students (p < 0.001), and who resided in the Northern region (p < 0.001). A positive association between fish consumption and healthy behavior (breakfast consumption: OR=1.16; 95%CI 1.10-1.22; meals with family members: lunch: OR = 1.07; 95%CI 1.01-1.13; dinner: OR = 1.13; 95%CI 1.04-1.23; physical activity: OR = 1.14; 95%CI 1.02-1.28) and an inverse association with hypertriglyceridemia (OR = 0.84; 95%CI 0.73-0.98) remained significant even after adjustment for possible confounding factors. This study demonstrated that fish consumption was associated with lower cardiovascular risk and may represent a marker of a healthy lifestyle in Brazilian adolescents.


Gender differences in suicidal ideation, suicide attempts, and suicide death among people living with HIV: A systematic review and meta-analysis.
Although excess mortality, especially suicide, is a critical trait in people living with HIV, consensus about gender differences in these areas is lacking. We conducted meta-analyses to examine gender differences in suicidal ideation, suicide attempts, and suicide death among people living with HIV. We systematically searched PubMed and Web of Science for studies written in English. In this review, suicide among people living with HIV includes suicide death, suicidal ideation, and suicide attempts. Studies reporting the suicide prevalence among males and females living with HIV were eligible for inclusion in our review. Odds ratios (ORs) and 95% confidence intervals (CIs) served as the effect size index. Fixed-effects or random-effects meta-analyses were chosen based on the size of the heterogeneity. A total of 27 studies comprising 801 017 participants from 11 countries were included in the meta-analysis. The overall prevalence of suicidal ideation was 18.0% (95% CI 13.3%-22.8%) in males and 20.8% (95% CI 16.4%-25.1%) in females, and there was a statistically significant higher risk of suicidal ideation in females living with HIV (OR 1.30; 95% CI 1.09-1.56; p < 0.05). The overall prevalence of suicide attempts was 16.8% (95% CI 9.0%-24.5%) in males and 24.7% (95% CI 12.4%-37.1%) in females, and there was a statistically significant higher risk of suicide attempts in females living with HIV (OR 1.34; 95% CI 1.02-1.75; p < 0.05). The pooled prevalence of suicide death was 1.2% (95% CI 0.5%-1.9%) among males and 0.2% (95% CI 0.1%-0.3%) among females, and the risk of suicide death between genders was not statistically significant (OR 0.78; 95% CI 0.50-1.24; p = 0.298). There were gender differences in suicidal ideation and suicide attempts among people living with HIV. Females living with HIV were more likely to experience suicidal ideation and make suicide attempts, but there were no statistically significant gender differences in suicide death. Appropriate initiatives to optimize the recognition, treatment, and management suicide behaviours of males and females living with HIV may narrow this gender gap.


Status and conservation challenges of the second-largest seagrass bed in India: Chilika lagoon.
Studies related to seagrass ecology, conservation, and management are sparse and highly biased in India. Even though the geographical distribution of seagrass is diverse in India, about 74% of the scientific publications have been made from Palk Bay and the Gulf of Mannar from Tamilnadu. Chilika, the largest lagoon in Asia, harbors the second largest seagrass meadow in India 22% of the total. The lagoon acts as a potential blue carbon stock and helps in thriving a rich floral and faunal biodiversity. However, the critical role of seagrass in this unique lagoon ecosystem is still poorly understood. This review is aimed at synthesizing the published literature about seagrass in Chilika. We believe this information would encourage more in-depth and diverse seagrass studies in the region and identify future priority areas for research. A total of seven species have been recorded from 169.2 sq. km of seagrass patch in Chilika. For the last two decades, no significant signs of decline in seagrass beds from this lagoon have been reported. Still, various natural and anthropogenic stressors could put this unique ecosystem under severe stress. Moreover, lax enforcement of existing legislation and a general lack of knowledge among the stakeholders about their ecosystem services can be significant impediments to their conservation. More targeted research on Chilika seagrass in changing climate regimes and their sustainable intensification is the need of the hour.


The paracrine stimulation of MCF-7 cells by MDA-MB-231 cells: possible role in antiestrogen failure.
The influence of paracrine factors secreted by the hormone independent breast cancer cell line MDA-MB-231 on the growth of the hormone dependent breast cancer cell line MCF-7 was examined in the absence of estradiol and in the presence of inhibitory concentrations of antiestrogens. MDA-MB-231 cells were grown on transwell membranes and co-cultured with MCF-7 cells (50,000) plated in 6-well tissue culture dishes. Growth was maximally increased (80%) when MCF-7 cells were grown in the presence of 150,000 or more MDA-MB-231 cells for 4 days. The non-steroidal antiestrogens tamoxifen (10(-10) to 10(-6) M) and 4-hydroxytamoxifen (10(-11) to 10(-7) M), the steroidal antiestrogens ICI 164384 (10(-10) to 10(-6) M) and Ru39411 (10(-11) to 10(-7) M) all inhibited estradiol (10(-11) M) stimulated MCF-7 cell growth in a dose related manner when cultured for 4 days. However, the paracrine stimulation of MCF-7 cells produced by co-culture with 250,000 MDA-MB-231 cells was not inhibited by any of these antiestrogens. These data suggest that in heterogeneous tumors, paracrine factors from hormone independent cells may reverse the growth inhibitory action that antiestrogens have on estrogen-dependent breast cancer cell growth.


Trichostatin A induces a unique set of microRNAs including miR-129-5p that blocks cyclin-dependent kinase 6 expression and proliferation in H9c2 cardiac myocytes.
The pan-histone deacetylase inhibitor (HDACI), trichostatin A (TSA), was shown to normalize interleukin-18-induced cardiac hypertrophy in vivo and in vitro; evidently, this occurred via epigenetic mechanisms that profoundly altered cardiac gene expression (Majumdar et al. in, Physiol Genom, 43: 1392, 2011; BMC Genom, 13: 709, 2012). Here, we tested the hypothesis that TSA-induced changes in chromatin architecture also led to altered expression of microRNAs that in turn, contributed to the unique transcriptome of cardiac myocytes exposed to the HDACI. Using miRCURY LNA™ Universal microRNA PCR system, we demonstrate that H9c2 cells exposed to TSA for 6 and 24 h elicited differential expression of 19 and 16 microRNAs, respectively. H9c2 cells incubated in medium-containing 100 nM of TSA elicited a rapid and robust induction of miR-129-5p. Enhanced expression of miR-129-5p was also observed in the hearts of TSA-treated mice. Induction of miR-129-5p in H9c2 cells was accompanied by reduced expression of its direct target, cyclin-dependent kinase 6 (CDK6) that is a key regulator of cell cycle. Using cell division-dependent dilution of Cell Trace™ violet measurements we showed that concomitant induction of miR-129-5p and reduced CDK6 expression were mechanistically involved in TSA-induced inhibition of proliferation of H9c2 cells. Consistent with this scenario, cells expressing an antagomiR of miR-129-5p were resistant to the anti-proliferative actions of TSA. These data indicate that although TSA treatment led to altered expression of several microRNAs, the overarching action of TSA (i.e., inhibition of cell division) in H9c2 cells was achieved via miR-129-5p.


Ozone decreases sperm quality in systemic lupus erythematosus patients.
To investigate the deleterious effects of air pollutants exposure in the Sao Paulo metropolitan region on semen quality in systemic lupus erythematosus (SLE). A seven-years longitudinal repeated-measures panel study was performed at the Laboratory of Experimental Air Pollution and Rheumatology Division. Two semen samples from 28 post-pubertal SLE patients were analyzed. Daily concentrations of air pollutants exposure: PM10, SO2, NO2, ozone, CO, and meteorological variables were evaluated on 90 days before each semen collection dates using generalized estimating equation models. Intravenous cyclophosphamide (IVCYC) and ozone had an association with a decrease in sperm quality of SLE patients. IVCYC was associated with decreases of 64.3 million of spermatozoa/mL (95% CI 39.01-89.65; p=0.0001) and 149.14 million of spermatozoa/ejaculate (95% CI 81.93-216.38; p=0.017). With regard to ozone, the most relevant adverse effects were observed from lags 80-88, when the exposure to an interquartile range increase in ozone 9-day moving average concentration led to decreases of 22.9 million of spermatozoa/mL (95% CI 5.8-40.0; p=0.009) and 70.5 million of spermatozoa/ejaculate (95% CI 12.3-128.7; p=0.016). Further analysis of 17 patients that never used IVCYC showed association between exposure to ozone (80-88 days) and decrease of 30.0 million of spermatozoa/mL (95% CI 7.0-53.0; p=0.011) and 79.0 million of spermatozoa/ejaculate (95% CI 2.1-155.9; p=0.044). Ozone and IVCYC had a consistent adverse effect on semen quality of SLE patients during spermatogenesis. Minimizing exposure to air pollution should be taken into account, especially for patients with chronic systemic inflammatory diseases living in large cities.


Enhanced expression of the P2X4 receptor in Duchenne muscular dystrophy correlates with macrophage invasion.
Following molecular and immunohistochemical analysis of the purinergic P2X4 receptor subunit in dystrophin-deficient muscle we have identified a distinct subpopulation of P2X4-positive cells infiltrating the dystrophic fibres. These cells were absent from normal muscle and rarely present in the dystrophic muscle taken before and after the onset of degeneration. We have identified these P2X4-positive cells as macrophages, demonstrating for the first time that human and mouse tissue macrophages express P2X4 in addition to P2X7 receptor subunits both in vitro and in situ. Moreover, we have demonstrated that the increase in the P2X4 expression is yet another feature of an inflammatory response identified in DNA arrays of dystrophic muscle. Immunohistochemical analysis failed to localise discernible expression of P2X4 protein in adult skeletal or cardiac muscle fibres, whilst myoblasts in culture expressed low levels of this subunit, as detected by RT-PCR and Western blotting. In light of the involvement of macrophages in the dystrophic process, the function of P2X receptors and their role in the Duchenne pathology as well as their potential role in therapeutic applications are discussed.


Initial experiences with percutaneous endovascular repair of popliteal artery lesions using a new PTFE stent-graft.
To report the percutaneous endovascular management of popliteal artery lesions with a new polytetrafluoroethylene (PTFE) stent-graft. Three patients have undergone endovascular treatment in the popliteal region owing to: (1) an iatrogenic false aneurysm with arteriovenous fistula, (2) entrapment syndrome, and (3) an isolated arterial aneurysm. In each case, arterial occlusion was treated initially with thrombolysis. The underlying vascular lesions then were stented using the new Hemobahn endograft, a nitinol stent covered internally with PTFE. Each percutaneous procedure was completed successfully without major complications. During early follow-up (6 to 7 months), no signs of intimal hyperplasia, occlusion, or stent migration have been observed. Based on these early experiences, percutaneous treatment of stenotic and aneurysmal lesions in the popliteal artery using the Hemobahn endograft appears feasible and effective. Further experience and longer follow-up are required to evaluate this new prosthesis.


Preleukemic change in the bone marrow of whole-body irradiated RFM/Up mice.
In the whole-body irradiated mouse, various late effects of radiation are observed after the recovery from acute radiation injury. Some of these account for the familiar proneness of certain mouse strains to develop leukemias. The two experiments described below were designed to (a) identify such preleukemic changes in blood-forming tissues and (b) to find ways to manipulate them experimentally with the purpose of preventing leukemia. Preleukemic change of the bone marrow appears to be a mere quantitative departure from normal in a qualitatively non-malignant tissue. It entails increased proneness of immature (precursor) cells to react with latent virus. Our data are consistent with the assumption that this proneness is enhanced (or brought about) by removal of a controlling influence exerted by the mature cells over their precursors (feed-back inhibition). Re-irradiation combined with intravenous bone marrow substitution offsets the leukemogenic influence of an earlier radiation exposure. The effect of re-irradiation on bone marrow displaying preleukemic lesions corroborates conclusions from earlier experiments on the nature of these lesions.


Recurrent hypoglycemia from insulin autoimmune syndrome.
Insulin autoimmune syndrome (IAS) is an uncommon cause of hyperinsulinemic hypoglycemia characterized by autoantibodies to endogenous insulin in individuals without previous exposure to exogenous insulin. IAS is the third leading cause of spontaneous hypoglycemia in Japan, and is increasingly being recognized worldwide in non-Asian populations. We report a case of IAS in a Caucasian woman with recurrent complaints of hypoglycemia, with laboratory findings of serum glucose 2.5 mmol/L (45 mg/dL), insulin 54,930 pmol/L (7,909 μIU/mL), connecting peptide (C-peptide) 4,104 pmol/L (12.4 ng/mL), and a corresponding insulin to C-peptide molar ratio of 13.4 during a spontaneous hypoglycemic event. Autoantibodies to insulin were markedly elevated at > 50 kU/L (> 50 U/mL). IAS should be considered in the differential diagnosis of hypoglycemia in non-diabetic individuals. Distinction from insulinoma is especially crucial to prevent unwarranted invasive procedures and surgical interventions in hypoglycemic patients.


Total ischemic burden: effect of vasoactive agents.
Treatment of the total ischemic burden is dependent on adequate documentation of both painful and painless episodes of myocardial ischemia, an understanding of the pathophysiologic mechanisms involved, and knowledge of prognosis for affected patients. Because a vasoconstrictive component appears to be an important element in the genesis of many episodes of myocardial ischemia, those vasoactive drugs that produce increased flow in the coronary circulation should be clinically useful. Nitrates and calcium blockers--especially nifedipine--have been found to be particularly valuable in this regard in both experimental and clinical trials.


Trans oral robotic surgery for oropharyngeal cancer: A multi institutional experience.
Trans Oral Robotic Surgery (TORS) has proved to be a safe and feasible treatment for oropharyngeal squamous cell carcinoma (OPSCC). The aim of this study is to analyse oncological outcomes of OPSCC patients treated with TORS. This study involved 139 patients with OPSCC, treated with TORS between 2008 and 2020. Clinicopathological characteristics, treatment details and oncological outcomes were evaluated retrospectively. The management strategies included TORS alone in 42.5%, TORS-RT in 25.2% and TORS-CRT in 30.9%. The ENE was noted in 28.8% of neck dissections. In 19 patients clinically classified as unknown primaries, the primary was found in 73.7%. Rates of local, regional relapses and distant metastasis were 8.6%, 7.2%, and 6.5%, respectively. The 5 year- Overall Survival and Disease Free Survival were 69.6% and 71.3%, respectively. TORS fits well in the modern management of OPSCC. Although definitive CRT remains a milestone, TORS is proving to be a valid and safe treatment option. The choice of the therapeutic strategy requires evaluation by a multidisciplinary team.


Remodelling the upper eyelid in the management of orbitopalpebral neurofibromatosis.
Neurofibromatosis of the orbitopalpebral complex is a debilitating disease. The development of safe craniofacial surgical technique has greatly improved outcome in the surgical palliation of this progressive condition, and various centres have reported their experience. Extensive upper eyelid neurofibroma creates a functional and aesthetic problem, which is inadequately addressed in the literature. The experience of the senior author (DM) representing the treatment of 27 such patients is currently reported. The stigmata of eyelid neurofibromatosis were scored retrospectively by a novel scale applied to standardised photographs at patient presentation. A numeric severity score was given to each of: upper eyelid ptosis, canthal malposition, and oculo-palpebral diastasis. Disease progression and post-operative result were similarly scored at various stages follow-up photography. All patients underwent a full thickness, transverse or transverse-oblique resection of upper eyelid disease, with immediate levator reconstruction and canthopexies as necessary. A mean improvement of 3.33 points was achieved in the majority of patients (median one operation per patient; mean 1.5, range 1-3). Secondary eyelid procedures (n=9 pts) represented tumour debulking, canthopexy, or procedures to deepen the fornix in patients with prostheses. This series demonstrates that a tailored, often radical palpebral remodelling in orbitopalpebral neurofibromatosis may provide functional and aesthetic benefit.


Long-term performance validation of NH3 concentration prediction model for virtual sensor application in livestock facility.
Livestock facilities commonly generate NH3, a hazardous substance that may also harm livestock. Therefore, monitoring of NH3 concentrations in livestock facilities is necessary to ensure proper control. However, NH3 is alkaline and toxic, causing corrosion inside detection sensors and making monitoring difficult. This study proposes a virtual sensor concept to complement the durability of NH3 physical sensors. The study also conducts a long-term performance validation of a data-driven NH3 concentration prediction model. Results indicate that the model's prediction performance declines sharply when the data generation pattern inside the livestock facility changes due to changes in outdoor conditions and facility operation. Furthermore, the prediction performance of the model differed depending on the training data period settings when updating the model. Hence, the model needs versioning and update management to respond to the data generation pattern in the livestock facility when operating the NH3 concentration virtual sensor. The virtual sensor is expected to enhance monitoring and reduce sensor management costs in livestock facilities.


Leuprorelin acetate depot: results of a multicentre Japanese trial. TAP-144-SR Study Group.
The clinical efficacy and safety of 3.75 or 7.5 mg leuprorelin acetate depot given subcutaneously once every 4 weeks was evaluated in a collaborative study of 81 patients with untreated prostatic cancer. Efficacy of treatment was assessed using criteria based on a meeting of the Prostatic Cancer Study Group funded by the Japanese Ministry of Health and Welfare and using National Prostatic Cancer Project criteria. Japanese criteria enabled evaluation of individual parameters, unlike the National Prostatic Cancer Project system which classified a patient as unevaluable if one evaluation parameter was unavailable. Leuprorelin acetate depot suppressed serum luteinizing hormone, follicle stimulating hormone and testosterone concentrations. Objective response rates of the prostate, bone metastases, serum prostatic acid phosphatase and soft tissue metastases, and subjective dysuria and pain responses were comparable to those found with conventional hormone therapy. Leuprorelin acetate depot was well tolerated, with no significant differences in response to the two doses.


Chitosan-Ocimum basilicum nanocomposite as a dietary additive in Oreochromis niloticus: Effects on immune-antioxidant response, head kidney gene expression, intestinal architecture, and growth.
Several studies have looked into the use of basil, Ocimum basilicum (L.) in aquaculture as a dietary additive; however, more research is needed to see the possibility of it's including in nanocarriers in aquafeeds. An experiment was undertaken to highlight the efficacy chitosan-Ocimum basilicum nanocomposite (COBN), for the first time, on Nile tilapia (Oreochromis niloticus) growth, stress and antioxidant status, immune-related parameters, and gene expression. For 60 days, fish (average weight: 23.55 ± 0.08 g) were fed diets provided with different concentrations of COBN (g/kg): 0 g [COBN0], 1 g [COBN1], 2 g [COBN2], and 3 g [COBN3], where COBN0 was kept as control diet. Following the trial, the fish were challenged with pathogenic bacteria (Aeromonas sobria) and yeast (Candida albicans) infection. In comparison to the control (COBN0), a notable increase in growth parameters (weight gain, feed intake, and specific growth rate) and intestinal morphometric indices (average intestinal goblet cells count, villous width, and length) in all COBN groups was observed, where COBN2 and COBN3 groups had the highest values. The COBN diets significantly (p < 0.05) declined levels of serum triglycerides, glucose, cholesterol, and hepatic malondialdehyde. Moreover, the higher levels of serum biochemical biomarkers (growth hormone, total protein, globulin, and albumin), immunological parameters (phagocytic activity%, nitric oxide, and lysozyme), and hepatic antioxidant parameters (superoxide dismutase, total antioxidant capacity, and glutathione peroxidase) were obvious in the COBN2 and COBN3 groups followed by COBN1. The immune-antioxidant genes (TNF-α, IL-10, IL-1β, TGF-β, GPx, and SOD) were found to be considerably up-regulated in all COBN groups (COBN2 and COBN3 followed by COBN1). Fifteen days post-challenge with A. sobria and C. albicans, the highest survival rate was recorded in the COBN2 group (83.33 and 91.67%) followed by the COBN3 group (75 and 83.33%), respectively. The findings showed that a dietary intervention with COBN can promote growth, intestinal architecture, immunity, and antioxidant markers as well as protect O. niloticus against A. sobria and C. albicans infection. As a result, the COBN at a dose of 2 g/kg could be used as a food additive for the sustainable aquaculture industry.


Peptide inhibitors of protein kinases-discovery, characterisation and use.
Protein kinases are now the second largest group of drug targets, and most protein kinase inhibitors in clinical development are directed towards the ATP-binding site. However, these inhibitors must compete with high intracellular ATP concentrations and they must discriminate between the ATP-binding sites of all protein kinases as well the other proteins that also utilise ATP. It would therefore be beneficial to target sites on protein kinases other than the ATP-binding site. This review describes the discovery, characterisation and use of peptide inhibitors of protein kinases. In many cases, the development of these peptides has resulted from an understanding of the specific protein-binding partners for a particular protein kinase. In addition, novel peptide sequences have been discovered in library screening approaches and have provided new leads in the discovery and/or design of peptide inhibitors of protein kinases. These approaches are therefore providing exciting new opportunities in the development of ATP non-competitive inhibitors of protein kinases.


Slow maturation of IgG1 avidity and persistence of specific IgM in congenital rubella: implications for diagnosis and immunopathology.
Without appropriately timed specimens, serological confirmation of congenital rubella infection may be a problem. We have compared the persistence of specific IgM and low avidity specific IgG1 in 141 sera from 120 cases of serologically confirmed congenital rubella infection with the known time scales for postnatal primary rubella. The results demonstrate that the maturation of the immune response to the rubella virus is abnormally slow in congenital rubella cases both in terms of the isotype switch and especially the development of high avidity specific IgG1. Thus avidity studies may permit serological confirmation of congenital rubella for longer than is possible with tests currently in use. The pathological implications of prolonged low avidity antibody production are discussed.


Upregulation of bfl-1 is a potential mechanism of chemoresistance in B-cell chronic lymphocytic leukaemia.
B-cell chronic lymphocytic leukaemia (B-CLL) is characterised by the progressive accumulation of monoclonal CD5(+) B cells. In a previous study, we have analysed the expression profile of apoptosis-regulating genes using a cDNA-based microarray and found overexpression of the antiapoptotic bcl-2 family member, bfl-1, in B-CLL cells with an apoptosis-resistant phenotype. In this study, bfl-1 mRNA levels have been determined by competitive PCR in an extended population of B-CLL patients to characterise its role in disease progression and development of chemoresistance. bfl-1 levels were significantly higher in patients with no response (NR) to last chemotherapy than in patients responding (partial response (PR)) to last chemotherapy (P<0.05) and in patients who had not required treatment (P<0.05). We found no correlation between bfl-1 mRNA levels and disease progression, IGHV mutational status or other clinical parameters. In addition, bfl-1 mRNA levels were inversely correlated with apoptotic response to in vitro fludarabine treatment of B-CLL cells. Specific downregulation of bfl-1 using siRNA induced apoptosis in resistant cells. Our data suggest that bfl-1 contributes to chemoresistance and might be a therapeutic target in B-CLL.


Computational intelligence modeling of hyoscine drug solubility and solvent density in supercritical processing: gradient boosting, extra trees, and random forest models.
This work presents the results of using tree-based models, including Gradient Boosting, Extra Trees, and Random Forest, to model the solubility of hyoscine drug and solvent density based on pressure and temperature as inputs. The models were trained on a dataset of hyoscine drug with known solubility and density values, optimized with WCA algorithm, and their accuracy was evaluated using R2, MSE, MAPE, and Max Error metrics. The results showed that Gradient Boosting and Extra Trees models had high accuracy, with R2 values above 0.96 and low MAPE and Max Error values for both solubility and density output. The Random Forest model was less accurate than the other two models. These findings demonstrate the effectiveness of tree-based models for predicting the solubility and density of chemical compounds and have potential applications in determination of drug solubility prior to process design by correlation of solubility and density to input parameters including pressure and temperature.


Bronchiolitis obliterans after bone marrow transplantation.
Bronchiolitis obliterans occurred in the setting of chronic graft-versus-host disease 1 year after allogeneic bone marrow transplantation for chronic myelogenous leukemia. The severe obstructive pulmonary disease followed an episode of interstitial pneumonitis. The etiology and possible relationship to graft-versus-host disease of this rare pulmonary lesion following bone marrow transplantation are discussed.


[CDKN2/p16 gene cloning and its probe preparation and application in lung cancer].
To clone CDKN2/p16(INK4a) gene, prepare its probe, and to study the change of CDKN2/p16(INK4a) gene in lung cancers. Total RNA of normal lung tissue was extracted, CDKN2/p16(INK4a) gene cDNA synthesized, and CDKN2/p16(INK4a) gene recombinant vector, constructed. Southern blot was used to study CDKN2/p16(INK4a) gene in 46 cases of lung cancers, 3 cases of normal lung tissues, 6 cases of lung tissues near cancer, and 3 cases of lymph nodes with lung cancer metastasis. Cloned CDKN2/p16(INK4a) cDNA was proved by enzyme digestion and sequencing. Southern blot showed 4.3 kb band in normal lung tissues and lung tissues near cancers, and deletion of CDKN2/p16(INK4a) gene in cancer tissues and lymph nodes with lung cancer metastasis, with a deletion rate of 17.4% (8/46). CDKN2/p16(INK4a) gene may play a role to some extent in progression of lung cancers.


The formation of mono-N-acetylhexosamine derivatives of dolichol diphosphate by pig liver microsomal fractions.
Incubation of pig liver microsomal preparations with UDP-N[U-14C]acetylglucosamine yields a 14C-labelled lipid. The requirement for Mn2+, the pH optimum, time-dependence and the reversibility by UMP of the transferase are reported. Evidence is presented in favour of the lipid being a mixture of dolichol diphosphate N-[14C]acetylglucosamine and dolichol diphosphate N-[14C]acetylmannosamine. Available data suggest that the epimerization takes place while the hexosamine is bound in this lipid-soluble form. The N-acetylmannosamine appeared not be be released into the medium. The subfractionation of the microsomal fraction to separate transferase activity from membrane-bound beta-N-acetylglucosaminidase activity is also reported.


Immunohistochemical study of the expression of drug-resistant proteins in large cell neuroendocrine carcinoma of the lung.
In 1999, the World Health Organization categorized pulmonary large cell neuroendocrine carcinoma as a variant of large cell carcinoma. However, an optimal treatment for large cell neuroendocrine carcinoma has not been established yet. Recently, multimodality therapy combining both surgery and adjuvant chemotherapy has been reported as a useful treatment for large cell neuroendocrine carcinoma, but the effect of chemotherapy on it has not yet been fully investigated. Thus, we evaluated immunohistochemical data of the expression of drug-resistant proteins in large cell neuroendocrine carcinoma. We identified 10 large cell neuroendocrine carcinomas (1.2%) out of 850 primary lung cancers that had been surgically resected. We examined the immunohistochemical staining of three drug-resistant proteins, namely, P-glycoprotein, metallothionein and glutathione S-transferase-pi to compare large cell neuroendocrine carcinoma with other histological types of lung cancer. The mean tumor cell positivity rates for P-glycoprotein, metallothionein and glutathione S-transferase-pi in large cell neuroendocrine carcinoma were 0%, 2.4 +/- 3.6% and 35.0 +/- 37.5%, respectively. The positivity rates for P-glycoprotein and glutathione S-transferase-pi were significantly lower than those in adenocarcinoma (P = 0.0003, P = 0.0009). The positivity rate for glutathione S-transferase-pi was also lower than that in squamous cell carcinoma (P = 0.0387). These drug-resistant proteins showed similar expression pattern in both large cell neuroendocrine carcinoma and small cell carcinoma except glutathione S-transferase-pi. Immunohistochemical expression of drug-resistant proteins in large cell neuroendocrine carcinoma was lower than that in adenocarcinoma and squamous cell carcinoma, and differences exist in drug-resistance between large cell neuroendocrine carcinoma and small cell carcinoma.


Cellular Composition of the Spleen and Changes in Splenic Lysosomes in the Dynamics of Dyslipidemia in Mice Caused by Repeated Administration of Poloxamer 407.
We studied the effect of dyslipidemia induced by poloxamer 407 (300 mg/kg twice a week for 30 days) on cellular composition of the spleen and splenocyte lysosomes in mice. Changes in blood lipid profile included elevated concentrations of total cholesterol, aterogenic LDL, and triglycerides most pronounced in 24 h after the last poloxamer 407 injection; gradual normalization of lipid profile was observed in 4 days (except triglycerides) and 10 days. The most pronounced changes in the spleen (increase in organ weight and number of cells, inhibition in apoptosis, and reduced accumulation of vital dye acridine orange in lysosomes) were detected on day 4; on day 10, the indices returned to normal. Cathepsin D activity in the spleen also increased at these terms. The relationship between changes in the cellular composition of the spleen and dynamics of serum lipid profile in mice in dyslipidemia caused by repeated administrations of relatively low doses of poloxamer 407 is discussed.


Noncommunicating accessory uterine cavity.
To report a case of a noncommunicating accessory uterine cavity. Case report. University-affiliated reproductive endocrinology practice. A 15-year-old nulligravida with increasing dysmenorrhea. Pelvic ultrasound, intravenous pyelogram, hysterosalpingogram, laparoscopy, laparotomy, and resection of noncommunicating accessory uterine cavity. Results of imaging studies, surgical examination, and resection of anomaly. Complete resection of accessory cavity and resolution of dysmenorrhea. The patient had a müllerian anomaly in which the uterus contained two uterine cavities. One normal uterine cavity with communication to both fallopian tubes was present along with a noncommunicating, accessory uterine cavity.


Enzymatic oxidation of the dopaminergic neurotoxin, 1(R), 2(N)-dimethyl-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline, into 1,2(N)-dimethyl-6,7-dihydroxyisoquinolinium ion.
The dopamine-derived alkaloid, 1(R),2(N)-dimethyl-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline [N-methyl(R)salsolinol] was found to be oxidized enzymatically into the more cytotoxic species, 1,2(N)-dimethyl-6,7-dihydroxyisoquinolinium ion, using enzyme sample prepared from human brain cortex. The values of the Michaelis constant and of the maximum velocity were 912 microM and 1,368 pmol/min/mg protein, respectively. The enzyme was not inhibited by inhibitors of monoamine oxidase, but was sensitive to semicarbazide. The oxidation is discussed in relation to the dopaminergic neurotoxicity of N-methyl-(R)salsolinol.


Superselective embolization for the management of postpancreatectomy hemorrhage: a single-center experience in 25 patients.
To evaluate the efficacy of superselective embolization in patients with late postpancreatectomy hemorrhage (PPH). Between January 2002 and July 2010, 25 patients (19 men) with suspected late PPH (> 24 hours after the operation) were evaluated. The primary study endpoint was technical success, defined as complete angiographic occlusion of the site of hemorrhage. Secondary study endpoints were multidetector computed tomography (CT) and angiographic findings regarding accurate detection of the site of hemorrhage, persistence of hemorrhage, or occurrence of rebleeding during clinical follow-up. Multidetector CT was performed before the intervention in 17 (68%) patients with detection of hemorrhage in 15 (88%) patients. The site of hemorrhage was detected in 23 (92%) of 25 patients by angiography. Four (17%) patients required a superselective catheter position. Embolization was attempted in all 23 patients with angiographically visible hemorrhage. In three (13%) patients, embolization could not be performed because a superselective catheter position was not achievable. Technical success was 83% (19 patients). In one patient, hemostasis was not achieved by embolization. Minor complications occurred in three (13%) patients. No major complications occurred. Three patients with technically successful embolization (16%) had a second episode of bleeding during follow-up and required repeat embolization 5-23 days after the procedure. The 30-day mortality rate was 20%. Superselective embolization is a technically and clinically effective procedure in patients with late PPH. Diagnostic angiography should be performed with a superselective microcatheter position to detect the bleeding site effectively.


The development and phase 1 evaluation of a Decision Aid for elective egg freezing.
Elective egg freezing decisions are complex. We developed a Decision Aid for elective egg freezing and conducted a phase 1 study to evaluate its acceptability and utility for decision-making. The online Decision Aid was developed according to International Patient Decision Aid Standards and evaluated using a pre/post survey design. Twenty-six Australian women aged 18-45 years, interested in receiving elective egg freezing information, proficient in English, and with access to the internet were recruited using social media and university newsletters. Main outcomes were: acceptability of the Decision Aid; feedback on the Decision Aid design and content; concern raised by the Decision Aid, and; utility of the Decision Aid as measured by scores on the Decisional Conflict Scale and on a study-specific scale assessing knowledge about egg freezing and age-related infertility. Most participants found the Decision Aid acceptable (23/25), balanced (21/26), useful for explaining their options (23/26), and for reaching a decision (18/26). Almost all reported satisfaction with the Decision Aid (25/26) and the level of guidance it provided (25/26). No participant reported serious concerns about the Decision Aid, and most would recommend it to other women considering elective egg freezing (22/26). Median Decisional Conflict Scale score decreased from 65/100 (Interquartile range: 45-80) pre-Decision Aid to 7.5/100 (Interquartile range: 0-37.5) post-Decision Aid review (p < 0.001). Median knowledge score increased from 8.5/14 (Interquartile range: 7-11) pre-Decision Aid to 11/14 (Interquartile range: 10-12) post-Decision Aid review (p = 0.01). This elective egg freezing Decision Aid appears acceptable and useful for decision-making. It improved knowledge, reduced decisional conflict and did not raise serious concerns. The Decision Aid will be further evaluated using a prospective randomised control trial. ACTRN12618001685202 (retrospectively registered: 12 October 2018).


Bleeding and ovulation control with use of a small contraceptive vaginal ring releasing levonorgestrel and estradiol.
Levonorgestrel and estradiol releasing contraceptive vaginal rings (CVR) with an outer diameter of 50 mm were used by twenty women. The treatment was given in three-week cycles followed by one treatment-free week. The treatment was planned to cover six cycles. All subjects kept records of bleeding and were controlled clinically in the course of treatment. Three subjects were followed by blood sampling. Plasma levonorgestrel, estradiol, progesterone and gonadotropins were determined. The subjects experienced no difficulties in using the CVR and 90 per cent continued the treatment through the whole experimental period of six months. One subject discontinued after three cycles because of irregular bleedings and one subject after five cycles because of urinary discomfort. Regular bleedings were observed only in three subjects and in nine cases the bleeding started with the CVR in situ during the last days of the three-week treatment period. Breakthrough bleeding occurred in the remaining eight subjects. The subjective side-effects were as follows: weight gain in four subjects, oedema in one subject and urinary discomfort in one subject. Pituitary function was not generally suppressed as judged by plasma gonadotropins. Out of the three subjects studied, two experienced ovulatory plasma progesterone concentrations.


Potentiation of morphine analgesia after pretreatment with probenecid or sulfinpyrazone.
Pretreatment with uricosuric agents probenecid or sulfinpyrazone potentiate the analgesic effects of morphine sulfate as ascertained using the phenylquinone (PQ)-induced writhing test. Doses of either uricosuric agent at 50 mg/kg had no effect on the number of PQ-induced writhes in test animals while potentiating the analgesic effects of morphine. High doses of probenecid or sulfinpyrazone alone did produce decreases in PQ-induced writing. Probenecid (50 mg/kg) did not alter hot water tail flick latency nor did it influence morphine analgesia. Attempts to uncover the underlying mechanisms in the uricosuric agent plus morphine attenuation of PQ-induced writhing were directed towards a possible displacement of morphine from plasma binding sites. However, administration of N-methyl-H3-morphine and estimation of plasma and brain morphine concentrations indicate no differences in the uricosuric drug pretreated groups compared to controls. The conflicting results in the PQ writhing test vs. hot water tail flick might indicate a false positive response in the former test. On the other hand this might be indicative of differing analgesic mechanisms for different types of pain. If the latter is true, this drug interaction may prove clinically useful.


Decreasing the pancreatic leak rate after pancreaticoduodenectomy.
Although pancreaticoduodenectomy has become a safe and effective procedure for benign and malignant pancreatic diseases in recent years, leakage of pancreaticoenteric anastomosis still remains a major cause of morbidity and even mortality. Various methods have been used to prevent pancreatic fistula with either pharmacologic or technical approaches. Based on meta-analysis of results from European and American trials, prophylactic use of octreotide to inhibit pancreatic secretion cannot be recommended for routine use in pancreaticoduodenectomy. Further randomized trials are required to clarify the role of selective use of octreotide in patients at high risk for pancreatic leakage. Technical improvement by surgeons is probably the most important approach to reduce pancreatic anastomotic leakage rate. Various technical modifications for pancreaticoenteric anastomosis have been suggested; some have been tested in randomized controlled trials, but data from randomized trials are generally scarce. Use of PG instead of PJ anastomosis, internal stenting of PJ anastomosis, pancreatic duct occlusion, and fibrin glue have not been shown to be effective in reducing pancreatic leakage rate after pancreaticoduodenectomy. One randomized trial recently showed significant reduction of pancreatic leakage rate using an external diverting stent after PJ anastomosis, and another randomized trial showed significant reduction in PJ anastomosis leakage using the binding PJ anastomosis technique. Nonetheless, further high-quality randomized controlled trials are needed to evaluate the benefit of these technical modifications in decreasing the pancreatic leakage rate after pancreaticoduodenectomy.


Clinical implication of blood glucose monitoring in general dental offices: the Ehime Dental Diabetes Study.
We examined whether general dentists can contribute to the detection of patients with undiagnosed diabetes and prediabetes by monitoring blood glucose in dental clinics. A total of 716 patients who visited clinics for dental treatment were enrolled and classified into 3 groups (mild, moderate, and severe) according to Kornman's criteria for periodontitis. The correlations between the casual blood glucose level, presence or absence of the history of diabetes, and/or severity of periodontitis were evaluated. 68 patients (9.5%) had hyperglycemia (blood glucose ≥200 mg/dL). Of these patients, 20 (29.4%) did not have a history of diabetes. Blood glucose tended to be higher with greater periodontitis severity. Of the 3 groups, the severe periodontitis group had the highest proportion of patients with hyperglycemia (p<0.0001). Patients with dental problems could be screened for diabetes, especially undiagnosed diabetes. General dentists could function as practitioners to screen for diabetes. UMIN-CTR 000014877.


Molecular dynamics simulation of dimeric and monomeric forms of human prion protein: insight into dynamics and properties.
A central theme in prion protein research is the detection of the process that underlies the conformational transition from the normal cellular prion form (PrP(C)) to its pathogenic isoform (PrP(Sc)). Although the three-dimensional structures of monomeric and dimeric human prion protein (HuPrP) have been revealed by NMR spectroscopy and x-ray crystallography, the process underlying the conformational change from PrP(C) to PrP(Sc) and the dynamics and functions of PrP(C) remain unknown. The dimeric form is thought to play an important role in the conformational transition. In this study, we performed molecular dynamics (MD) simulations on monomeric and dimeric HuPrP at 300 K and 500 K for 10 ns to investigate the differences in the properties of the monomer and the dimer from the perspective of dynamic and structural behaviors. Simulations were also undertaken with Asp178Asn and acidic pH, which is known as a disease-associated factor. Our results indicate that the dynamics of the dimer and monomer were similar (e.g., denaturation of helices and elongation of the beta-sheet). However, additional secondary structure elements formed in the dimer might result in showing the differences in dynamics and properties between the monomer and dimer (e.g., the greater retention of dimeric than monomeric tertiary structure).


Meiosis-activating sterols: background, discovery, and possible use.
Several years ago we discovered that spent media from cultured human and bull testes contain components that initiate meiosis in germ cells from fetal mouse testes which have been cultured for 6 days in the spent medium. The active substance(s) was termed meiosis-inducing substance. We later found that human follicular fluid harvested after stimulation with gonadotropins has a similar effect. These meiosis-activating substances have now been identified and characterized in extracts from bull testes and human preovulatory follicular fluid as naturally occurring sterols (meiosis-activating sterols, MAS). MAS are intermediates in the cholesterol biosynthetic pathway and are thus present in all cells which produce cholesterol de novo and from lanosterol. However, MAS accumulate only in the gonads. We discuss the possible physiological role of these sterols in initiating meiosis and in oocyte resumption of meiosis, and their potential use in promoting and preventing fertility.


Association of CYP2C9, CYP2A6, ACSM2A, and CPT1A gene polymorphisms with adverse effects of valproic acid in Chinese patients with epilepsy.
To explore the influence of CYP2C9, CYP2A6, ACSM2A, CPT1A gene polymorphisms on valproic acid (VPA) and its role in metabolism-related liver dysfunction in order to guide the clinical safety and rational use of VPA. One hundred two patients taking sodium valproate oral solution were genotyped. To assess the genotypes of relevant genes, the CYP2C9 gene was directly sequenced; for polymorphism classification, multiple Long-PCR electrophoresis was conducted for CYP2A6; and imLDR method was used for ACSM2A and CPT1A. GC-MS-SIM was used to determine the levels of VPA and 2-propyl-4-pentenoic acid (4-ene-VPA) in human plasma simultaneously. CYP2C9 mutations had a significant impact on 4-ene-VPA concentration, in patients with wild-type CYP2C9 (CYP2C9*1), which has a greater capacity for VPA metabolism than the mutant type (CYP2C9*3), liver dysfunction was substantially higher. Patients with an ACSM2A polymorphism had higher levels of ALT and AST compared with wild-type (p<0.05), but the mutations had no effect on the VPA-related liver dysfunction (p>0.05). Among different CYP2A6 and CPT1A genotype groups, there was no significant correlation in the levels of VPA, 4-ene-VPA, ALT, AST or TB (p>0.05). The content of 4-ene-VPA had no direct correlation with the incidence of liver dysfunction. Early detection of CYP2C9 gene polymorphisms may help to predict or prevent liver dysfunction caused by VPA. While the concentration of 4-ene-VPA was not suitable as an early warning index, the results provide clear theoretical guidance for the rational and safe clinical use of VPA.


GelRed/[G3T]5/Tb3+ hybrid: A novel label-free ratiometric fluorescent probe for H2O2 and oxidase-based visual biosensing.
A novel label-free ratiometric fluorescent probe is developed for the detection of H2O2 based on GelRed/[G3T]5/Tb3+ hybrid, in which GelRed (a nucleic acid dye) intercalated into the designed single-stranded DNA [G3T]5 (i.e. GelRed/[G3T]5) acts as a stable build-in reference with red emission and Tb3+ sensitized by [G3T]5 (i.e. [G3T]5/Tb3+) as a sensitive response signal with green fluorescence. With the successive addition of Hg2+ and cysteine (Cys) to GelRed/[G3T]5/Tb3+, the fluorescence of [G3T]5/Tb3+ can be effectively quenched and recovered respectively, while the fluorescence of GelRed/[G3T]5 remains unchangeable. Combined with these properties, we have demonstrated its application for label-free ratiometric fluorescence detection of H2O2 with self-calibration. The sensing mechanism is based on the specific reaction between H2O2 and Cys, the resulting disulfide reverses the Cys-mediated fluorescence changes of [G3T]5/Tb3+. This method is further applied to the monitoring of oxidase-related reactions. As the glucose oxidase (GOx)-biocatalyzed oxidation of glucose and the acetylcholine esterase/choline oxidase (AChE/ChOx) cascade yield H2O2, such two biocatalytic processes are successfully examined utilized our proposed method, indicating its potential broad applications in biomedical analysis. In addition, we have also demonstrated its feasibility for oxidase-based visual biosensing, in which only a handheld UV lamp is used. Importantly, coupled with our proposed visual biosensing, a realtime quantitative scanometric assays has been also demonstrated by using a smartphone with easy-to-access color-scanning APP as the detection platform. Compared to traditional methods, the proposed design is cost-effective, simple to prepare and easy-to-use without fluorescence labeling or chemical modification.


Spore photoproduct within DNA is a surprisingly poor substrate for its designated repair enzyme-The spore photoproduct lyase.
DNA repair enzymes typically recognize their substrate lesions with high affinity to ensure efficient lesion repair. In UV irradiated endospores, a special thymine dimer, 5-thyminyl-5,6-dihydrothymine, termed the spore photoproduct (SP), is the dominant DNA photolesion, which is rapidly repaired during spore outgrowth mainly by spore photoproduct lyase (SPL) using an unprecedented protein-harbored radical transfer process. Surprisingly, our in vitro studies using SP-containing short oligonucleotides, pUC 18 plasmid DNA, and E. coli genomic DNA found that they are all poor substrates for SPL in general, exhibiting turnover numbers of 0.01-0.2min-1. The faster turnover numbers are reached under single turnover conditions, and SPL activity is low with oligonucleotide substrates at higher concentrations. Moreover, SP-containing oligonucleotides do not go past one turnover. In contrast, the dinucleotide SP TpT exhibits a turnover number of 0.3-0.4min-1, and the reaction may reach up to 10 turnovers. These observations distinguish SPL from other specialized DNA repair enzymes. To the best of our knowledge, SPL represents an unprecedented example of a major DNA repair enzyme that cannot effectively repair its substrate lesion within the normal DNA conformation adopted in growing cells. Factors such as other DNA binding proteins, helicases or an altered DNA conformation may cooperate with SPL to enable efficient SP repair in germinating spores. Therefore, both SP formation and SP repair are likely to be tightly controlled by the unique cellular environment in dormant and outgrowing spore-forming bacteria, and thus SP repair may be extremely slow in non-spore-forming organisms.


Extensive genic and allelic heterogeneity underlying inherited retinal dystrophies in Mexican patients molecularly analyzed by next-generation sequencing.
Retinal dystrophies (RDs) are one of the most genetically heterogeneous monogenic disorders with ~270 associated loci identified by early 2019. The recent application of next-generation sequencing (NGS) has greatly improved the molecular diagnosis of RD patients. Genetic characterization of RD cohorts from different ethnic groups is justified, as it would improve the knowledge of molecular basis of the disease. Here, we present the results of genetic analysis in a large cohort of 143 unrelated Mexican subjects with a variety of RDs. A targeted NGS approach covering 199 RD genes was employed for molecular screening of 143 unrelated patients. In addition to probands, 258 relatives were genotyped by Sanger sequencing for familial segregation of pathogenic variants. A solving rate of 66% (95/143) was achieved, with evidence of extensive loci (44 genes) and allelic (110 pathogenic variants) heterogeneity. Forty-eight percent of the identified pathogenic variants were novel while ABCA4, CRB1, USH2A, and RPE65 carried the greatest number of alterations. Novel deleterious variants in IDH3B and ARL6 were identified, supporting their involvement in RD. Familial segregation of causal variants allowed the recognition of 124 autosomal or X-linked carriers. Our results illustrate the utility of NGS for genetic diagnosis of RDs of different populations for a better knowledge of the mutational landscape associated with the disease.


Reduction and catalytic properties of cytochrome P-450 LM2 in reconstituted system containing monomeric carriers.
The membrane microsomal monooxygenase system can be reconstituted in solution from NADPH-specific flavoprotein and cytochrome P-450 which exist in the monomeric state in the presence of Emulgen 913 at molar ratio of the proteins and detergent of 1:1:300. Oxidized and dithionite-reduced monomers of cytochrome P-450 were much less thermostable than its initial aggregates, while thermal stability of NADPH-specific flavoprotein did not depend on its aggregation state. Binding spectra of cytochrome P-450 monomers with benzphetamine were atypical and had an absorbance minimum at 422 nm only. The addition of benzphetamine and/or flavoprotein to cytochrome P-450 monomers did not cause the spin equilibrium shift and the low-spin form content was higher than 85% in all cases. Investigation of the dependence of the initial rates of NADPH-dependent cytochrome P-450 reduction and benzphetamine oxidation on the stoichiometry of the flavoprotein and cytochrome P-450 at their constant total concentration showed that the molar ratio of 1:1 was required for maximal activity. Thus this system works in full accordance with the mass action law.


Corpus callosum diffusion abnormalities in refractory epilepsy associated with hippocampal sclerosis.
To detect by diffusion tensor imaging (DTI) the extent of microstructural integrity changes of the corpus callosum (CC) in patients with hippocampal sclerosis (HS) and to evaluate possible association with clinical characteristics. Fourty-two patients with temporal lobe epilepsy (TLE) and HS and 30 control subjects were studied with DTI. We grouped patients according to lesion side (left or right) HS. Mean diffusivity (MD), fractional anisotropy (FA), radial (RD) and axial diffusivity (AD) were extracted from five segments in CC midsagittal section obtained by automatic segmentation. CC DTI findings were compared between groups. We also evaluated association of DTI changes and clinical characteristics. HS patients displayed decreased FA and increased MD and RD in the anterior, mid-posterior and posterior CC segments, compared to controls. No differences were observed in AD. Patients reporting febrile seizure as the initial precipitating event presented more intense diffusion changes. No differences were seen comparing left and right HS. Age at epilepsy onset, disease duration and seizure frequency were not associated with DTI findings. This is one of the largest series of TLE-HS patients evaluating CC white matter fiber integrity by DTI, which allowed us to study how some clinical characteristics, such as seizure frequency, disease duration and lesion side, are related to CC integrity. Occurrence of febrile seizure was the only factor that had significant impact on tract integrity. Diffusion changes were not restricted to the posterior part of the CC; we observed the same changes for the anterior part of the CC. Diffusion changes were characterized by an increase in RD, while the AD remained intact for all regions of the CC.


Young adults with hemophilia in the U.S.: demographics, comorbidities, and health status.
Improvements in hemophilia care over the last several decades might lead to expectations of a near-normal quality of life for young adults with hemophilia. However, few published reports specifically examine health status indicators in this population. To remedy this knowledge gap, we examined the impact of hemophilia on physical and social functioning and quality of life among a national US cohort of 141 young men with hemophilia aged 18-34 years of age who received care at 10 geographically diverse, federally funded hemophilia treatment centers in 11 states between 2005 and 2013 and enrolled in the Hemophilia Utilization Group Studies. Indicators studied included educational achievement, employment status, insurance, health-related quality of life, and prevalence of the following comorbidities: pain, range of motion limitation, overweight/obesity, and viral status. The cohort was analyzed to compare those aged 18-24 to those aged 25-34 years. When compared to the general US adult population, this nationally representative cohort of young US adults with hemophilia experienced significant health and social burdens: more liver disease, joint damage, joint pain, and unemployment as well as lower high-school graduation rates. Nearly half were overweight or obese. Conversely, this cohort had higher levels of health insurance and equivalent mental health scores. While attention has typically focused on newborns, children, adolescents, and increasingly, on older persons with hemophilia, our findings suggest that a specific focus on young adults is warranted to determine the most effective interventions to improve health and functioning for this apparently vulnerable age group.


Intramedullary Screw Fixation Comprehensive Technique Guide for Metacarpal and Phalanx Fractures: Pearls and Pitfalls.
Phalangeal and metacarpal fractures are the second and third most common upper extremity fractures after distal radius fractures with varying methods of fixation techniques. Intramedullary screw fixation is an increasingly preferred method of fixation. Benefits include early range of motion, faster recovery, limited dissection, and reduced complications. Improper technique, which is readily avoidable, can lead to suboptimal results. A review of recent literature on current techniques aims to summarize the biomechanics of intramedullary screw fixation and outline appropriate technique of placing headless compression screws for metacarpal and phalangeal fractures. We discuss through images and videos the indications, preoperative workup, and technical pearls and pitfalls to encourage surgeons to add this technique to their armamentarium and improve outcomes. intrameduallry screw fixation is a powerful option for metacarpal and phalanx fixation that allows rigid stability, enabling early return of function with excellent rates of union and total active motion. With knowledge of technical pearls presented in this article, common mistakes can be avoided to improve efficiency of screw placement and optimize patient outcomes.


A Prospective Observational Comparison of Two Approaches to Anesthetizing the Trachea for Awake Intubation.
Multiple techniques have been described for anesthetizing the lower glottis and trachea prior to awake fiberoptic intubation. The primary aim of this study is to evaluate whether direct application of local anesthetic to the lower airway via an epidural catheter under direct vision is equally efficacious when compared to use of a transtracheal block in adult patients with an anticipated difficult airway. Patients age >18 years requiring awake fiberoptic intubation who underwent upper and lower airway topicalization were observed prospectively. Following topicalization of the upper airway, patients underwent either a transtracheal block or had their trachea and lower glottis anesthetized under direct vision via dispersion of local anesthetic through a multi-orifice epidural catheter. Choice of technique was at the discretion of the attending anesthesiologist. The primary outcome was defined as the degree of coughing observed at the time of intubation based on a 4-point ordinal scale. Awake intubations in 88 patients were observed with 44 patients undergoing transtracheal block and 44 patients undergoing the epidural catheter technique. Degree of coughing with intubation was similar for each approach with a coughing score of (0, IQR (0,1)) versus (0, IQR (0,1)) in the epidural catheter and transtracheal groups respectively (p = 0.385). Duration of procedure was less in the transtracheal group (1.35 ± 1.54 min) vs. epidural catheter approach (2.86 ± 2.20 min) (p< 0.001). The epidural catheter and transtracheal approach appear to be equally effective at preventing coughing with intubation during awake fiberoptic intubation.


The Sonographic Appearance of Endometrial Intraepithelial Neoplasia.
To describe the sonographic findings of endometrial intraepithelial neoplasia (EIN), a precursor of endometrial cancer. Cases were found by word search of pathology database 1/2013 to 6/2019. One hundred and seventy-eight patients with ultrasound <1 year prior to biopsy were included. Medical records were searched for patient data. Two radiologists blindly classified images. Differences of opinion were decided by clinical report. Univariate and multivariate analyses were performed. Median time between ultrasound and first sampling procedure was 49 days. Median age was 55 (range 28-85) years. Endometrial thickness ranged from 2 to 90 mm. Mean endometrial thickness was 13 ± 6 mm in the noncancer group and 16 ± 11 mm in the cancer group (P = .02). The endometrium was almost always heterogeneous 175/178 (98%). Cysts were almost always multiple (89/109, 82%) and >1 mm (72/109, 66%). Masses were most often >5 mm (56/105, 55%) and ill-defined (41/105, 39%). Vascularity was present in 93/178 examinations (52%) and always associated with cysts and/or mass. There were 92 cancers, 25 with invasion (including 4 with tumor extension into adenomyosis). In 47 cases, the endometrial-myometrial interface was graded as ill-defined, 39 of whom had hysterectomy. There was macroscopic cancer in 11, microscopic cancer in 4, and invasive carcinoma in 12 patients (P for invasive cancer versus other outcomes = .02). Depth of invasion was 5- >95%, with 6 cancers >50%. Multivariate analysis showed thickness, polyps, and type of bleeding as the best set of independent variables for cancer (area under the receiver operating characteristic (ROC) curve [AUC] = .75). Replacing type of bleeding with age or menopausal status had AUC of .73 and .74, respectively. EIN has a variety of sonographic appearances with thickened endometrium with cysts and masses being common. Ill-definition of the endometrial-myometrial interface is a poor prognostic finding when seen in the absence of adenomyosis.


The absence of reporting standards and a lack of objective, performance-based outcomes following intramedullary nailing of tibial shaft fractures: findings from a scoping review into 179 articles.
This scoping review was conducted to summarise the outcome tools reported in the assessment of tibial shaft fractures treated with intramedullary (IM) nailing, with a key focus on knee pain and function, and performance-based outcomes. PubMed and Embase databases were searched on May 31, 2023. All study designs and populations were included, including ex vivo studies without fracture. Studies with only open or intra-articular fractures, or other fracture fixation, were excluded. Reported outcome tools and pertinent study characteristics were extracted and summarised. Of 488 articles identified, 179 met the inclusion criteria. For in vivo studies (n = 152), there were 13,705 fractures; the IM nailing approach not described for 30% of these. There were 133 unique patient outcomes, with a binary assessment of knee pain (29% of studies) and Lysholm score (21%) most common. Only 10/152 (7%) in vivo studies included an objective, performance-based measure of knee function. Fracture union was most frequent (52%) of 81 different clinical outcomes. For ex vivo studies (n = 29), there were 408 tibias included, with nail insertion location most prevalent (66% of studies) of 34 reported outcomes. The heterogeneity of outcome tools reported limits comparison between studies and the most commonly reported patient outcomes may not be the most appropriate. Future studies should report the IM nailing approach and consider capturing both patient-reported and performance-based outcomes to help inform surgical decision making.


Red blood cells differentiated in vitro using sequential liquid and semi-solid culture as a pre-clinical model.
The in vitro erythrocyte differentiation model remains a strong, clinically relevant tool to model erythroid development in normal and disease related hematopoiesis. This model also has application to developing therapeutics for diseases related to red blood cells such as sickle cell anemia where targeting increased expression of fetal hemoglobin has been a major emphasis. Since the original protocol's publication in 2002, many groups have published modified methodologies to address issues in efficiency of maturation and terminal enucleation, as well as in scalability. While all reports have merit and show efficient enucleation, the methodologies used vary widely in technique and cytokine content. Additionally, despite the strengths in these methods, reproducibility of efficient differentiation to the point of differentiation is difficult. To address these limitations, we developed a streamlined process where total PBMCs are first primed using the original liquid culture expansion phase (published in 2002) before being differentiated with minimal input via standardized, commercially purchased semi-solid medium culture pre-supplemented with erythropoietin. Our data show this methodology to produce similar levels of CD235/CD71 positivity as previous methods but with enhanced CD235 solo positivity and evidence of enucleated cells in comparison with other widely used methods. Given the difficulty and wide variation in in vitro differentiation techniques, we present this methodology as a streamlined methodology for production of mature erythroid cells with minimal input using easily purchased reagents.


Radiological investigations at the "Taiga" nuclear explosion site, part II: man-made γ-ray emitting radionuclides in the ground and the resultant kerma rate in air.
Samples of soil and epigeic lichens were collected from the "Taiga" peaceful nuclear explosion site (61.30°N 56.60°E, the Perm region, Russia) in 2009 and analyzed using high resolution γ-ray spectrometry. For soil samples obtained at six different plots, two products of fission ((137)Cs and (155)Eu), five products of neutron activation ((60)Co, (94)Nb, (152)Eu, (154)Eu, (207)Bi) and (241)Am have been identified and quantified. The maximal activity concentrations of (60)Co, (137)Cs, and (241)Am for the soils samples were measured as 1650, 7100, and 6800 Bq kg(-1) (d.w.), respectively. The deposit of (137)Cs for the top 20 cm of soil on the tested plots at the "Taiga" site ranged from 30 to 1020 kBq m(-2); the maximal value greatly (by almost 3 orders of magnitude) exceeded the regional background (from global fallout) level of 1.4 kBq m(-2). (137)Cs contributes approximately 57% of the total ground inventory of the man-made γ-ray emitters for the six plots tested at the "Taiga" site. The other major radionuclides -(241)Am and (60)Co, constitute around 40%. Such radionuclides as (60)Co, (137)Cs, (241)Am, and (207)Bi have also been determined for the epigeic lichens (genera Cladonia) that colonized certain areas at the ground lip produced by the "Taiga" explosion. Maximal activity concentrations (up to 80 Bq kg(-1) for (60)Co, 580 Bq kg(-1) for (137)Cs, 200 Bq kg(-1) for (241)Am, and 5 Bq kg(-1) for (207)Bi; all are given in terms of d.w.) have been detected for the lower dead section of the organisms. The air kerma rates associated with the anthropogenic sources of gamma radiation have been calculated using the data obtained from the laboratory analysis. For the six plots tested, the kerma rates ranged from 50 to 1200 nGy h(-1); on average, 51% of the dose can be attributed to (137)Cs and 45% to (60)Co. These estimates agree reasonably well with the results of the in situ measurements made during our field survey of the "Taiga" site in August 2009.


Agonist and antagonist effects of histamine H3 receptor ligands on 5-HT3 receptor-mediated ion currents in NG108-15 cells.
The ability of histamine H3 receptor ligands to interact with 5-HT3 receptors in NG108-15 cells was studied using the whole cell patch clamp recording technique. Imetit, a histamine H3 receptor agonist, generated inward currents and exhibited weak partial agonist activity at the 5-HT3 receptor (EC50 = 11.8 microM). Imetit-induced currents were slow to desensitize and at a high concentration reduced in size. The histamine H3 receptor antagonists iodophenpropit and thioperamide did not generate inward currents but were able to inhibit 5-hydroxytryptamine (5-HT) responses with an IC50 of 1.57+/-0.3 microM and 13.7+/-3.5 microM, respectively. Thioperamide is probably a non-competitive antagonist which may have more than one binding site on the receptor.


Arterial blood flow occlusion by high intensity focused ultrasound and histologic evaluation of its effect on arteries and surrounding tissues.
High intensity focused ultrasound (HIFU) has been used in a number of clinical studies. High intensity focused ultrasound could become the minimally invasive tumor therapy of choice if it could be used to occlude the feeding arteries of tumors by exposure from outside the body. We used high intensity focused ultrasound to insonate the femoral arteries of both thighs of 18 male and female Sprague-Dawley (SD) rats, at frequencies of 1 or 3 MHz. Duration of exposure to high intensity focused ultrasound was 5 or 10 seconds. The femoral artery that was exposed to high intensity focused ultrasound was resected immediately after or 24 hours, 48 hours, or 12 days (at 1 MHz) after insonation. Arterial blood flow occlusion was achieved at 3 MHz, but not at 1 MHz. Free-field spatial peak intensity was 800 W/cm(2) at 1 MHz and 10 kW/cm(2) at 3 MHz. Histologic examination of the resected specimen showed that changes varied with ultrasound frequency and time course after exposure to high intensity focused ultrasound. We suggest that high intensity focused ultrasound might prove useful in occluding arteries when treating tumors in the clinical setting.


Structure, function, and metabolism in the lung.
Research into the molecular basis of cellular injury and repair will lead to advances in our understanding of the pathogenesis of infectious and noninfectious respiratory disease in cattle. In order to appreciate these advances fully, it is necessary to define the basics of structure, function, and metabolism in the respiratory system.


Automated QuEChERS tips for analysis of pesticide residues in fruits and vegetables by GC-MS.
This paper reports the development of an automated method of QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) using pipet tips fitted with filtration screens and containing primary-secondary amine, magnesium sulfate, and graphitized carbon black. These tips are referred to as "QuEChERS Tips". Using loosely contained sorbent, dispersive solid phase extraction (dSPE) cleanup was performed with the QuEChERS Tips and automation. The main advantage of the QuEChERS Tips is that they are readily automated because this dSPE method does not require centrifugation. High recoveries (70-117%) and good reproducibilities (<12%) are shown for over 200 pesticides using automated QuEChERS Tips and GC-MS in various sample matrices.


Temperament and character personality profiles and personality disorders in chronic pain patients.
In his psychobiological model of personality, Cloninger developed a novel approach concerning the relationships between psychopathological syndromes and personality. We investigated 207 chronic pain patients (CPPs) and compared them to 105 pain-free control subjects. Participants were assessed using the Temperament and Character Inventory (TCI), the Structured-Clinical-Interview-II, the Beck Depression Inventory and the Spielberger Anxiety Inventory. The CPPs scored higher on the depression and state anxiety scales and 41% fulfilled the criteria of having at least one personality disorder (PD). We used a covariance analysis to control for depression and state anxiety and found that the CPPs scored higher on the Harm Avoidance Temperament Dimension and lower on the Self-Directedness and Cooperativeness Character Dimensions. In CPPs, the symptom counts of all PD subtypes were significantly related to low Self-Directedness and, to a lesser degree, low Cooperativeness. The PD symptoms in Cluster A were related to low Reward Dependence, those in Cluster B were related to high Novelty Seeking and the PD symptoms in Cluster C were related to high Harm Avoidance. In multiple hierarchical regression analyses, controlling for age, gender, depression and state anxiety, TCI scales predicted on average 23% in PD symptom counts. The Self-Directedness and Cooperativeness personality traits appeared to be significant predictors in determining the presence or absence of a PD by correctly classifying 75.8% of CPPs. The TCI provides further insight into the mechanisms underlying the development of chronic pain. This useful diagnostic instrument helps to economically and validly facilitate the identification of core PD features.


Evidence of adipocere in a burial pit from the foot and mouth epidemic of 1967 using gas chromatography-mass spectrometry.
Gas-chromatography-mass spectrometry was used to characterise the fatty acids from soils and associated tissues excavated from a 1967 Foot and Mouth burial pit. Subcutaneous fats were mainly comprised of 55-75% palmitic acid, 17-22% stearic acid and 3-16% oleic acid as well as 5-7% myristic acid. The distribution of fatty acids confirmed that the tissues were decayed to adipocere. The loss of oleic acid to <3% in two of the decayed fats suggested advanced stages of adipocere. However, adipocere formation was limited in a third tissue sample recovered from greater depth. Inductively coupled plasma atomic emission spectrometry of the pore waters revealed a decrease in Ca concentration and concurrent increase in Na concentrations this suggested that insoluble calcium salt had formed through displacement of sodium. The use of fatty acid profiles from soils and soil interstitial pore waters provide complementary evidence of adipocere formation in foot and mouth burial pits.


Giant lipoma in superior vena cava: A case report and literature review.
Intravascular lipomas are rare occurrences, especially in major vessels. This tumour is composed of adipocytes in a fibrous capsule that has a slow growth rate and usually shows no symptoms. There were only eight reports in the literature regarding intravascular lipoma located in the superior vena cava. A 54-year-old man had episodes of supraventricular tachycardia and atrial flutter for over a year. Preoperative radiological findings showed a giant mass that arose from the superior vena cava to the right atrium and a biopsy catheter showed that there were no signs of malignancy. The patient then underwent surgery through median sternotomy and the mass was extirpated on the highest part of the stalk that could be reached. The patient was stable and remained to show no symptoms or evidence of residual mass or stalk in 2 years follow-up. The surgical approach in excising lipoma in SVC should be considered wisely with the support of adequate preoperative diagnosis. Since lipoma is a very slow-growing tumour, extensive manipulation that could increase surgical technique difficulty or postoperative morbidity and mortality is not necessary.


Pulsatilla chinensis saponins ameliorated murine depression by inhibiting intestinal inflammation mediated IDO1 overexpression and rebalancing tryptophan metabolism.
Current antidepressant therapy remains unsatisfactory due to the complex pathogenesis. Emerging evidence suggested that depression is associated with inflammatory bowel disease (IBD), intestinal inflammation is an increasingly accepted factor that influences depression, but the mechanism is unclear. In the current study, we determined whether Pulsatilla chinensis saponins (PRS), a phytomedicine from Pulsatilla chinensis (Bunge) Regel with excellent anti-IBD effect, could improve the depression. Furthermore, we investigated the mechanisms to explore the relationship between IBD and depression and provide new source for the urgent development of antidepressants from phytomedicine. The antidepressant activity of PRS was accessed by behavioral test and multichannel technology in depression mice induced by Chronic Unpredictable Mild Stress (CUMS). 16S rDNA-based microbiota and RNA-seq in colon was used to explore potential intestinal metabolism affected by PRS. To illustrate the underlying mechanisms of anti-depression effect of PRS, targeted metabolomics, ELISA assay, immunofluorescence staining, Western Blot, and qPCR were carried out. The results clarified that CUMS induced depression with tryptophan (Trp) metabolism and intestinal inflammation. PRS effectively suppressed the depression and acted as a regulator of Trp/kynurenine (Kyn) metabolic and intestinal inflammation confirmed by analysis of microflora and colon RNA. Meanwhile PRS reduced interferon gamma (IFN-γ), inhibited JAK1-STAT1 phosphorylation, decreased IDO1 levels to protect against the overactivity of Trp/kyn path, suggesting that IFN-γ activated IDO1 probably a significant target for PRS to exert anti-depression effects. To further confirm the mechanism, this research expressed that PRS improved IDO1 activity and depressive behavior in mice with IFN-γ-induced depression. Furthermore, the therapeutic effect of 1-methyl-tryptophan (1-MT) well known as an IDO1 inhibitor in depression and clinically used anti-UC drug Mesalazine (MS) was demonstrated to confirm the potential mechanism. The study is the first to reveal the antidepressant effect of PRS and further demonstrate its potential therapeutic targets. In addition, it also clarifies that the Trp/kyn pathway is the crosstalk between IBD and depression and provides new choice for depression treatment. And it also provides an important basis for the follow-up development and exploration of anti-intestinal antidepressants.


Effect of adrenalectomy on acceleration of gluconeogenesis by calcium ions, adenosine 3':5'-cyclic monophosphate and adrenaline in rat kidney tubules.
1. Gluconeogenesis from pyruvate was measured in renal-cortical-tubules fragments prepared from fed male rats 6-8 days after adrenalectomy or sham adrenalectomy. The response of this process to 3':5'-cyclic AMP and adrenaline was compared in these two states at two Ca2+ concentrations. 2. Adrenalectomy decreased the percentage stimulation of gluconeogenesis by 3':5'-cyclic AMP, but increased percentage stimulation by adrenaline. Cortisol treatment of adrenalectomized rats (50 mg/kg, twice daily for 2 days) did not reverse the change in responsiveness to 3':5'-cyclic AMP and adrenaline. 3. Stimulation of gluconeogenesis by 1 micron-adrenaline was unaffected by 10 micron-propranolol (beta-blocker) in either state. Phentolamine (alpha-blocker; 10 micron) totally blocked stimulation of gluconeogenesis by 1 micron-adrenaline in the sham-operated condition, but was only partially effective in this respect after adrenalectomy.


[The diagnosis and treatment procedure in periodic disease].
The main forms of periodic disease are presented, clinical cases with the different forms are described. A special attention is paid to the problems of diagnosis and treatment in the emergency surgery of the abdominal organs is abdominal form of the disease.


Acanthamoeba myosin IA, IB, and II heavy chains are synthesized in vitro from Acanthamoeba messenger RNA.
Addition of a myosin IB polyclonal antibody, which reacts with the heavy chains of both myosins IA and IB, to the [35S]methionine-labeled proteins synthesized in vitro using Acanthamoeba messenger RNA specifically immunoprecipitated radioactive polypeptides that migrated on sodium dodecyl sulfate-polyacrylamide gels exactly with the heavy chains of purified myosins IA and IB. A myosin II polyclonal antibody specifically immunoprecipitated a radioactive polypeptide which migrated exactly with the heavy chain of myosin II. These results provide strong evidence that Acanthamoeba myosins IA, IB, and II are separate gene products and that the purified proteins contain native, undegraded heavy chains. These results are especially important for myosins IA and IB, because they possess unusually small heavy chains for myosins.


Biodegradable implants versus standard metal fixation for displaced radial head fractures. A prospective, randomized, multicenter study.
This multicenter, prospective, randomized study compares the use of biodegradable polylactide pins with standard metal mini-fragment implants for the treatment of displaced radial head fractures. It compares complication rates and clinical outcomes of both treatment methods. At 2 years, 135 (82%) of 164 patients were available for evaluation. Equivalence of treatment method was defined as a difference of 10% or less in the number of complication-free patients. Functional status was assessed by using the Broberg and Morrey Elbow Score and compared by an unpaired t test. Good or excellent clinical results were achieved by 92% (56/61) of the control patients and 96% (71/74) of the polylactide patients. The incidence of complication-free patients was 3.7% less in the polylactide group than in the control group. The 1-sided 95% confidence interval for the treatment difference between the 2 groups was more than -6.1%. Biodegradable polylactide pins have at least comparable outcomes as standard metal implants for the internal fixation of reconstructable displaced radial head fractures.