Insulin resistance but not hypertriglyceridemia per se is associated with endothelial dysfunction in chronic hypertriglyceridemia.
To infer the relative impact of elevated triglyceride levels and insulin resistance on endothelial dysfunction in patients with chronic hypertriglyceridemia (HTG). Endothelial function was studied in 11 HTG patients and 16 normolipidemic controls. Cumulative-dose infusions of 5-hydroxytryptamine (5HT) and sodium nitroprusside were infused locally into the brachial artery to study endothelium-dependent and endothelium-independent vasodilation, respectively. Data of the HTG patients were dichotomized around the median of insulin resistance, calculated as HOMA-index, forming HTG groups with mild (HTG-MIR) and severe insulin resistance (HTG-SIR). HTG patients had higher triglyceride levels and smaller LDL particle size than controls (both P< or =0.001), whereas these parameters did not differ between both HTG groups. Insulin resistance was higher in both HTG groups than in controls (11.1 (7.0-14.5) and 4.9 (4.0-6.7) vs. 2.4 (4.9-5.2), respectively, both P<0.001). Similarly, free fatty acid levels, another indicator of insulin resistance, were highest in the HTG-SIR group, followed by those in the HTG-MIR and control group (0.7 (0.6-0.8), 0.5 (0.4-0.6) and 0.4 (0.3-0.4) mmol/l, respectively, all P<0.05). Endothelial-dependent vasodilation was similar in HTG-MIR and controls. In contrast, the response to 5HT was attenuated in the HTG-SIR group compared to controls (low and high dose by, respectively, -60 and -44%, both P<0.01), and tended to be lower than in the HTG-MIR group (-43%, P=0.068 and -41%, P=0.100, respectively). Endothelium-independent vasodilation did not differ between the three groups. These findings indicate that chronic hypertriglyceridemia per se is not associated with endothelial dysfunction. In contrast, the presence of insulin resistance, characterized by hyperinsulinemia and FFA elevation, contributes to the induction of endothelial dysfunction in chronic HTG.


Identification of three novel HLA-DRB1 alleles, HLA-DRB1*07:150, HLA-DRB1*13:345 and HLA-DRB1*14:256.
Three novel HLA-DRB1 alleles, HLA-DRB1*07:150, HLA-DRB1*13:345 and HLA-DRB1*14:256, were identified.


Low-dose intravenous recombinant tissue-type plasminogen activator therapy for patients with stroke outside European indications: Stroke Acute Management with Urgent Risk-factor Assessment and Improvement (SAMURAI) rtPA Registry.
The purpose of this study was to determine the safety and efficacy of intravenous recombinant tissue-type plasminogen activator (0.6 mg/kg alteplase) within 3 hours of stroke onset in Japanese patients outside the indications in the European license. Of the 600 patients who were treated with recombinant tissue-type plasminogen activator, 422 met the inclusion criteria of the European license (IN group) and 178 did not (OUT group). The OUT group was inversely associated with any intracerebral hemorrhage (adjusted OR, 0.50; 95% CI, 0.29-0.84), positively associated with an unfavorable outcome (2.48; 1.55-3.94) and mortality (2.04; 1.02-4.04), and not associated with symptomatic intracerebral hemorrhage (0.53; 0.11-1.79) or complete independency (0.65; 0.40-1.03) after multivariate adjustment. Functional and vital outcomes 3 months after low-dose recombinant tissue-type plasminogen activator in patients outside the European indications were less favorable compared with those included in the indications; however, the risk of intracerebral hemorrhage was not.


Mitochondria bridge HIF signaling and ferroptosis blockage in acute kidney injury.
Ferroptosis, a form of regulated cell death, plays an important role in acute kidney injury (AKI). Previous studies have shown that prolyl hydroxylase domain protein (PHD) inhibitors that activate HIF signaling provide strong protection against AKI, which is characterized by marked cell death. However, the relationship between PHD inhibition/HIF signaling and ferroptosis in AKI has not been elucidated. Here, we review recent studies to explore the issue. First, we will review the literature concerning the functions of HIF in promoting mitophagy, suppressing mitochondrial respiration and modulating redox homeostasis. Second, we will describe the current understanding of ferroptosis and its role in AKI, particularly from the perspective of mitochondrial dysfunction. Finally, we will discuss the possibility that mitochondria link PHD inhibition/HIF signaling and ferroptosis in AKI. In conclusion, we propose that HIF may protect renal cells against ferroptosis in AKI by reducing mitochondrial oxidative stress and damage.


Clinical utility of topiramate extended-release capsules (USL255): Bioequivalence of USL255 sprinkled and intact capsule in healthy adults and an in vitro evaluation of sprinkle delivery via enteral feeding tubes.
The objectives of these two studies were to determine if beads from extended-release topiramate capsules sprinkled onto soft food are bioequivalent to the intact capsule and if beads from the capsule can be passed through enteral gastrostomy (G-) and jejunostomy (J-) feeding tubes. Bioequivalence of 200-mg USL255 (Qudexy XR [topiramate] extended-release capsules) sprinkled onto soft food (applesauce) versus the intact capsule was evaluated in a phase 1, randomized, single-dose, crossover study (N=36). Pharmacokinetic evaluations included area under the curve (AUC), maximum plasma concentration (Cmax), time to Cmax (Tmax), and terminal elimination half-life (t1/2). If 90% confidence intervals (CI) of the ratio of geometric least-squares means were between 0.80 and 1.25, AUC and Cmax were considered bioequivalent. In separate in vitro experiments, 100-mg USL255 beads were passed through feeding tubes using gentle syringe pressure to develop a clog-free bead-delivery method. Multiple tube sizes (14- to 18-French [Fr] tubes), dilutions (5 mg/15 mL-25 mg/15 mL), and diluents (deionized water, apple juice, Ketocal, sparkling water) were tested. Area under the curve and Cmax for USL255 beads sprinkled onto applesauce were bioequivalent to the intact capsule (GLSM [90% CI]: AUC0-t 1.01 [0.97-1.04], AUC0-∞ 1.02 [0.98-1.05]; Cmax 1.09 [1.03-1.14]). Median Tmax was 4h earlier for USL255 sprinkled versus the intact capsule (10 vs 14 h; p=0.0018), and t1/2 was similar (84 vs 82 h, respectively). In 14-Fr G-tubes, USL255 beads diluted in Ketocal minimized bead clogging versus deionized water. Recovery of USL255 beads diluted in deionized water was nearly 100% in 16-Fr G-, 18-Fr G-, and 18-Fr J-tubes. For patients with difficulty swallowing pills, USL255 sprinkled onto applesauce offers a useful once-daily option for taking topiramate. USL255 beads were also successfully delivered in vitro through ≥14-Fr G- or J-tubes, with tube clogging minimized by portioning the dose and using glidant diluents for smaller tubes.


Management of Gastrointestinal Bleeding and Resumption of Oral Anticoagulant Therapy in Patients with Atrial Fibrillation: A Multidisciplinary Discussion.
Direct oral anticoagulants (DOACs) are recommended for the prevention of thromboembolism in patients with atrial fibrillation (AF), and are now preferred over vitamin K antagonists due to their beneficial efficacy and safety profile. However, all oral anticoagulants carry a risk of gastrointestinal (GI) bleeding. Although the risk is well documented and acute bleeding well codified, there is limited high-quality evidence and no guidelines to guide physicians on the optimal management of anticoagulation after a GI bleeding event. The aim of this review is to provide a multidisciplinary critical discussion of the optimal management of GI bleeding in patients with AF receiving oral anticoagulants to help physicians provide individualized treatment for each patient and optimize outcomes. It is important to perform endoscopy when a patient presents with bleeding manifestations or hemodynamic instability to determine the bleed location and severity of bleeding and then perform initial resuscitation. Administration of all anticoagulants and antiplatelets should be stopped and bleeding allowed to resolve with time; however, anticoagulant reversal should be considered for patients who have life-threatening bleeding or when the bleeding is not controlled by the initial resuscitation. Anticoagulation needs to be timely resumed considering that bleeding risk outweighs thrombotic risk when anticoagulation is resumed early after the bleeding event. To prevent further bleeding, physicians should prescribe anticoagulant therapy with the lowest risk of GI bleeding, avoid medications with GI toxicity, and consider the effect of concomitant medications on potentiating the bleeding risk.


[The production of high-yielding strain of rubella virus from wild type virus extracted from a patient in the Western Siberia in 2006].
The study was targeted to investigate the propagation of rubella virus in the cell cultures of various origins and with different cultivation methods. The high-yielding strain of rubella virus was produced. The "spinner-culture" cultivation method was applied and the strain's RNA was detected in 10-8 dilution in real time mode. This strain is supposed to be used in preparation of the standard antigen to implement in the development of immune enzyme test system targeted to the rubella virus specific antibodies.


Structural characterization of chlorophyll-a by high resolution tandem mass spectrometry.
A high resolution Fourier transform ion cyclotron resonance (FTICR) mass spectrometer is used for characterizing the fragmentation of chlorophyll-a. Three tandem mass spectrometry (MS/MS) techniques, including electron-induced dissociation (EID), collisionally activated dissociation (CAD), and infrared mutiphoton dissociation (IRMPD) are applied to the singly protonated chlorophyll-a. Some previously unpublished fragments are identified unambiguously by utilizing high resolution and accurate mass value provided by the FTICR mass spectrometer. According to this research, the two long aliphatic side chains are shown to be the most labile parts, and favorable cleavage sites are proposed. Even though similar fragmentation patterns are generated by all three methods, there are much more abundant peaks in EID and IRMPD spectra. The similarities and differences are discussed in detail. Comparatively, cleavage leading to odd electron species and H(•) loss both seem more common in EID experiments. Extensive loss of small side groups (e.g., methyl and ethyl) next to the macrocyclic ring was observed. Coupling the high performance FTICR mass spectrometer with contemporary MS/MS techniques, especially IRMPD and EID, proved to be very promising for the structural characterization of chlorophyll, which is also suitable for the rapid and accurate structural investigation of other singly charged porphyrinic compounds.


Management of Hypertelorism.
To summarize the experience of the author with the treatment of hypertelorism. The author has been heading a high-caseload department of craniofacial surgery for 38 years; the research is based on his experience with this pathology by this time. The charts of 38 patients were used for this research. No statistic was used; the author has just given his personal insights as the result of a professional life devoted to the problem. Most of the hypertelorism cases requiring surgical correction are rare interorbital clefts (Tessier's 14-12). Among the syndromic ones, cranio-fronto orbital dysplasia is the most demanding for surgery because it is associated with craniosynostosis, which has to be addressed at the same time. Among the technics published for hypertelorism correction, craniofacial bipartition has our preference for several reasons: easily done and redone when necessary, safer to the vascularization, and trophicity of displaced parts of the skeleton. Complex craniofacial conditions such as hypertelorism have to be treated only in specialized craniofacial centers by a multidisciplinary team. The caseload has to be high, and the follow-up is very strict to get the benefits of experience to improve the results.


Decrease in the mechanical strength of bones of rats administered cadmium.
The mechanical properties of the bones of young, adult and old rats administered various concentrations of cadmium were measured to prove the direct effect of cadmium on the bones of young rats. The young rats were divided into three subgroups, which were administered 0 (control), 5 and 10 ppm cadmium, respectively. The adult rats were subdivided into six groups, administered 0, 10, 20, 40, 80 and 160 ppm cadmium, respectively. The old rats were divided into three subgroups, which were administered 0, 80, and 160 ppm cadmium, respectively. The length of the administration was 4 weeks in every group. The decrease in the mechanical strengths of bones of young rats administered with cadmium was observed. On the other hand, no change in mechanical strength of bones was observed in the case of adult and old rats, administered up to 160 ppm cadmium. The correlation between the cadmium in bones and the decrease in the strength of the bone shows that cadmium directly affects the mechanical properties of bones of young rats.


Periodontal and systemic health of morbidly obese patients eligible for bariatric surgery: a cross-sectional study.
In obese patients, periodontitis might be associated with deprived systemic health. Edmonton obesity staging system (EOSS) is a new tool for classification of obesity that considers the metabolic, physical, and psychological health. The cross-sectional study aimed to evaluate the periodontal status of morbidly obese patients eligible for bariatric surgery and the association between periodontitis, obesity-related comorbidities, and EOSS. Morbidly obese patients eligible for bariatric surgery underwent detailed periodontal examination and were divided into the periodontitis group (PG) and the non-periodontitis group (NPG). The medical and demographic data were obtained from medical files, while behavioural data were obtained by the interview. Descriptive statistics and simple statistical tests were used to summarise the characteristics of the sample and the differences between PG and NPG. The logistic regression models were used to calculate the association (odds ratio (OR)) between periodontitis and obesity-related diseases and EOSS. The study included 79 patients, with an average BMI of 44.6 kg/m2 (SD = 7.2). The prevalence of periodontitis was 65% (CI 95% 53%-75%). PG patients (n = 51) were older, more often smokers and were more often hypertensive than NPG patients (n = 28) (p < 0.05). Hypertension was positively associated with periodontitis with adjusted OR 3.98 (95% CI 1.23-12.8; p = 0.021)) and age with adjusted OR 1.06, (95% CI 1.01-1.13; p = 0.038)), while other tested conditions (diabetes, dyslipidaemia, and smoking habits) did not show significant association with periodontitis. Periodontitis did not correlate with EOSS or other obesity-related comorbidities (p > 0.05). The morbidly obese patients eligible for bariatric surgery show a high prevalence of periodontitis and, therefore, are advised to be examined by a dentist before undergoing surgery. They have higher odds of hypertension but not of other obesity-related diseases or higher stages of EOSS. The medical personnel should raise awareness among obese patients on the potential association of poor periodontal health with hypertension. NCT04653714.


Putative function of TAP63α during endochondral bone formation.
P63, a member of the P53 tumor suppressor family, is known to play important functions in cancer and development. Interestingly, previous studies have shown that p63 null mice are absent or have truncated limbs, while mutations in human P63 cause several skeletal syndromes that also show limb and digit abnormalities, suggesting its essential role in long bone development. Indeed, we detected increased level of p63 transcript in hypertrophic MCT cells (an established cell model of chondrocyte maturation) than in proliferative MCT cells. To investigate the in vivo role of P63 upon endochondral bone formation, we have established transgenic mouse lines in which HA- and Flag-tagged TAP63α (the longest P63 isoform) is driven by the hypertrophic chondrocyte-specific Col10a1 regulatory elements. Skeletal staining of Col10a1-TAP63α transgenic mice at either embryonic day 17.5 (E17.5) or postnatal day 1 (P1) observed accelerated ossification in long bone, digit and tail bones compared to their wild-type littermates, suggesting a putative function of P63 during skeletal development. We also detected decreased level of Sox9 and Bcl-2 transcripts, while Alp and Ank are slightly upregulated in Col10a1-TAP63α transgenic mouse limbs. Further immunohistochemical analysis confirmed the decreased Sox9 expression in the proliferative and hypertrophic zone of these mice. Von Kossa staining suggests increased mineralization in hypertrophic zone of transgenic mice compared to littermate controls. Together, our results suggest a role of TAP63α upon skeletal development. TAP63a may promote endochondral ossification through interaction with genes relevant to matrix mineralization and chondrocyte maturation or apoptosis.


Comparative effectiveness of cladribine tablets versus fingolimod in the treatment of highly active multiple sclerosis: A real-world study.
Cladribine tablets and fingolimod have similar marketing authorisations in Europe for the treatment of patients with highly active relapsing multiple sclerosis (HA-RMS). In the absence of direct head-to-head studies, real-world data are important to assess the comparative effectiveness of these oral disease-modifying therapies (DMTs). The primary objective of the present study was to compare relapse rates between patients who received either cladribine tablets or fingolimod. This multicentre retrospective study conducted in the United Kingdom and Germany assessed non-inferiority in relapse rates of cladribine tablets versus fingolimod in HA-RMS patients over a 12-month period. Eligible patients who initiated treatment with cladribine tablets or fingolimod at least 12 months prior to the screening date were sampled consecutively until the target sample size was reached. Patients were censored at discontinuation of study treatment, commencement of another DMT, death, loss to follow-up, or at 12 months post-baseline, whichever happened earliest. The primary analytic timeframe for physician-confirmed relapse outcomes was the study effectiveness period (nine months of follow-up after an initial 12 weeks of treatment). Propensity score analysis was applied based on the inverse probability of treatment weighting approach. The primary analytic cohort consisted of 1,095 HA-RMS patients: 610 (55.7%) receiving cladribine tablets and 485 (44.3%) receiving fingolimod. Fewer patients discontinued cladribine tablets and/or switched to another DMT compared with fingolimod (0.2% versus 3.5%, respectively). The primary endpoint, adjusted annualised relapse rate (ARR), was 0.10 (95% confidence interval [CI]: 0.07-0.14) for cladribine tablets and 0.14 (95% CI: 0.10-0.20) for fingolimod. The adjusted ARR ratio of cladribine tablets versus fingolimod was 0.68 (95% CI: 0.42-1.11). Given the entire 95% CI was less than the non-inferiority margin of 1.2, cladribine tablets was non-inferior to fingolimod. In this real-world retrospective cohort study, cladribine tablets demonstrated comparable effectiveness to fingolimod at one year following treatment initiation. The full treatment dosage of cladribine tablets is completed over two years and so these results may be conservative.


[Protective stoma after deep anterior rectal resection: pro].
There is still controversy about the necessity of a diverting stoma after deep anterior resection with total mesorectal excision for rectal cancer. Recent results of randomized controlled trials and from systematic reviews have improved the currently available data. A significant benefit was shown for patients with diverting stoma in terms of clinically relevant anastomotic leakage and re-operation rates. The influence on mortality is not as clear. However, analysis of the data of 19 prospective studies within a systematic review including more than 9,000 patients, revealed a significant benefit for stoma creation. Furthermore, the rate of patients with stoma 5 years after primary resection was lower in the group of patients with diverting stoma. The purpose of this manuscript is to show the necessity of a diverting stoma based on the currently available data.


Response to and toxicity of gemcitabine for recurrent ovarian cancer according to number of previous chemotherapy regimens.
Gemcitabine is used not only as a second-line, but also as a third-line or higher regimen for taxane/platinum-resistant recurrent ovarian cancer. The purpose of this study was to clarify the response to and toxicity of gemcitabine for recurrent ovarian cancer according to the number of previous chemotherapy regimens. The subjects were patients with taxane/platinum-resistant recurrent ovarian cancer on gemcitabine treatment at the present hospital between June 2007 and September 2013. We retrospectively reviewed the medical records. Response and adverse events were assessed using the Response Evaluation Criteria in Solid Tumors version 1.1. and the Common Terminology Criteria for Adverse Events v4.0, respectively. The subjects consisted of 65 patients. The median number of previous chemotherapy regimens was 3 (range, 1-7). Overall response rate was 4.6%, and disease control rate (DCR) was 40.0%. DCR versus one, two, three, and ≥four previous chemotherapy regimens was 83.3%, 45.0%, 36.4%, and 23.5%, respectively. Grade 3/4 neutropenia, anemia, and thrombocytopenia occurred in 52.3%, 9.2%, and 9.2% of patients, respectively. Prevalence of grade 3/4 neutropenia according to one, two, three, and ≥four previous chemotherapy regimens was 66.7%, 55.0%, 54.5%, and 41.2%, respectively. Prevalence of anemia, thrombocytopenia, and almost all the non-hematological toxicities also did not increase with an increase in the number of previous chemotherapy regimens. Although DCR decreased as the number of previous chemotherapy regimens increased, the toxicities did not increase. Gemcitabine may be relatively safe in heavily pretreated ovarian cancer patients.


Prochlorperazine in childhood: side-effects.
The side-effects of prochlorperazine (CompazineR StemetilR) were traced in thirty-six children up to the age of sixteen years and compared with those collected from literature. These side-effects appeared to be predominantly neurological and were independent of the dosage. Impaired consciousness, dyskinesia, pyramidal signs and hypertonus were the main neurological manifestations. Dyskinesia was the most frequent sign. These side-effects disappear spontaneously after discontinuation of the drug. Instant cure can be acquired by intra-muscular administration of orphenadrine (DisipalR).


Cryopreservation of fetal rat brain tissue later used for intracerebral transplantation.
Intracerebral grafting of immature brain tissue is now widely used as a tool to study neuronal development and regeneration in the brain and spinal cord. This has stimulated the interest in methods for storage of such tissue before transplantation. In this study a method for cryopreservation of immature rat central nervous tissue is presented and discussed in relation to current cryobiological principles. The method was applied to brain tissue from 16- and 17-day-old fetal rats, including the neocortex, habenula, septum and basal forebrain, cerebellum, and retina. After storage in liquid nitrogen from 6 to 52 days the tissue was grafted into the brain of adult rats. The recipients survived for 23 to 673 days before their brains were processed by current neuroanatomical, histological methods. The presence of graft tissues was recorded and their cellular and connective organization was examined, including their exchange of nerve connections with the host brain. The results obtained were comparable with results from other studies where the same tissues were grafted immediately after removal from the donor, and a study of cryopreservation of developing hippocampal tissue. We conclude that cryopreservation is a reliable method for storage of immature neural tissue later to be used for intracerebral grafting.


Age related differences in functional synchronization of EEG activity as evaluated by means of TMS-EEG coregistrations.
It was recently demonstrated that the characteristics of EEG rhythms preceding a transcranial magnetic stimulation (TMS) of the motor cortex (M1) influence the motor-evoked potential (MEP) amplitude with a peculiar pattern, thus reflecting the M1 functional state. As physiological aging is related to a decrease in motor performance and changes in excitability and connectivity strength within cerebral sensorimotor circuits, we aimed to explore whether aging affects EEG-MEP interactions. Using MRI-navigated TMS and multichannel EEG, we compared the EEG-MEP interactions observed in healthy aged subjects with those observed in young volunteers. We divided the MEPs amplitude into two different subgroups consisting of "high" and "low" MEPs, based on the 50th percentile of their amplitude distribution. Then we analysed the characteristics of the pre-stimulus EEG from M1 and correlated areas separately for the "high" and "low" MEPs, comparing the two conditions. In both young and old subjects, significantly larger MEPs were evoked when the stimulated M1 was coupled in the beta-2 band with the homolateral prefrontal cortex. Conversely, only in young participants was the MEP size modulated when the M1 and homolateral parieto-occipital cortices were coupled in the delta band. The elderly didn't show this kind of pattern. Importantly, this coupling was significantly higher in elderly brains than in young brains, both for high and low MEPs. Our results suggest an age-related significant influence of time-varying coupling of spatially patterned EEG rhythms on motor cortex excitability in response to TMS.


The bi-directional regulation of filamin on the ATPase activity of smooth muscle myosin.
The aim of this study is to investigate the functional relationship between filamin, a known actin binding protein, and myosin and the effects of filamin on the interaction between myosin and actin. Ultra-centrifugation method was used to investigate the binding of filamin to both phosphorylated and unphosphorylated myosins. Mg-ATPase activities of both phosphorylated and unphosphorylated myosins in the presence and absence of actin were measured to observe the effects resulted from filamin-actin and filamin-myosin interactions. It was found that filamin is also a myosin binding protein. Filamin inhibited the actin activated Mg-ATPase activity of phosphorylated myosin and stimulated Mg-ATPase of phosphorylated myosin in the absence of actin; in addition, filamin stimulated Mg-ATPase activity of unphosphorylated myosin in both the presence or absence of actin. The result suggest that the effects of filamin on the myosin Mg-ATPase activities are bi-directional, i.e., stimulatory via binding to myosin and inhibitory via binding to actin.


Molecular characterization and phylogenetic analysis of Plasmodium vivax, Plasmodium falciparum, Plasmodium ovale, Plasmodium malariae and Plasmodium cynomolgi.
18S ribosomal RNA gene sequences of different species of Plasmodium were aligned and analyzed to determine the molecular diversity among different species of Plasmodium. AT content of P. cynomolgi, P. ovale, P. falciparum, P. vivax and P. malariae ranged from 62.30 to 63.15, 63.90 to 65.29, 66.67 to 68.40, 61.66 to 63.25 and 64.09 to 76.36 in case respectively. GC content of P. cynomolgi, P. ovale, P. falciparum, P. vivaxand P. malariae ranged from 36.85 to 37.70, 34.71 to 36.43, 31.60 to 33.27, 23.64 to 35.91 and 36.75 to 38.34 respectively. Molecular weight (Da) of P. cynomolgi, P. ovale, P. falciparum, P. vivax and P. malariae ranged from 1,160,127.00 to 1,400,332.00, 437,273.00 to 481,438.00, 668,021.00 to 685,243.00, 540,378.00 to 1,176,962.00 and 132,788.00 to 305,211.00 respectively. Relative melting temperature (Tm) of P. cynomolgi and P. ovale varied from 0.36799 to 0.36837 and 0.36712 to 0.36814 respectively. In P. falciparum and P. vivax relative Tm ranged from 0.36545 to 0.36626 and 0.36802 to 0.36866 respectively. Relative Tm of P. malariae varied from 0.36174 to 0.36790.


Respiratory muscle work influences locomotor convective and diffusive oxygen transport in human heart failure during exercise.
It remains unclear if naturally occurring respiratory muscle (RM) work influences leg diffusive O2 transport during exercise in heart failure patients with reduced ejection fraction (HFrEF). In this retrospective study, we hypothesized that RM unloading during submaximal exercise will lead to increases in locomotor muscle O2 diffusion capacity (DM O2 ) contributing to the greater leg VO2 . Ten HFrEF patients and 10 healthy control matched participants performed two submaximal exercise bouts (i.e., with and without RM unloading). During exercise, leg blood flow was measured via constant infusion thermodilution. Intrathoracic pressure was measured via esophageal balloon. Radial arterial and femoral venous blood gases were measured and used to calculate leg arterial and venous content (CaO2 and CvO2 , respectively), VO2 , O2 delivery, and DM O2 . From CTL to RM unloading, leg VO2 , O2 delivery, and DM O2 were not different in healthy participants during submaximal exercise (all, p > .15). In HFrEF, leg VO2 (CTL: 0.7 ± 0.3 vs. RM unloading: 1.0 ± 0.4 L/min, p < .01), leg O2 delivery (CTL: 0.9 ± 0.4 vs. RM unloading: 1.4 ± 0.5 L/min, p < .01), and leg DM O2 (CTL: 31.5 ± 11.4 vs. RM unloading: 49.7 ± 18.6 ml min-1 mmHg-1 ) increased from CTL to RM unloading during submaximal exercise (all, p < .01), whereas CaO2 -CvO2 was not different (p = .51). The degree of RM unloading (i.e., % decrease in esophageal pressure-time integral during inspiration) was related to the % increase in leg DM O2 with RM unloading (r = -.76, p = .01). Our data suggest RM unloading leads to increased leg VO2 due to greater convective and diffusive O2 transport during submaximal exercise in HFrEF patients.


Insulin inhibits cellular and herpes simplex virus DNA synthesis in transformed lymphoblastoid cell lines.
Treatment of uninfected and herpes simplex virus-infected lymphoblastoid cells with 4 units of insulin per milliliter resulted in the inhibition of both cellular and viral DNA synthesis.


Bacterial community composition and structure in an Urban River impacted by different pollutant sources.
Microbial communities in terrestrial fresh water are diverse and dynamic in composition due to different environmental factors. The goal of this study was to undertake a comprehensive analysis of bacterial composition along different rivers and creeks and correlate these to land-use practices and pollutant sources. Here we used 454 pyrosequencing to determine the total bacterial community composition, and bacterial communities that are potentially of fecal origin, and of relevance to water quality assessment. The results were analyzed using UniFrac coupled with principal coordinate analysis (PCoA) to compare diversity, abundance, and community composition. Detrended correspondence analysis (DCA) and canonical correspondence analysis (CCA) were used to correlate bacterial composition in streams and creeks to different environmental parameters impacting bacterial communities in the sediment and surface water within the watershed. Bacteria were dominated by the phyla Proteobacteria, Bacteroidetes, Acidobacteria, and Actinobacteria, with Bacteroidetes significantly (P<0.001) higher in all water samples than sediment, where as Acidobacteria and Actinobacteria where significantly higher (P<0.05) in all the sediment samples than surface water. Overall results, using the β diversity measures, coupled with PCoA and DCA showed that bacterial composition in sediment and surface water was significantly different (P<0.001). Also, there were differences in bacterial community composition between agricultural runoff and urban runoff based on parsimony tests using 454 pyrosequencing data. Fecal indicator bacteria in surface water along different creeks and channels were significantly correlated with pH (P<0.01), NO2 (P<0.03), and NH4N (P<0.005); and in the sediment with NO3 (P<0.015). Our results suggest that microbial community compositions were influenced by several environmental factors, and pH, NO2, and NH4 were the major environmental factors driving FIB in surface water based on CCA analysis, while NO3 was the only factor in sediment.


Basidiosins A and B: Cyclopentapeptides from the entomophthoralean fungus Basidiobolus meristosporus.
Two new cyclopentapeptides, basidiosins A and B (1 and 2) were isolated from the mycelia extracts of entomophthoralean fungus Basidiobolus meristosporus RCEF 4516. The structures were determined based on spectroscopic methods, and the absolute config urations were assigned by Marfey's method on their acid hydrolyzates. Compounds 1 and 2 were identified as cyclo(L-Thr-L-Leu- L-Ile-D-Tyr-D-Thr) and cyclo(L-Thr-L-Leu-L-Val-D-Val-D-Ser), respectively. They were evaluated for the biological activities including antibacterial, antifungal and antioxidative activities. Furthermore, the biosynthetic pathway of 1 was proposed by bioinformatic analysis. This is the first study on the isolation of natural products from Basidiobolus fungus.


BmBR-C Z4 is an upstream regulatory factor of BmPOUM2 controlling the pupal specific expression of BmWCP4 in the silkworm, Bombyx mori.
20-hydroxyecdysone (20E)-induced expression of the wing disc cuticle protein gene BmWCP4 was mediated by the transcription factor BmPOUM2, which binds to the cis-response elements (CREs) of BmWCP4 gene in Bombyx mori. In this study we report the regulation of BmPOUM2. RT-PCR analysis indicated that in response to 20E, BmPOUM2 was expressed at higher levels in the wing discs during the pre-pupal and mid-pupal stages than other stages and the expression pattern of BmBR-C Z1, BmBR-C Z2 and BmBR-C Z4 was in tandem with the expression of BmPOUM2. BmBR-C Z4 was induced by 20E in the wing discs, whereas BmBR-C Z1 and BmBR-C Z2 were not. Three potential BR-C Z4 cis-response elements (CREs) were identified in the promoter region of BmPOUM2. The expression of BmPOUM2 mRNA and protein was increased by the over-expression of BmBR-C Z4 in BmN cells, which acted at the promoter of BmPOUM2. Electrophoretic mobility shift assay (EMSA) and the luciferase activity analysis under the control of wild-type and mutants of the BR-C Z4 CREs suggested that BmBR-C Z4 protein bound to the predicted BRC-Z4 CRE C (-684 ∼ -660). Taken together the data suggest that BmBR-C Z4 is a direct upstream regulator of BmPOUM2 and regulates the pupal-specific expression of BmWCP4 through BmPOUM2.


Mechanism of the action of the inhibitor of reverse transcriptase.
An inhibitor of reverse transcriptase of Moloney leukemia virus was reported previously in the cytoplasm of the cultured cells (1). In this report, the mechanism of the inhibition was examined. The inhibitory activity was completely abrogated by treatment at 55 degrees C for 20 min. Destruction of the reaction products of reverse transcriptase was not observed. Strong inhibition was observed by preincubation of poly rA oligo dT used as a template-primer, together with the inhibitor, but was not by incubation of poly rA or oligo dT with the inhibitor separately. It is suggested that the inhibitor reacts with poly rA oligo dT complex and interferes with attachment of the reverse transcriptase to the template-primer complex.


Application of various genetic analysis techniques for detecting two rare cases of 9p duplication mosaicism during prenatal diagnosis.
The identification of genetic mosaicism and the genetic counseling needed following its discovery have been challenging problems in the field of prenatal diagnosis. Herein, we describe the clinical phenotypes and various prenatal diagnostic processes used for two rare cases of 9p duplication mosaicism and review the prior literature in the field to evaluate the merits of different methods for diagnosing mosaic 9p duplication. We recorded ultrasound examinations, reported the screening and diagnosis pathways, and analyzed the mosaic levels of the two cases of 9p duplication using karyotype analysis, chromosomal microarray analysis (CMA), and fluorescence in situ hybridization analysis (FISH). Case 1 had a normal clinical phenotype for tetrasomy 9p mosaicism, and Case 2 showed multiple malformations caused by both trisomy 9 and trisomy 9p mosaicism. Both cases were initially suspected after non-invasive prenatal screening (NIPT) based on cell-free DNA. The mosaic ratio of 9p duplication found via karyotyping was lower than what was discovered by CMA and FISH, in both cases. Contrary to previous findings, the mosaic level of trisomy 9 found by karyotype analysis was greater than what was found by CMA, in terms of complex mosaicism involving trisomy 9 and trisomy 9p, in Case 2. NIPT can indicate 9p duplication mosaicism during prenatal screening. Different strengths and limitations existed in terms of diagnosing mosaic 9p duplication by karyotype analysis, CMA, and FISH. The combined use of various methods may be capable of more accurately determining break-points and mosaic levels of 9p duplication during prenatal diagnosis.


MicroRNA-30c-5p arrests bladder cancer G2/M phase and suppresses its progression by targeting PRC1-mediated blocking of CDK1/Cyclin B1 axis.
MicroRNAs (miRNAs) play a critical role in cancer development and progression, the dis-regulation of miR-30c-5p has been observed in various malignant tumors but no research was done in bladder cancer (BCa). This study aims to investigate the downregulation of miR-30c-5p in BCa, and examine its mechanism and prognostic significance. Bioinformatics analyses and clinical specimens were employed to investigate the relationship between miR-30c-5p and clinical information in BCa patients. The expression levels of miR-30c-5p and its target gene were assessed by real-time PCR and western blot. Cell viability was evaluated through clonogenic capacity, CCK-8, and EdU assays. Cell cycle distribution and cell apoptosis were determined by flow cytometry. The anti-tumor effect of miR-30c-5p was also validated in animal models. The expression levels of miR-30c-5p were significantly decreased in both bladder tumor tissue and BCa cell lines. Low miR-30c-5p expression was found to be correlated with unfavorable TNM stages and poor prognosis. Over-expressing miR-30c-5p was observed to hinder BCa cell growth, migration, and invasion abilities and causing cell cycle arrest. Mechanistically, miR-30c-5p directly binds and suppresses PRC1, thereby blocking the CDK1/Cyclin B1 axis in BCa, thus impairing BCa cell viability and inducing cell cycle arrest at G2/M phase. Down-regulated miR-30c-5p promotes BCa through its target gene PRC1, miR-30c-5p is a favorable biomarker for predicting clinical outcomes in BCa patients and has the potential to be a therapeutic target.


Synthesis of Red-Light-Responsive Pheophorbide-a Tryptamine Conjugated Photosensitizers for Photodynamic Therapy.
Six methyl pheophorbide-a derivatives were prepared by linking a tryptamine side chain at the C-131 , C-152 and C-173 positions of pheophorbide-a. Prepared conjugates were characterized and evaluated for their photocytotoxicity against A549 cells. The conjugate 6 a with strong absorption at 413 nm (Soret band), 663-671 nm (Q bands) and comparable fluorescence quantum yield (0.26) was found to exhibit significant cytotoxicity (659 nM). Molecular integration of pheophorbide-a and tryptamines showed synergistic effects as the most potent conjugate 6 a was identified with enhanced photocytotoxicity when compared to methyl pheophorbide-a. The conjugate 6 a was smoothly taken up by A549 cells and exhibited intracellular localization predominantly to lysosome in the cytoplasm. Upon photoirradiation 6 a generated singlet oxygen to show potent cytotoxicity toward A549 cells.


Hepatic dysfunction in patients with extrahepatic portal venous obstruction.
Extrahepatic portal venous obstruction (EHPVO) developing due to thrombotic occlusion of the portal vein in children is generally considered a benign disease. Whether hepatic dysfunction develops in these patients in the absence of a gastrointestinal bleed has not been well studied. Forty-three patients with EHPVO who had not bled in the last 3 months were studied. Patients were divided into those with (group I) or without ascites (group II). Matched cirrhotic patients with ascites (group III) served as controls. Clinical, biochemical, ultrasonographic, and histopathological evaluation was carried out. Portal biliopathy was assessed in five patients in group I and in 12 patients in group II by cholangiography. Of 43 EHPVO patients, ascites was seen in nine (21%) patients (group I). Thirty-four patients had no ascites (group II). Serum ALT (54 +/- 24 vs. 34 +/- 10 IU/l, P < 0.01), albumin (3.2 +/- 0.3 vs. 3.7 +/- 0.4 g/dl, P < 0.01), and prothrombin time difference (9.0 +/- 4.5 vs. 2.4 +/- 1.9 s, P < 0.05) were deranged in patients in group I compared with group II. Patients in group I were 4 years older, and the duration of portal hypertension was longer than in group II (11.5 vs. 5.6 year, P < 0.05). Portal biliopathy changes were significantly more severe in group I than in group II patients. Ascites was high gradient in all the patients in group I and the serum-ascitic albumin gradient was comparable between groups I and III. None of the EHPVO patients, but four cirrhotic patients, developed spontaneous bacterial peritonitis during a follow-up of 11 +/- 4 months. Hepatic dysfunction in the form of ascites and deranged liver functions is not uncommon in patients with EHPVO, more so in patients with prolonged portal hypertension. Based on our data it would be worthwhile to study whether prolonged portal vein thrombosis in EHPVO patients could lead to progressive liver disease.


Mimicry and inhibition of nerve growth factor effects: interactions of staurosporine, forskolin, and K252a in PC12 cells and normal rat chromaffin cells in vitro.
The structurally similar compounds staurosporine and K252a are potent inhibitors of protein kinases. K252a has previously been reported to inhibit most or all of the effects of nerve growth factor (NGF) on PC12 pheochromocytoma cells, and staurosporine has been reported both to inhibit and to mimic NGF-induced neurite outgrowth from a PC12 cell subclone in a dose-dependent manner. We have studied the interactions of these agents with each other, with NGF, and with forskolin, an activator of adenylate cyclase, on the parent PC12 cell line and on normal neonatal and adult rat chromaffin cells. Staurosporine alone or in conjunction with forskolin induces outgrowth of short neurites from PC12 cells but does not substitute for NGF in promoting cell survival. It does not abolish NGF-induced neurite outgrowth but does reverse the effects of NGF on catecholamine synthesis. K252a abolishes NGF-induced neurite outgrowth but only partially decreases outgrowth induced by NGF plus forskolin. It does not inhibit neurite outgrowth produced by staurosporine or staurosporine plus forskolin. These findings with PC12 cells suggest that staurosporine might act downstream from K252a and NGF on components of one or more signal transduction pathways by which NGF selectively affects the expression of certain traits. Both neonatal and adult rat chromaffin cells show dramatic flattening and extension of filopodia in response to staurosporine, an observation suggesting that some of the same pathways might remain active in cells that do not exhibit a typical NGF response. Only a small amount of neurite outgrowth is observed, however, and only in neonatal cultures.(ABSTRACT TRUNCATED AT 250 WORDS)


Variations in the 5-hydroxy-indoleacetic acid concentration in the cerebrospinal fluid during the sleep-wake cycle in man.
A variation in the ventricular fluid 5-HIAA concentration has been observed during the phases of the sleep-wake cycle in man. This variation affects the NREM phase, during which 5-HIAA increases markedly, rather than the waking state and the REM phase. The results are discussed in relation to the possible relevance of serotonin to the sleep mechanism.


The challenges to public health nurse practice in rural Ireland.
The primary objective was to understand the contemporary public health nurse's (PHN) role and the issues that they face working in rural Ireland. This study was based on an ethnographic approach with 13 PHNs working in rural areas in the South West of Ireland. A combination of solicited diaries and semi-structured interviews referred to as the diary/interview method were employed. Diaries were used by the PHNs to record their working day on a staggered basis from February to April 2017 with the subsequent interviews carried out in June and July 2017. Working as a PHN in a rural area presented a number of issues such as time spent on traveling as a result of geographical disparity & poor road networks; client transport issues; a sense of working in isolation; communication issues with respect to computer/tablet hardware availability, mobile phone and broadband connectivity and the availability of, and the physical access to services. PHNs operating in rural communities face a distinct set of challenges that they have a limited ability to address. Legislators, health care providers and policymakers need to create a supportive environment that helps address these challenges in Ireland.


Non-negative Independent Factor Analysis disentangles discrete and continuous sources of variation in scRNA-seq data.
Single-cell RNA-seq analysis has emerged as a powerful tool for understanding inter-cellular heterogeneity. Due to the inherent noise of the data, computational techniques often rely on dimensionality reduction (DR) as both a pre-processing step and an analysis tool. Ideally, DR should preserve the biological information while discarding the noise. However, if the DR is to be used directly to gain biological insight it must also be interpretable-that is the individual dimensions of the reduction should correspond to specific biological variables such as cell-type identity or pathway activity. Maximizing biological interpretability necessitates making assumption about the data structures and the choice of the model is critical. We present a new probabilistic single-cell factor analysis model, Non-negative Independent Factor Analysis (NIFA), that incorporates different interpretability inducing assumptions into a single modeling framework. The key advantage of our NIFA model is that it simultaneously models uni- and multi-modal latent factors, and thus isolates discrete cell-type identity and continuous pathway activity into separate components. We apply our approach to a range of datasets where cell-type identity is known, and we show that NIFA-derived factors outperform results from ICA, PCA, NMF and scCoGAPS (an NMF method designed for single-cell data) in terms of disentangling biological sources of variation. Studying an immunotherapy dataset in detail, we show that NIFA is able to reproduce and refine previous findings in a single analysis framework and enables the discovery of new clinically relevant cell states. NFIA is a R package which is freely available at GitHub (https://github.com/wgmao/NIFA). The test dataset is archived at https://zenodo.org/record/6286646. Supplementary data are available at Bioinformatics online.


Association of beta-defensin 1 single nucleotide polymorphisms with Crohn's disease.
It has been suggested that deficient defensin expression is associated with the chronic inflammation of Crohn's disease. The regional localization of Crohn's disease, ileal or colonic disease can be linked to different defensin profiles. As constitutive beta-defensin 1 has a colonic expression, we considered it of interest to investigate single nucleotide polymorphisms (SNPs) of the beta-defensin 1 gene (DEFB1) in Crohn's disease. Three SNPs of the DEFB1 gene, DEFB1 G-20A (rs11362), DEFB1 C-44G (rs1800972) and DEFB1 G-52A (rs1799946), were genotyped either by Custom TaqMan SNP genotyping assays or by restriction fragment length polymorphisms (RFLP) in 190 patients with Crohn's disease and 95 Hungarian controls. It was found that the G-20A and C-44G SNPs had a strong association with the colonic and ileocolonic localizations of the disease, respectively, but no association was detected for the ileal localization. A significantly higher frequency of the GA genotype of G-20A was observed among patients with colonic localization (60%) as compared with healthy controls (39%), with an odds ratio (OR) of 2.39. The GG genotype of C-44G SNP, which is regarded as a protective genotype, was much less frequent (4%) among patients than among controls (12%), OR 3.367. These results indicate that genetic variations in the DEFB1 gene encoding constitutive human beta-defensin 1 may be associated with the risk for Crohn's disease and may determine disease phenotype, e.g. colonic localization.


Genetic causes of primary aldosteronism.
Primary aldosteronism is characterized by at least partially autonomous production of the adrenal steroid hormone aldosterone and is the most common cause of secondary hypertension. The most frequent subforms are idiopathic hyperaldosteronism and aldosterone-producing adenoma. Rare causes include unilateral hyperplasia, adrenocortical carcinoma and Mendelian forms (familial hyperaldosteronism). Studies conducted in the last eight years have identified somatic driver mutations in a substantial portion of aldosterone-producing adenomas, including the genes KCNJ5 (encoding inwardly rectifying potassium channel GIRK4), CACNA1D (encoding a subunit of L-type voltage-gated calcium channel CaV1.3), ATP1A1 (encoding a subunit of Na+/K+-ATPase), ATP2B3 (encoding a Ca2+-ATPase), and CTNNB1 (encoding ß-catenin). In addition, aldosterone-producing cells were recently reported to form small clusters (aldosterone-producing cell clusters) beneath the adrenal capsule. Such clusters accumulate with age and appear to be more frequent in individuals with idiopathic hyperaldosteronism. The fact that they are associated with somatic mutations implicated in aldosterone-producing adenomas also suggests a precursor function for adenomas. Rare germline variants of CYP11B2 (encoding aldosterone synthase), CLCN2 (encoding voltage-gated chloride channel ClC-2), KCNJ5, CACNA1H (encoding a subunit of T-type voltage-gated calcium channel CaV3.2), and CACNA1D have been reported in different subtypes of familial hyperaldosteronism. Collectively, these studies suggest that primary aldosteronism is largely due to genetic mutations in single genes, with potential implications for diagnosis and therapy.


The effect of prostaglandin I2 and 6a-Carba-PGI2 on the motility of isolated osteoclasts.
We have recently found that calcitonin (CT), a hormone which inhibits osteoclastic bone resorption, completely abolishes the normally intense cytoplasmic movement of isolated osteoclasts. We have also found that prostaglandin (PG)I2 causes an identical change in behaviour. In this paper we extend our investigations into the mode of action of PGI2 and a stable analogue, 6a-Carba-PGI2. We found that, unlike CT which causes prolonged immotility in osteoclasts, the effect of PGI2 and 6a-Carba-PGI2 were transient. Our results suggest that the transient nature of the inhibition was neither caused by inactivation of these compounds, nor was it due to production in the cultures of an osteoclastic stimulator. CT, PGI2 and 6a-Carba-PGI2 all appear to operate by increasing the intracellular cyclic AMP level. We found no refractoriness to either CT, dibutyryl cyclic AMP or 8-bromo cyclic AMP, and neither PGI2 nor 6a-Carba-PGI2 affected the sensitivity of osteoclasts to CT or dibutyryl cyclic AMP. This implies that refractoriness of osteoclasts to PGI2 and 6a-Carba-PGI2 develops at some stage in the interaction between PG and cell proximal to cyclic AMP production. We also found that there was cross-tachyphylaxis between PGI2 and 6a-Carba-PGI2, and this suggests that these two compounds share a receptor site on osteoclasts.


Serosurvey of spotted fever group Rickettsia in equids from western Pará, Amazon, Brazil.
From a previous large epidemiological survey, we randomly selected 474 serum samples (463 horses and 11 mules) distributed among four municipalities of Pará state, Amazon region, Brazil, and from three types: farm animal, urban carthorse, and sport horse. Samples were tested by indirect fluorescence antibody test (IFAT ≥ 64) for antibodies reactive to spotted fever group (SFG) rickettsiae using Rickettsia rickettsii as crude antigens. From the 474 equids tested, 149 (31.4%) had ticks attached during sampling, belonging mostly to the species Dermacentor nitens. The overall seroprevalence for SFG rickettsiae was 31.4% (95% confidence interval: 27.3-35.9%) with 149 seropositive animals out of 474 screened. Notably, 77 equids (16.2%) had high endpoint titers ranging from 512 to 16,384, indicating that they had been exposed to SFG rickettsiae not long before sampling. Animal type affected rickettsial seroprevalence, with significantly higher values among farm horses when compared with urban and sport animals. Presence of dogs and tick infestation were negatively associated with equid seropositivity to R. rickettsii. This is the first report of SFG rickettsiae-reactive antibodies in equids from Pará state, Brazilian Amazon.


Synthesis, cytotoxicity and QSAR study of N-tosyl-1,2,3,4-tetrahydroisoquinoline derivatives.
1-Substituted-N-tosyl-1,2,3,4-tetrahydroisoquinoline analogs (4a-4l) were synthesized using the modified Pictet-Spengler reaction and evaluated for cytotoxicity. All tetrahydroisoquinolines displayed cytotoxicity against MOLT-3 cell lines, except for p-methoxy analog 4d. Interestingly, the o-hydroxy derivative 4k was shown to be the most potent cytotoxic against HuCCA-1, A-549 and MOLT-3 cell lines. The lowest IC₅₀ value of 1.23 μM was observed for MOLT-3 cells. Trimethoxy analog 4f exerted the most potent activity against HepG2 with an IC₅₀ of 22.70 μM, which is lower than the reference drug, etoposide. QSAR studies showed that total symmetry index (Gu), 3D-MoRSE (Mor31v and Mor32u) and 3D Petitjean index (PJI3) were the most important descriptors accounting for the observed cytotoxicities. The most potent cytotoxic compound (4k) against MOLT-3 had the highest Gu value, correspondingly the inactive p-methoxy analog (4d) had the lowest Gu value. On the other hand, the highest molecular mass compound (4f) was shown to be the most potent cytotoxic against HepG2 cells. The studies disclose that tetrahydroisoquinolines 4f and 4k are potentially interesting lead pharmacophores that should be further explored. The QSAR models provided insights into the physicochemical properties of the investigated compounds.


Psychrophilic anaerobic digestion biotechnology for swine mortality disposal.
The objective of this study was to investigate the feasibility of using psychrophilic anaerobic digestion in sequencing batch reactors (PADSBRs) to co-digest grinded swine carcasses and swine manure slurry at 20 degrees C and 25 degrees C. The PADSBRs were operated on two-week and four-week treatment cycle lengths, which included the fill, react, and draw phases. Two carcass loading rates (CLRs) were tested, that is 20 and 40g of carcass per litre of manure, which were equivalent to 4 and 8 times, respectively, the normal mortality rate on commercial farms. The PADSBR performance was compared to that of PADSBRs operated at 25 degrees C and fed manure only. The addition of swine carcass to PADSBR feed did not affect the stability of the bioreactors at both CLRs. The performance of the PADSBRs co-digesting swine carcasses was not statistically different from the control in terms of biogas production and quality. There was no accumulation of volatile fatty acids in the bioreactors at the end of the treatment cycle. The mixed-liquor pH and alkalinity remained within acceptable ranges for the anaerobic microflora. Also, there was no operational problem caused by the formation of foam and scum in the system.


Separation and isolation of BTV dsRNA segments and viral proteins.
Bluetongue virus (BTV) genome contains ten double-stranded RNA segments. The sequence of the plus strand of each of the BTV genomic double-stranded RNAs is the same as that of its mRNA, which encodes for a single viral protein, except the smallest S4 segment which can encode for two nonstructural proteins, primarily for the release assistance of the viral progeny. The separation and isolation of each BTV dsRNA segment and viral protein have provided extensive data related to its viral infection, pathology, suppression of host cellular functions, and eventual apoptosis of the infected host cells. This cytoplasmic virus is also an animal killer that costs the U.S. livestock industry at least $125 million yearly. However, this virus has no known effect on humans. Thus, it is very safe to carry out investigation with the virus, preferably in a BSL-2 laboratory.


Acupuncture induced pneumothorax: A case report.
Various forms of alternative medicinal practices are gaining popularity. With this, there will be rise in the complications arising from these practices. Acupuncture is also such practice which though safe can rarely cause life-threatening complications.


Use of β-galactosidase liposome model as a novel method to screen freeze-drying cryoprotectants.
This study developed a novel method of screening cryoprotectants used to improve the survivability of lyophilized Lactobacillus helveticus. To develop a liposome encapsulated β-galactosidase (β-gal) as a cell membrane model, the β-gal liposome was characterized in terms of mean size, poly dispersity index, zeta potential, along with transmission electron microscopy. 800 W of ultrasonic power and 10 min of sonication time were the optimal experimental conditions to obtain the desirable β-gal liposome. Subsequently, different cryoprotectants were mixed with the β-gal liposome during freeze-drying. After freeze-drying, liposomes were hydrolized, and the protective effect of cryoprotectants was assessed as the release rate of encapsulated β-gal. The lowest release rate of β-gal was obtained using 10 mg/100 ml trehalose and 0.2 mg/100 ml hyaluronic acid.


Expression of rat liver heme oxygenase gene during development.
We studied the genetic expression during fetal development of heme oxygenase, the rate-limiting enzyme in the oxidation of heme to bilirubin. The transcription of the heme oxygenase gene in livers of fetal and neonatal rats (9 days before birth to 28 days after birth) was examined. Hybridization analyses of total RNA from livers of these animals using cDNA for rat heme oxygenase as the probe revealed a single mRNA species of approximately 18 S in every sample examined. The mRNA level was above the adult level throughout the course of study and reached a maximum 24 h after birth. The high level of heme oxygenase mRNA in fetuses was unaffected when Sn-protoporphyrin, a potent inhibitor of heme oxygenase, was administered to their mothers. On the other hand, the mRNA levels in the mothers treated with this heme analog were substantially increased, possibly by the same mechanism as for the induction by heme. Sn-protoporphyrin potentiates induction of heme oxygenase mRNA in cobalt chloride-pretreated rats, and also acts as a potent inhibitor of heme oxygenase enzyme activities. Our results also indicate that high heme oxygenase levels during fetal maturation are due to an increase in transcription of the gene. Thus, Sn-protoporphyrin which crosses the placenta controls fetal hyperbilirubinemia by direct enzyme inhibition.


Protective effects of various antioxidants during ischemia-reperfusion in the rat retina.
Oxidative stress during ischemia-reperfusion (I/R) is thought to be a major cause of retinal injury after I/R. The present study was aimed at investigating the protective role of antioxidant application. Four commonly used antioxidants (vitamin E=alpha tocopherol, lutein, fenugreek=Trigonella foenum-graecum and germander=Teucrium multicaule) were applied in ischemia-reperfusion (I/R) injury of the right retinae of 51 adult pigmented rats. Each of the antioxidants was administered every 6 h, beginning 6 h before the ischemia. After 60 min ischemia and 24 h reperfusion, we assayed (1) oxidative damage by measuring malondialdehyde (MDA), (2) apoptosis by measuring activated caspase-3 (using immunoblots), and (3) intrinsic antioxidative capacity by measuring glutathione (GSH) levels in the retinae. In the order of lutein>Trigonella>vitamin E>Teucrium, all four compounds were effective in preventing retinal damage by I/R, as (1) they significantly decreased the formation of MDA (8.83, 16.48, 17.24, 18.5 nmol/100 mg tissue wet weight, respectively) compared with I/R without protection (23.29 nmol/100 mg tissue wet weight; controls: 8.0 nmol/100 mg tissue wet weight); (2) they significantly inhibited the activation of caspase-3 [0.01, 0.02, 0,02, and 0.04 arbitrary units (AU), respectively, versus control, 0.0, and I/R, 0.08 AU]; and (3) they significantly decelerated the loss of GSH (from control levels of 36.04 nmol/100 mg tissue wet weight) to 30.4, 15.98, 18.1, 15.02 nmol/100 mg tissue wet weight, respectively (lutein, Trigonella, vitamin E, Teucrium), compared with unprotected I/R (12.84 nmol/100 mg tissue wet weight). Our study shows that lutein, Trigonella, Teucrium and vitamin E exert protection against in vivo retinal I/R injury in rats; this may recommend these compounds (in particular, lutein) for clinical use in patients with different types of ocular I/R injuries.


The Role of Religiosity in Families at High Risk for Depression.
About forty years ago we began a study of the offspring of depressed (high-risk) and not depressed (low-risk) parents, matched for age and gender, from the same community. We interviewed all of their biological children, blind to the clinical status of the parents. Over the years, we returned to re-interview the families at baseline, 2, 10, 20, 25 30, and 35 years. As the years went by and the sample grew up, we also interviewed the third generation, the grandchildren. As technology became available, we included measures of electrophysiology and magnetic resonance imaging in order to better understand the mechanisms of risk. At the 10-year follow up, we included measures of religion and spirituality - namely, personal religious/spiritual importance and frequency of religious service attendance. We included these measures in all subsequent waves including a more extensive follow up of religious beliefs at the 35-year follow up. This paper describes the study design and highlights the key findings of the role of religious/spiritual belief in the transmission and endurance of depression using clinical and biological approaches. We describe study findings based on clinical measures, as well as physiological measures that employed electrophysiology and magnetic resonance imaging. Taken together, the findings suggest that religiosity/spirituality is protective against depression in high-risk individuals at both clinical and physiological levels. The findings suggest religiosity interacts with both culture and biology in its impact on depression.


[Relationship between the magnitude of the inflowing current of isolated Limnea stagnalis mollusk neurons and the external concentration of the current-bearing calcium and strontium ions].
The dose-inward current relationships for current-carrying ions Ca2+ and Sr2+ were studied by means of intracellular dialysis and voltage clamp in isolated neurons of the mollusc Limnea stagnalis. For Ca2+ and Sr2+ ions these relationships had ceiling. The data obtained are explained in terms of activation theory using two-barrier model of calcium channel. Values of velocity constants vx for ion x2+ transition over the second energetic barrier and of constants Kx corresponding to effectiveness of binding in the channel's entrance were determined: vca:vsr = 1:2; KCa = 1400 M-1; KSr = 270 M-1.


Seed germination as a thermobiological problem.
Thermal effects on seed germination are considered through the changes brought about by temperature in the germination capacity, in the germination rate, and in the distribution of the relative frequency of germination along the incubation times. A number of questions of general thermobiological interest are thus raised, entailing the need of an analysis of the temperature dependence of the seed germination rate. A treatment of these rates by the activation-energy approach cannot be general, for their Arrhenius plots are not always linear. Moreover, it is shown that any process displaying a temperature optimum (as happens in the germination of most seed species) cannot follow one of the fundamental tenets of the collision rate theory. The need of a theoretical treatment stressing the essential role of the partition of energy within the seed system has led to an anlysis using the absolute reaction rate theory. New experimental prospects for the physiology of seed germination are thus raised, concerning the meaning of the temperature cardinal points, the growth pattern of the embryo in germinating seeds, the dual effect of protein thermodenaturation, the effects of high hydrostatic pressures, and a whole pharmacological line of work. The cybernetic counterpart of the thermodynamic view of seed germination appears in the study of the distribution of the relative frequency of germination along the isothermal incubation time. In some species of seeds the thermal communication between the environment and the seed growth effector can be shown to proceed by molecular collisions at all germination isotherms. In the seeds of Dolichos biflorus this communication through random thermal noise prevails only at temperatures close to both extreme limits of germination. Both in this species and in Calotropis procera there is a temperature range (encompassing the optimum) within which a temperature signal is superimposed upon the gaussian noise. An interpretation is proposed according to which the temperature signal is transduced in a protein-conformation code.


[Appendicitis and pregnancy: 21 cases].
Describe the epidemiological, diagnostic and therapeutic features of acute appendicitis in pregnant women. Retrospective analysis of a series of 21 cases of appendicitis in pregnant women who underwent surgery between January 1997 and June 2001. The association of acute appendicitis with pregnancy was noted in 0.2% of the pregnant women. Ten patients were in the first trimester of pregnancy, ten in the second and one in the third. Abdominal pain was noted in all patients. The localization varied with gestational age. Abdominal ultrasound contributed to the diagnosis in twenty patients who underwent the examination. Diagnoses retained were: acute uncomplicated appendicitis (n=18), acute appendicitis with focal induration (n=1) and acute diffuse peritonitis (n=2). Appendectomy was performed in all cases. Intensive care, peritoneal cleaning and drainage were associated in cases with appendicitis. In cases with focal induration, medical treatment was followed by appendectomy performed after delivery. Tocolysis was instituted in all cases. Twelve pregnancies continued to delivery: one fetal death, one premature delivery and ten term deliveries of live infants. Pregnancy makes it difficult to confirm the diagnosis of appendicitis. Appendectomy should be performed in patients presenting a highly suggestive clinical and ultrasonographic picture, preferably by laparoscopy in order to avoid more severe complications which could be life-threatening for the mother or infant.


Hyperthermophilic redox chemistry: a re-evaluation.
The redox chemistry of Pyrococcus furiosus rubredoxin and ferredoxin has been studied as a function of temperature in direct voltammetry and in EPR monitored bulk titrations. The Ems of both proteins, measured with direct voltammetry, have a normal (linear) temperature dependence and show no pH dependence. EPR monitoring is not a reliable method to determine the temperature dependence of the Em: upon rapid freezing the proteins take their conformation corresponding to the freezing point of the solution.


High-yield synthesis of wax esters catalysed by modified Candida rugosa lipase.
Candida rugosa lipase (CRL) was applied in a non-solvent esterification reaction to yield twelve wax esters. All products were obtained in nearly 100% yield for 10 h at 50 degrees C when immobilized PEG(2000)-activated C. rugosa lipase was added to the reaction mixture. The surfactant had also a beneficial effect on the stability of the biocatalytic preparation with 83% of its activity conserved after the seventh run of repeated batch reactions.


Suprasellar mass lesion with unusual magnetic resonance imaging characteristics.
Our case emphasizes the complex nature of signal abnormalities observed with MRI and underscores that a given histopathologic lesion (such as craniopharyngioma) may have variable appearances upon imaging with this modality.


Role of polyphenols in improving endothelial dysfunction in diabetes.
Endothelial cells covering the luminal surface of all blood vessels have a pivotal role in the control of vascular homeostasis mostly via the activation of protective mechanisms. These mechanisms include the endothelial formation of nitric oxide (NO) in response to the activation of endothelial NO synthase, endothelium-dependent hyperpolarization involving endothelial SKCa and IKCa, which is subsequently transmitted to the underlying vascular smooth muscle via myoendothelial junctions, and in some blood vessels, also prostacyclin. These endothelial protective mechanisms will promote vasodilatation, inhibit platelet activation, and also prevent the expression of pro-atherothrombotic factors. In most types of cardiovascular diseases including type 1 and 2 diabetes (T2D), an endothelial dysfunction as indicated by blunted endothelium-dependent vasorelaxations and often also by the appearance of endothelium-dependent cyclooxygenase-mediated contractile responses is observed early in the development of the pathology and has been suggested to contribute to the development of both macro-vascular and micro-vascular complications in T2D. The impaired endothelium-dependent relaxations in T2D has indicated the involvement of reduced NO and endothelium-dependent hyperpolarization components and is associated with increased oxidative stress in the arterial wall involving superoxide anion and hydrogen peroxide predominantly due to an up-regulation of NADPH oxidase and possibly also uncoupling of endothelial NO synthase. Investigations in experimental animals and humans have also suggested that the angiotensin system contributes to the impaired endothelial function in T2D. Several sources of polyphenols (i.e., cocoa and plant extracts) have been shown to retard the induction of endothelial dysfunction in TD2 in part by preventing oxidative stress in the arterial wall most likely by normalizing the expression of NADPH oxidase, cyclooxygenase-1 and 2, eNOS uncoupling, and the angiotensin system. Both pre-clinical and clinical data will be presented and discussed. In addition, polyphenols may possibly also improve the endothelial function by preventing the induction of endothelial senescence to high glucose and angiotensin II. Thus, polyphenols by improving the endothelial dysfunction might help to retard the development of both macro- and micro-vascular complications in diabetes.


Chronic pouchitis is not related to small intestine bacterial overgrowth.
Restorative ileal pouch-anal anastomosis (IPAA) potentially may lead to upper gastrointestinal tract motility disturbances. In addition, a bacterial etiology of IPAA complication-pouchitis-has been suggested. The oro-anal transit time is significantly reduced in this patient group. Therefore, we investigated the hypothesis if IPAA constitutes a significant risk for small intestine bacterial overgrowth (SIBO). Twenty-eight patients age 23-71 years with IPAA operated due to ulcerative colitis without subjective symptoms of pouchitis were evaluated as outpatients according to the prescheduled follow-up after operation and included in the study. The modified Pouchitis Disease Activity Index (PDAI) was determined in all IPAA patients, including clinical, endoscopic, and histopathological (Moskowitz criteria) parameters. In addition, anorectal manometry was performed. The presence of SIBO was determined with the use of a glucose breath test (GBT). In 1 subject (3.6%) an abnormal GBT result was recorded consistent with SIBO. In addition, 2 borderline values (7.1%) were documented. Both patients with SIBO as subjects with borderline values presented with low PDAI values. All patients with PDAI >7 had normal GBT results. In patients with SIBO the maximal tolerated rectal volume was significantly higher than in subjects without SIBO (P < 0.007). Similarly, the PDAI value was significantly lower (P < 0.014). Asymptomatic chronic pouchitis is not related to SIBO. However, excessive colonization of the small intestine does occur in some IPAA patients and needs to be kept in the differential diagnosis.


Redox proteomics analysis of oxidatively modified proteins in G93A-SOD1 transgenic mice--a model of familial amyotrophic lateral sclerosis.
Amyotrophic lateral sclerosis (ALS) is a fatal motor neuron degenerative disease characterized by the loss of neuronal function in the motor cortex, brain stem, and spinal cord. Familial ALS cases, accounting for 10-15% of all ALS disease, are caused by a gain-of-function mutation in Cu,Zn-superoxide dismutase (SOD1). Two hypotheses have been proposed to explain the toxic gain of function of mutant SOD (mSOD). One is that mSOD can directly promote reactive oxygen species and reactive nitrogen species generation, whereas the other hypothesis suggests that mSODs are prone to aggregation due to instability or association with other proteins. However, the hypotheses of oxidative stress and protein aggregation are not mutually exclusive. G93A-SOD1 transgenic mice show significantly increased protein carbonyl levels in their spinal cord from 2 to 4 months and eventually develop ALS-like motor neuron disease and die within 5-6 months. Here, we used a parallel proteomics approach to investigate the effect of the G93A-SOD1 mutation on protein oxidation in the spinal cord of G93A-SOD1 transgenic mice. Four proteins in the spinal cord of G93A-SOD1 transgenic mice have higher specific carbonyl levels compared to those of non-transgenic mice. These proteins are SOD1, translationally controlled tumor protein (TCTP), ubiquitin carboxyl-terminal hydrolase-L1 (UCH-L1), and, possibly, alphaB-crystallin. Because oxidative modification can lead to structural alteration and activity decline, our current study suggests that oxidative modification of UCH-L1, TCTP, SOD1, and possibly alphaB-crystallin may play an important role in the neurodegeneration of ALS.


Development and Validation of Blood-Based Predictive Biomarkers for Response to PD-1/PD-L1 Checkpoint Inhibitors: Evidence of a Universal Systemic Core of 3D Immunogenetic Profiling across Multiple Oncological Indications.
Unprecedented advantages in cancer treatment with immune checkpoint inhibitors (ICIs) remain limited to only a subset of patients. Systemic analyses of the regulatory 3D genome architecture linked to individual epigenetic and immunogenetic controls associated with tumour immune evasion mechanisms and immune checkpoint pathways reveal a highly prevalent molecular profile predictive of response to PD-1/PD-L1 ICIs. A clinical blood test based on a set of eight (8) 3D genomic biomarkers has been developed and validated on the basis of an observational trial to predict response to ICI therapy. The predictive eight biomarker set is derived from prospective observational clinical trials, representing 280 treatments with Pembrolizumab, Atezolizumab, Durvalumab, Nivolumab, and Avelumab in a broad range of indications: melanoma, lung, hepatocellular, renal, breast, bladder, colon, head and neck, bone, brain, lymphoma, prostate, vulvar, and cervical cancers. The 3D genomic eight biomarker panel for response to immune checkpoint therapy achieved a high accuracy of 85%, sensitivity of 93%, and specificity of 82%. This study demonstrates that a 3D genomic approach can be used to develop a predictive clinical assay for response to PD-1/PD-L1 checkpoint inhibition in cancer patients.


Intraventricular nicardipine for refractory cerebral vasospasm after subarachnoid hemorrhage.
Delayed ischemic deficit from vasospasm is a leading cause of morbidity and mortality after aneurysmal subarachnoid hemorrhage. Although several treatment modalities have been used to reverse the deleterious effects of vasospasm, alternative therapies are needed, as conventional therapies are often ineffective or contraindicated. Intrathecal nicardipine has been suggested for the prevention of vasospasm. We report our clinical experience with intraventricular nicardipine for refractory vasospasm in eight patients in whom conventional therapies were ineffective, contraindicated, or technically not feasible. Retrospective case series performed at a tertiary care university hospital. Eight patients (median Hunt-Hess grade = 2, median Fisher score = 4) with refractory vasospasm received intraventricular nicardipine (4 mg every 12 h) for a total of 5-17 days. One patient died in the intensive care unit. Seven patients had moderate to good outcomes with 6 being discharged to home (median Rankin Score = 2). Intraventricular nicardipine was well tolerated with minimal side effects. Our preliminary observations suggest that intraventricular nicardipine could be considered as a safe and effective treatment modality to treat vasospasm refractory to conventional management. A randomized, controlled trial to verify the efficacy and safety of intrathecal nicardipine in the prevention and treatment of cerebral vasospasm is warranted.


Modeling and cleaning RNA-seq data significantly improve detection of differentially expressed genes.
RNA-seq has become a standard technology to quantify mRNA. The measured values usually vary by several orders of magnitude, and while the detection of differences at high values is statistically well grounded, the significance of the differences for rare mRNAs can be weakened by the presence of biological and technical noise. We have developed a method for cleaning RNA-seq data, which improves the detection of differentially expressed genes and specifically genes with low to moderate transcription. Using a data modeling approach, parameters of randomly distributed mRNA counts are identified and reads, most probably originating from technical noise, are removed. We demonstrate that the removal of this random component leads to the significant increase in the number of detected differentially expressed genes, more significant pvalues and no bias towards low-count genes. Application of RNAdeNoise to our RNA-seq data on polysome profiling and several published RNA-seq datasets reveals its suitability for different organisms and sequencing technologies such as Illumina and BGI, shows improved detection of differentially expressed genes, and excludes the subjective setting of thresholds for minimal RNA counts. The program, RNA-seq data, resulted gene lists and examples of use are in the supplementary data and at https://github.com/Deyneko/RNAdeNoise .


Polymethylcarborane as a novel bioactive moiety: derivatives with potent retinoid antagonistic activity.
4[(Deca-B-methyl-1,12-dicarba-closo-dodecaboran-1-yl)c arbamoyl]benzoic acid and its congeners showed potent antagonistic activity at concentrations of 10(-7)-10(-8) M on the differentiation-inducing action of retinoids towards human promyelocytic leukemia HL-60 cells. This is the first example of derivatives of polymethylcarborane, which resembles C60 in size, with biological activity.


Engagement of CD99 induces apoptosis through a calcineurin-independent pathway in Ewing's sarcoma cells.
Programmed cell death (PCD) is a prominent feature of the development of the immune and nervous systems. In both systems, widespread PCD occurs in primitive progenitor cells during development. In this study, we demonstrated that Ewing's sarcoma (ES) cells, undifferentiated neural precursors, underwent apoptosis upon engagement of CD99 with anti-CD99 monoclonal antibody. Apoptosis via CD99 occurred only in the undifferentiated state of ES cells, but not in differentiated ES cells. CD99-induced apoptosis in ES cells appeared to require de novo synthesis of RNA and protein as well as caspase activation. Cyclosporin A, known to be a potent inhibitor of both calcineurin activation and mitochondrial permeability transition pore opening, inhibited CD99-mediated apoptosis, whereas FK-506, a specific calcineurin inhibitor, did not, indicating the induction of CD99-mediated apoptosis through a calcineurin-independent pathway. Furthermore, the dying cells displayed the reduction of mitochondrial transmembrane potential (delta psi m). These results suggest that CD99 engagement induce CsA-inhibitable mitochondrial permeability transition pore opening, followed by a reduction of delta psi m and caspase activation, thereby leading to apoptosis. Based on these results, we suggest the possible involvement of CD99 in the apoptotic processes that occur during nervous system development and also its application in immunotherapeutic trials for ES cases.


Fixation and permeabilization protocol is critical for the immunolabeling of lipid droplet proteins.
The number of proteins known to be associated with lipid droplets (LDs) is increasing. However, the reported distribution of a given protein in the LDs was, in some cases, found not reproduced by other groups. We report here that the choice of the fixation and permeabilization method is important in order to observe LD proteins using immunofluorescence microscopy. Formaldehyde fixation followed by treatment with Triton X-100, one of the most frequently used protocols for the immunolabeling of cultured cells, was not appropriate to label adipocyte differentiation-related protein (ADRP), TIP47, or Rab18 in LDs. Formaldehyde fixation followed by treatment with digitonin or saponin, allowed the visualization of all these proteins in LDs. When cells were fixed with glutaraldehyde, permeabilization by Triton X-100 could also be used for ADRP. These observations suggest that LD proteins are likely to be solubilized by some detergents, and strong cross-linkage to the surrounding protein matrix or mild permeabilization is necessary for their retention on the LD surface.


Superficial ulnar artery terminating in a normal ulnar artery.
Routine dissection of the left upper limb of an 86-year-old male cadaver showed a superficial ulnar artery that anastomosed with the ulnar artery. The superficial ulnar artery arose from the third part of the axillary artery, coursed distally over the flexor muscles of the forearm, and terminated by anastomosing with the ulnar artery in the distal third of the forearm. Arterial and neural variations were also observed on the contralateral side. The presence of a superficial ulnar artery is important clinically when raising forearm flaps in reconstructive surgery.


Nonoperative Management or 'Watch and Wait' for Rectal Cancer with Complete Clinical Response After Neoadjuvant Chemoradiotherapy: A Critical Appraisal.
There is increasing interest in nonoperative management (NOM) for rectal cancer with complete clinical response (cCR) after neoadjuvant chemoradiation (nCRT). The aim of this systematic review was to summarize the available data on NOM, with the intention of formulating standardized protocols on which to base future investigations. A systematic review following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines was conducted. A highly sensitive literature search identified all relevant studies published between January 2004 and December 2016. Data extraction and quality assessment was performed independently by two authors, and resolved by consensus with a third reviewer. In total, 15 studies, including 920 patients, met the inclusion criteria; 575 (62.5%) of these patients underwent NOM after cCR, with the remaining patients forming a surgical control group. The weighted mean follow-up was 39.4 (12.7) months in the NOM group and 39.8 (5.1) months in the surgery group. The pooled regrowth rate in the NOM group was 21.3% at a mean of 15.6 (7.0) months. Surgical salvage was possible and was undertaken in 93.2% of these patients. Overall survival in the NOM group was 91.7%, while disease-free survival was 82.7%. For the comparison proctectomy group, pooled rates of local recurrence, overall survival, and disease-free survival were 8.4, 92.4, and 87.5%, respectively. NOM may be a feasible option for surgically eligible rectal cancer patients with cCR after nCRT. Before such a strategy can be widely implemented, further prospective data are required with standardized definitions, diagnostic criteria, and management protocols, with an emphasis on shared patient-provider decision making and patient-centered outcomes.


Directional growth of quasi-2D Cu2O monocrystals on rGO membranes in aqueous environments.
The preparation technology of unconventional low-dimensional Cu2O monocrystals, which exhibit specific crystal planes and present significantly unique interfacial and physicochemical properties, is attracting increasing attention and interest. Herein, by integrating a high-temperature oxidation process under vacuum and a pure-water incubation process under ambient conditions, we propose the self-assembled growth and synthesis of quasi-two-dimensional Cu2O monocrystals on reduced graphene oxide (rGO) membranes. The prepared Cu2O crystals have a single (110) crystal plane, regular rectangular morphology, and potentially well conductivity. Experimental and theoretical results suggest that this assembly is attributed to the pre-nucleation clusters aggregation and directional attachment of Cu and O on the rGO membranes in aqueous environment and cation-π interactions between the (110) crystal plane of Cu2O and rGO surface. Our findings offer a potential avenue for the discovery and design of advanced low-dimensional single-crystal materials with specific interfacial properties in a pure aqueous environment.


The codon for the methionine at position 129 (M129) in the human prion protein provides an alternative initiation site for translation and renders individuals homozygous for M129 more susceptible to prion disease.
Single amino-acid substitutions in the prion protein have been found to lead to resistance or susceptibility to amyloid fibril formation. In humans, the presence of methionine at position 129 in the prion protein results in increased susceptibility to prion disease, while the presence of valine at that position appears to be protective. It is hypothesized that the codon for M129 is an alternative initiation site for translation, which results in a truncated molecule that is missing the first 128 amino acids, including the signal peptide. This N-terminal truncated form of the prion molecule will not be transported to the extracellular space and thus will accumulate in the cytosol where it is more susceptible to fibril formation and aggregation; this aggregation could hinder normal degradation processes and cause disease. The results of experimental studies on truncated prion molecules support this hypothesis. To test the hypothesis, a gene segment, which when transcribed would result in a prion molecule starting at methionine 129, could be introduced into a convenient experimental animal to see if there is increased incidence of prion disease. Or, fibrils from the brains of affected M129/M129 homozygous individuals could be isolated and the molecules in the fibrils analyzed to determine the identity of the N-terminal amino acid(s). We predict that those isolates will have a preponderance of molecules that start with the methionine at position 129 in the intact protein.


Impact of patient characteristics and comorbidity profile on activation of patients on the kidney transplantation waiting list.
The aim of this study was to examine which demographic and comorbidity factors affected the activation of patients with end-stage renal disease on the national kidney transplantation waiting list. This was a prospective cohort study across 13 transplantation centers in the United Kingdom from October 1, 2006 to September 30, 2007. Data were collected for all new adult patients (n = 1530) referred to the renal transplantation assessment clinic. The proportion of patients who were activated to the waiting list after a minimum one year follow-up was estimated. Factors influencing activation of patients on the waiting list were examined. A total of 872 (58.9%) patients were activated to the transplantation waiting list. The likelihood of activation to the transplantation waiting list was lower in patients older than 65 years (P = .021), nonwhite ethnicity (P < .0001), smokers (P < .0001), and those in whom diabetes was the cause of renal failure (P = .004). Multivariate analysis showed that there was an adverse impact of comorbidity such as ischemic heart disease (P = .003), diabetes (P = .006), and peripheral vascular disease (P = .007) on the likelihood of activation to the waiting list. Patient characteristics and comorbidity are associated with the probability of activation of patients to the waiting list.


Comprehensive bioactivity and chemical characterization of the endemic plant Scorzonera hieraciifolia Hayek extracts: A promising source of bioactive compounds.
Members of Scorzonera genus are known for their potential application in both food and pharmaceutical industries. The present study reports the chemical component and bioactivity of the dichloromethane, ethyl acetate, hexane, methanol and infused extracts of Scorzonera hieraciifolia Hayek aerial parts and roots. The methanolic extracts of both aerial parts and roots were good sources of total phenolic and flavonoid content. For the aerial parts, the infused and methanolic extracts showed highest (p < 0.05) free radical scavenging (2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS)), reducing (cupric reducing antioxidant capacity (CUPRAC))and ferric reducing antioxidant power (FRAP)) and metal chelating capacities. Whilst, for the root extracts, overall most remarkable free scavenging and reducing activity was observed for the methanolic extracts (p < 0.05) while the hexane extracts showed highest metal chelating activity. The inhibitory activity of the extracts was evaluated against key enzymes. Overall, the hexane extracts of both aerial parts (acetylcholinesterase (AChE): 2.27 ± 0.04 and butyrlcholinesterase (BChE): 5.75 ± 0.48 mg galantamine equivalent (GALAE)/g) and roots (AChE: 2.44 ± 0.35 and BChE: 53.17 ± 0.22 mg GALAE/g) were most potent against AChE and BChE, suggesting a role for the more lipophilic compounds in this regard. The highest inhibitory activity against tyrosinase was displayed by methanolic extracts of aerial parts (64.70 ± 0.26 mg kojic acid equivalent (KAE)/g) and roots (63.63 ± 0.20 mg KAE/g). Modest activity was observed against α-amylase and α-glucosidase, two major enzymes implicated in the pathology of diabetes. Liquid chromatography-mass spectrometry (LC-MS) and nuclear magnetic resonance (NMR) analysis showed the presence of 40 compounds such as caffeic, ursolic, betulinic and oleanolic acid and their derivatives in the studied extracts. Multivariate analysis showed that the biological activities were dependent on the plant parts used as well as on the type of solvent used. Overall, the results showed that S. hieraciifolia is a valuable source of phytochemicals for both the nutraceutical and pharmaceutical sectors.


Plasma cholecystokinin-octapeptide like immunoreactivity in patients with hepatic cirrhosis.
Molecular forms of cholecystokinin (CCK) in the peripheral circulation were studied in normal subjects and cirrhotic patients. Fractionation of plasma extract collected 20 min after intraduodenal infusion of fat revealed four major peaks by Sephadex G-50 column chromatography in normal subjects. Peak I eluted at a position similar to CCK-33, peaks II and III eluted between CCK-33 and CCK-14, and peak IV eluted between CCK-14 and CCK-8. In cirrhotic patients, there was a prominent peak (peak V) eluted at a position similar to CCK-8, in addition to those four peaks. These findings are consistent with the previous observations of hepatic elimination of CCK-8, and suggest that smaller forms of CCK similar in size to CCK-8 are not major forms of CCK in plasma in normal subjects but circulate substantially in cirrhotic patients.


Wall shear stress as measured in vivo: consequences for the design of the arterial system.
Based upon theory, wall shear stress (WSS), an important determinant of endothelial function and gene expression, has been assumed to be constant along the arterial tree and the same in a particular artery across species. In vivo measurements of WSS, however, have shown that these assumptions are far from valid. In this survey we will discuss the assessment of WSS in the arterial system in vivo and present the results obtained in large arteries and arterioles. In vivo WSS can be estimated from wall shear rate, as derived from non-invasively recorded velocity profiles, and whole blood viscosity in large arteries and plasma viscosity in arterioles, avoiding theoretical assumptions. In large arteries velocity profiles can be recorded by means of a specially designed ultrasound system and in arterioles via optical techniques using fluorescent flow velocity tracers. It is shown that in humans mean WSS is substantially higher in the carotid artery (1.1-1.3 Pa) than in the brachial (0.4-0.5 Pa) and femoral (0.3-0.5 Pa) arteries. Also in animals mean WSS varies substantially along the arterial tree. Mean WSS in arterioles varies between about 1.0 and 5.0 Pa in the various studies and is dependent on the site of measurement in these vessels. Across species mean WSS in a particular artery decreases linearly with body mass, e.g., in the infra-renal aorta from 8.8 Pa in mice to 0.5 Pa in humans. The observation that mean WSS is far from constant along the arterial tree implies that Murray's cube law on flow-diameter relations cannot be applied to the whole arterial system. Because blood flow velocity is not constant along the arterial tree either, a square law also does not hold. The exponent in the power law likely varies along the arterial system, probably from 2 in large arteries near the heart to 3 in arterioles. The in vivo findings also imply that in in vitro studies no average shear stress value can be taken to study effects on endothelial cells derived from different vascular areas or from the same artery in different species. The cells have to be studied under the shear stress conditions they are exposed to in real life.


Enhanced productivity of Protein C by recombinant human cells in automated fed-batch cultures.
Recombinant human kidney cells, 293, were cultivated in serum-free fed-batch cultures for the production of Protein C. By coupling the feeding of concentrated medium to pH control based on established stoichiometric relations, the titer of Protein C increased by more than ten fold as compared to batch culture, even though the total cell concentration increased only by less than two fold. Such a fed-batch culture is a simple system for enhancing the productivity of mammalian cells in culture.


lncRNA MIR600HG induces the proliferation and invasion of colorectal cancer cells via regulating miR-144-3p/KIF3A.
Colorectal cancer (CRC) is a common digestive system malignancy characterized by the high morbidity and mortality rates. Due to the limited therapeutic options, the majority of CRC patients do not receive timely treatment, which has thus increased the risk of tumor recurrence. Long noncoding RNAs (lncRNAs), as the essential regulators, are involved in numerous pivotal physiological and pathological processes in CRC. By qRT-PCR, it could be discovered in this study that MIR600HG expression obviously increased in CRC tissues and cell lines compared with the normal group. Moreover, the high expression of MIR600HG was correlated with tumor size, tumor volume and TNM stage. Overexpression of MIR600HG affected the expression of downstream molecules, such as miR-144-3p and KIF3A, providing the possible mechanisms of MIR600HG in the development and progression of CRC.


Antiferromagnetism in Ni-Based Superconductors.
Due to the lack of any magnetic order down to 1.7 K in the parent bulk compound NdNiO2 , the recently discovered 9-15 K superconductivity in the infinite-layer Nd0.8 Sr0.2 NiO2 thin films has provided an exciting playground for unearthing new superconductivity mechanisms. Herein, the successful synthesis of a series of superconducting Nd0.8 Sr0.2 NiO2 thin films ranging from 8 to 40 nm is reported. The large exchange bias effect is observed between the superconducting Nd0.8 Sr0.2 NiO2 films and a thin ferromagnetic layer, which suggests the existence of the antiferromagnetic order. Furthermore, the existence of the antiferromagnetic order is evidenced by X-ray magnetic linear dichroism measurements. These experimental results are fundamentally critical for the current field.