At-home use of parasacral transcutaneous electrical nerve stimulation for pediatric voiding dysfunction: a randomized controlled trial to assess its safety and feasibility.
Treating pediatric voiding dysfunction involves behavioral changes that require significant time or medications that are often avoided or discontinued due to side effects. Using parasacral transcutaneous electrical nerve stimulation (PTENS) has shown to have reasonable efficacy, but the safety and feasibility of its off-label use for pediatric voiding dysfunction are not well-established. Concerns have also been raised over treatment adherence. In-home therapy might improve adherence compared with office-based therapy; however, no studies have evaluated in-home feasibility to date. This study aims to assess the safety and feasibility of off-label use of PTENS for pediatric voiding dysfunction. A single-institution prospective, randomized controlled study was conducted from March 2019 to March 2020. Participants aged 6-18 years diagnosed with voiding dysfunction, overactive bladder, or urinary incontinence were eligible for the study. Those with known neurologic disorders, implanted electrical devices, anatomic lower urinary tract abnormality, and recurrent urinary tract infections and those taking bladder medications were excluded. Children with primary monosymptomatic nocturnal enuresis were also excluded due to previous work suggesting a lack of efficacy. Participants were randomly assigned to receive 12 weeks of urotherapy alone (control) or urotherapy plus at-home PTENS treatment. Families were contacted weekly to assess for adverse events (AEs) and treatment adherence. The primary and secondary outcomes were safety, defined as the absence of AEs and treatment adherence, respectively. A total of 30 eligible participants were divided into two groups, with 15 participants in each arm. The median age was 9.4 years (interquartile range: 7.7-10.6). In total, 60% were male. Baseline demographics and urotherapy compliance were similar between the two groups. With PTENS use, two AEs were reported, including mild pruritus at the pad site and discomfort when removing pads, while no AEs were noted in the control group. In total, 60% of patients completed three 30-min sessions per week, and all participants were able to complete treatment sessions for at least 10 weeks, involving 30 min of PTENS treatment each time. This randomized controlled study confirms that at-home use of PTENS is feasible with reasonable treatment adherence and minimal AEs. Future collaborative, multi-institutional studies may better determine the efficacy of this treatment modality.


What is the clinical and functional effect of performing suspension arthroplasty with abductor pollicis longus tendon slip to carpometacarpal joint osteoarthritis of the thumb?
This study aims to investigate the clinical, radiological, and functional results of trapeziectomy, ligament reconstruction and suspensionplasty surgery with abductor pollicis longus (APL) tendon slip autograft to thumb carpometacarpal (CMC) osteoarthritis. Between January 2011 and June 2017, a total of 25 hands of 25 patients (4 males, 21 females; mean age: 63.6±5.8 years; range, 54 to 76 years) were included. All patients underwent trapeziectomy and ligament reconstruction and suspensionplasty procedure due to the diagnosis of thumb CMC joint osteoarthritis. The patients were administered the Patient-Rated Wrist Evaluation Questionnaire (PRWE), Quick Disabilities of the Arm, Shoulder, and Hand (Q-DASH), and Visual Analog Scale (VAS). Scaphometacarpal distance (SMD), Kapandji opposition score, thumb range of motion (ROM), grip strength, tip pinch, lateral pinch, and palmer pinch values were measured for both hands. The values calculated for the operated and contralateral hands of the patients were compared. The ROM measurements, Kapandji opposition scores, and pinch values of the operated hands of the patients were found to be statistically significant lower compared to the contralateral hands (p<0.001, p<0.001, p=0.002; respectively). The grip strength values were similar for both hands (p=0.147). The median SMD in the operated hand was found to be 7.5 mm and 12.1 mm in the contralateral hand (p<0.001). The ROM, strength and functional results were reached a satisfactory level, particularly in the patients with a follow-up period of ≥36 months. In patients with thumb CMC joint osteoarthritis, ligament reconstruction and suspensionplasty using APL tendon slip are considered to be a useful and preferable surgical technique modification.


Induction of Liver Tumors in Mice with N-Ethyl-N-Nitrosourea or N-Nitrosodiethylamine.
Since the groundbreaking studies in the middle part of the last century showing liver cancer in rodents exposed to aromatic amines, the liver has been widely used as a model target organ of chemical carcinogenesis. This protocol describes a method for inducing liver tumors by injecting mice with the widely used alkylating agents N-ethyl-N-nitrosourea (ENU) and N-nitrosodiethylamine (DEN). ENU does not require metabolic activation and readily induces tumors in a number of tissues, including the lungs, stomach, and ovaries, as well as inducing lymphomas. Mice injected with DEN can also develop other tumors, including those of the gastrointestinal tract, skin, lungs, and lymphocytes, but because DEN is metabolized in the liver, it is most effective at inducing liver tumors.


Immunogenic proteins of Leishmania major during mouse infection.
Mice of different strains (C3H/He, A/Jax, DBA/2, BALB/c and CBA/Ca) were infected intracutaneously with Leishmania major. Antibody production was analysed on the antigens of the promastigote form of the parasite which had previously been separated by SDS-polyacrylamide gel electrophoresis and transferred to nitrocellulose sheets. The kinetics of IgG antibody production were determined and the major immunogenic proteins of the parasite were identified for each strain of mouse. It was found that: 1 Antibody directed against parasite proteins appeared in the blood in all mouse strains tested soon after the first week of infection and rose steadily for 5 to 12 weeks. 2 Qualitative individual variation within a given strain was commonly observed in the response to the various antigens. Each mouse strain synthesized IgG directed against a somewhat different spectrum of parasite proteins. Homologies in antigen recognition by pooled sera of five mice ranged from 54% to 79% from one mouse strain to another. Hybrid mice from a BALB/c X CBA cross (F1 generation) recognized essentially all antigens recognized by the two parents. Proteins of L. major with apparent molecular weights of 19 kD, 60 kD, 65 kD, 70 kD and greater than 100 kD were immunogenic in all strains of mice tested.


Unenhanced Cone Beam Computed Tomography and Fusion Imaging in Direct Percutaneous Sac Injection for Treatment of Type II Endoleak: Technical Note.
To evaluate safety, feasibility, technical success, and clinical success of direct percutaneous sac injection (DPSI) for the treatment of type II endoleaks (T2EL) using anatomical landmarks on cone beam computed tomography (CBCT) and fusion imaging (FI). Eight patients with T2EL were treated with DPSI using CBCT as imaging guidance. Anatomical landmarks on unenhanced CBCT were used for referencing T2EL location in the first five patients, while FI between unenhanced CBCT and pre-procedural computed tomography angiography (CTA) was used in the remaining three patients. Embolization was performed with thrombin, glue, and ethylene–vinyl alcohol copolymer. Technical and clinical success, iodinated contrast utilization, procedural time, fluoroscopy time, and mean radiation dose were registered. DPSI was technically successful in all patients: the needle was correctly positioned at the first attempt in six patients, while in two of the first five patients the needle was repositioned once. Neither minor nor major complications were registered. Average procedural time was 45 min and the average administered iodinated contrast was 13 ml. Mean radiation dose of the procedure was 60.43 Gy cm2 and mean fluoroscopy time was 18 min. Clinical success was achieved in all patients (mean followup of 36 months): no sign of T2EL was reported in seven patients until last CT follow-up, while it persisted in one patient with stability of sac diameter. DPSI using unenhanced CBCT and FI is feasible and provides the interventional radiologist with an accurate and safe alternative to endovascular treatment with limited iodinated contrast utilization.


Pentacyclic Triterpene Bioavailability: An Overview of In Vitro and In Vivo Studies.
Pentacyclic triterpenes are naturally found in a great variety of fruits, vegetables and medicinal plants and are therefore part of the human diet. The beneficial health effects of edible and medicinal plants have partly been associated with their triterpene content, but the in vivo efficacy in humans depends on many factors, including absorption and metabolism. This review presents an overview of in vitro and in vivo studies that were carried out to determine the bioavailability of pentacyclic triterpenes and highlights the efforts that have been performed to improve the dissolution properties and absorption of these compounds. As plant matrices play a critical role in triterpene bioaccessibility, this review covers literature data on the bioavailability of pentacyclic triterpenes ingested either from foods and medicinal plants or in their free form.


Improved siRNA delivery efficiency via solvent-induced condensation of micellar nanoparticles.
Efficient delivery of short interfering RNA (siRNA) remains one of the primary challenges of RNA interference therapy. Polyethylene glycol (PEG)ylated polycationic carriers have been widely used for the condensation of DNA and RNA molecules into complex-core micelles. The PEG corona of such nanoparticles can significantly improve their colloidal stability in serum, but PEGylation of the carriers also reduces their condensation capacity, hindering the generation of micellar particles with sufficient complex stability. This presents a particularly significant challenge for packaging siRNA into complex micelles, as it has a much smaller size and more rigid chain structure than DNA plasmids. Here, we report a new method to enhance the condensation of siRNA with PEGylated linear polyethylenimine using organic solvent and to prepare smaller siRNA nanoparticles with a more extended PEG corona and consequently higher stability. As a proof of principle, we have demonstrated the improved gene knockdown efficiency resulting from the reduced siRNA micelle size in mice livers following intravenous administration.


Effect of the nucleotides surrounding the start codon on the translation of foot-and-mouth disease virus RNA.
As for the alternative AUGs in foot-and-mouth disease virus (FMDV), nucleotide bias of the context flanking the AUG(2nd) could be used as a strong signal to initiate translation. To determine the role of the specific nucleotide context, dicistronic reporter constructs were engineered to contain different versions of nucleotide context linking between internal ribosome entry site (IRES) and downstream gene. The results indicate that under FMDV IRES-dependent mechanism, the nucleotide contexts flanking start codon can influence the translation initiation efficiencies. The most optimal sequences for both start codons have proved to be UUU AUG(1st) AAC and AAG AUG(2nd) GAA.


Activated MEK cooperates with Cdkn2a and Pten loss to promote the development and maintenance of melanoma.
The development of targeted inhibitors, vemurafenib and dabrafenib, has led to improved clinical outcome for melanoma patients with BRAFV600E mutations. Although the initial response to these inhibitors can be dramatic, sometimes causing complete tumor regression, the majority of melanomas eventually become resistant. Mitogen-activated protein kinase kinase (MEK) mutations are found in primary melanomas and frequently reported in BRAF melanomas that develop resistance to targeted therapy; however, melanoma is a molecularly heterogeneous cancer, and which mutations are drivers and which are passengers remains to be determined. In this study, we demonstrate that in BRAFV600E melanoma cell lines, activating MEK mutations drive resistance and contribute to suboptimal growth of melanoma cells following the withdrawal of BRAF inhibition. In this manner, the cells are drug-addicted, suggesting that melanoma cells evolve a 'just right' level of mitogen-activated protein kinase signaling and the additive effects of MEK and BRAF mutations are counterproductive. We also used a novel mouse model of melanoma to demonstrate that several of these MEK mutants promote the development, growth and maintenance of melanoma in vivo in the context of Cdkn2a and Pten loss. By utilizing a genetic approach to control mutant MEK expression in vivo, we were able to induce tumor regression and significantly increase survival; however, after a long latency, all tumors subsequently became resistant. These data suggest that resistance to BRAF or MEK inhibitors is probably inevitable, and novel therapeutic approaches are needed to target dormant tumors.


Is it time to re-evaluate our use of acetaminophen in certain sub-groups of pregnant women?
New insight into the pharmacological mechanisms of acetaminophen raises questions about its possible safety in certain subgroups of pregnant women. Recent focus on acetaminophen's ability to modulate prostaglandin synthesis may require modification of its ubiquitous use in pregnancy. Further basic science, epidemiologic and clinical studies are required to explore the interaction of acetaminophen with placentation leading to vascular pathologic conditions during pregnancy.


Impact of Surgical Route in Influencing the Risk of Lymphatic Complications After Ovarian Cancer Staging.
Lymphatic complications are a common occurrence after staging surgery for early-stage ovarian cancer (eEOC). We investigated whether the introduction of minimally invasive surgery influences the risk of developing lymphoceles and lymphorrhea in patients undergoing staging for eEOC. For this purpose, data of consecutive patients affected by eEOC undergoing staging surgery between January 1980 and January 2016 were retrospectively reviewed, and a systematic review and meta-analysis was performed. This systematic review was registered in the International Prospective Register of Systematic Review. Among 341 patients included in the present study, 47 severe postoperative complications occurred (13.7%), including 40 lymphatic complications: 31 symptomatic lymphoceles (9%) and 9 cases of lymphorrhea (2.6%), respectively. Laparoscopic staging correlated with a lower risk of developing any severe lymphatic complications in comparison with open surgery (p = .02). In particular, the laparoscopic approach and para-aortic node involvement were associated with a trend toward lower lymphoceles (odds ratio, .13; 95% confidence interval, .07-2.20; p = .05) and a trend toward higher risk of lymphorrhea developing (odds ratio, 4.02; 95% confidence interval, .93-17.3; p = .06), respectively. In conclusion, the implementation of a minimally invasive approach might result in a slight reduction of lymphatic complications after eEOC staging.


Nitroarginine, an inhibitor of nitric oxide synthase, prevents changes in superoxide radical and antioxidant enzymes induced by ammonia intoxication.
Injection of large doses of ammonium salts leads to the rapid death of animals. However, the molecular mechanisms involved in ammonia toxicity remain to be clarified. We reported that injecting ammonium acetate (7 mmol/kg) to rats increases the production of superoxide and reduces the activities of some antioxidant enzymes in rat liver and brain. We proposed that these effects induced by ammonia intoxication would be mediated by formation of nitric oxide. To test this possibility we tested whether injection of nitroarginine, an inhibitor of nitric oxide synthase, prevents the effects of ammonia intoxication on antioxidant enzymes and superoxide formation. Following injection of ammonia, glutathione peroxidase, superoxide dismutase and catalase activities were decreased in liver by 42%, 54% and 44%, respectively. In brain these activities were reduced by 35%, 46% and 65%, respectively. Glutathione reductase remained unchanged. Superoxide production in submitochondrial particles from liver and brain was increased by more than 100% in both tissues. Both reduction of activity of antioxidant enzymes and increased superoxide radical production were prevented by previous injection of 45 mg/kg of nitroarginine, indicating that ammonia induces increased formation of nitric oxide, which in turn reduces the activity of antioxidant enzymes, leading to increased formation of superoxide.


Deep learning strategy for small dataset from atomic force microscopy mechano-imaging on macrophages phenotypes.
The cytoskeleton is involved during movement, shaping, resilience, and functionality in immune system cells. Biomarkers such as elasticity and adhesion can be promising alternatives to detect the status of cells upon phenotype activation in correlation with functionality. For instance, professional immune cells such as macrophages undergo phenotype functional polarization, and their biomechanical behaviors can be used as indicators for early diagnostics. For this purpose, combining the biomechanical sensitivity of atomic force microscopy (AFM) with the automation and performance of a deep neural network (DNN) is a promising strategy to distinguish and classify different activation states. To resolve the issue of small datasets in AFM-typical experiments, nanomechanical maps were divided into pixels with additional localization data. On such an enlarged dataset, a DNN was trained by multimodal fusion, and the prediction was obtained by voting classification. Without using conventional biomarkers, our algorithm demonstrated high performance in predicting the phenotype of macrophages. Moreover, permutation feature importance was employed to interpret the results and unveil the importance of different biophysical properties and, in turn, correlated this with the local density of the cytoskeleton. While our results were demonstrated on the RAW264.7 model cell line, we expect that our methodology could be opportunely customized and applied to distinguish different cell systems and correlate feature importance with biophysical properties to unveil innovative markers for diagnostics.


Detection of antibodies to co-trimoxazole (preservative drug) interfering with routine red cell antibody screening.
Drug-dependent antibodies can rarely cause interference in pretransfusion antibody screening. The diluents for commercial reagent red blood cells contain different antibiotics, such as chloramphenicol, neomycin sulfate, and gentamycin as a preservative. The presence of antibodies to a given drug in patient may lead to positive results when performing antibody identification. We present a rare case of detection of anti-co-trimoxazole antibody during routine antibody screening in a female patient undergoing neurosurgery. These antibodies mimicked as antibody against high-frequency red cell antigens reacting in both saline phase as well as antiglobulin phase. Anti-co-trimoxazole antibody was confirmed by repeating antibody screen using reagent red cells of different manufacturers with and without co-trimoxazole drug as preservative as well as using washed red cell panels. There were no associated clinical or laboratory evidence of hemolysis.


What, if anything, is a Tilapia?-mitochondrial ND2 phylogeny of tilapiines and the evolution of parental care systems in the African cichlid fishes.
We estimated a novel phylogeny of tilapiine cichlid fish (an assemblage endemic to Africa and the Near East) within the African cichlid fishes on the basis of complete mitochondrial NADH dehydrogenase subunit 2 (ND2) gene sequences. The ND2 (1,047 bp) gene was sequenced in 39 tilapiine cichlids (38 species and 1 subspecies) and in an additional 14 nontilapiine cichlid species in order to evaluate the traditional morphologically based hypothesis of the respective monophyly of the tilapiine and haplochromine cichlid fish assemblages. The analyses included many additional cichlid lineages, not only the so-called tilapiines, but also lineages from Lake Tanganyika, east Africa, the Neotropics and an out-group from Madagascar with a wide range of parental care and mating systems. Our results suggest, in contrast to the historical morphology-based hypotheses from Regan (1920, 1922 ), Trewavas (1983), and Stiassny (1991), that the tilapiines do not form a monophyletic group because there is strong evidence that the genus Tilapia is not monophyletic but divided into at least five distinct groups. In contrast to this finding, an allozyme analysis of Pouyaud and Agnèse (1995), largely based on the same samples as used here, found a clustering of the Tilapia species into only two groups. This discrepancy is likely caused by the difference in resolution power of the two marker systems used. Our data suggest that only type species Tilapia sparrmanii Smith (1840) should retain the genus name TILAPIA: One particular group of tilapiines (composed of genera Sarotherodon, Oreochromis, Iranocichla, and Tristramella) is more closely related to an evolutionarily highly successful lineage, the haplochromine cichlids that compose the adaptive radiations of cichlid species flocks of east Africa. It appears that the highly adaptable biology of tilapiines is the ancestral state for all African cichlids and that the more stenotypic lifestyle of the haplochromine cichlids is derived from this condition. We reconstructed the evolution of the highly variable parental care systems on the basis of the most inclusive composite phylogeny to date of the African, Neotropical, and Madagascan cichlids with special emphasis on a group of tilapiines comprising the substrate-spawning genus Tilapia, and the mouthbrooding genera Sarotherodon and OREOCHROMIS: We demonstrate several independent origins of derived mouthbrooding behaviors in the family Cichlidae.


Psychopathology associated with suicide attempts among rural adolescents of China.
This case control study examined the relationship between psychopathological profiles and self-reported suicide attempts among rural adolescents of China. Cases consisted of 142 adolescents who reported a suicide attempt in the past 6 months in a questionnaire survey (n=1365). An equal number of adolescents without reporting a suicide attempt, matched on age, gender, and school class, were selected from the same survey sample as controls. The Youth Self Report (YSR) was used to assess psychopathological profiles for suicide attempters and matched controls. Results indicated that 73% of suicide attempters had at least one behavioral/emotional syndrome, significantly higher than in controls (30%). Logistic regression analyses showed that anxiety/depression (OR = 3.68) and aggression (OR = 5.02) were each significantly associated with increased risk of suicide attempts after controlling for other behavioral problems. These findings demonstrated the independent contribution of anxiety/depression and aggression to suicide attempts in Chinese adolescents.


Occurrence of multinucleated hepatocytes in cynomolgus monkeys (Macaca fascicularis) from different geographical regions.
Multinucleated hepatocytes (MNHs) have been occasionally reported in macaques, as well as chimpanzees and gorillas, as an incidental finding. However, information is sparse on variations in incidence in the cynomolgus macaque (Macaca fascicularis). A survey was conducted to assess the occurrence of MNHs in the liver of stock (nonstudy) animals from SNBL SRC (Alice, TX) and SNBL USA (Everett, WA) submitted for diagnostic purposes. A total of 215 cynomolgus monkeys originally from Cambodia (61), China (5), Indonesia (125), and Mauritius (24) were used for this investigation. From each animal, usually 2 liver samples were processed for histopathology with 2 sections in each slide. An MNH was defined as a hepatocyte with 3 or more nuclei. A threshold of 3 MNHs was selected for the Multinucleated Hepatocyte Grading System: 0 = not remarkable (≤3 MNHs counted from 2-4 liver sections), minimal = 4 to 15 MNHs, mild = 16 to 30 MNHs, moderate = 31 to 59 MNHs, and severe ≥60 MNHs. The incidence of MNHs was 60 of 86 (70%) in males and 72 of 129 (56%) in females for a total overall incidence of 132 of 215 animals (61%). Affected hepatocytes were frequently observed close to the capsule and generally had 3 to 8 nuclei per hepatocyte but as many as 15 occurred in a single cell. Awareness of the incidence of MNHs in cynomolgus monkeys is important for potential use as background data in preclinical safety and toxicity evaluation studies.


3D visualization and quantification of microvessels in the whole ischemic mouse brain using solvent-based clearing and light sheet microscopy.
The visualization of cerebral microvessels is essential for understanding brain remodeling after stroke. Injection of dyes allows for the evaluation of perfused vessels, but has limitations related either to incomplete microvascular filling or leakage. In conventional histochemistry, the analysis of microvessels is limited to 2D structures, with apparent limitations regarding the interpretation of vascular circuits. Herein, we developed a straight-forward technique to visualize microvessels in the whole ischemic mouse brain, combining the injection of a fluorescent-labeled low viscosity hydrogel conjugate with 3D solvent clearing followed by automated light sheet microscopy. We performed transient middle cerebral artery occlusion in C57Bl/6j mice and acquired detailed 3D vasculature images from whole brains. Subsequent image processing, rendering and fitting of blood vessels to a filament model was employed to calculate vessel length density, resulting in 0.922 ± 0.176 m/mm3 in healthy tissue and 0.329 ± 0.131 m/mm3 in ischemic tissue. This analysis showed a marked loss of capillaries with a diameter ≤ 10 µm and a more moderate loss of microvessels in the range > 10 and ≤ 20 µm, whereas vessels > 20 µm were unaffected by focal cerebral ischemia. We propose that this protocol is highly suitable for studying microvascular injury and remodeling post-stroke.


Communication of HIV viral load to guide sexual risk decisions with serodiscordant partners among San Francisco men who have sex with men.
The objective of this study was to estimate frequency and correlates of discussing HIV viral load (VL) with serodiscordant sex partners to guide decisions about sexual activities among men who have sex with men (MSM). We conducted a cross-sectional survey of 573 San Francisco MSM. Among 507 who knew their HIV status, 397 (78%) were familiar with the term 'viral load', and half (n=199) had a serodiscordant partner in the prior year. These 199 respondents (n=130 [65%] HIV-positive; n=69 [35%] HIV-negative) were the focus of this analysis. A majority (n=111, 56%) discussed VL in the prior year with serodiscordant partners specifically to guide decisions about sexual risk behaviour. Discussion was more common among HIV-positive than HIV-negative participants (adjusted odds ratio [AOR], 3.5; 95% confidence interval [CI], 1.6-7.6), and African Americans compared to whites (AOR, 3.7; 95% CI, 1.5-9.5). HIV-negative men who discussed VL were more concerned about becoming infected, but also more willing to engage in risky behaviour with a partner whose VL is undetectable, than men not discussing VL. Some HIV-negative men may be discussing VL to engage in higher risk behaviour upon learning of an HIV-positive partner's undetectable VL. Interventions targeting MSM should explain that while risk of transmission is likely reduced with a low blood plasma VL, it is not necessarily eliminated.


Sexual harassment in online communications: effects of gender and discourse medium.
Differences in the perception of sexual harassment depending on discourse medium (traditional classroom setting versus online) and gender were examined via survey with 270 undergraduate participants. It was hypothesized that (1) participants would differ in their evaluations of similar behaviors when they were described as occurring in a traditional classroom setting as opposed to an online environment, and (2) males and females would differ in their evaluations of online behaviors. Eight potentially harassing acts were examined--including sexually explicit pictures, content, jokes, misogyny, the use of nicknames, requests for company, sexual favors, and comments about dress. Misogynist comments were seen as more harassing online than in traditional settings, as well as using nicknames and comments about dress. In a traditional setting, only requests for company were seen as more harassing. In terms of gender differences, women rated online pictures and jokes as significantly more harassing than men.


Applied Sciences: Absence of CD44-standard in human neuroblastoma correlates with histological dedifferentiation, N-myc amplification and reduced survival probability.
Expression of CD44 was examined by immunohistochemistry in 205 primary neuroblastomas together with histological grading according to the Shimada classification at the time of diagnosis. In addition, Southern blot analysis to determine N-myc gene amplification was carried out in the same tissue. When compared with clinical data such as stage, age and event-free survival probability it was found that CD44 expression characterizes well differentiated tumours and thus correlates with prognosis (event-free survival probability in CD44 positive patients 0.6 (n = 129) vs. 0.0 (n = 21) in CD44 negative patients). In tumours with N-myc = 1, CD44 positivity was found in 91% of patients as compared to 57% of patients with N-myc > 1 in tumours. All tumours of 13 patients with metastatic stage 4s (with good prognosis) showed CD44s expression. Thus, detection of CD44s expression might serve as a prognostic indicator which can be rapidly detected at diagnosis.


Crystallization and preliminary X-ray diffraction analysis of the ArsC protein from the Escherichia coli arsenical resistance plasmid, R773.
Diffraction data to 3.0 A resolution were collected on crystals of ArsC protein from the conjugative resistance factor R773 which mediates arsenical resistance in the Gram negative bacterium Escherichia coli. The crystal system is tetragonal, a = 116.2 A, c = 145.0 A and the space group is either P4(1)2(1)2 or P4(3)2(1)2. The most probable range for the contents of the asymmetric unit is four to eight ArsC molecules (M(r) = 15,811).


Characterizing T-cell response in low-grade and high-grade vulval intraepithelial neoplasia, study of CD3, CD4 and CD8 expressions.
The objective of our study was to compare immunocyte infiltrates in vulval epithelium from low-grade and high-grade vulval intraepithelial neoplasia (VIN) lesions to determine if difference in T-cell presence reflected the grade of VIN. Thirty-six vulval specimens were obtained from 24 patients who had previously undergone vulval biopsies for VIN, 14 high-grade diseases (VIN 3 with or without HPV) and 14 low-grade diseases (VIN 1 and VIN 2 with or without HPV). Eight samples of normal vulval tissue were selected from the excision margins of resected vulval biopsies. The lymphocyte surface markers included CD3 (Pan T-cell marker), CD4 (T helper cells), and CD8 (T cytotoxic cells). Each tissue section was visualized under high power magnification and cells were counted in 10 random areas at the dermo-epidermal junction. A significantly higher number of total mean T lymphocytes were detected in VIN specimens compared to normal vulval tissue (P = 0.002). In low-grade VIN, there were significantly more CD8 cells than CD4 when compared to high-grade VIN. This difference in CD4/CD8 ratio was significant (P = 0.001). This study suggests that increased CD8 response in VIN is a feature of low-grade disease and we speculate that this may be a protective mechanism. In high-grade disease, both CD4 cells and CD8 cells are equally present with preservation of normal CD4/CD8 ratio.


Herpes simplex virus infections of the central nervous system. A review.
Herpes simplex virus (HSV) infections of the central nervous system are a significant cause of mortality and morbidity. The introduction of antiviral therapy has improved the outcome for patients with life-threatening disease. Neonatal HSV infection is usually acquired at the time of delivery by contact of the fetus with infected maternal genital secretions resulting in disease that can be localized to the skin, eye, and mouth, and can lead to encephalitis or become disseminated. A total of 291 babies with neonatal HSV infection have been evaluated over a period of 14 years with mortality and morbidity rates determined at one year. Vidarabine therapy decreased the incidence of mortality and improved morbidity rates; however, further improvement in mortality rates with acyclovir therapy has not been apparent. No significant clinical toxicity appeared in either treatment group. In order to improve outcome, earlier intervention and prophylactic strategies must be developed. For patients with herpes simplex encephalitis, acyclovir therapy is superior to vidarabine therapy for biopsy-proven disease. When outcome is compared for 136 vidarabine- and 46 acyclovir-treated, biopsy-proven patients, mortality rates are decreased to 20 percent with acyclovir, and approximately 40 percent of survivors are evaluated as normal at one year after therapy. Despite better outcome with antiviral therapy for the treatment of biopsy-proven herpes simplex encephalitis, further improvement is required.


[A Case of Hemodialysis-Associated Pelvic Amyloidoma Mimicking Renal Cancer-Associated Etiology].
Patients with renal insufficiency receiving long-term hemodialysis often develop so-called hemodialysis amyloidosis characterized by systemic β 2-microglobulin amyloid lesions, while patients with renal cell carcinoma may develop amyloid A(AA) amyloidosis. Herein, we present a 67-year-old man on thirty-yearlong hemodialysis who was diagnosed to have left renal cell carcinoma coincident with a large spaceoccupying lesion adjacent to the psoas muscle in the pelvic cavity. An ultrasound-guided percutaneous needle biopsy was performed at the time of laparoscopic radical nephrectomy. The pathological work-up on the needle biopsy specimen revealed that the lesion was not an AA amyloidoma but a β2-microglobulin amyloidoma, which is a rare manifestation of hemodialysis amyloidosis.


Developing a one-stop clinic for leg ulcer patients.
Leg ulceration may be caused by a variety of different factors and specialist assessment is required. Patients with complex leg ulcers in Nottingham are assessed at a one-stop dermatology/vascular clinic. Nurses undertake assessments, provide education and support and liaise with the community nursing services.


Structure of external aldimine of Escherichia coli CsdB, an IscS/NifS homolog: implications for its specificity toward selenocysteine.
Escherichia coli CsdB is a pyridoxal 5'-phosphate (PLP)-dependent enzyme that catalyzes both cysteine desulfuration and selenocysteine deselenation. The enzyme has a high specific activity for L-selenocysteine relative to L-cysteine. On the other hand, its paralog, IscS, exhibits higher activity for L-cysteine, which acts as a sulfur donor during the biosynthesis of the iron-sulfur cluster and 4-thiouridine. The structure of CsdB complexed with L-propargylglycine was determined by X-ray crystallography at 2.8 A resolution. The overall polypeptide fold of the complex is similar to that of the uncomplexed enzyme, indicating that no significant structural change occurs upon formation of the complex. In the complex, propargylglycine forms a Schiff base with PLP, providing the features of the external aldimine formed in the active site. The Cys364 residue, which is essential for the activity of CsdB toward L-cysteine but not toward L-selenocysteine, is clearly visible on a loop of the extended lobe (Thr362-Arg375) in all enzyme forms studied, in contrast to the corresponding disordered loop (Ser321-Arg332) of the Thermotoga maritima NifS-like protein, which is closely related to IscS. The extended lobe of CsdB has an 11-residue deletion compared with that of the NifS-like protein. These facts suggest that the restricted flexibility of the Cys364-anchoring extended lobe in CsdB may be responsible for the ability of the enzyme to discriminate between selenium and sulfur.


High-Throughput Screening Methodology to Identify Alpha-Synuclein Aggregation Inhibitors.
An increasing number of neurodegenerative diseases are being found to be associated with the abnormal accumulation of aggregated proteins in the brain. In Parkinson's disease, this process involves the aggregation of alpha-synuclein (α-syn) into intraneuronal inclusions. Thus, compounds that inhibit α-syn aggregation represent a promising therapeutic strategy as disease-modifying agents for neurodegeneration. The formation of α-syn amyloid aggregates can be reproduced in vitro by incubation of the recombinant protein. However, the in vitro aggregation of α-syn is exceedingly slow and highly irreproducible, therefore precluding fast high throughput anti-aggregation drug screening. Here, we present a simple and easy-to-implement in-plate method for screening large chemical libraries in the search for α-syn aggregation modulators. It allows us to monitor aggregation kinetics with high reproducibility, while being faster and requiring lower protein amounts than conventional aggregation assays. We illustrate how the approach enables the identification of strong aggregation inhibitors in a library of more than 14,000 compounds.


Receptor regulation of senile phenoptosis.
Here we present a concept that considers organism aging as an additional facultative function promoting evolution, but counterproductive for an individual. We hypothesize that aging can be inhibited or even arrested when full mobilization of all resources is needed for the survival of an individual. We believe that the organism makes such a decision based on the analysis of signals of special receptors that monitor a number of parameters of the internal and external environment. The amount of available food is one of these parameters. Food restriction is perceived by the organism as a signal of coming starvation; in response to it, the organism inhibits its counterproductive programs, in particular, aging. We hypothesize that the level of protein obtained with food is estimated based on blood concentration of one of the essential amino acids (methionine), of carbohydrates - via glucose level, and fats - based on the level of one of the free fatty acids. When the amount of available food is sufficient, these receptors transmit the signal allowing aging. In case of lack of food, this signal is cancelled, and as a result aging is inhibited, i.e. age-related weakening of physiological functions is inhibited, and lifespan increases (the well-known geroprotective effect of partial food restriction). In Caenorhabditis elegans, lowering of the ambient temperature has a similar effect. This geroprotective effect is removed by the knockout of one of the cold receptors, and replacement of the C. elegans receptor by a similar human receptor restores the ability of low temperature to increase the lifespan of the nematode. A chain of events linking the receptor with the aging mechanism has been discovered in mice - for one of the pain receptors in neurons, the nerve endings of which entwine pancreas β-cells. Age-related activation of these receptors inhibits the work of insulin genes in β-cells. Problems with insulin secretion lead to oxidative stress, chronic inflammation, and type II diabetes, which can be regarded as one of the forms of senile phenoptosis. In conclusion, we consider the role of some psychological factors in the regulation of the aging program.


Excitation regeneration in delay-coupled oscillators.
A network of phase-coupled oscillators constitutes a system in which excitable 2pi phase slips can be induced by noise. We show that such an excitation in one of the oscillators can be regenerated by subsequent oscillators and become self-sustained for certain topologies. We focus on the simplest such topology: two mutually coupled oscillators.Our analysis is bolstered [corrected] by an experimental confirmation of the phenomenon via a pair of mutually delay coupled quantum-dot lasers. Both the intensities and phases of the laser outputs were measured confirming the interpretation.


[Clinical usefulness of adrenal gland computed tomography in the etiologic diagnosis of Addison's disease].
The aim of this study was to evaluate adrenal gland morphology by computerized tomography (CT) in the etiologic diagnosis of Addison's disease. Twenty-two patients were grouped according to their etiology based on the study of antiadrenal antibodies at diagnosis of the disease: 7 were positive (autoimmune etiology or EAA), 11 were negative (tuberculous etiology or EAT) and in four serologic study was not available (undetermined etiology or EAI). Adrenal gland CT was performed with contiguous sections every 5 mm. In eight cases the examination was carried out upon diagnosis of the disease (initial stage) and in 14 between 2 and 17 years following diagnosis (evolutive stage). In all the patients of the EAA group, examined in either the initial or evolutive stages, the adrenal glands appeared atrophic without calcifications. In the patients of the EAT group adrenal masses of proven tuberculous origin were found in five who were studied in the initial stage and with atrophic glands with calcifications in the remaining patients examined in the evolutive stage, with the exception of two cases with atrophic adrenal glands without calcifications which could serologically be considered as false negatives having autoimmune etiology. The patients in the EAI groups were studied in the evolutive stage with three having atrophic glands with calcifications and one atrophic adrenal glands without calcifications. Adrenal gland computerized tomography is an useful examination to differentiate between autoimmune or tuberculous origin in Addison's disease. Clinical application may be justified in all patients of recent diagnosis when the study of the antiadrenal antibodies is negative or not possible.


Fully Automated Prediction of Geographic Atrophy Growth Using Quantitative Spectral-Domain Optical Coherence Tomography Biomarkers.
To develop a predictive model based on quantitative characteristics of geographic atrophy (GA) to estimate future potential regions of GA growth. Progression study and predictive model. One hundred eighteen spectral-domain (SD) optical coherence tomography (OCT) scans of 38 eyes in 29 patients. Imaging features of GA quantifying its extent and location, as well as characteristics at each topographic location related to individual retinal layer thickness and reflectivity, the presence of pathologic features (like reticular pseudodrusen or loss of photoreceptors), and other known risk factors of GA growth, were extracted automatically from 118 SD OCT scans of 38 eyes from 29 patients collected over a median follow-up of 2.25 years. We developed and evaluated a model to predict the magnitude and location of GA growth at given future times using the quantitative features as predictors in 3 possible scenarios. Potential regions of GA growth. In descending order of out-of-bag feature importance, the most predictive SD OCT biomarkers for predicting the future regions of GA growth were thickness loss of bands 11 through 14 (5.66), reflectivity of bands 11 and 12 (5.37), thickness of reticular pseudodrusen (5.01), thickness of bands 5 through 11 (4.82), reflectivity of bands 7 through 11 (4.78), GA projection image (4.73), increased minimum retinal intensity map (4.59), and GA eccentricity (4.49). The predicted GA regions in the 3 tested scenarios resulted in a Dice index mean ± standard deviation of 0.81±0.12, 0.84±0.10, and 0.87±0.06, respectively, when compared with the observed ground truth. Considering only the regions without evidence of GA at baseline, predicted regions of future GA growth showed relatively high Dice indices of 0.72±0.18, 0.74±0.17, and 0.72±0.22, respectively. Predictions and actual values of GA growth rate and future GA involvement in the central fovea showed high correlations. Experimental results demonstrated the potential of our predictive model to predict future regions where GA is likely to grow and to identify the most discriminant early indicator (thickness loss of bands 11 through 14) of regions susceptible to GA growth.


Effects of altered epididymal sperm transit time on sperm quality.
The epididymal sperm transit time seems to have an important role in the process of sperm maturation, and it seems that alterations to the transit can harm the process. The aim of the present work was to evaluate the influence of altered sperm transit time through the epididymis on sperm parameters and fertility of rats, as well as the role of testosterone in the alterations. Sprague-Dawley adult male rats were randomly assigned to four different groups and were treated for 12 days: (i) 10 microg/rat/day DES, to accelerate the transit; (ii) 6.25 mg/kg/day guanethidine sulphate, to delay the transit; (iii) same treatment as group 1, plus androgen supplementation; (iv) control animals received the vehicles. Guanethidine treatment delayed the sperm transit time through the epididymal cauda, provoking increased sperm reserves in this region. Animals exposed to DES showed an acceleration of sperm transit time in the epididymis, and consequently decreased sperm density in both epididymal regions, the caput-corpus and cauda, and diminished sperm motility. In both cases sperm production was not altered. Testosterone supplementation was able to restore the transit time to values close to normality, as they were higher than in the control rats. The same occurred in relation to sperm motility. Rats exposed to DES presented lower fertility after in utero artificial insemination using sperm collected from the proximal cauda epididymis. Therefore, it was concluded that the acceleration of rat sperm transit time appeared to harm normal sperm maturation, thus decreasing sperm quality and fertility capacity, in an androgen-dependent way.


Characterization of a Multimodular Endo-β-1,4-Glucanase (Cel9K) from Paenibacillus sp. X4 with a Potential Additive for Saccharification.
An endo-β-1,4-glucanase gene, cel9K, was cloned using the shot-gun method from Paenibacillus sp. X4, which was isolated from alpine soil. The gene was 2,994 bp in length, encoding a protein of 997 amino acid residues with a predicted signal peptide composed of 32 amino acid residues. Cel9K was a multimodular enzyme, and the molecular mass and theoretical pI of the mature Cel9K were 103.5 kDa and 4.81, respectively. Cel9K contains the GGxxDAGD, PHHR, GAxxGG, YxDDI, and EVxxDYN motifs found in most glycoside hydrolase family 9 (GH9). The protein sequence showed the highest similarity (88%) with the cellulase of Bacillus sp. BP23 in comparison to the enzymes with reported properties. The enzyme was purified by chromatography using HiTrap Q, CHT-II, and HiTrap Butyl HP. Using SDS-PAGE/activity staining, the molecular mass of Cel9K was estimated to be 93 kDa, which is a truncated form produced by the proteolytic cleavage of its C-terminus. Cel9K was optimally active at pH 5.5 and 50°C and showed a half-life of 59.2 min at 50°C. The CMCase activity was increased to more than 150% in the presence of 2 mM Na⁺, K⁺, and Ba²⁺, but decreased significantly to less than 50% by Mn²⁺ and Co²⁺. The addition of Cel9K to a commercial enzyme set (Celluclast 1.5L + Novozym 188) increased the saccharification of the pretreated reed and rice straw powders by 30.4 and 15.9%, respectively. The results suggest that Cel9K can be used to enhance the enzymatic conversion of lignocellulosic biomass to reducing sugars as an additive.


Effects of insulin on ribonucleic acid synthesis in toad urinary bladder.
We previously found that the sustained rise in active sodium transport by the toad urinary bladder after exposure to insulin is associated with the synthesis of a membrane protein with a molecular weight of 25,000. Studying the effects of insulin on the incorporation of uridine into RNA of epithelial cells of the bladder, we now report that insulin causes a 39% increase in the incorporation of [3H]uridine into poly(A)-containing RNA extracted from granular cells, the predominant epithelial cell type. The increased accumulation of label in RNA, most notably species ranging from 5-10S, could be demonstrated whether cells were pulsed with [3H]uridine or continuously exposed to the label. Using a reticulocyte lysate system, we compared the translation products of RNA from untreated and insulin-treated granular mucosal cells. Exposure to insulin was associated with the expression of a mRNA coding for a protein with molecular weight of approximately 25,000, similar in size to the insulin-induced membrane protein we previously identified in the intact tissue.


Genetic variants for long QT syndrome among infants and children from a statewide newborn hearing screening program cohort.
Autosomal recessive long QT syndrome (LQTS), or Jervell and Lange-Nielsen syndrome (JLNS), can be associated with sensorineural hearing loss. We aimed to explore newborn hearing screening combined with electrocardiograms (ECGs) for early JLNS detection. In California, we conducted statewide, prospective ECG screening of children ≤ 6 years of age with unilateral or bilateral, severe or profound, sensorineural or mixed hearing loss. Families were identified through newborn hearing screening and interviewed about medical and family histories. Twelve-lead ECGs were obtained. Those with positive histories or heart rate corrected QT (QTc) intervals ≥ 450 ms had repeat ECGs. DNA sequencing of 12 LQTS genes was performed for repeat QTc intervals ≥ 450 ms. We screened 707 subjects by ECGs (number screened/number of responses = 91%; number of responses/number of families who were mailed invitations = 54%). Of these, 73 had repeat ECGs, and 19 underwent gene testing. No subject had homozygous or compound heterozygous LQTS mutations, as in JLNS. However, 3 individuals (with QTc intervals of 472, 457, and 456 ms, respectively) were heterozygous for variants that cause truncation or missplicing: 2 in KCNQ1 (c.1343dupC or p.Glu449Argfs*14; c.1590+1G>A or p.Glu530sp) and 1 in SCN5A (c.5872C>T or p.Arg1958*). In contrast to reports of JLNS in up to 4% of children with sensorineural hearing loss, we found no examples of JLNS. Because the 3 variants identified were unrelated to hearing, they likely represent the prevalence of potential LQTS mutations in the general population. Further studies are needed to define consequences of such mutations and assess the overall prevalence.


Exploring the abundance of oleate hydratases in the genus Rhodococcus-discovery of novel enzymes with complementary substrate scope.
Oleate hydratases (Ohys, EC 22.214.171.124) are a class of enzymes capable of selective water addition reactions to a broad range of unsaturated fatty acids leading to the respective chiral alcohols. Much research was dedicated to improving the applications of existing Ohys as well as to the identification of undescribed Ohys with potentially novel properties. This study focuses on the latter by exploring the genus Rhodococcus for its plenitude of oleate hydratases. Three different Rhodococcus clades showed the presence of oleate hydratases whereby each clade was represented by a specific oleate hydratase family (HFam). Phylogenetic and sequence analyses revealed HFam-specific patterns amongst conserved amino acids. Oleate hydratases from two Rhodococcus strains (HFam 2 and 3) were heterologously expressed in Escherichia coli and their substrate scope investigated. Here, both enzymes showed a complementary behaviour towards sterically demanding and multiple unsaturated fatty acids. Furthermore, this study includes the characterisation of the newly discovered Rhodococcus pyridinivorans Ohy. The steady-state kinetics of R. pyridinivorans Ohy was measured using a novel coupled assay based on the alcohol dehydrogenase and NAD+-dependent oxidation of 10-hydroxystearic acid.


CD40 ligand (CD40L) does not stimulate proliferation of vascular smooth muscle cells.
The present study investigates the effects of CD40 ligand (CD40L) on mitogenic signalling, proliferation, and migration of cultured bovine coronary artery smooth muscle cells (SMC). A time- and concentration-dependent phosphorylation of the extracellular signal-regulated kinases-1/2 (ERK-1/2) and the mitogen-activated protein kinase p38 (p38-MAPK) was observed upon stimulation with soluble CD40L (sCD40L). This phosphorylation was inhibited by neutralizing antibodies against the CD40 and CD40L, respectively. Activation of the phosphatidylinositol-3-phosphate (PI-3) kinase pathway by sCD40L, as determined by the measurement of Akt phosphorylation, was not detected. However, there was evidence for direct activation of the NFkappaB system (degradation of IkappaBalpha and nuclear translocation of the p65 NFkappaB subunit) by sCD40L. Accordingly, sCD40L caused a small but significant increase in DNA synthesis. However, sCD40L-induced DNA synthesis was not followed by proliferation (increase in cell number). Furthermore, sCD40L did not potentiate SMC mitogenesis induced by known mitogens such as platelet-derived growth factor-BB, thrombin or serum. The lack of cell proliferation was not caused by a concomitant induction of SMC apoptosis by sCD40L. The possible role of membrane-bound CD40L in SMC mitogenesis was also studied using different membrane preparations (platelets, lymphocytes). However, no mitogenic effects of membrane-bound CD40L were detected. Finally, sCD40L did not induce SMC migration. From these data it is concluded that CD40L activates mitogenic signalling and DNA synthesis but does not contribute to proliferation or migration of vascular SMC.


Two-dimensional tension tests in plant biomechanics--sweet cherry fruit skin as a model system.
Splitting of fruits is a function of two-dimensional tension caused by different growth rates of tissues and turgor, especially water uptake shortly before harvest. In order to analyse the mechanical properties of spheroid plant material close to stress-strain conditions in vivo, a new hydraulic two-dimensional testing device was set up. Sweet cherry (Prunus avium L.) fruit skin was chosen as a model system. The recorded pressure-deflection curves were non-linear, with a considerable initial "lag phase" and a distinct increasing end part. Taking into account the special geometry, these curves could be modelled with a newly developed analytical approach based on linear elastic material behaviour. The results demonstrated good correlation if a modulus of elasticity ranging from 160 to 250 MPa for the cherry fruit skin was chosen. In addition, a mean strength value of 47 MPa was calculated based on the theory of thin shells and spheres. The results are compared with mechanical data found for fruits and other plant material. In order to test the theoretical approach, two- and one-dimensional tension tests were performed on packaging PE foil, revealing a mean modulus of 171 MPa in bi-axial tension, and 193 and 242 MPa in uni-axial tension, depending on the test speed. The results demonstrate that it seems to be feasible to use this method to analyse the two-dimensional stress-strain conditions of spheroid plant materials such as cherry fruit skins. It may be applied as a tool for crop testing to elucidate the mechanical basis of cracking susceptibility of fruits.


Ultrathin 1T-MoS2 Nanoplates Induced by Quaternary Ammonium-Type Ionic Liquids on Polypyrrole/Graphene Oxide Nanosheets and Its Irreversible Crystal Phase Transition During Electrocatalytic Nitrogen Reduction.
Ultrathin nanoplates of metastable 1T-MoS2 have been successfully stabilized and uniformly distributed on the surface of n-butyl triethyl ammonium bromide functionalized polypyrrole/graphene oxide (BTAB/PPy/GO) by a very simple hydrothermal method. BTAB as a typical kind of quaternary ammonium-type ionic liquids (ILs) played a crucial role in the formation of the obtained 1T-MoS2/BTAB/PPy/GO. It was covalently linked with PPy/GO and arranged in a highly ordered order at the solid-liquid interface of PPy/GO and H2O due to Coulombic interactions and other intermolecular interactions, which would induce and stabilize ultrathin 1T-MoS2 nanoplates by morphosynthesis. The good electrocatalytic activity toward nitrogen reduction reaction (NRR) with strong durability and good stability can be achieved by 1T-MoS2/BTAB/PPy/GO due to their excellent inorganic/organic hierarchical lamellar micro-/nanostructures. Especially, after the long-term electrocatalysis for NRR at a negative potential, metastable 1T-MoS2 as the catalytic center undergoes two types of irreversible crystal phase transition, which was converted to 1T'-MoS2 and Mo2N, caused by the competitive hydrogen evolution reaction (HER) process and the electrochemical reaction between the electroactive 1T-MoS2 and N2, respectively. The new N-Mo bonding prevents Mo atoms from binding to other N atoms in N2, resulting in the deactivation of the electrocatalysts to NRR after being used for 18 h. Even so, quaternary ammonium-type ILs would induce the crystal structures of transition-metal dichalcogenides (TMDCs), which might provide a new thought for the reasonable design of electrocatalysts based on TMDCs for electrocatalysis.


Modeling a snap-action, variable-delay switch controlling extrinsic cell death.
When exposed to tumor necrosis factor (TNF) or TNF-related apoptosis-inducing ligand (TRAIL), a closely related death ligand and investigational therapeutic, cells enter a protracted period of variable duration in which only upstream initiator caspases are active. A subsequent and sudden transition marks activation of the downstream effector caspases that rapidly dismantle the cell. Thus, extrinsic apoptosis is controlled by an unusual variable-delay, snap-action switch that enforces an unambiguous choice between life and death. To understand how the extrinsic apoptosis switch functions in quantitative terms, we constructed a mathematical model based on a mass-action representation of known reaction pathways. The model was trained against experimental data obtained by live-cell imaging, flow cytometry, and immunoblotting of cells perturbed by protein depletion and overexpression. The trained model accurately reproduces the behavior of normal and perturbed cells exposed to TRAIL, making it possible to study switching mechanisms in detail. Model analysis shows, and experiments confirm, that the duration of the delay prior to effector caspase activation is determined by initiator caspase-8 activity and the rates of other reactions lying immediately downstream of the TRAIL receptor. Sudden activation of effector caspases is achieved downstream by reactions involved in permeabilization of the mitochondrial membrane and relocalization of proteins such as Smac. We find that the pattern of interactions among Bcl-2 family members, the partitioning of Smac from its binding partner XIAP, and the mechanics of pore assembly are all critical for snap-action control.


Neurochemical evaluation of chlordecone toxicity in the mouse.
Plasma and brain levels of chlordecone in chlordecone-induced motor impairment, lethal levels of chlordecone, and cumulative LD50 of chlordecone in the mouse are presented. In terms of biochemical events, the possible contributory role of neurotransmitters in chlordecone-induced neurotoxicity is also discussed. On the basis of the data obtained so far, it is suggested that the dopaminergic pathway and its interaction with other neurotransmitter systems in the basal ganglia may be primarily involved in chlordecone-induced neurotoxicity.


[Effects of hypertonic saline on erythrocyte adherence function and bacterial infection of hemorrhagic shock rabbits].
To explore the effect of hypertonic saline on the erythrocyte adherence function and bacterial infection of hemorrhagic shock rabbits. 60 Japanese rabbits were randomly divided into 6 groups, 10 for each group. Artery catheterization and heparin were given to the rabbits in group 1 (sham shock group). Hemorrhagic shock model was set up by bleeding resulting from carotid artery catheter in group 2 (normal saline group )and group 3 (hypertonic saline group). 30 minutes after shock, the rabbits in group 1 and group 2 were treated with normal saline and balanced salt solution containing 1 x 10(9)/kg E.coli, respectively. And the rabbits in group 3 were treated with 75 g/L NaCl solution and balanced salt solution containing 1 x 10(9)/kg E.coli. Then the survival rates of the rabbits in group 1-3 were observed. Rabbits in group 4-6 were same treatment as received, group 1-3, respectively, except that there was no E.coli in balanced salt solution. The erythrocyte immune adherence function of rabbits in group 4-6 were detected 5 hours after shock by RBC-C3bR and RBC-IC rosette forming assays. The survival rate of rabbits in hypertonic saline group was significantly higher than that in normal saline group. The RBC-C3bR rosette forming rate of the normal saline treated rabbits were pronouncedly decreased, while RBC-IC rosette forming rate was notably elevated, as compared with those of either sham shock group or hypertonic saline group(P<0.01). Hypertonic saline markedly increased RBC C3bR rosette forming rate. The above findings suggest that hypertonic salt solution can remarkably improve the depressed erythrocyte immune adherence function and enhance the rabbit's resistance to E.coli challenge after hemorrhagic shock.


[Influence of admission glucose level on long-term prognosis in patients with acute coronary syndrome].
Hyperglycemia can increase the risk of death or a poor outcome following myocardial infarction. Our objective was to investigate the value of the admission glucose level in predicting long-term outcome in patients with acute coronary syndrome. The study population comprised 565 patients admitted with acute coronary syndrome within 24 hours of the start of symptoms. The final diagnosis was myocardial infarction in 56% and unstable angina in 44%. The patients' mean glucose level was 143 (77) mg/dL. During follow-up (42 [6] months), 55 (9.7%) patients died. The area under the receiver operating characteristic curve for the optimum cut point for predicting death from the glucose level was 0.67; the cut point was 128 mg/dL, with a sensitivity of 85% and a specificity of 62%. Patients were divided into 2 groups according to blood glucose level: in group 1 (36.8%), it was > or = 128 mg/dL; in group 2, <128 mg/dL. There were differences between the groups in the incidence of diabetes (47.2% vs 12.6%; P< .001), systolic blood pressure (138 [33] mm Hg vs 133 [33] mm Hg; P< .001), and ejection fraction (48.3 [0.9]% vs 55.2 [12.4]%; P=.004). At 4 years, the survival rates were 40% and 77% in groups 1 and 2, respectively (log rank test P< .001). The following were independent predictors of mortality: admission glucose level > or =128 mg/dL (hazard ratio [HR= 2.41; P=.021), admission systolic blood pressure (HR= 0.97; P< .001), admission troponin-T level (HR=4.88; P< .001), and the development of heart failure (HR=1.04; P=.001). A rise of 10 mg/dL in glucose level was associated with a 2.56-fold increase in the risk of death (P=.012). In patients with acute coronary syndrome, hyperglycemia at admission (cut point > or =128 mg/dL) was associated with increased long-term risk and, in addition, was a strong independent predictor of mortality.


Immunomodulatory capacity of the serotonin receptor 5-HT2B in a subset of human dendritic cells.
Serotonin is a monoamine neurotransmitter that signals through a wide array of receptors (5-HT1-7) many of which are also involved in immune processes. Dendritic cells (DCs) are crucial players in immune defense by bridging innate and adaptive immune responses via their vast repertoire of pattern recognition receptors and antigen-presenting capability. Although serotonin is known to influence immunity at many levels, cell type-specific expression and function of its receptors remains poorly understood. Here we aimed to study 5-HT1-7 expression and function in CD1a- and CD1a+ human monocyte-derived DCs (moDCs). We found that the 5-HT2B receptor-subtype is solely expressed by the inflammatory CD1a+ moDC subset. Specific 5-HT2B activation potently inhibited TLR2, TLR3, and TLR7/8-induced proinflammatory cytokine and chemokine (TNF-α, IL-6, IL-8, IP-10, IL-12) but not type I interferon-β responses. 5-HT2B agonism also interfered with the polarization of CD1a+ moDC-primed CD4+ T cells towards inflammatory Th1 and Th17 effector lymphocytes. Here we report the subset-specific expression and immunomodulatory function of 5-HT2B in human moDCs. Our results expand the biological role of 5-HT2B which may act not only as a neurotransmitter receptor, but also as an important modulator of both innate and adaptive immune responses.


Using model texts as a type of feedback in EFL writing.
Recent work has established that model texts could be employed as a useful feedback technique. However, few studies have employed argumentative writing tasks and analyzed draft quality, and little is known about the role played by the language analytic ability in using model texts. The current study aimed to investigate what Chinese EFL learners (n = 60) noticed at the composition and comparison (comparing their texts with model texts) processes in a four-stage argumentative writing task and explore to what degree model texts can enhance the learners' subsequent writing. The four stages were: (1) writing (pre-test); (2) comparing (treatment); (3) rewriting (immediate post-test); (4) delayed writing (delayed post-test). The findings showed that learners primarily noticed lexical features in the composition and comparison stages. Higher language analytic ability (LAA) learners and guided noticing learners could notice and elicit more information from the model texts. Overall, the use of model texts was effective in improving learners' writing by providing alternative elements associated with lexis, grammar, content, and organization. In addition, the beneficial effect of model texts on writing could be maintained after 1 week. Some pedagogical implications are put forward to help teachers make better use of model texts to improve learners' writing. This study also provides new insights into how language analytic ability affects the effectiveness of using models and provides more information on the type of learner most likely to benefit from model texts.


Poly[[diaqua-(μ(2)-1,4-dioxane-κO:O')(μ(2)-2,3,5,6-tetra-fluoro-benzene-1,4-dicarboxyl-ato-κO:O)copper(II)] 1,4-dioxane disolvate dihydrate].
In the title complex, {[Cu(C(8)F(4)O(4))(C(4)H(8)O(2))(H(2)O)(2)]·2C(4)H(8)O(2)·2H(2)O}(n), the Cu(II) ion is six-coordinated by two oxygen donors from two trans 2,3,5,6-tetra-fluoro-1,4-dicarboxyl-ate (BDC-F(4)) ligands, two O atoms from two chair 1,4-dioxane ligands and two O atoms from two terminal water mol-ecules, adopting a distorted octa-hedral coordinated geometry. Each BDC-F(4) anion bridges two Cu(II) ions in a bis-monodentate fashion, forming a [Cu(BDC-F(4))](n) chain. These chains are further linked by bridging 1,4-dioxane ligands, generating a two-dimensional net with approximately recta-ngular grids of 11.253 × 7.654 Å. Such adjacent parallel layers are connected by O-H⋯O hydrogen bonds between guest water mol-ecules and the uncoordinated carboxyl-ate O atoms and coordinated water mol-ecules into the final three-dimensional supra-molecular network.


[Analysis of the HLA-DPA1 and HLA-DPB1 polymorphism of Zhejiang Han population by PCR-sequence based typing].
To investigate the polymorphisms of HLA-DPA1 and HLA-DPB1 loci of Han population in Zhejiang province of China. The alleles of HLA-DPA1 and HLA-DPB1 loci in 100 unrelated healthy individuals were analyzed using polymerase chain reaction-sequence based typing. Eight HLA-DPA1 alleles and 19 HLA-DPB1 alleles were found in the population. The HLA-DPA1 alleles with higher frequencies were DPA1*020202 (47.0%), DPA1*010301 (38.5%) and DPA1*020101(10.5%). The HLA-DPB1 alleles with higher frequencies were DPB1*0501, DPB1*020102 and DPB1*040101. The frequencies were 39.5%, 13.5% and 13.0%, respectively. A total of 44 estimated DPA1-DPB1 haplotypes were detected. The HLA-DPA1*020202-DPB1*0501(29.5%) was the most frequent haplotype. The polymorphism data of the HLA-DPA1 and -DPB1 were obtained in Han population in Zhejiang province of China. There was linkage disequilibrium between the two loci.


Sex differences of gray matter morphology in cortico-limbic-striatal neural system in major depressive disorder.
Sex differences are observed in both epidemiological and clinical aspects of major depressive disorder (MDD). The cortico-limbic-striatal neural system, including the prefrontal cortex, amygdala, hippocampus, and striatum, have shown sexually dimorphic morphological features and have been implicated in the dysfunctional regulation of mood and emotion in MDD. In this study, we utilized a whole-brain, voxel-based approach to examine sex differences in the regional distribution of gray matter (GM) morphological abnormalities in medication-naïve participants with MDD. Participants included 29 medication-naïve individuals with MDD (16 females and 13 males) and 33 healthy controls (HC) (17 females and 16 males). Gray matter morphology of the cortico-limbic-striatal neural system was examined using voxel-based morphometry analyzes of high-resolution structural magnetic resonance imaging scans. The main effect of diagnosis and interaction effect of diagnosis by sex on GM morphology were statistically significant (p < 0.05, corrected) in the left ventral prefrontal cortex, right amygdala, right hippocampus and bilateral caudate when comparing the MDD and HC groups. Posthoc analyzes showed that females with MDD had significant GM decreases in limbic regions (p < 0.05, corrected), compared to female HC; while males with MDD demonstrated significant GM reduction in striatal regions, (p < 0.05, corrected), compared to HC males. The observed sex-related patterns of abnormalities within the cortico-limbic-strial neural system, such as predominant prefrontal-limbic abnormalities in MDD females vs. predominant prefrontal-striatal abnormalities in MDD males, suggest differences in neural circuitry that may mediate sex differences in the clinical presentation of MDD and potential targets for sex-differentiated treatment of the disorder.


Questionnaire survey on lifestyle of patients with nonalcoholic steatohepatitis.
Lack of exercise and excessive food intake are known to be the important causes of nonalcoholic steatohepatitis (NASH). To elucidate the relationship between lifestyle and NASH, we surveyed exercise and dietary habits, comparing them among 171 biopsy-proven NASH patients, 29 nonalcoholic fatty liver (NAFL) patients and 49 normal subjects. Dietary habits including the duration of dinner time, amount of rice at dinner, and weekly frequencies of meat, fries, Chinese noodles, sweets, and instant food consumption were significantly different in male NASH patients compared to normal male subjects. In women, differences were seen in the amount of rice at dinner, frequency of eating out, and proclivity for sweets. In male NASH patients, the frequency of physical exercise was significantly lower. The lifestyle tendencies of NASH were almost similar to those of NAFL. In the comparison between obese NASH and non-obese NASH, no clear lifestyle differences were found. In conclusion, the most striking result of this survey was that the lifestyle of males contributed significantly to the development of NASH. These results point to treatment of NASH in males. In female NASH patients, lifestyle differences were minimal, and the effects of other factors such as genetic background will need to be investigated.


SUMOylation facilitates the assembly of a Nuclear Factor-Y complex to enhance thermotolerance in Arabidopsis.
Heat stress (HS) has serious negative effects on plant development and has become a major threat to agriculture. A rapid transcriptional regulatory cascade has evolved in plants in response to HS. Nuclear Factor-Y (NF-Y) complexes are critical for this mechanism, but how NF-Y complexes are regulated remains unclear. In this study, we identified NF-YC10 (NF-Y subunit C10), a central regulator of the HS response in Arabidopsis thaliana, as a substrate of SUMOylation, an important post-translational modification. Biochemical analysis showed that the SUMO ligase SIZ1 (SAP AND MIZ1 DOMAIN-CONTAINING LIGASE1) interacts with NF-YC10 and enhances its SUMOylation during HS. The SUMOylation of NF-YC10 facilitates its interaction with and the nuclear translocation of NF-YB3, in which the SUMO interaction motif (SIM) is essential for its efficient association with NF-YC10. Further functional analysis indicated that the SUMOylation of NF-YC10 and the SIM of NF-YB3 are critical for HS-responsive gene expression and plant thermotolerance. These findings uncover a role for the SIZ1-mediated SUMOylation of NF-YC10 in NF-Y complex assembly under HS, providing new insights into the role of a post-translational modification in regulating transcription during abiotic stress responses in plants.


The Exchange of Informational Support in Online Health Communities at the Onset of the COVID-19 Pandemic: Content Analysis.
Online health communities (OHCs) provide social support for ongoing health-related problems. COVID-19, the disease caused by SARS-CoV-2, has been an acute and substantial stressor worldwide. The disease and its impact, especially in the beginning phases, left many people with questions about the nature, treatment, and prevention of COVID-19. Unlike typical chronic ailments discussed on OHCs, which are more established, COVID-19, at least at the onset of the pandemic, is distinct in that it lacks a consensus of clinical diagnosis and an existing community foundation. The study aims to investigate a newly formed OHC for COVID-19 to determine the topics and types of information exchange as well as the sources of information this community referenced during the early phases of the COVID-19 pandemic in the United States. A total of 357 posts from a COVID-19 OHC on the MedHelp platform were annotated according to an open-coding process. Participants' engagement patterns, topics of posts, and sources of information were quantified. Participants who offered informational support had a significantly higher percentage of responding more than once than those seeking information (P<.001). Among the information-seeking topics, symptoms and public health practice and psychological impacts were the most frequently discussed, with 26% (17/65) and 15% (10/65) of posts, respectively. Most informational support was expressed through feedback/opinion (181/220, 82.3%). Additionally, the most frequently referenced source of information was news outlets/websites, at 55% (11/20). Governmental websites were referenced less frequently. The trends of this community could be useful in prioritizing public health responses to address the most common questions asked by the public during crisis communication and in identifying which venue of communication is most effective in reaching a public audience during such times.


Bioinformatic analysis of bacteria and host cell dual RNA-sequencing experiments.
Bacterial pathogens subvert host cells by manipulating cellular pathways for survival and replication; in turn, host cells respond to the invading pathogen through cascading changes in gene expression. Deciphering these complex temporal and spatial dynamics to identify novel bacterial virulence factors or host response pathways is crucial for improved diagnostics and therapeutics. Dual RNA sequencing (dRNA-Seq) has recently been developed to simultaneously capture host and bacterial transcriptomes from an infected cell. This approach builds on the high sensitivity and resolution of RNA sequencing technology and is applicable to any bacteria that interact with eukaryotic cells, encompassing parasitic, commensal or mutualistic lifestyles. Several laboratory protocols have been presented that outline the collection, extraction and sequencing of total RNA for dRNA-Seq experiments, but there is relatively little guidance available for the detailed bioinformatic analyses required. This protocol outlines a typical dRNA-Seq experiment, based on a Chlamydia trachomatis-infected host cell, with a detailed description of the necessary bioinformatic analyses with currently available software tools.


Transcatheter edge-to-edge repair of anterior mitral flail through a dehisced annuloplasty ring.
Recurrent mitral regurgitation (MR) due to mitral annuloplasty ring dehiscence is an uncommon late complication of surgery, and is associated with an increased risk of heart failure and mortality. We report the first application of MitraClip for recurrent primary MR and annuloplasty ring dehiscence in a patient with prohibitive surgical risk.


Transcriptional profiling of lipopolysaccharide-induced acute lung injury.
Mortality associated with acute lung injury (ALI) induced by lipopolysaccharide (LPS) remains high in humans, warranting improved treatment and prevention strategies. ALI is characterized by the expression of proinflammatory mediators and extensive neutrophil influx into the lung, followed by severe lung damage. Understanding the pathogenesis of LPS-induced ALI is a prerequisite for designing better therapeutic strategies. In the present study, we used microarrays to gain a global view of the transcriptional responses of the lung to LPS in a mouse model of ALI that mimics ALI in humans. A total of 71 inflammation-associated genes were up-regulated in LPS-treated lungs, including a chemokine, LPS-induced CXC chemokine (LIX), whose role in the induction of ALI is unknown. Most of the inflammatory genes peaked at 2 h post-LPS treatment. Real-time reverse transcription-PCR confirmed the LPS-induced up-regulation of selected genes identified by microarray analysis, including LIX. The up-regulation of LIX, tumor necrosis factor alpha, and macrophage inflammatory protein 2 was confirmed at the protein level by enzyme-linked immunosorbent assays. To determine the role of LIX in the induction of ALI, we used both exogenous LIX and a LIX blocking antibody. Exogenous LIX alone elicited a neutrophil influx in the lungs, and the anti-LIX antibody attenuated the LPS-induced neutrophil accumulation in the lungs. Taken together, the results of our study demonstrate for the first time the temporal expression of inflammatory genes during LPS-induced ALI and suggest that early therapeutic intervention is crucial to attenuate lung damage. Moreover, we identified a role for LIX in the induction of ALI, and therefore LIX may serve as a novel therapeutic target for the minimization of ALI.


GmSK1, an SKP1 homologue in soybean, is involved in the tolerance to salt and drought.
In plants, various proteins are regulated by the ubiquitin-mediated system in response to different environmental stresses, such as drought, cold and heat. The Skp1-Cullin-F-box (SCF) complex, one of the multisubunit E3 ligases, has been shown to be involved in abiotic response pathways. In this study, Glycine max SKP1-like 1 (GmSK1), which had the typical characteristics of an SKP1 protein, with an alpha/beta structure, targeted to the cytoplasm and nucleus, was isolated from soybean [Glycine max (L.)]. GmSK1 was constitutively expressed in all the tested tissues, especially in the roots. Furthermore, the expression of GmSK1 was simultaneously induced by abscisic acid (ABA), jasmonic acid (JA), salicylic acid (SA), NaCl, low temperatures and drought, which suggests important roles for GmSK1 in plant responses to hormone treatments and abiotic stress. GmSK1-overexpressing transgenic tobacco (Nicotiana tobacum cv. Samsun) plants showed enhanced tolerance to high salinity and drought stress; exhibited significantly reduced inhibition of growth, greenness and water loss; and exhibited increased MDA accumulation compared with wild-type controls. Our results suggest that GmSK1 might play a role in the crosstalk between ubiquitination and abiotic stress responses in plants.


A microdevice for the creation of patent, three-dimensional endothelial cell-based microcirculatory networks.
Microvascular network formation is a significant and challenging goal in the engineering of large three-dimensional artificial tissue structures. We show here the development of a fully patent, 3D endothelial cell (microvascular) microfluidic network that has a single inlet and outlet, created in only 28 h in a microdevice involving fluid flow equivalent to natural vasculature. Our microdevice features a tailored "multi-rung ladder" network, a stylized mimic of an arterial-to-venous pedicle, designed to also allow for systematic and reproducible cell seeding. Immunofluorescence staining revealed a highly contiguous endothelial monolayer (human umbilical vein endothelial cells) throughout the whole network after 24 h of continuous perfusion. This network persisted for up to 72 h of culture, providing a useful template from which the effects of surface chemistry, fluid flow, and environmental conditions on the development of artificial vascular networks ex vivo may be rapidly and robustly evaluated.


Synthesis of Functionalized Benzo[b]furans via Oxidative Cyclization of o-Cinnamyl Phenols.
Disclosed herein is an efficient synthetic route for the synthesis of functionalized 2-benzyl benzo[b]furans via a regioselective 5-exo-trig intramolecular oxidative cyclization of ortho-cinnamyl phenols using [PdCl2(CH3CN)2] as catalyst and benzoquinone as an oxidant. Further, a sequential ortho-cinnamylation of phenols using cinnamyl alcohols catalyzed by Re2O7, followed by an oxidative cyclization using the above Pd catalyst, is performed. The reaction showed broad substrate scope with good to excellent yields.


JNK Signaling Pathway Suppresses LPS-Mediated Apoptosis of HK-2 Cells by Upregulating NGAL.
To explore the role of the c-Jun N-terminal kinase (JNK) signaling pathway in upregulated NGAL expression and its antiapoptotic mechanism in lipopolysaccharide (LPS)-mediated renal tubular epithelial cell injury. In vitro, HK-2 cells were divided into five groups (Con, LPS 1 h, LPS 3 h, LPS 6 h, and LPS 12 h groups) based on the time of LPS (10 μM) treatment. NGAL and caspase-3 gene expression levels were detected by RT-PCR to assess dynamic changes. HK-2 cells were pretreated with SP600125 (20 μM) for 2 hours, followed by LPS (10 μM) stimulation for 3 hours. NGAL and caspase-3 gene expression levels were then determined. NGAL mRNA was increased significantly within 6 hours, and caspase-3 mRNA was increased within 3 hours after treatment (P < 0.05). Correlation analysis showed a high correlation between their expression (r = 0.448, P < 0.05). After pretreatment with SP600125, mRNA expression of NGAL in the LPS group was inhibited, while that of caspase-3 was increased significantly. The NGAL mRNA expression level in the SB + LPS group was decreased significantly compared with that in the LPS group, but it was slightly higher than that in the SP group (∼1.5 times of that in the Con group). However, caspase-3 mRNA expression was increased significantly in the SB + LPS group (P < 0.001) (3.5 times of that in the Con group). It also showed a significant increase compared with SP and LPS groups (P < 0.001 vs. SB group; P < 0.05 vs. LPS group). We also found that NGAL and caspase 3 proteins were increased significantly in LPS and SP + LPS groups, but SP600125 decreased the NGAL level by almost 35% and increased the caspase 3 level by 50% in the SP + LPS group compared with the LPS group (P < 0.05). The JNK signaling pathway inhibits LPS-mediated apoptosis of renal tubular epithelial cells by upregulating NGAL.


Greater development of 1,2-dimethylhydrazine-induced colon cancer in a rat model of type 2 diabetes mellitus.
Several clinical cohort and case-control studies have suggested a link between diabetes and colon cancer. Otsuka Long-Evans Tokushima Fat (OLETF) rats spontaneously develop type 2 diabetes mellitus and Long-Evans Tokushima Otsuka (LETO) rats are non-diabetic. The relationship between type 2 diabetes mellitus and colon cancer was examined in these rats. The carcinogen 1,2-dimethylhydrazine was administered subcutaneously once weekly for 10 weeks, and the animals were killed and necropsied in week 29. All OLETF rats and 80% of the LETO rats developed cancer. The number of colon cancers per rat was significantly greater in the diabetic than in the non-diabetic rats. Although the tumours tended to be larger in diabetic rats, the difference was not statistically significant. No significant differences were observed in the depth of invasion or histological type of cancer in the two groups. Type 2 diabetes mellitus may enhance the generation and growth of colon cancer.


Inhaled Insulin - Current Direction of Insulin Research.
Diabetes Mellitus (DM) is a metabolic disorder characterized by relative or absolute deficiency of insulin, resulting in hyperglycemia. Subcutaneous insulin and Oral Hypoglycaemic Agents (OHA) constitute the main treatment option for DM. Insulin is administered by injection or continuous infusion to control glucose levels mainly in Type I diabetes. Newer routes both oral and non oral, for insulin administration are current direction of insulin research as insulin injection therapy is burdensome and painful for many patients. Inhalational insulin is an attractive alternative for systemic administration of insulin given its accessibility and large alveolar-capillary network of lungs for drug absorption. Afrezza, inhaled insulin has been recently approved by Food and Drug Administration (FDA). It is a new, quicker acting inhalable insulin with a different and safer pharmacokinetic profile in comparison to previously failed inhaled form of insulin.


Collaborating sites for community-oriented integrated care and health promotion.
London Journal of Primary Care wishes to develop a network of collaborating sites to better understand how to achieve community-oriented integrated care and health promotion in different contexts. A collaborating site can do more than submit papers. It can develop its own domain on the LJPC website, contribute to the development of LJPC policy, and stimulate discussions with other collaborating sites. At any time a collaborating site can opt out. In addition to securing papers for publication, a site might nurture a network of supporters, teach people to use multiple research and quality improvement methods, develop a system of governance for locally led inquiries, develop case studies of community-oriented integrated care and health promotion and facilitate within-site and between-site learning and change.


Impact of laboratory treatment with coloring and fluorescent liquids on the optical properties of zirconia before and after accelerated aging.
Laboratory procedures, such as dipping in coloring and fluorescent liquids, can be used to improve the optical properties of zirconia. However, information is lacking on the effect of these liquids. The purpose of this in vitro study was to evaluate the color differences and degree of fluorescence of zirconia (3Y-TZP) treated with coloring and fluorescent liquids before and after an accelerated aging protocol. Forty disk-shaped specimens of 3Y-TZP were fabricated by milling and separated according to the laboratory treatment performed: white zirconia (control group); zirconia treated with coloring liquid (A2 group); zirconia treated with fluorescent liquid (fluorescent group); and zirconia treated with both liquids (A2 fluorescent group). The L*a*b* coordinates before aging (T0) were obtained with a spectrophotometer, and the degree of fluorescence was measured. The disks were subjected to accelerated aging for 1 hour (T1) and 5 hours (T2). Measurements were made before and after each time interval. Color differences (ΔE00) were calculated using the CIEDE2000 formula and analyzed by 2-way ANOVA. Lightness (ΔL'), chroma (ΔC'), and hue differences (ΔH') were analyzed by multivariate ANOVA. Degrees of fluorescence were obtained as percentages and were analyzed by 2-way ANOVA. Multiple comparisons were performed by the Tukey HSD test (α=.05). Color differences were observed when 3Y-TZP disks were treated with coloring (7.91 ΔE00), with fluorescent liquid (5.81 ΔE00), and with both liquids (5.52 ΔE00). Accelerated aging resulted in color differences in the T2 A2 group (6.74 ΔE00) and at both times evaluated in the fluorescent group (T1=8.59 ΔE00 and T2=8.47 ΔE00) (P<.001). In the A2 fluorescent group, the degree of fluorescence was not influenced significantly (P>.05). The use of fluorescent liquid influenced the degree of fluorescence in the fluorescent group (T0=20%). Significant differences in color, lightness, chroma, and hue were achieved in all tested groups before and after aging. The degree of fluorescence was statistically different only in the fluorescent group and was not influenced by accelerated aging.


Characteristics and risk factors of UCS fracture subtypes in periprosthetic fractures around the hip.
Periprosthetic fractures (PPFs) following hip arthroplasty are complex injuries. This study evaluates patient demographic characteristics, management, outcomes, and risk factors associated with PPF subtypes over a decade. Using a multicentre collaborative study design, independent of registry data, we identified adults from 29 centres with PPFs around the hip between January 2010 and December 2019. Radiographs were assessed for the Unified Classification System (UCS) grade. Patient and injury characteristics, management, and outcomes were compared between UCS grades. A multinomial logistic regression was performed to estimate relative risk ratios (RRR) of variables on UCS grade. A total of 1,104 patients were included. The majority were female (57.9%; n = 639), ethnically white (88.5%; n = 977), used mobility aids (67%; n = 743), and had a median age of 82 years (interquartile range (IQR) 74 to 87). A total of 77 (7%) had pain prior to the PPF. The most common UCS grade was B2 (33%; n = 368). UCS type D fractures had the longest length of stay (median 19 days (IQR 11 to 26)), highest readmission to hospital (21%; n = 9), and highest rate of discharge to step-down care (52%; n = 23). Multinomial regression suggests that uncemented femoral stems are associated with a reduced risk of UCS C (RRR 0.36 (95% confidence interval (CI) 0.2 to 0.7); p = 0.002) and increased risk of UCS A (RRR 3.3 (95% CI 1.9 to 5.7); p < 0.001), compared to UCS B fracture. The most common PPF type in elderly frail patients is UCS B2. Uncemented stems have a lower risk of UCS C fractures compared to cemented stems. A national PPF database is needed to further identify correlation between implants and fracture subtypes.


Emerging roles for phospholipase A2 enzymes in cancer.
Phospholipase A(2) (PLA(2)) enzymes (EC22.214.171.124) regulate the release of biologically active fatty acids and lysophospholipids from membrane phospholipid pools. These lipids are also substrates for intracellular biochemical pathways that generate potent autocrine and paracrine lipid mediators such as the eicosanoids and platelet activating factor. These factors, in turn, regulate cell proliferation, survival, differentiation, motility, tissue vascularisation, and immune surveillance in virtually all tissues, functions that are subverted by cancer cells for tumour growth and metastasis. Thus the relevance of PLA(2)-dependent pathways to the genesis and progression of cancer has been of interest since their discovery and with recent technological advances, their role in tumourigenesis has become more tractable experimentally. Limited human genetic studies have not yet identified PLA(2) enzymes as classical mutated oncogenes or tumour suppressor genes. However, there is strong evidence that of the 22 identified human PLA(2) enzymes, ten of which have been studied in cancer to date, most are aberrantly expressed in a proportion of tumours derived from diverse organs. Correlative and functional studies implicate the expression of some secreted enzymes (sPLA(2)s), particularly the best studied enzyme Group IIA sPLA(2) in either tumour promotion or inhibition, depending on the organ involved and the biochemical microenvironment of tumours. As in immune-mediated inflammatory pathologies, genetic deletion studies in mice, supported by limited studies with human cells and tissues, have identified an important role for Group IVA PLA(2) in regulating certain cancers. Pharmacological intervention studies in prostate cancer suggest that hGIIA-dependent tumour growth is dependent on indirect regulation of Group IVA PLA(2). Group VI calcium-independent PLA(2) enzymes have also been recently implicated in tumourigenesis with in vitro studies suggesting multiple possible roles for these enzymes. Though apparently complex, further characterization of the regulatory relationships amongst PLA(2) enzymes, lipid mediator biosynthetic enzymes and the lipid mediators they produce during tumour progression is required to define the biochemical context in which the enzymes modulate cancer growth and development.


Helicobacter pylori primary resistance to metronidazole and clarithromycin in Brazil.
Helicobacter pylori resistance to metronidazole was detected in 107 (52.97%) of 202 strains. Twenty (9.85%) strains, 18 of them harboring 23S ribosomal DNA mutations, were resistant to clarithromycin. Metronidazole resistance was associated with female gender. Resistance to metronidazole and resistance to clarithromycin were associated. Increasing clarithromycin resistance rates were observed over time.


Use of cefoperazone MacConkey agar for selective isolation of Laribacter hongkongensis.
A new selective medium, cefoperazone MacConkey agar (CMA), was developed for primary isolation of Laribacter hongkongensis from stool. Its performance in quantitative recovery and in a clinical evaluation of 4,741 human diarrheal stool specimens was superior to that of charcoal cefoperazone deoxycholate agar. In addition, with CMA, Arcobacter butzleri was unexpectedly isolated from the stools of six patients.


The role of onsite Implanon insertion training for HEWs for sustainable FP programs in Ethiopia: a mixed-method study.
Training health extension workers on Implanon insertion offsite, or away from the workplace, can be cost-intensive, can depend on the human and financial resources of partners, and can compromise routine health services by taking health workers off the job. To address these limitations, the USAID Transform Primary Health Care Activity in Ethiopia designed an onsite Implanon insertion training at the primary health care level. This study compared and documented the implementation experience of onsite vs offsite Implanon insertion training for health extension workers. In a mixed-method study conducted in March 2020, the team collected training data from 468 participants-half trained onsite and half offsite-and conducted key informant interviews with 20 purposively sampled individuals. The team analyzed this data, summarizing the data in tables and figures and performing a t test with p value < 0.05 using SPSS v.20. Qualitative data were analyzed manually in Excel and summarized in Word based on emerging themes. Health extension workers trained onsite were away from routine work an average of 3 days compared to 8 days for those trained offsite (P < 0.001). The difference in average per-trainee cost of onsite (2707 Birr = 87.3 USD) and offsite (6006 Birr = 193.7USD) training was significant (P < 0.001). There was no significant difference in mean scores of onsite and offsite trainees on the knowledge pre-test (P < 0.947) and post-test (P < 0.220) or in simulated practice on an arm model (p < 0.202). Onsite trainees, assigned to their own health post for clinical practice, performed Implanon insertions on an average of 10 clients: offsite trainees on an average of 5 clients. Most interview participants reported that the onsite Implanon training was better organized, conducted, followed up, and monitored by health centers to ensure community-level access to Implanon services, with quality and continuity. Onsite training is a promising approach and minimizes service interruption. It is a likely strategy for on-demand training of health extension workers and immediate assignment of skilled providers to ensure access to and continuity of quality community-level Implanon care. Trial registration N/A.


Time Trials Versus Time-to-Exhaustion Tests: Effects on Critical Power, W', and Oxygen-Uptake Kinetics.
To investigate single-day time-to-exhaustion (TTE) and time-trial (TT) -based laboratory tests values of critical power (CP), W prime (W'), and respective oxygen-uptake-kinetic responses. Twelve cyclists performed a maximal ramp test followed by 3 TTE and 3 TT efforts interspersed by 60 min recovery between efforts. Oxygen uptake ( V ˙ O 2 ) was measured during all trials. The mean response time was calculated as a description of the overall [Formula: see text]-kinetic response from the onset to 2 min of exercise. TTE-determined CP was 279 ± 52 W, and TT-determined CP was 276 ± 50 W (P = .237). Values of W' were 14.3 ± 3.4 kJ (TTE W') and 16.5 ± 4.2 kJ (TT W') (P = .028). While a high level of agreement (-12 to 17 W) and a low prediction error of 2.7% were established for CP, for W' limits of agreements were markedly lower (-8 to 3.7 kJ), with a prediction error of 18.8%. The mean standard error for TTE CP values was significantly higher than that for TT CP values (2.4% ± 1.9% vs 1.2% ± 0.7% W). The standard errors for TTE W' and TT W' were 11.2% ± 8.1% and 5.6% ± 3.6%, respectively. The [Formula: see text] response was significantly faster during TT (~22 s) than TTE (~28 s). The TT protocol with a 60-min recovery period offers a valid, time-saving, and less error-filled alternative to conventional and more recent testing methods. Results, however, cannot be transferred to W'.