Ataxia telangiectasia mutated kinase inhibition promotes irradiation-induced PD-L1 expression in tumour-associated macrophages through IFN-I/JAK signalling pathway.
Tumour-associated macrophages (TAMs) are one of the primary sources of PD-L1 expression in the tumour microenvironment (TME). Ionizing radiation (IR) promotes PD-L1 expression in tumour cells. However, the effect of IR on macrophage PD-L1 expression and the underlying mechanisms remain unclear. ATM kinase, as the key kinase for initiating DNA damage repair (DDR) process, is associated with innate immune STING axis activation. Here, we explored the molecular mechanism implicated in macrophage PD-L1 expression regulated by IR as well as the role of ATM kinase in this process. IR-regulated PD-L1 expression in macrophages and associated signalling pathways were explored by in vitro studies using murine and human macrophage cell lines. A colorectal xenograft murine model was employed to demonstrate the impact of targeting ATM and PD-L1 expression in TAMs following IR on growth of tumour in vivo. IR up-regulated PD-L1 expression in macrophages, which was further augmented by ATM kinase inhibition. ATM inhibition increased IR-induced DNA damage, which activated STING/interferon regulatory factor 3 (IRF3) signalling pathway and up-regulated type I interferon (IFN-I) expression in macrophages. IFN-I bound to the IFN α receptor 1 on macrophages, activated the downstream JAK1 and STAT1/3 signalling and eventually led to PD-L1 up-expression. ATM inhibition augmented IR-induced PD-L1 expression in macrophages and CD8+ T cell infiltration, and promoted anti-tumour efficacy in vivo. These results suggested that ATM inhibition promoted IR-induced PD-L1 expression through the activation of innate immunity in TAMs, which provided a novel approach to enhance the anti-tumour efficacy of RT.


Inhibition of Histone Deacetylase 6 by Tubastatin A Attenuates the Progress of Osteoarthritis via Improving Mitochondrial Function.
Because chondrocytes are the only resident cells in articular cartilage, the steady state of these cells is important for the maintenance of joint function. In various osteoarthritis diseases, chondrocytes undergo a series of pathophysiologic changes, leading to the loss of chondrocytes and the degradation of extracellular matrix (ECM). This study found that Cytoplasmic localized histone deacetylase 6 (HDAC6) is up-regulated on the articular surface in a destabilization of the medial meniscus-induced mouse osteoarthritis model. Because HDAC6 is highly related to the acetylation of tubulin and the function of the microtubule system is closely related to material transport and signal transduction, the relationship between the expression level or activity of HDAC6 and the fate of chondrocytes in vitro and in vivo were confirmed. Primary chondrocytes overexpressing DNA-HDAC6 with plasmid were constructed in vitro, and HDAC6 inhibitor Tubastatin A was selected to inhibit HDAC6 enzyme activity in vivo and in vitro. Subsequently, mitochondrial spatial arrangement, degradation of ECM, and pathological changes in joint were defined. The results indicate that overexpression of HDAC6 causes mitochondrial dysfunction and promotes reactive oxygen species production, leading to degradation of ECM. Tubastatin A treatment after osteoarthritis ameliorates the degradation of cartilage and improves the microenvironment and function of the joint. HDAC6 may be targeted to treat osteoarthritis.


Stability indicating high-performance thin-layer chromatographic determination of gatifloxacin as bulk drug and from polymeric nanoparticles.
A simple, sensitive, selective, precise and stability indicating high-performance thin-layer chromatographic method for determination of gatifloxacin both as a bulk drug and from polymeric nanoparticles was developed and validated as per the International Conference on Harmonization (ICH) guidelines. The method employed thin-layer chromatography (TLC) aluminium plates precoated with silica gel 60F-254 as the stationary phase and the mobile phase consisted of n-propanol-methanol-concentrated ammonia solution (25%) (5:1:0.9, v/v/v). This solvent system was found to give compact spots for gatifloxacin (R(f) value of 0.60+/-0.02). Densitometric analysis of gatifloxacin was carried out in the absorbance mode at 292 nm. The linear regression analysis data for the calibration plots showed good linear relationship with r=0.9953 with respect to peak area in the concentration range of 400-1200 ng spot(-1). The mean value (+/-S.D.) of slope and intercept were 9.66+/-0.05 and 956.33+/-27.67, respectively. The method was validated for precision, accuracy, ruggedness and recovery. The limits of detection and quantitation were 2.73 and 8.27 ng spot(-1), respectively. Gatifloxacin was subjected to acid and alkali hydrolysis, oxidation, photodegradation and dry heat treatment. The drug undergoes degradation under acidic and basic conditions and upon wet and dry heat treatment. The degraded products were well separated from the pure drug. The statistical analysis proves that the developed method for quantification of gatifloxacin as bulk drug and from polymeric nanoparticles is reproducible and selective. As the method could effectively separate the drug from its degradation products, it can be employed as stability-indicating one.


Antithrombotic activities of pellitorine in vitro and in vivo.
Pellitorine (PLT), an active amide compound, is well known to possess insecticidal, antibacterial and anticancer properties. However, the anti-coagulant functions of PLT are not studied yet. Here, the anticoagulant activities of PLT were examined by monitoring activated partial thromboplastin time (aPTT), prothrombin time (PT), and the activities of cell-based thrombin and activated factor X (FXa). Furthermore, the effects of PLT on the expressions of plasminogen activator inhibitor type 1 (PAI-1) and tissue-type plasminogen activator (t-PA) were tested in tumor necrosis factor (TNF)-α activated human umbilical vein endothelial cells (HUVECs). Treatment with PLT resulted in prolonged aPTT and PT and inhibition of the activities of thrombin and FXa, and PLT inhibited production of thrombin and FXa in HUVECs. And PLT inhibited thrombin-catalyzed fibrin polymerization and platelet aggregation. In accordance with these anticoagulant activities, PLT elicited anticoagulant effects in mouse. In addition, treatment with PLT resulted in the inhibition of TNF-α-induced production of PAI-1 and in the significant reduction of the PAI-1 to t-PA ratio. Collectively, PLT possesses antithrombotic activities and offers bases for development of a novel anticoagulant.


Differential Associations of 6 Adiposity Indices With Dementia in Older Adults: The MIND-China Study.
The cross-sectional association between late-life obesity and dementia is often explained by the obesity paradox. We assessed the potential differential associations of various adiposity indices with dementia and subtypes of dementia in rural Chinese older adults. A population-based cross-sectional study. A total of 5277 participants (age ≥60 years; 57.23% female) who were living in rural communities and were examined in March-September 2018 for MIND-China. We used weight, height, and waist circumference (WC) to calculate 6 adiposity indices: body mass index (BMI), waist-to-height ratio (WHtR), weight-adjusted-waist index (WWI), A Body Shape Index (ABSI), body roundness index (BRI), and Conicity Index (ConI). Dementia, Alzheimer's disease (AD), and vascular dementia (VaD) were clinically diagnosed following the international criteria. Data were analyzed with logistic regression models. Of the 5277 participants, 303 were diagnosed with dementia, including 193 with AD and 99 with VaD. The multivariable-adjusted odds ratio (95% CI) of dementia associated with the highest (vs lowest) quintile of adiposity index was 2.32 (1.40-3.85) for WWI, 1.56 (1.03-2.36) for ABSI, and 1.40 (0.92-2.11) for ConI. Similarly, higher levels of these 3 adiposity indices were significantly associated with an increased likelihood of AD, whereas a higher BMI was associated with a decreased likelihood of AD. None of the 6 examined adiposity indices was significantly associated with VaD when adjusting for multiple confounders. The adiposity index WWI is linearly associated with the likelihood of dementia and AD. An increased WWI may be a clinical marker for the dementia syndrome and Alzheimer's dementia.


Hearing loss and cognitive decline in the general population: a prospective cohort study.
Previous studies identifying hearing loss as a promising modifiable risk factor for cognitive decline mostly adjusted for baseline age solely. As such a faster cognitive decline at a higher age, which is expected considering the non-linear relationship between cognition and age, may have been overlooked. Therefore it remains uncertain whether effects of hearing loss on cognitive decline extend beyond age-related declines of cognitive function. 3,590 non-demented participants were eligible for analysis at baseline, and a maximum of 837 participants were eligible for the longitudinal analysis. Hearing loss was defined at baseline. Cognitive function was measured at baseline and at follow-up (4.4 years [SD: 0.2]). Multivariable linear regression analysis was used for the cross-sectional analysis. Linear mixed models were used to assess the longitudinal association between hearing loss and cognitive decline over time while adjusting for confounders and the interaction of age and follow-up time. Hearing loss was associated with lower cognitive function at baseline. Moreover, hearing loss was associated with accelerated cognitive decline over time on a memory test. After additionally adjusting for the interaction between age and follow-up time, we found that hearing loss did not accelerate cognitive decline anymore. Hearing loss was associated with lower cognitive function at baseline and accelerated cognitive decline on a memory test. The association between hearing loss and accelerated cognitive decline was non-significant after additional adjustment for non-linear age effects. More evidence is needed to ensure the role of hearing loss as a modifiable risk factor for cognitive decline.


DNA damage in germ cells induces an innate immune response that triggers systemic stress resistance.
DNA damage responses have been well characterized with regard to their cell-autonomous checkpoint functions leading to cell cycle arrest, senescence and apoptosis. In contrast, systemic responses to tissue-specific genome instability remain poorly understood. In adult Caenorhabditis elegans worms germ cells undergo mitotic and meiotic cell divisions, whereas somatic tissues are entirely post-mitotic. Consequently, DNA damage checkpoints function specifically in the germ line, whereas somatic tissues in adult C. elegans are highly radio-resistant. Some DNA repair systems such as global-genome nucleotide excision repair (GG-NER) remove lesions specifically in germ cells. Here we investigated how genome instability in germ cells affects somatic tissues in C. elegans. We show that exogenous and endogenous DNA damage in germ cells evokes elevated resistance to heat and oxidative stress. The somatic stress resistance is mediated by the ERK MAP kinase MPK-1 in germ cells that triggers the induction of putative secreted peptides associated with innate immunity. The innate immune response leads to activation of the ubiquitin-proteasome system (UPS) in somatic tissues, which confers enhanced proteostasis and systemic stress resistance. We propose that elevated systemic stress resistance promotes endurance of somatic tissues to allow delay of progeny production when germ cells are genomically compromised.


Risk factors for cardiovascular diseases in patients with systemic lupus erythematosus: an umbrella review.
Cardiovascular disease in patients with systemic lupus erythematosus (SLE) remains one of the most common causes of death and is caused by several factors, including both traditional and disease-specific risk factors. We aimed to systematically appraise the evidence of cardiovascular disease risk factors focusing on the SLE population. The protocol for this umbrella review is registered in PROSPERO (registration no. CRD42020206858). A systematic literature search was conducted in PubMed, Embase, and the Cochrane Library from database inception to June 22, 2022, for systematic reviews and meta-analyzes that examined cardiovascular disease risk factors in patients with SLE. Two reviewers independently extracted data and assessed the quality of the included studies using the "Assessing the Methodological Quality of Systematic Reviews 2 (AMSTER 2)" tool. Of the 102 identified articles, nine systematic reviews were included in this umbrella review. All included systematic reviews were assessed as critically low quality according to the AMSTER 2 tool. The traditional risk factors identified in this study were older age, male sex, hypertension, dyslipidemia, smoking, and a family history of cardiovascular disease. SLE-specific risk factors were long-term disease duration, lupus nephritis, neurological disorders, high disease activity, organ damage, use of glucocorticoids, azathioprine, and antiphospholipid antibodies, including anticardiolipin antibodies and lupus anticoagulant. This umbrella review identified some cardiovascular disease risk factors in patients with SLE; however, the study quality of all included systematic reviews was critically low. Key Points • We examined the evidence of cardiovascular disease risk factors focusing on patients with systemic lupus erythematosus. • We found that long-term disease duration, lupus nephritis, neurological disorders, high disease activity, organ damage, use of glucocorticoids, azathioprine, and antiphospholipid antibodies, including anticardiolipin antibodies and lupus anticoagulant, were cardiovascular disease risk factors among patients with systemic lupus erythematosus. • The review indicates the need for well-validated and high-quality future reviews that assess major adverse cardiovascular events as an outcome in patients with systemic lupus erythematosus.


Rapid determination of capsaicinoids by colorimetric method.
Capsaicinoids, the pungent component of chili peppers, are generally analyzed by precise analytical techniques, such as gas chromatography and high-performance liquid chromatography (HPLC), but these are not practical for the mass analyses of samples. To analyze mass samples rapidly, a colorimetric method was suggested. In this work, pigments and capsaicinoids were efficiently separated from chili pepper extract by sequential solid-liquid extraction and liquid-liquid extraction in test tubes followed by a colorimetric analysis on the capsaicinoids by a selective chromogenic reaction with Gibbs reagent (2,6-dichloroquinone-4-chloroimide). In the comparison of the capsaicinoid content by the colorimetric method and HPLC using acetone extracts of fresh pepper and dry red pepper as samples, R2 was 0.9973 and 0.9816, respectively, which shows a high linear correlation. In addition, a minimum of 1 μg/mL capsaicinoids can be detected and it was therefore determined that the method can efficiently analyze a great quantity of samples in a short time.


Wide Area Transepithelial Sampling with Computer-Assisted Analysis (WATS3D) Is Cost-Effective in Barrett's Esophagus Screening.
Wide area transepithelial sampling with three-dimensional computer-assisted analysis (WATS3D) is an adjunct to the standard random 4-quadrant forceps biopsies (FB, "Seattle protocol") that significantly increases the detection of Barrett's esophagus (BE) and associated neoplasia in patients undergoing screening or surveillance. To examine the cost-effectiveness of adding WATS3D to the Seattle protocol in screening patients for BE. A decision analytic model was used to compare the effectiveness and cost-effectiveness of two alternative BE screening strategies in chronic gastroesophageal reflux disease patients: FB with and without WATS3D. The reference case was a 60-year-old white male with gastroesophageal reflux disease (GERD). Effectiveness was measured by the number needed to screen to avert one cancer and one cancer-related death, and quality-adjusted life years (QALYs). Cost was measured in 2019 US$, and the incremental cost-effectiveness ratio (ICER) was measured in $/QALY using thresholds for cost-effectiveness of $100,000/QALY and $150,000/QALY. Cost was measured in 2019 US$. Cost and QALYs were discounted at 3% per year. Between 320 and 337 people would need to be screened with WATS3D in addition to FB to avert one additional cancer, and 328-367 people to avert one cancer-related death. Screening with WATS3D costs an additional $1219 and produced an additional 0.017 QALYs, for an ICER of $71,395/QALY. All one-way sensitivity analyses resulted in ICERs under $84,000/QALY. Screening for BE in 60-year-old white male GERD patients is more cost-effective when WATS3D is used adjunctively to the Seattle protocol than with the Seattle protocol alone.


Medical specialty certification in the United States-a false idol?
Recent changes to medical specialty certification in the USA have prompted the process to come under intense scrutiny. We review the history of board certification and the changes made to the process. As part of this review, we examine both literature and public record to examine the motives behind the changes made. We then review the legal challenges and changes under way to modify the current ABMS board re-certification process. In 1917, the first board certification was a lifetime designation, voluntary, and managed by unpaid board members with a focus to enhance quality for patients. Corresponding to the implementation of time-limited certification, $55 million of physician testing fees were transferred from the American Board of Internal Medicine to its Foundation between 1989 and 1999. From 2000 through 2007, and additional $20.66 million were transferred from the ABIM to its Foundation culminating in the purchase of a $2.3 million luxury condominium in December 2007. Significant financial conflicts of interest for the implementation of time-limited specialty certification exited and continue to plague the medical profession. The specialty boards and the organizations that created them should remove all requirements for time-limited board certification and resort to conventional self-selected ACGME-approved CME programs for ongoing education.


Food label accuracy of common snack foods.
Nutrition labels have raised awareness of the energetic value of foods, and represent for many a pivotal guideline to regulate food intake. However, recent data have created doubts on label accuracy. We tested label accuracy for energy and macronutrient content of prepackaged energy-dense snack food products. We measured "true" caloric content of 24 popular snack food products in the U.S. and determined macronutrient content in 10 selected items. Bomb calorimetry and food factors were used to estimate energy content. Macronutrient content was determined according to Official Methods of Analysis. Calorimetric measurements were performed in our metabolic laboratory between April 20th and May 18th and macronutrient content was measured between September 28th and October 7th of 2010. Serving size, by weight, exceeded label statements by 1.2% [median] (25th percentile -1.4, 75th percentile 4.3, P = 0.10). When differences in serving size were accounted for, metabolizable calories were 6.8 kcal (0.5, 23.5, P = 0.0003) or 4.3% (0.2, 13.7, P = 0.001) higher than the label statement. In a small convenience sample of the tested snack foods, carbohydrate content exceeded label statements by 7.7% (0.8, 16.7, P = 0.01); however fat and protein content were not significantly different from label statements (-12.8% [-38.6, 9.6], P = 0.23; 6.1% [-6.1, 17.5], P = 0.32). Carbohydrate content explained 40% and serving size an additional 55% of the excess calories. Among a convenience sample of energy-dense snack foods, caloric content is higher than stated on the nutrition labels, but overall well within FDA limits. This discrepancy may be explained by inaccurate carbohydrate content and serving size.


A pig model for investigation of muscle and myocutaneous flaps.
An experimental model in the pig is described which readily enables studies to be made on musculocutaneous and muscle flaps. The model allows easy, reliable control over the pedicle blood supply at any time in the postoperative period.


'From soil to chocolate bar': identifying critical steps in the journey of cadmium in a Colombian cacao plantation.
Regulation of maximum levels of cadmium in chocolate is an issue for cacao exportation from many parts of Latin America, including Colombia. These limits are related to the final product, but buyers often request maximum levels of Cd in the beans. However, to date, there is neither a clear understanding of the relationship between the specified levels of Cd in chocolate and cocoa derivatives and levels in harvested beans or soil nor of the effect of post-harvest processes on the levels of Cd in the final product. To address this, the fate of Cd concentration from soil to chocolate bar was followed in a single farm in Santander district, Colombia. The concentration of Cd in soils was measured using ICP-OES and correlated with soil pH, soil organic matter (SOM), and the use of P-based fertilisers. Cd concentrations were also measured in unfermented seeds, fermented and dried beans, shell, nibs, and chocolate. SOM (2.93-3.78%), soil pH (4.7-4.9), soil P concentration (120-132 mg kg-1) affect Cd availability. However, it is still unclear whether Cd concentration of P-based fertilisers (3-30 mg kg-1) is important or not. While post-harvest treatments did not affect the Cd concentration of beans (4.17 ± 0.8 mg kg-1 on average), the removal of the shell (6.57 mg kg-1) from the nibs (3.28 mg kg-1), as well as the percentage of cocoa mass used contributes to a reduction in Cd concentration in the chocolate bar (1.60 mg kg-1). This study provides clear indications on where research into mitigation measures should be focussed, as well as indicating the importance of carrying out analyses for Cd in the nib or cacao mass, rather than the whole bean, reducing Cd concentration by up to 40%.


Evidence of neurophysiological improvement of early manifestations of small-fiber dysfunction after liver transplantation in a patient with familial amyloid neuropathy.
Small fiber polyneuropathy (SFP) is a common heralding clinical manifestation of damage to the nervous system in patients with familial amyloidosis. The diagnosis of SFP is a significant factor in the decision to treat a previously asymptomatic gene carrier, as treatment would prevent irreversible nerve damage. This requires detection of the earliest but unequivocal signs of peripheral nerve involvement. We present the case of a young female who was diagnosed of SFP, supported by data from quantitative sensory testing. She had preserved sensory nerve action potentials in the distalmost nerves of her feet and recordable nociceptive evoked potentials. She was successfully transplanted the liver from a previously healthy donor, and recovered fully of her symptoms and signs. Improvement was documented with repeated psychophysical and electrodiagnostic testing in the course of 4 years after transplantation. This case illustrates the utility of psychophysical testing to support the diagnosis of SFP.


Gitelman syndrome in pregnancy: a case series.
To evaluate maternal and fetal outcomes in women with Gitelman syndrome (GS). Retrospective analysis of the clinical data of five patients with the clinical diagnosis of GS during pregnancy, who were admitted to Beijing Shijitan Hospital, Capital Medical University between 2013 and 2019, was conducted. Five women with GS during pregnancy who finally gave birth to a total of eight newborns have been included. Three cases were primiparas and two cases were multiparas. Two cases were diagnosed before pregnancy and three cases were diagnosed in first or second trimester. The primary treatment was oral or intravenous electrolytes supplement. Three patients delivered through the vagina, and shoulder dystocia occurred in one patient. Two patients delivered by cesarean section, with one because of symptom of limb weakness during the course of labor and the other owing to gestational diabetes with fetal macrosomia. Postpartum hemorrhage and urinary retention were not reported in these cases. In perinatal period all the infants had good outcome. The children, aged between six months and five years, were healthy and well-developed during follow-up. The maternal and perinatal outcome is usually favorable. We should pay attention to electrolyte examination in the first trimester in order to diagnose and manage the GS efficiently. Well-controlled patients with Gitelman syndrome can deliver through the vagina.


Motivational dynamics and impact on performances: The case of physicians of university polyclinics in the NHS scenario.
The principle of inseparability of teaching, academic research and assistance to patients, to which university physicians are subject in carrying out their work, invites reflection, aimed at identifying a combination of the three activities characterised, at the same time, by efficiency and effectiveness. The present paper illustrates a paradigm based on the balance of intrinsic and extrinsic motivations with the purpose of identifying an efficient solution in the performance of the duties of university doctors. The analysis refers to Italy in terms of the main scenario, or, conversely, the achievement of more profit-related goals. Depending on the prevailing goal, it is possible to recognise the superiority of the constitutional principles.


Underestimation of impaired kidney function with serum creatinine.
Serum creatinine (SCr) levels are frequently used as a screening test to assess impaired renal function; however, patients can have significantly decreased glomerular filtration rate (GFR) with normal SCr values and making the recognition of kidney dysfunction more difficult. Hence, this study was designed to determine the extent of misclassification of the patients who have significantly reduced GFR as calculated by reexpressed four variable modification of diet in renal disease (MDRD) equation but, normal range of SCr. The study included 1040 in and out patients referred by physicians for serum creatinine measurement. When an exclusion criterion was applied 928 patients were qualified for the study. SCr was measured in 928 patients by a Roche kinetic compensated Jaffe's assay. GFR was calculated using reexpressed four variable MDRD study equation. Of the 928 patients 270 (29.1%) had renal dysfunction on the basis of eGFR (<60 ml/min/1.73 m(2)). However, with SCr only 162 (17.5%) patients had abnormal renal function (>1.5 mg/dl) and SCr values misrepresented (108) 11.6% patients with impaired kidney function. In addition, more females, about 15% were failed to detect by SCr method in contrast to males of 9%. This study documented that, a large proportion of patients with impaired renal function are not diagnosed if clinicians rely solely on normal SCr as evidence of normal renal function. Inclusion of eGFR calculated by re-expressed 4 variable MDRD equation may facilitates the early identification and intervention of patients with renal impairment.


Rheumatic manifestations of diseases associated with substance abuse.
Rheumatic manifestations of substance abuse are uncommon but recognized complications. Repeated injections of drugs and adulterants represent repeated antigenic challenge. The population at greatest risk is that of young males, although females with eating disorders are more apt to develop myopathy, clubbing, or periostitis. Alcohol, the most common substance abused, is associated more often with myopathy. In IV drug abusers, hepatitis B viral infection, bacterial endocarditis, primary skeletal infections, and venous complications are most common in that order. However, the spectrum may evolve as the pattern of substance abuse changes. First, the frequency of cocaine dependence is rapidly approaching that of alcohol. Two regular cocaine users are reported as having Raynaud's phenomenon and abnormal serologies. Second, synthesis of lookalike drugs may produce new associations, such as parkinsonism after IV N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine. Third, the increased use of ipecac and purgatives in eating disorders is an important consideration in young women with otherwise unexplained myopathy or arthritis. Finally, patients with AIDS are presenting with autoimmune phenomena or a spectrum of unusual infections that change as the epidemic evolves. Increased recognition of these symptom complexes may lead to earlier, more accurate diagnoses and avoidance of unnecessary diagnostic evaluations and delays in treatment.


Wandering Ozurdex(®) implant.
To report the behavior of intravitreal Ozurdex(®) implant in eyes with post-lensectomy-vitrectomy (PLV) aphakia. Retrospective chart review of three eyes with PLV aphakia (three patients with uveitis) who received intravitreal injection of Ozurdex(®) for cystoid macular edema (one eye), persistent inflammation (one eye), and ocular hypotony (one eye). Final outcome was assessed in terms of effectiveness, stability, and tolerance of the implant. Following the implant, an initial improvement was seen in all the three eyes. However, the implant migrated into the anterior chamber (AC) at 1 week in two eyes and at 5 weeks in one eye, and wandered between the AC and vitreous cavity with changing postures of the patient. Two eyes developed corneal edema, of which one eye underwent implant removal from the AC. Ozurdex(®) implant should be contraindicated in eyes with PLV aphakia to avoid its deleterious effect on the corneal endothelium.


Insulin-dependent diabetes--associated HLA-D region encoded determinants.
We have studied the relative frequency of Dw specificities (defined with homozygous typing cells or primed LD (lymphocyte defined) typing reagents) associated with DR4 and DR2 in the normal and insulin-dependent diabetic population. Our findings demonstrate that there is a highly significantly increased frequency of Dw4 in DR4 positive diabetics as compared with normals and a significantly decreased frequency of Dw2 and Dw12 in the few DR2 positive insulin-dependent diabetics that we have found. In addition, we have used PLT reagents to define a new LD specificity, LD-MN2, that is associated with DR2 and is found significantly more frequently in DR2+ IDD patients than in DR2+ normals. These results suggest that determinants of import in the association between HLA-D and IDD may be more closely related to Dw than to DR.


Proteomic analysis reveals that Polygonatum cyrtonema Hua polysaccharide ameliorates mice muscle atrophy in chemotherapy-induced cachexia.
Polygonatum cyrtonema Hua polysaccharide (PCP) is the main bioactive compound derived from the herb Polygonati Rhizoma, known for its anti-fatigue, antioxidant, immunomodulatory, and anti-inflammatory properties. However, its effectiveness on alleviating chemotherapy-induced muscle atrophy has been unclear. In this study, we utilized proteomic analysis to investigate the effects and mechanisms of PCP on gemcitabine plus cisplatin (GC) induced muscle atrophy in mice. Quality control analysis revealed that the functional PCP, rich in glucose, is a heterogeneous polysaccharide comprised of nine monosaccharides. PCP (64 mg/kg) significantly alleviated body muscle, organ weight loss, and muscle fiber atrophy in chemotherapy-induced cachectic mice. Moreover, PCP suppressed the decrease in serum immunoglobulin levels and the increase in pro-inflammatory factor interleukin-6 (IL-6). Proteomic analysis demonstrated that PCP contributed to the homeostasis of protein metabolism in gastrocnemius muscle. Diacylglycerol kinase (DGKζ) and cathepsin L (CTSL) were identified as primary PCP targets. Furthermore, the IL-6/STAT3/CTSL and DGKζ/FoxO/Atrogin1 signaling pathways were validated. Our findings suggest that PCP exerts an anti-atrophy effect on chemotherapy-induced muscle atrophy by regulating the autophagy-lysosome and ubiquitin-proteasome systems.


KR-31831 improves survival and protects hematopoietic cells and radiosensitive tissues against radiation-induced injuries in mice.
This study explored the radioprotective effects and possible underlying mechanisms of KR-31831 against radiation-induced injury in a mouse model. KR-31831 (30 and 50 mg/kg) was administered to mice 24 h and 30 min before exposure to a single lethal or sublethal dose of whole-body irradiation (WBI) (7 or 4 Gy, respectively). These animals were then evaluated for changes in mortality, various hematological and biochemical parameters, and histological features in response to these treatments. In addition, RNA sequencing was used to profile the radiation-induced transcriptomic response in the bone marrow cells. The results showed that KR-31831 dose-dependently prolonged the 30-day survival period and prevented damage to radiation-sensitive organs, such as the intestine and testis, in response to WBI. Damage to the hematopoietic system was also notably improved in the KR-31831-treated mice, as evidenced by an increase in bone marrow and peripheral blood cells, as well as recovery of the histopathological characteristics of the bone marrow. These protective effects were achieved, at least in part, via the suppression of radiation-induced increases in apoptotic cell death and erythropoietin levels in the plasma. Furthermore, the gene expression profiles of the bone marrow cells of the WBI-treated mice suggested that KR-31831 upregulates the expression of the genes involved in regulating apoptosis and modulating the immune response, both of which are required for protecting the bone marrow. These results suggest the potential therapeutic efficacy of KR-31831 for protection against radiation-induced injury.


Photoreceptor coupling and boundary detection.
Electrical coupling between photoreceptors results in the extensive spreading of output potentials along the syncytium of photoreceptor terminals. This smoothing of output potentials seems to make spatial resolution worse. However, the photoreceptor noise that is considered to be non-correlated both in space and time is smoothed to the greater extent than the correlated potential difference across the boundaries between areas of different brightness. This improves the signal-to-noise ratio more for more extended boundaries and favours lowering the threshold so that they can be detected more easily during the subsequent processing. The results have a striking parallel with a well known dependence of contrast threshold on stimulus size as measured psychophysically.


Induction of labour with unfavourable local conditions for suspected fetal growth restriction after 36 weeks of gestation: Factors associated with the risk of caesarean.
Induction of labour in women with an unfavourable cervix is associated with a risk of caesarean delivery. When a diagnosis of fetal growth restriction (FGR) is also involved, the risk of intrapartum fetal acidosis increases. The main objective was to identify prognostic factors for the risk of caesarean delivery after induction for suspected FGR after 36 weeks of gestation with an unripe cervix. This was a retrospective, single-centre (Port Royal, Paris, France) study of women with a singleton fetus in cephalic presentation, with labour induced at or after 36 weeks for suspected FGR diagnosed during second or third trimester of pregnancy with an unripe cervix (Bishop score under 6) who gave birth between 1 January 2015 and 31 December 2019. A multivariable analysis was performed to identify the factors related to an increased risk of caesarean section. Of the 146 women included, 56 (38.4 %) had caesarean deliveries. After adjustment, the factors significantly associated with the risk of caesarean were maternal age greater than 39 years (ORa = 4.33 [1.22-17.2], reference: 25-39 years), nulliparity (ORa = 3.49 [1.25-11.2]), and an abnormal fetal umbilical artery Doppler velocimetry (ORa = 3.50 [1.47-8.70]). The risk of poor neonatal condition did not differ significantly between women with vaginal and caesarean deliveries (2.3 % vs 7.3 %, P = 0.21). When FGR is suspected at 36 weeks of gestation and later, induction of labour is a reasonable option, even if the cervix is unripe, as the risk of caesarean delivery appears acceptable and neonatal status is good and similar with both modes of delivery.


Applications of Microfluidics in Stem Cell Biology.
Stem cell research can significantly benefit from recent advances of microfluidics technology. In a rationally designed microfluidics device, analyses of stem cells can be done in a much deeper and wider way than in a conventional tissue culture dish. Miniaturization makes analyses operated in a high-throughput fashion, while controls of fluids help to reconstruct the physiological environments. Through integration with present characterization tools like fluorescent microscope, microfluidics offers a systematic way to study the decision-making process of stem cells, which has attractive medical applications. In this paper, recent progress of microfluidics devices on stem cell research are discussed. The purpose of this review is to highlight some key features of microfluidics for stem cell biologists, as well as provide physicists/engineers an overview of how microfluidics has been and could be used for stem cell research.


Glial progenitor heterogeneity and key regulators revealed by single-cell RNA sequencing provide insight to regeneration in spinal cord injury.
Recent studies have revealed the heterogeneous nature of astrocytes; however, how diverse constituents of astrocyte-lineage cells are regulated in adult spinal cord after injury and contribute to regeneration remains elusive. We perform single-cell RNA sequencing of GFAP-expressing cells from sub-chronic spinal cord injury models and identify and compare with the subpopulations in acute-stage data. We find subpopulations with distinct functional enrichment and their identities defined by subpopulation-specific transcription factors and regulons. Immunohistochemistry, RNAscope experiments, and quantification by stereology verify the molecular signature, location, and morphology of potential resident neural progenitors or neural stem cells in the adult spinal cord before and after injury and uncover the populations of the intermediate cells enriched in neuronal genes that could potentially transition into other subpopulations. This study has expanded the knowledge of the heterogeneity and cell state transition of glial progenitors in adult spinal cord before and after injury.


Expression of thymosin beta-4 in human periodontal ligament cells and mouse periodontal tissue and its role in osteoblastic/cementoblastic differentiation.
A recent report showed that thymosin beta-4 (Tβ4) is expressed during the development of tooth germ, but its effect on osteoblastic/cementoblastic differentiation is a controversial topic. Furthermore, the precise expression and function of Tβ4 in periodontal tissue remains unclear. Therefore, the purpose of this study was to investigate the immunolocalization of Tβ4 in the developing periodontium of mouse, the function of Tβ4 in osteoblastic/cementoblastic differentiation, and the underlying mechanism regulating periodontal regeneration in human periodontal ligament cells (hPDLCs), cementoblasts, and osteoblasts. Tβ4 expression was observed in differentiating hPDLCs, osteoblasts of the periodontium during development, as well as in mature tissue. Higher Tβ4 expression was observed in hPDLCs than in cementoblasts and osteoblasts in the developing periodontium. The expression of Tβ4 mRNA and protein gradually increased during PDL cell differentiation. The downregulation of Tβ4 expression by Tβ4 siRNA transfection inhibited osteoblastic differentiation by decreasing calcium nodule formation, alkaline phosphatase (ALP) activity, and mRNA expression of differentiation markers in hPDLCs, cementoblasts, and osteoblasts. In contrast, Tβ4 activation using a Tβ4 peptide, promoted these processes by activation of Akt, p38, ERK MAPKs, and the NF-κB pathway. The expression of nuclear NFATc1 was upregulated by Tβ4 peptide in hPDLCs. Inhibition of the calcineurin/NFATc1 pathway by cyclosporin A and FK506, attenuated Tβ4-induced osteoblastic differentiation and activation of Wnt-related genes, as well as nuclear β-catenin in hPDLCs. In conclusion, this study demonstrates, for the first time, that Tβ4 is expressed in developing periodontal tissue and that its expression is associated with osteoblastic/cementoblastic differentiation. These results suggests that Tβ4 is a potential therapeutic target for periodontal regeneration or bone disease.


Flow cytometric estimation of 'labile iron pool' in human white blood cells reveals a positive association with ageing.
A small part of cellular iron, usually called 'labile iron pool' (LIP), is not securely stored and has the potential to catalyse the formation of highly reactive oxygen species. The present work estimated LIP levels in human white cells by using the analytical power of flow cytometry. The method relies essentially on already established principles but has the added value of monitoring LIP in different subpopulations of human blood cells concurrently in a single sample. Examination of 41 apparently healthy individuals revealed a positive correlation between LIP levels and the age of the donors (r=0.656, 0.572 and 0.702 for granulocytes, lymphocytes and monocytes, respectively, p<0.0001), indicating that cells of older individuals are prone to oxidations in conditions of oxidative stress. It is suggested that LIP estimation may represent a valuable tool in examinations searching for links between iron and a variety of oxidative stress-related pathological conditions.


Building evidence to reduce inequities in youth physical activity and obesity: Introduction to the Physical Activity Research Center (PARC) Special Section.
This guest editorial introduces the rationale and goals of the Physical Activity Research Center. It provides an overview of the five papers in this Special Section plus six commissioned studies intended to inform advocacy efforts.


Triptolide Promotes the Clearance of α-Synuclein by Enhancing Autophagy in Neuronal Cells.
Parkinson's disease (PD) is an aging-associated neurodegenerative disease with a characteristic feature of α-synuclein accumulation. Point mutations (A53T, A30P) that increase the aggregation propensity of α-synuclein result in familial early onset PD. The abnormal metabolism of α-synuclein results in aberrant level changes of α-synuclein in PD. In pathological conditions, α-synuclein is degraded mainly by the autophagy-lysosome pathway. Triptolide (T10) is a monomeric compound isolated from a traditional Chinese herb. Our group demonstrated for the first time that T10 possesses potent neuroprotective properties both in vitro and in vivo PD models. In the present study, we reported T10 as a potent autophagy inducer in neuronal cells, which helped to promote the clearance of various forms of α-synuclein in neuronal cells. We transfected neuronal cells with A53T mutant (A53T) or wild-type (WT) α-synuclein plasmids and found T10 attenuated the cytotoxicity induced by pathogenic A53T α-synuclein overexpression. We observed that T10 significantly reduced both A53T and WT α-synuclein level in neuronal cell line, as well as in primary cultured cortical neurons. Excluding the changes of syntheses, secretion, and aggregation of α-synuclein, we further added autophagy inhibitor or proteasome inhibitor with T10, and we noticed that T10 promoted the clearance of α-synuclein mainly by the autophagic pathway. Lastly, we observed increased autophagy marker LC3-II expression and autophagosomes by GFP-LC3-II accumulation and ultrastructural characterization. However, the lysosome activity and cell viability were not modulated by T10. Our study revealed that T10 could induce autophagy and promote the clearance of both WT and A53T α-synuclein in neurons. These results provide evidence of T10 as a promising mean to treat PD and other neurodegenerative diseases by reducing pathogenic proteins in neurons.


Acrylamide gel analysis of sucrose gradient-derived RNA B fractions transferring delayed sensitivity in vitro.
Sucrose-derived RNA fractions transferring specific delayed sensitivity in vitro were extracted from mono-(p-azobenzenearsonate)-N-chloracetyl-L-tyrosine (ARSNAT)- or keyhole limpet hemocyanin (KLH)-sensitized guinea pigs. Fractions having biological activity were assessed by acrylamide gel analysis to enumerate the number of RNA species in active fractions, and to compare and examine the banding patterns of each RNA fraction. Each isolated B fraction of RNA exhibited multiple bands of RNA with molecular weights ranging from 4.0 X 10(5) to 8.5 X 10(5). Depending on the source and antigen sensitivity of the RNA donor, several differences were observed among the analyzed fractions. These were bands of varying intensity, presence of additional RNA bands, absence of bands in certain positions, and RNA bands migrating in different positions on the gels. Acrylamide gel analysis separation, and resolution of B fractions with specific immunobiological activity now offers an approach for further isolation and resolution of the active species.


Progesterone for preterm birth prevention.
Preterm birth—delivery before 37 weeks of gestation—is the second leading cause of infant mortality in the United States after congenital malformations. Spontaneous preterm birth, due to either preterm labor or preterm premature membrane rupture, encompasses approximately 75% of all preterm births, almost 400,000 births per year. Since the 1960s, different formulations of progesterone have been investigated for preterm birth prevention. This article addresses the use of progesterone for the prevention of preterm birth, including selection of candidates for progesterone, pharmacokinetics, dosing, and formulations. This article aims to provide a practical guide for using progesterone in clinical practice.


Loss of heterozygosity of BRCA1, TP53 and TCRD markers analysed in sporadic endometrial cancer.
Genetic alterations of tumour suppressor genes, for which loss of heterozygosity (LOH) is one mechanism of gene inactivation, are important steps in the development of endometrial cancer. To investigate the clinical relevance of LOH of BRCA1 (17q21), TP53 (17p13) and TCRD (14q11) in endometrial cancer, polymerase chain reaction (PCR)-based fluorescent DNA technology for the detection of microsatellite polymorphisms was applied. One hundred and thirteen archival endometrial cancer samples with matched normal tissues were examined. Allele loss at three loci were correlated with age, tumour size, lymph node status, metastases, stage, histological types, grade, expression of oestrogen receptor (ER) and progesterone receptor (PgR), family history of cancer, previous history of cancer or precursor lesions, and previous history of hormone replacement therapy (HRT). LOH for BRCA1 was detected in 18.1%, of TP53 in 26.9%, and of TCRD in 26.3% of informative cases. LOH of BRCA1 correlated with medium grade, positive ER status, and family history of cancer; LOH of TP53 correlated with younger age, high grade, positive PgR status, and with tumours from patients without HRT; LOH of TCRD correlated only with family history of cancer. LOH at all three loci correlated only with grade and positive family history. Allele loss of one of the three tumour suppressor loci did not correlate with disease-free survival (DFS), but LOH of BRCA1 correlated significantly with decreased overall survival (OS). The latter, together with the correlation of LOH of BRCA1 locus with steroid hormone receptor expression, might give a hint to the potential involvement of the co-localised 17 beta-hydroxysteroid dehydrogenase (HSD) gene in the development of endometrial cancer.


Control of CSF-1 induced inflammation in teleost fish by a soluble form of the CSF-1 receptor.
The colony-stimulating factor-1 (CSF-1) is the principal regulator of the survival, proliferation, differentiation, and function of macrophages and their precursors, and has been shown to play a role in the etiology of inflammation. We recently identified a novel mechanism for the control of CSF-1 activity in teleost fish, through the production of an inhibitory soluble form of the CSF-1 receptor (sCSF-1R). Primary goldfish kidney macrophages selectively expressed sCSF-1R during the senescence phase, which corresponds to a defined stage of in vitro culture development where inhibition of macrophage proliferation and apoptotic cell death are prominent. In contrast, primary macrophage cultures undergoing active proliferation displayed low levels of sCSF-1R expression. Addition of purified recombinant sCSF-1R to developing primary macrophage cultures leads to a dose-dependent decrease in macrophage proliferation and inhibits macrophage antimicrobial functions including chemotaxis, phagocytosis, and production of reactive oxygen intermediates. Using a goldfish in vivo model of self-resolving peritonitis, we found that sCSF-1R plays a role in the inhibition of inflammation, following an initial acute phase of antimicrobial responses within an inflammatory site. Soluble CSF-1R inhibits pro-inflammatory cytokine production, inhibits leukocyte recruitment to the inflammatory site and decreases ROS production in a dose-dependent manner. This sCSF-1R-dependent regulation of inflammation appears to be an elegant mechanism for the control of macrophage numbers at inflammatory sites of lower vertebrates. Overall, our results provide new insights into the evolutionary origins of the CSF-1 immune regulatory axis.


Treatment options for choroidal malignant melanoma: a case report featuring transpupillary thermotherapy.
Choroidal melanoma is the most common primary ocular tumor in adults. These tumors are almost always unilateral and develop spontaneously or arise from pre-existing nevi. Historically, treatment for choroidal melanoma was enucleation. New therapies have been developed to treat choroidal melanoma and to preserve the eye. Currently, these treatment options include brachytherapy and transpupillary thermotherapy. A 51-year-old man came to the eye clinic with symptoms of mildly decreased fluctuating vision and floaters with no flashes in the temporal visual field of the right eye. Best-corrected visual acuities were 20/30 O.D. and 20/20 O.S. A small-to-medium choroidal lesion was found in the right eye. Ultrasonography and A-scan showed this lesion to be a choroidal malignant melanoma. The patient's surgical options included enucleation, brachytherapy, and transpupillary thermotherapy. The patient opted to have transpupillary thermotherapy to preserve the eye, and subsequently underwent two procedures that eventually obliterated the melanoma, resulting in best-corrected visual acuity of 20/40. Transpupillary thermotherapy is an excellent option for treatment of small-to-medium choroidal melanomas. Brachytherapy is also an option for treatment, but with increased ocular side effects and complications. Enucleation is still commonly performed on medium and/or large choroidal melanomas.


Influence of temperature and surfactant on Escherichia coli inactivation in aqueous suspensions treated by moderate pulsed electric fields.
This research employed a conductometric technique to estimate the inactivation kinetics of Escherichia coli cells in aqueous suspensions (1 wt.%) during simultaneous pulsed electric fields (PEF) and thermal treatments. The electric field strength was E=5 kV/cm, the effective PEF treatment time t(PEF) was within 0-0.2 s, the pulse duration t(i) was 10(-3) s, the medium temperature was 30-50 degrees C, and the time of thermal treatment t(T) was within 0-7000 s. The damage of E. coli was accompanied by cell size decrease and release of intracellular components. The synergy between PEF and thermal treatments on E. coli inactivation was clearly present. The non-ionic surfactant Triton X-100 additionally improved its inactivation. The characteristic damage time followed the Arrhenius law within the temperature range 30-50 degrees C with activation energies W=94+/-2 kJ mol(-1) and W=103+/-5 kJ mol(-1) with and without the presence of surfactant, respectively. Relations between cell aggregation, cell zeta-potentials and presence of surfactant were analysed.


Goat embryonic stem-like cell derivation and characterization.
Embryonic stem (ES) cells are derived from the inner cell masses of preimplantation embryos. ES cells are pluripotent cells with the capacity for long-term propagation and broad differentiation plasticity. These cells have an exceptional functional feature in that they can differentiate into all tissues and organs, including germ cells. Established ES cell lines have been generated in mouse, human, and nonhuman primate but derivation of ES cells in farm animals has been problematic. Several ES-like cell lines from farm animals have been reported to exhibit properties of pluripotency in vitro. However, only a few of them morphologically resemble ES cells, or express markers that are associated with established ES cell lines from mouse and humans. Methods for derivation, propagation, and differentiation of ES cells from domestic animals have not been fully established. In this chapter, we describe methods for isolation of goat ES (gES) cell lines from in vivo-derived blastocysts and characterization of markers indicative of pluripotency. In addition, we outline differentiation of gES cells into all three germ layers in vivo by forming teratomas as a hallmark of pluripotency.


Urothelial carcinoma within the prostatic utricle of an adult with hypospadias and Fanconi anemia.
Prostatic utricles are rare in the general population and are often otherwise unremarkable anatomic variants. These structures are contiguous with the prostatic urethra and are nevertheless susceptible to urothelial carcinoma. This case report discusses the first reported patient with Fanconi anemia with urothelial carcinoma within an enlarged prostatic utricle.


Role of novel DSP_p.Q986X genetic variation in arrhythmogenic right ventricular cardiomyopathy.
Arrhythmogenic right ventricular cardiomyopathy is an inherited disease characterized by a progressive myocardium fibrofatty replacement. This abnormality disrupts electrical transmission causing ventricular arrhythmias and sudden cardiac death. This genetic disease is transmitted mainly with an autosomal dominant pattern. Our aim was to identify the genetic defect responsible for the pathology in a Spanish family, and to perform its phenotype connotations. A total of 15 individuals in a three-generation Spanish family were screened after the sudden cardiac death of one family member. All they underwent a complete physical examination, 12-lead electrocardiogram, 2-dimensional echocardiography, magnetic resonance imaging, exercise stress test, 24-h Holter and genetic testing. Autopsy revealed the presence of biventricular arrhythmogenic dysplasia in deceased member. Six family members showed clinical symptoms but only three of them fulfilled definite diagnostic criteria of the disease. Genetic analysis showed a novel nonsense genetic variation in nine family members. All family members with clinical symptoms carried the genetic variation. Genetic testing in families affected by arrhythmogenic right ventricular cardiomyopathy helps to identify the genetic cause responsible for the disease. The incomplete penetrance and variable phenotypic expression highlights the need of comprehensive genetic analysis and further phenotype implications of genetics to clarify the pathophysiology of the disease.


Immunocapture isotope dilution mass spectrometry in response to a pandemic influenza threat.
As a result of recent advances in mass spectrometry-based protein quantitation methods, these techniques are now poised to play a critical role in rapid formulation of pandemic influenza vaccines. Analytical techniques that have been developed and validated on seasonal influenza strains can be used to increase the quality and decrease the time required to deliver protective pandemic vaccines to the global population. The emergence of a potentially pandemic avian influenza A (H7N9) virus in March of 2013, prompted the US public health authorities and the vaccine industry to initiate production of a pre-pandemic vaccine for preparedness purposes. To this end, we evaluated the feasibility of using immunocapture isotope dilution mass spectrometry (IC-IDMS) to evaluate the suitability of the underlying monoclonal and polyclonal antibodies (mAbs and pAbs) for their capacity to isolate the H7 hemagglutinin (HA) in this new vaccine for quantification by IDMS. A broad range of H7 capture efficiencies was observed among mAbs tested by IC-IDMS with FR-545, 46/6, and G3 A533 exhibiting the highest cross-reactivity capabilities to H7 of A/Shanghai/2/2013. MAb FR-545 was selected for continued assessment, evaluated by IC-IDMS for mAb reactivity against H7 in the H7N9 candidate vaccine virus and compared with/to reactivity to the reference polyclonal antiserum in allantoic fluid, purified whole virus, lyophilized whole virus and final detergent-split monovalent vaccine preparations for vaccine development. IC-IDMS assessment of FR-545 alongside IC-IDMS using the reference polyclonal antiserum to A/Shanghai/2/2013 and with the regulatory SRID method showed strong correlation and mAb IC-IDMS could have played an important role in the event a potential surrogate potency test was required to be rapidly implemented.


Posterior hamstring harvest improves aesthetic satisfaction and decreases sensory complications as compared to the classic anterior approach in anterior cruciate ligament reconstruction surgery.
The use of the posterior approach for harvesting hamstring grafts has recently become popular thanks to new all-inside techniques and retrograde drills. This study aims to compare the classic anterior approach with the posterior approach in the popliteal fossa. Retrospective comparative study of 100 consecutive cases of primary ligamentoplasty performed using ipsilateral semitendinosus autograft with at least one year of follow-up. 50 patients with anterior approach (group A) and 50 patients with posterior approach (P). Ratio men/women: 9/1. Mean age: 32 ± 13 years. Mean operative time: 64.88 ± 12.28 min. Graft harvest time; intraoperative complications (semitendinous [ST] tendon cut); postoperative neurological complications (allodynia, paresthesia, pain) or hematoma in the donor area; atrophy of the operated thigh compared to the contralateral thigh, postoperative VAS score, aesthetic satisfaction and overall satisfaction. Graft harvest time of 9.5 min in group A versus 5.25 min in group P (p < 0.05). Sensory complications: 16% in group A versus 2% in group P (p < 0.05). Regarding the patient's evaluation of the aesthetic result of the surgery, 80% in group A and 92% in group P were very satisfied, 16% in group A and 8% in group P were satisfied and 4% in group A and no patients in group P not very satisfied (p < 0.05). No significant differences were found in terms of total operative time, postoperative joint movement, atrophy of the operated thigh, postoperative VAS, or overall patient satisfaction. The posterior approach to harvesting the ipsilateral hamstring graft obtained better results than the anterior approach in terms of aesthetic satisfaction of the patient, lower rate of neurological complications (allodynia, paresthesias and hypoesthesia in the anterior region of the knee and leg) and shorter hamstring harvest time. IV.


The role of protein phosphatases in synaptic transmission, plasticity and neuronal development.
In the past year significant advances have been made in our understanding of the role of protein dephosphorylation in the control of neuronal function. Molecular cloning has identified a large number of serine/threonine and tyrosine protein phosphatases in the nervous system. Many of these enzymes are selectively enriched in the nervous system, some are localized to specific neurons, and yet others are expressed only during specific periods of neuronal development. The availability of purified protein phosphatases and selective inhibitors has facilitated the analysis of these enzymes and their role in the regulation of neurotransmitter receptors and ion channels.


Mutations in the catalytic domain of human coagulation factor IX: rapid characterization by direct genomic sequencing of DNA fragments displaying an altered melting behavior.
Deficiency in coagulation factor IX, a plasma glycoprotein constituent of the clotting cascade, results in hemophilia B, an inherited recessive X-linked bleeding disorder. Some affected individuals, referred to as antigen positive or CRM+, express an inactive factor IX gene product at normal levels and are expected to have natural mutations altering domains of the molecule that are critical for its correct function. The serine protease catalytic domain of activated factor IX, encoded by exons VII and VIII of the gene, is a possible target for such mutations. We designed a strategy allowing rapid analysis of this region through enzymatic amplification of genomic DNA, analysis of the amplification products by denaturing gradient gel electrophoresis, and direct sequencing of the fragments displaying an altered melting behavior. This procedure permitted us to characterize two previously undescribed mutations. Factor IX Angers is a G-to-A substitution generating an Arg in place of a Gly at amino acid 396 of the mature factor IX protein. Factor IX Bordeaux is an A-to-T substitution introducing a nonsense codon in place of the normal codon for Lys at position 411. Moreover, the already described factor IX Vancouver defect was found in three apparently independent families. These results provide further insight into the molecular heterogeneity of hemophilia B. In addition, we demonstrate the usefulness of this rapid screening procedure, which has broad applications in human genetics and can be used as an alternative to RFLP analysis in carrier detection or prenatal diagnosis studies.


Intersexual differences in phenology and damage by herbivores and pathogens in dioecious Rubus chamaemorus L.
Shoot development and damage by herbivores and pathogens to male, female and non-floral ramets of the dioecious, perennial herb Rubus chamaemorus were studied in the field during three consecutive years. Leaves on male ramets were usually consumed more by herbivores and attacked more by fungi than were leaves on female ramets. Male ramets unfolded their leaves later than did female ramets. In 1983, when the level of herbivory was comparatively high, ramets that were fully developed in early June were more damaged by herbivores than were ramets that unfolded their leaves later. Non-floral ramets usually showed intermediate levels of damage compared to male and female flowering ramets.Defoliation caused a greater increase in ramet mortality among females than among males in a field experiment. It is suggested that the different reproductive roles of males and females differently constrain the evolution of defense against herbivores and pathogens in dioecious plants. This may result in intersexual differences both in palatability to herbivores and susceptibility to pathogens.


Immunohistochemical localization of laminin, neural cell adhesion molecule, collagen type IV and T-61 antigen in the embryonic retina of the Japanese quail by in vivo injection of antibodies.
Antibodies against laminin (LN), fibronectin (FN), collagen type IV (Col IV), neural cell adhesion molecule (N-CAM), T-61 antigen, actin, tubulin and neurofilament protein were injected into the eyes of quail embryos (Coturnix coturnix japonica) of different ages. Twenty h after injection, the heads of the embryos were fixed and the antibodies visualized in sections with the use of fluorescein-isothiocyanate (FITC) or peroxidase-labeled second antibodies by light- and electron microscopy. Antibodies against cell surface molecules, such as N-CAM, LN, Col IV and T 61, labeled matrix and membrane components of the retinal cells in different antigen-specific patterns. Antibodies against intracellular antigens, such as actin, tubulin and neurofilament protein labeled nonspecifically the vitreous body and the inner basal lamina of the retina, but resulted in only a very weak and diffuse labeling of retinal cells. N-CAM was detected in high concentration in the optic fiber layer on the surface of axons and on the membranes of all retinal cells. Col IV, LN and T 61 antigen were found predominantly in the optic fiber layer. LN and Col IV were located on the surface of axons and the endfeet of ventricular (neuroepithelial) cells in a patchy distribution. The T-61 antigen was found in early stages in the cell-free space of the optic fiber layer, on the surface of ventricular cells and axons, and at later stages also in high-density patches between nerve fibers. The distribution of LN and T-61 antigen together with data from in vitro experiments suggests a crucial role of these proteins in axon extension in the avian retina during early development of the optic fiber layer.


β Cell function after Roux-en-Y gastric bypass surgery or reduced energy intake alone in people with obesity.
BackgroundThe effects of diet-induced weight loss (WL) and WL after Roux-en-Y gastric bypass (RYGB) surgery on β cell function (BCF) are unclear because of conflicting results from different studies, presumably because of differences in the methods used to measure BCF, the amount of WL between treatment groups, and baseline BCF. We evaluated the effect of WL after RYGB surgery or reduced energy intake alone on BCF in people with obesity with and without type 2 diabetes.MethodsBCF (insulin secretion in relationship to plasma glucose) was assessed before and after glucose or mixed-meal ingestion before and after (a) progressive amounts (6%, 11%, 16%) of WL induced by a low-calorie diet (LCD) in people with obesity without diabetes, (b) ~20% WL after RYGB surgery or laparoscopic adjustable gastric banding (LAGB) in people with obesity without diabetes, and (c) ~20% WL after RYGB surgery or LCD alone in people with obesity and diabetes.ResultsDiet-induced progressive WL in people without diabetes progressively decreased BCF. Marked WL after LAGB or RYGB in people without diabetes did not alter BCF. Marked WL after LCD or RYGB in people with diabetes markedly increased BCF, without a difference between groups.ConclusionMarked WL increases BCF in people with obesity and diabetes but not in people with obesity without diabetes. The effect of RYGB-induced WL on BCF is not different from the effect of matched WL after LAGB or LCD alone.trial registrationNCT00981500, NCT02207777, NCT01299519.FundingNIH grants R01 DK037948, P30 DK056341, P30 DK020579, UL1 TR002345.


Inequalities in access to pediatric ESRD care: a global health challenge.
The increasing attention paid to chronic kidney disease (CKD) as a major cause of mortality and disability, as well as the advances in management of CKD in children, have created a growing demand for pediatric renal replacement therapy (RRT) worldwide. A study by Koch Nogueira and colleagues of children on the transplant waiting list showed large disparities in access to pediatric kidney transplantation between regions in Brazil. This finding raises a wider question about inequalities in access to CKD care in children. Here we review the available data on the global burden of end-stage renal disease in children, the need for pediatric RRT, and its actual provision worldwide. We focus on inequalities in access to renal care for children that currently exist between and within countries. Reduction in worldwide inequalities in access to RRT in children remains a challenge, which requires greater awareness and effective interventions and policies.


Extracellular secretion of anticoagulant peptide hirudin in Lactococcus lactis using SP310mut2 signal peptide.
Hirudin can be used as an oral anticoagulant and antithrombotic agent. The hirudin variant III gene, derived from the medicinal leech, Hirudo medicinalis, was fused to SP310mut2 signal sequence and expressed by a nisin-controlled gene expression system in Lactococcus lactis which was then grown in a 7 l fermenter. After induction with 8 ng nisin ml(-1), the product was secreted into the culture medium and accumulated up to ~2.7 mg l(-1). MALDI-TOF/MS and anticoagulant activity analyses on the purified product confirmed its authenticity. This is the first demonstration that hirudin can be extracellularly secreted and correctly processed in L. lactis.


[Clinical stage I seminoma. Is surveillance a new therapy standard?].
The therapy of stage I seminoma is under constant change. While surveillance, adjuvant radiotherapy and adjuvant chemotherapy were seen as equal therapeutic alternatives up to a few years ago, recently published studies make it necessary to adopt a more differentiated approach. In this review, recent data on the long-term effects of adjuvant radiotherapy and chemotherapy, the question of risk stratification as well as the advantages and limitations of a surveillance strategy are discussed.


Assessment of DNA fragmentation of spermatozoa that were surgically retrieved from men with obstructive azoospermia.
To determine the degree of DNA fragmentation in spermatozoa of men with obstructive azoospermia or anejaculation compared with that of ejaculated spermatozoa from fertile donors. Observational study. SETTING; University Medical Center St. Radboud, Nijmegen. The Netherlands. Forty-one patients with obstructive azoospermia or anejaculation and 10 fertile donors. Sperm samples were obtained surgically from the epididymis or testis of men with azoospermia or anejeculation and by ejaculation in fertile patients. DNA fragmentation was analyzed in the total sample and in a motile fraction that was isolated as in routine ICSI procedures. DNA breaks were measured by using the TdT-mediated dUTP nick-end labeling assay. A higher percentage of cells with DNA breaks was found in men with obstructive azoospermia or anejaculation compared with donors (mean, 18.9% vs. 6.2%). A significant lower degree of DNA fragmentation was observed in the motile fraction from patients compared with donors (0.4% vs. 0.6%). High percentages of cells with DNA damage were found in sperm samples from men with obstructive azoospermia or anejaculation, but a very low frequency of damage to the DNA was observed in the motile fraction. In an ICSI setting, the use of motile sperm retrieved from epididymis or testis of men with obstructive azoospermia does not seem to pose a higher genetic risk to the progeny than does use of motile ejaculated sperm.


Multiview robotic microscope reveals the in-plane kinematics of amphibian neurulation.
A new robotic microscope system, called the Frogatron 3000, was developed to collect time-lapse images from arbitrary viewing angles over the surface of live embryos. Embryos are mounted at the center of a horizontal, fluid-filled, cylindrical glass chamber around which a camera with special optics traverses. To hold them at the center of the chamber and revolve them about a vertical axis, the embryos are placed on the end of a small vertical glass tube that is rotated under computer control. To demonstrate operation of the system, it was used to capture time-lapse images of developing axolotl (amphibian) embryos from 63 viewing angles during the process of neurulation and the in-plane kinematics of the epithelia visible at the center of each view was calculated. The motions of points on the surface of the embryo were determined by digital tracking of their natural surface texture, and a least-squares algorithm was developed to calculate the deformation-rate tensor from the motions of these surface points. Principal strain rates and directions were extracted from this tensor using decomposition and eigenvector techniques. The highest observed principal true strain rate was 28 +/- 5% per hour, along the midline of the neural plate during developmental stage 14, while the greatest contractile true strain rate was--35 +/- 5% per hour, normal to the embryo midline during stage 15.


Forecasting length of stay in child residential treatment.
A sample of 126 consecutively admitted residential treatment children (mean age = 9.86, SD=1.84; 70.6% male; 42.1% African American; 50% Caucasian) were studied over a five-year period to identify predictors of length-of-stay. Cox regression was the primary statistical method used to analyze psychiatric and behavioral rating data for children assessed by teachers and treatment staff using the Devereux Scales of Mental Disorders (DSMD). Parental alcohol abuse, and children's age, medication status, race, initial DSMD total and critical pathology scores, were predictive of length-of-stay. Residential length-of-stay was strongly linked to initial levels of psychiatric symptomatology. Models that can help forecast length of stay are vital tools in helping to improve both clinical and utilization management strategies.


Multi-criteria evaluation in paradigmatic perspectives of agricultural environmental management.
Agriculture is one of the primary activities that affects the environment due to natural resources consumption. Therefore, systematic environmental management for the agricultural sector is required. This study was conducted to analyze the paradigmatic perspective and strategies of agricultural environmental management in Iran. Considering basic criteria of environmental management, three paradigms of frontier economics, eco-development and deep ecology were compared using Analytical Hierarchy Process (AHP). The AHP, is a multi-criteria decision making techniques which is useful when there are different alternatives or indicators in decision making. Comparisons were based on the viewpoints of 117 policy makers, superior managers and main elites and agriculture sector researchers. Environmental managerial strategies also have been studied. Findings revealed paradoxes among the paradigmatic perspectives and selected strategies of different agricultural stakeholders which reduce their effective interactions. Frontier economics is the dominant viewpoint of key agricultural policy makers and other governmental executives. They prefer independent reactive strategies to cope with environmental challenges. Agricultural researchers and private sector authorities believe in eco-development. They have selected cooperative proactive strategies in this regard. Finally, deep ecology has the highest priority according to environmental specialists, who endorse strategic maneuvering and believe in modifying, rethinking and redesigning previous strategies. A paradigm shift, as well as consistency between paradigmatic perspectives and executive strategies, is suggested.


Wear your health on your sleeve.
Wearable electronics could enable us to optimize every aspect of our personal health and performance. They might also have important ramifications for clinical use and theranostics.


Prevalence of Autism and its Comorbidities and the Relationship with Maternal Psychopathology: A National Population-Based Study.
There is no clear picture regarding the prevalence rates of autism and its comorbidities among Iranian children and adolescents. The present study aimed to estimate these rates as part of a large national population-based study on epidemiology of psychiatric disorders in Iranian children and adolescents. The total sample consisted of 31000 children and adolescents between 6 to 18 years of age. The Kiddie Schedule for Affective Disorders and Schizophrenia for School-Age Present and Life time version (K-SADS-PL) was used as the diagnostic tool. The prevalence of autism in the total sample is equal to 0.1% (10 per 10000), with a 2:1 male-to-female ratio. In total, 86% of people with autism had at least one comorbid condition. Intellectual disability, epilepsy, enuresis and attention deficit and hyperactivity disorder (ADHD) with prevalence rates of 70.3%, 29.7%, 27% and 21.62%, respectively, were the most prevalent comorbid conditions in people with autism. Maternal personality disorders were also shown to be associated with increasing risk of autism. The present study shows high prevalence rates for autism and its comorbid conditions among Iranian children and adolescents. It also reveals that there is a relationship between some maternal psychiatric disorders and the risk of autism.


Process evaluation of the Saskatoon breastfeeding center.
A process evaluation of the Breastfeeding Center in Saskatoon was conducted to assess clients' characteristics and satisfaction. Fifty women completed a questionnaire immediately after visiting the center and were interviewed by telephone 2 to 4 weeks later; another 25 women telephoned the center for information and were interviewed by telephone 2 to 4 weeks after initial contact. Four visits were observed and timed. Respondents were generally very satisfied with all aspects of the center. All said they would use the center again and recommend it to others. Most of the respondents were still breastfeeding when interviewed and felt that the center had helped them do so. Observation of the visits revealed a consistent sequence of activities, with visits taking longer on average than the time allotted. The evaluation confirmed that the center was meeting its process objectives. This evaluation could be used as a model for other community-based breastfeeding centers.


Vaginal dose de-escalation in image guided adaptive brachytherapy for locally advanced cervical cancer.
Vaginal stenosis is a major problem following radiotherapy in cervical cancer. We investigated a new dose planning strategy for vaginal dose de-escalation (VDD). Fifty consecutive locally advanced cervical cancer patients without lower or middle vaginal involvement at diagnosis from 3 institutions were analysed. External beam radiotherapy was combined with MRI-guided brachytherapy. VDD was obtained by decreasing dwell times in ovoid/ring and increasing dwell times in tandem/needles. The aim was to maintain the target dose (D90 of HR-CTV⩾85Gy EQD2) while reducing the dose to the surface of the vagina to <140% of the physical fractional brachytherapy dose corresponding to a total EQD2 of 85Gy. The mean vaginal loading (ovoid/ring) was reduced from 51% to 33% of the total loading with VDD, which significantly reduced the dose to the vaginal dose points (p<0.001) without compromising the target dose. The dose to the ICRU recto-vaginal point was reduced by a mean of 4±4Gy EQD2 (p<0.001), while doses to bladder and rectum (D2cm3) were reduced by 2±2Gy and 3±2Gy, respectively (p<0.001). VDD significantly reduces dose to the upper vagina which is expected to result in reduction of vaginal stenosis.


A positive association of peripheral arterial occlusive disease (PAD) and Chlamydophila (Clamydia) pneumoniae.
The relationship between Chlamydophila pneumoniae (Cp) infection and peripheral arterial occlusive disease (PAD) was studied by analyzing clinical samples from 95 patients with PAD and 100 controls. The following investigations were conducted: IgG and IgA against lipopolysaccharide (LPS) and against purified Cp-specific antigens from elementary bodies (EB) with ELISA; anti-EB IgG, with MIF; Cp DNA in arterial biopsy and peripheral blood mononuclear cells with heminested PCR; LPS with ELISA; and bacteria culture in HEp-2 cells from arterial biopsy. A significantly higher ratio of anti-EB IgG was detected in patients. There were no significant differences in anti-LPS IgG, anti-LPS IgA and anti-EB IgA between cases and controls. Cp DNA findings in the vascular wall biopsy showed significant differences between cases and controls. We obtained results that significantly involve Cp infection with PAD through the detection of anti-EB IgG from serum and bacterial DNA from arterial biopsy.


Concentration- and time-dependent toxicity of commonly encountered pesticides and pesticide mixtures to honeybees (Apis mellifera L.).
Honeybees are exposed to a wide range of pesticides for long periods via contaminated water, pollen and nectar. Some of those pesticides might constitute health hazards in a time- and dose-dependent manner. Time-dependent toxicity profiles for many applied pesticides are lacking, despite the fact that such profiles are crucial for toxicological evaluations. Therefore, we sought to determine the time-dependent toxicities of pesticides/pesticide metabolites frequently found in Israeli beehives, namely, amitraz metabolites, N'-(2,4-dimethylphenyl)-N-methylformamidine (DMPF) and N-(2,4-dimethylphenyl)-formamide (DMF), coumaphos, imidacloprid, thiacloprid, acetamiprid and dimethoate (toxic reference). By applying accepted methodological approaches such as the modified Haber's rule (product of concentration and exposure duration leads to a constant effect) and comparisons between cumulative doses at different time points, we determined the time-dependent toxicities of these pesticides. We also studied the mixture toxicities of frequently occurring pesticide combinations and estimated their potential contributions to the overall toxicities of neonicotinoids. Thiacloprid was the only pesticide that complied with Haber's rule. DMPF, dimethoate and imidacloprid exhibited time-diminished -toxicities. In contrast, DMF and acetamiprid exhibited time-reinforced toxicities. Neither the binary mixtures nor the tertiary mixtures of DMF, DMPF and coumaphos at 10 times their environmentally relevant concentrations potentiated the neonicotinoids' toxicities. DMPF and imidacloprid were found to present the greatest hazard to honeybees, based on their 50% lethal cumulative dose and 50% lethal time. Amitraz's instability, its low detection frequency and high toxicity profile of its metabolite, DMPF, lead us to the conclusion that DMPF constitutes the actual toxic entity responsible for amitraz's toxic effect.


A shotgun proteomic approach reveals novel potential salivary protein biomarkers for asthma.
The aim of this study was to determine if there is an association between the salivary protein profile and disease control in asthma. Thirty asthmatic patients (17 adults and 13 children) participated in this study. Saliva samples were collected from healthy subjects, controlled and uncontrolled asthmatics. Individual samples from each group were combined to form a pooled sample, from which proteomic analysis was performed using gel-based quantitative proteomics. Fourteen out of thirty asthmatics were classified to be controlled asthma. Most of asthmatics received inhaled corticosteroids as the controller medications. SDS-PAGE showed predominant bands at high molecular weight in asthmatic saliva compared to that of the controls. Shotgun proteomic analyses indicated that 193 salivary proteins were expressed in both controlled and uncontrolled asthmatics. They were predicted to associate with proteins involved in pathogenesis of asthma including IL-5, IL-6, MCP-1, VEGF, and periostin and asthma medicines (Cromolyn, Nedocromil, and Theophylline). Nucleoside diphosphate kinase (NME1-NME2) only expressed in controlled asthmatics whereas polycystic kidney and hepatic disease 1 (PKHD1)/fibrocystin, zinc finger protein 263 (ZNF263), uncharacterized LOC101060047 (ENSG00000268865), desmoglein 2 (DSG2) and S100 calcium binding protein A2 (S100A2) were only found in uncontrolled asthma. Therefore, the six proteins were associated with disease control in children and adults with asthma. Our findings suggest that NME1-NME2, PKHD1, ZNF 263, uncharacterized LOC101060047, DSG 2 and S100 A2 in saliva are associated with disease control in asthma.


Correlations Between Root Metabolomics and Bacterial Community Structures in the Phragmites australis Under Acid Mine Drainage-Polluted Wetland Ecosystem.
Despite root microecology playing critical role in plant growth and fidelity, relatively few studies have focused on the link between the microbial communities and root metabolome in the aquatic macrophytes under heavy metal (HM) pollution. Using high-throughput metagenomic sequencing, targeted metabolomics and community-level physiological profile analyses, we investigated the symbiotic associations between Phragmites australis with rhizospheric bacterial communities under differing acid mine drainage (AMD) pollution. Results indicated that AMD pollution and root localization significantly affected root metabolome profiles. Higher accumulation of adenosine monophosphate, inosine, methionine, carnitine and dimethylglycine were observed in the rhizosphere under AMD than non-AMD habitat. Overall, the bacterial diversity and richness, and functional (metabolic) diversity were lower under high-AMD pollution. While non-AMD site was enriched with members of phylum Firmicutes, Proteobacteria were the most abundant taxa in the rhizosphere and endosphere under AMD-polluted sites. Further, plant growth promoting rhizobacteria (Rhizobium, Delftia, Bradyrhizobium, and Mesorhizobium) and metal-tolerant bacteria (Bacillus, Arthrobacter, Massilia and Methylocystis) were most abundant in AMD-polluted than non-AMD habitat. Finally, pH, TDS (total dissolved solids), Cu, Cr, Fe, and Zn content were the key environmental factors that strongly contributed to the spatial perturbation of rhizospheric metabolites, proteobacterial and acidobacterial taxa. Overall, the study linked the differential endospheric and rhizospheric bacterial community and metabolite profiles in P. australis under AMD environment and provided insights into HM adaptability and phytoremediation potential.


Mental health in relation to changes in sleep, exercise, alcohol and diet during the COVID-19 pandemic: examination of four UK cohort studies.
Responses to the COVID-19 pandemic have included lockdowns and social distancing with considerable disruptions to people's lives. These changes may have particularly impacted on those with mental health problems, leading to a worsening of inequalities in the behaviours which influence health. We used data from four national longitudinal British cohort studies (N = 10 666). Respondents reported mental health (psychological distress and anxiety/depression symptoms) and health behaviours (alcohol, diet, physical activity and sleep) before and during the pandemic. Associations between pre-pandemic mental ill-health and pandemic mental ill-health and health behaviours were examined using logistic regression; pooled effects were estimated using meta-analysis. Worse mental health was related to adverse health behaviours; effect sizes were largest for sleep, exercise and diet, and weaker for alcohol. The associations between poor mental health and adverse health behaviours were larger during the May lockdown than pre-pandemic. In September, when restrictions had eased, inequalities had largely reverted to pre-pandemic levels. A notable exception was for sleep, where differences by mental health status remained high. Risk differences for adverse sleep for those with the highest level of prior mental ill-health compared to those with the lowest were 21.2% (95% CI 16.2-26.2) before lockdown, 25.5% (20.0-30.3) in May and 28.2% (21.2-35.2) in September. Taken together, our findings suggest that mental health is an increasingly important factor in health behaviour inequality in the COVID era. The promotion of mental health may thus be an important component of improving post-COVID population health.


Effect of progressive muscle relaxation exercise on anxiety among nursing students prior to critical care clinical training.
Intensive care units are considered stressful and emotionally risky situations by both professionals and patients and their families. The purpose was to assess the effect of progressive muscle relaxation exercise on anxiety among nursing students in intensive care units prior to clinical training. A randomized, controlled study design was used. The study composed of 80 nursing students from Arab American University. For 2 weeks, the experimental group's 40 participants were taught progressive muscle relaxation exercises to help with anxiety control, whereas the control group's 40 participants received no training. The results revealed that the experimental group was capable of reducing the anxiety (p < 0.05). Also, the experimental group had les anxiety (1.15 ± SD = 0.43) compared with the control group (2.83 ± SD = 0.40). The current study's findings confirmed the effect of progressive muscle relaxation exercise (PMRE) on anxiety reduction in intensive care units during clinical training among nursing students.


Reduction of 125I-angiotensin II binding sites in rat brain following monosodium glutamate treatment.
Adult Sprague-Dawley rats were pretreated with 6 g/kg of monosodium glutamate (MSG). After one month, specific binding of 125I-angiotensin II was measured in membrane preparations from the brain. A 40-45% decrease in the number of 125I-angiotensin II binding sites was observed in brain tissues containing circumventricular organs. No decrease in binding sites was observed in the thalamus/hypothalamus brain region, which resides totally within the blood-brain barrier. The results suggest that MSG can destroy angiotensin-sensitive neurons in circumventricular structures, which are located primarily outside the blood-brain barrier.


Relationships of curiosity to perceptual and verbal fluency in young children.
The relations of curiosity measured separately by teachers' rating and a revised object-curiosity task to verbal fluency and originality, measured by a work fluency test, and to perceptual fluency, obtained from the "Visual Closure" test of the Illinois Test of Psycholinguistic Abilities, were examined in 60 5- to 6-year-old boys. Although teachers' ratings of curiosity were contaminated by MA, they were significantly related to fluency in verbal recall and less strongly to fluency in visual exploration and verbal originality even when wht effect of MA was held constant. Object curiosity, which was negatively correlated with MA, showed at best negligible covariation with these three measures.


Egg distribution and survivorship in the pierid butterfly, Colias alexandra.
Patterns of egg distribution and survivorship were examined for six generations (1975-79 and 1981) of a univoltine population of the pierid butterfly Colias alexandra. Unlike some other of the so-called red-egged pierids, there is no evidence of egg-recognition in this Coliadine species. Two distribution patterns were observed: contagious or clumped in 1976-1979, but random in 1981. Generation to generation, survivorship to diapause of clumped individuals relative to all individuals shows no consistent pattern. Among clumped individuals, survivorship is two times greater for the first or previously-laid individual than the second or subsequently-laid individual, suggesting that if there is a genetic basis for the common contagious pattern, selection is acting against it. The importance of obtaining data which reveals inter-year variation in assessing or interpreting egg distribution is discussed.


Evaluation of the tightness of contact between limbal sclera tunnel and tube following Ahmed glaucoma valve implantation.
To investigate whether the tightness of contact between the tube and the limbal sclera tunnel can be evaluated with high-magnification anterior segment optical coherence tomography (OCT) imaging following Ahmed glaucoma valve implantation. Tightness between the tube and the limbal sclera tunnel was investigated with the CAM-L cornea lens adapter of the Optovue Fourier-domain OCT (RTVue-OCT) for 21 uncomplicated Ahmed glaucoma valves implanted in 20 eyes of 19 patients with glaucoma. Nineteen valves were implanted 4 to 124 months earlier (late postoperative cases) and 2 valves 1 day prior to the imaging (early postoperative cases). All valves were introduced into the anterior chamber via a limbal sclera tunnel. The limbal intratunnel part of the tube was successfully imaged in all but 2 cases where an additional full-thickness sclera patch was used. In 14 cases, the contact was tight without tube compression. In 5 cases, the tube was partially compressed but remained open in the limbal sclera tunnel, and redilated behind the limbus. No case with loose contact or peritubular filtration was seen. The posterior run of the tube was successfully imaged in all 19 cases without a full-thickness sclera patch. High-magnification imaging with the CAM- L anterior segment adapter of the RTVue-OCT allows detailed examination of the limbal insertion area of tubes in both the early and late postoperative periods. Therefore this method may potentially be applied for detection of complications related to tube insertion after glaucoma drainage device surgery.


Blood safety begins with safe donations: update among blood donors in Yaounde, Cameroon.
Blood safety continues to be an issue of concern in most countries of the African region. An important contributing factor to blood insecurity is the absence of regular, voluntary and nonremunerated blood donors. An update is provided on blood donors in Yaoundé, Cameroon. Raising the alarm through the dissemination of alarming statistics and continuous sensitization of the community remain strategies for attempting to provide a solution to a long-standing but dangerous problem.


Ruthenium-Catalyzed Peri- and Ortho-Alkynylation with Bromoalkynes via Insertion and Elimination.
The alkynylation of naphthols takes place with total regiocontrol at the peri position of the hydroxyl group in the presence of [RuCl2(p-cymene)]2 as the catalyst. This reaction features high functional group tolerance. The related ortho-alkynylation of benzoic acids proceeds under similar conditions and also shows wide functional group tolerance. Both reactions proceed through metalation, insertion of the alkyne, and bromide elimination.


An experimental study on corneocytes of acutely and chronically irritated skin.
An experimental study concerning the effect of acute and chronic irritation on corneocytes was made in relation to cell number, size, and shape. Corneocytes from skin acutely irritated with a rough towel differed from those of normal skin: There was about a 60% decrease in the count and the corneocytes were 14% smaller in size on day 1 of the experiment. These parameters became normal in count and size after 10 days and 5 days, respectively. Concerning the morphological classification of cell outlines, there was no significant difference between the experimental and control groups except for slightly increased numbers of irregular cells in the experimental group. For the chronic irritation study, subjects were scrub nurses who had worked in the operating room for more than 3 years; ward nurses were used as a control group. Corneocytes from scrub nurses differed from those of the nonirritated skin of ward nurses; the number was twice as high on day 1 of the experiment. Two and four days later, the cell number markedly decreased and was similar to that in the control group. Throughout the experimental period, the surface of the corneocytes was 15% smaller in the experimental group than in the control group. There was no significant difference between the two groups with regard to corneocyte morphology.


Cost-effectiveness analysis of trastuzumab to treat HER2-positive advanced gastric cancer based on the randomised ToGA trial.
We performed a cost-effectiveness analysis of trastuzumab plus chemotherapy for human epidermal growth factor type-2 (HER2)-positive advanced gastric cancer (GC) based on data obtained from the Trastuzumab for Gastric Cancer (ToGA) trial from a Japanese perspective. The following Japanese and Korean populations of the ToGA trial were analysed to obtain mean overall and progression-free survival times: (1) all HER2-positive populations, (2) immunohistochemical (IHC) 2+/fluorescence in situ hybridisation (FISH)+ or IHC 3+ populations, and (3) IHC 3+ only population. The effect of trastuzumab treatment on mean survival time was estimated by fitting a Weibull parametric function. Costs were calculated from the perspective of health-care payer. Neither costs nor outcomes were discounted because of short life expectancy. In the base-case analysis, the incremental cost-effectiveness ratio was (1) JPY 12 million (€110,000) per quality-adjusted life year (QALY) gained and JPY 8.9 million (€81,000) per life-year gained (LYG) for all HER2-positive populations, (2) JPY 9.1 million (€83,000) per QALY gained and JPY 6.6 million (€60,000) per LYG for the IHC 2+/FISH+ or IHC 3+ population, and (3) JPY 6.1 million (€55,000) per QALY gained and JPY 4.3 million (€39,000) per LYG for the IHC 3+ population. Trastuzumab treatment for IHC 3+ populations is cost effective. Our analysis can find a cost-effective subgroup when advanced GC is treated by trastuzumab.


Integrated pipeline for inferring the evolutionary history of a gene family embedded in the species tree: a case study on the STIMATE gene family.
Because phylogenetic inference is an important basis for answering many evolutionary problems, a large number of algorithms have been developed. Some of these algorithms have been improved by integrating gene evolution models with the expectation of accommodating the hierarchy of evolutionary processes. To the best of our knowledge, however, there still is no single unifying model or algorithm that can take all evolutionary processes into account through a stepwise or simultaneous method. On the basis of three existing phylogenetic inference algorithms, we built an integrated pipeline for inferring the evolutionary history of a given gene family; this pipeline can model gene sequence evolution, gene duplication-loss, gene transfer and multispecies coalescent processes. As a case study, we applied this pipeline to the STIMATE (TMEM110) gene family, which has recently been reported to play an important role in store-operated Ca2+ entry (SOCE) mediated by ORAI and STIM proteins. We inferred their phylogenetic trees in 69 sequenced chordate genomes. By integrating three tree reconstruction algorithms with diverse evolutionary models, a pipeline for inferring the evolutionary history of a gene family was developed, and its application was demonstrated.