Serotonin and dopamine in the retina of a lizard.
The objective of this paper is to report the presence and localization of serotonin and dopamine in the retina of the lizard Uta stansburiana. High performance liquid chromatography and electrochemical detection were used to identify and quantitate the two amines. Both compounds are present as endogenous molecules in this retina and are found in concentrations similar to those reported in other non-mammalian retinas. The same methods were employed to confirm, in the isolated retina, the synthesis of serotonin from precursor, tryptophan. Immunocytochemical methods were used to localize, in the neural retina, serotonin and the rate-limiting enzyme of dopamine synthesis, tyrosine hydroxylase. Serotonin immunoreactivity was observed in bistratified amacrine cells (ca. 7 micron dia.) with processes ramifying in sublayers 1, 4, and 5 of the inner plexiform layer. Immunoreactivity to tyrosine hydroxylase was observed in a different population of bistratified amacrine cells (ca. 11 micron dia.) that had processes ramifying in sublayers 1 and 5 (and perhaps 3) of the inner plexiform layer. The enzymes for further metabolism of dopamine were not found in the retina of this lizard by immunocytochemical methods. The results of this research suggest that only single classes of serotoninergic and dopaminergic neurons are present in the retina of U. stansburiana. This retina might, therefore be an appropriate place in which to investigate the functioning of these amines in visual information processing.


Relation between intra-articular temperature of the arthritic temporomandibular joint and presence of calcitonin gene-related peptide in the joint fluid. A clinical study.
Arthritic temporomandibular joints were investigated for intra-articular temperature and joint fluid content of calcitonin gene-related peptide. Eleven patients (16 joints) with rheumatoid arthritis, ankylosing spondylitis, psoriatic arthritis, or chronic unspecific polyarthritis or monarthritis participated in the study. The intra-articular temperature varied between 35.5 and 37.5 degrees C, with a mean of 36.5 degrees C. The concentration of calcitonin gene-related peptide varied between 7.5 and 749.0 pmol/l, with a mean of 108.6 pmol/l. There was a positive correlation between the intra-articular temperature and the joint fluid concentration of calcitonin gene-related peptide. The plasma level of the peptide was on an average 5% of the joint fluid level.


A Facebook-Based Obesity Prevention Program for Korean American Adolescents: Usability Evaluation.
Adolescent obesity is one of the most serious global public health challenges. Social networking sites are currently popular among adolescents. Therefore, the obesity prevention program for Korean American adolescents was developed on the most popular social networking site, Facebook. The purpose of this study was to evaluate the usability of a culturally tailored Facebook-based obesity prevention program for Korean American adolescents (Healthy Teens). An explorative descriptive design of usability testing was used. Usability testing employing one-on-one observation, the think-aloud method, audio taping, screen activity capture, and surveys was performed. Twenty participants were recruited from two Korean language schools (mean age, 15.40 ± 1.50 years). Recruitment and user testing was performed between February and April 2014. Content analysis, using the inductive coding approach, was performed by three coders to analyze transcriptions. Descriptive statistics were used to analyze quantitative data including demographic characteristics, perceived usability, eHealth literacy, and health behaviors. Testing revealed several usability issues in content, appearance, and navigation. Participants' comments regarding content were positive. Although the Facebook platform provided limited flexibility with respect to building the site, participants described the program's appearance as appropriate. Most participants did not experience difficulty in navigating the program. Our preliminary findings indicated that participants perceived the Healthy Teens program as usable and useful. This program could be used as a robust platform for the delivery of health education to adolescents. Further research is required to assess the effects of Facebook-based programs on adolescent obesity prevention.


Micro-CT and histological analysis of Ti6Al7Nb custom made implants with hydroxyapatite and SiO2-TiO2 coatings in a rabbit model.
Bone defect reconstruction in the maxillofacial area comes as a necessity after traumatic, oncological or congenital pathology. Custom made implant manufacturing, such as selective laser melting (SLM), is very helpful when bone reconstruction is needed. In the present study we assessed the osseointegration of custom made implants made of Ti6Al7Nb with two different coatings: SiO2-TiO2 and hydroxyapatite, by comparing the bone mineral density (BMD) measured on micro-CT and the histological mineralized bone surrounding the implants. Custom made - cylindrical type - implants were produced by selective laser melting, coated with SiO2-TiO2 and hydroxyapatite and implanted in the rabbit femur. The animals (divided into 3 groups) were sacrificed at 1, 3 and 6 months and the implants were removed together with the surrounding bone. Bone mineral density and histological examination of the bone-implant surface was performed for each group. BMD and histological examination of the samples determined the quantity of mineralized bone at the implant site, showing a good percentage of mineralized bone for the coated implants at 1, 3 and 6 months. The measurements for the implants without coating showed a significant lower quantity of mineralized bone at 3 months compared with the implants with coating, and a good quantity of mineralized bone at 6 months, showing a process of demineralization followed by remineralization in the last month. The measurements of BMD showed similar results with the histological examination. The use of micro-CT and the measurement of BMD are a reliable, minimally invasive and a quick method of osseointegration assessment.


Temporal and spatial factors in diethylstilbestrol-induced squamous metaplasia of the developing human prostate.
The effects of exogenous estrogen on the developing human prostate were examined in human fetal prostatic rudiments grown for 1 month as subcapsular renal grafts in athymic male nude mice treated with diethylstilbestrol (DES). Prostatic rudiments grown in untreated (control) hosts exhibited normal ductal morphogenesis, growth, and secretory cytodifferentiation. Squamous metaplasia was observed rarely in controls (2 of 22). Conversely, squamous metaplasia was observed in the prostatic utricle and urethra of all 17 specimens grown in DES-treated hosts. Proximal and distal prostatic ducts exhibited DES-induced squamous metaplasia in 13 and 12 of the specimens, respectively. When present, metaplastic changes were mild in the distal ducts. The distribution and intensity of metaplasia were related to the age of the specimen during the period of DES exposure. In specimens grown in DES-treated hosts to a gestational age equivalent of 16.5 weeks or less, squamous metaplasia was only seen in the prostatic utricle and urethra; however, in specimens grown to a gestational age equivalency of 17 weeks or more, squamous metaplasia was observed in the prostatic ducts as well. The nonuniform distribution of estrogen-induced squamous metaplasia within the developing prostatic ductal system emphasizes a regional and age-dependent susceptibility to exogenous estrogens in proximal ductal areas (near the urethra) as compared with distal ductal regions of the developing human prostate.


[Gastroduodenal polyposis in patients with familiar adenomatous polyposis after rectocolectomy].
The extra colonic manifestations, like upper gastrointestinal tract polyps and duodenal cancer are disorders that affect long-term morbidity and mortality of patients with familial adenomatous polyposis, after rectocolectomy. To describe the frequency of those disorders in patients with familial adenomatous polyposis and to review efficacy of upper gastrointestinal endoscopic surveillance. Between 1984 and 2005, 62 patients with familial adenomatous polyposis after rectocolectomy, were studied retrospectively, by Coloproctology Group, Medical Sciences Faculty, State University of Campinas, SP, Brazil. It was possible to analyze 53 patients (85,5%) in this study. Twenty seven (50,9%) of 53 patients in follow-up had upper gastrointestinal polyps. Eight (15,4%) had gastric adenomatous polyps, 14 (27%), duodenal polyps and 5 (9,6%) duodenal and gastric polyps. Two patients (3,8%) had adenomatous duodenal polyps with severe dysplasia, and one (1,9%) had adenocarcinoma of the duodenal papilla. The upper gastrointestinal endoscopic surveillance has importance and the aim is to detect as early as possible the occurrence of duodenal adenocarcinoma and upper gastrointestinal polyps with severe dysplasia.


Using ePROMs for follow-up after palliative radiotherapy: An exploratory study with patients and health care providers.
Patients treated with palliative radiotherapy may experience symptoms decreasing their quality of life. Electronic patient-reported outcome measures (ePROMs) could provide an opportunity to follow-up patients after treatment. A mixed-method study was performed using self-constructed questionnaires, focus groups and interviews with patients and health care professionals (HCP). A qualitative approach was used to code the data. Forty-two patients, 21 radiation-oncologists, 15 general practitioners (GPs) and 24 home-care nurses completed a questionnaire. Ten patients, 6 radiation-oncologists, 14 GPs and 5 nurses were interviewed or participated in a focus group. Although patients and HCP are satisfied with current care, they believe ePROMs could improve follow-up, communication, continuity of care and self-management of symptoms. An easy to use, versatile ePROM platform seems to be important for successful implementation. Self-care tips and contact information should be added to relevant ePROM-questions, on both physical and psychological symptoms. Patients and HCP agree that ePROMs could improve systematical clinical follow-up after palliative radiotherapy, with self-management support being the primary objective of such a system. Practice implications ePROMs after palliative radiotherapy seem feasible, the exact patient population that could benefit the most will need to be explored further; as the palliative population is very diverse.


Aspects of altered metabolism in children with cancer.
Severe weight loss associated with cancer continues to be a major cause of morbidity in cases of childhood malignancy. The etiology is not completely understood but is probably multifactorial, including reduced ingestion and altered metabolism of nutrients. Changes in the host metabolism of protein, fat and carbohydrate in the cancer-bearing host have been demonstrated both in animal models and in patients. Changes include increased protein turnover and loss of the normal compensatory mechanisms seen in starvation. Additionally, increased lipid breakdown results in depletion of lipid stores and changes in carbohydrate metabolism result in an energy-losing cycle. The increase in protein turnover seen in children with leukemia may be related to the tumor, the chemotherapy administered or to related conditions such as febrile neutropenia. The role of endogenous mediators of cancer cachexia has not yet been clearly elucidated, although tumor necrosis factor, interleukin I and interleukin 6 appear to be involved. Studies of energy expenditure in children with cancer have indicated that certain patients with a raised metabolic rate are at particular risk of severe weight loss. The challenge is to identify these vulnerable patients and to provide adequate nutritional support early in treatment and therefore avoid the deleterious effects of cachexia.


Changes in neurotransmitter metabolism following olfactory bulbectomy in the rat.
Bilateral ablation of the olfactory bulbs caused marked changes in the 'turnover' of several neurotransmitters in the amygdaloid cortex and the mid-brain areas of the rat brain. Following tyrosine and tryptophan hydroxylase inhibition, the decrease in the concentration of noradrenaline and serotonin respectively in the amygdaloid cortex was not so marked in the bulbectomized rats as in their controls. This suggests that the 'turnover' of these biogenic amines is reduced following bulbectomy. Following GABA transaminase inhibition, the increase in the concentration of GABA in this region was increased compared to the controls thereby suggesting that the 'turnover' of the inhibitory neurotransmitter was enhanced, glutamate decarboxylase activity was also increased in the amygdaloid cortex. No changes were found in the 'turnover' of noradrenaline or serotonin in the mid-brain but that of dopamine was decreased as was the activity of glutamate decarboxylase. It is concluded that changes in neurotransmitter 'turnover' in these brain regions are attributable to the destruction of the olfactory bulbs and may contribute to the behavioural deficits which we, and others, have reported elsewhere.


The outcome of control groups in clinical trials of conservative treatments for chronic mechanical neck pain: a systematic review.
Chronic neck pain is highly prevalent in Western societies, with about 15% of females and 10% of males suffering with it at any time. The course of untreated chronic neck pain patients in clinical trials has not been well-defined and the placebo effect has not been clarified. A systematic review of RCT's of conservative treatments for chronic mechanical neck pain was conducted. Studies were excluded if they did not include a control group, if they involved subjects with whiplash injuries, a predominance of headache or arm pain associated with chronic neck pain and if only one treatment was reported. Only studies scoring 3-5 out of 5 on the Jadad Scale for quality were included in the final analysis. Data on change in pain scores of subjects in both placebo (PL) as well as no-treatment (NT) control groups were analyzed. Mean changes in pain scores as well as effect sizes were calculated, summarized and compared between these groups. Twenty (20) studies, 5 in the NT group and 15 in the PL group, with outcome intervals ranging from 1-52 weeks were included in the final analysis. The mean [95% CI] effect size of change in pain ratings in the no-treatment control studies at outcome points up to 10 weeks was 0.18 [-0.05, 0.41] and for outcomes from 12-52 weeks it was 0.4 [0.12, 0.68]. In the placebo control groups it was 0.50 [0.10, 0.90] at up to 10 weeks and 0.33. [-1.97, 2.66] at 12-24 weeks. None of the comparisons between the no-treatment and placebo groups were statistically significant. It appears that the changes in pain scores in subjects with chronic neck pain not due to whiplash who are enrolled in no-treatment and placebo control groups were similarly small and not significantly different. As well, they do not appear to increase over longer-term follow-up.


Parent assessment of medical student's skills in ambulatory pediatrics.
Partnership with parents is a vital part of pediatric medical education, yet few studies have examined parent attitudes towards learners in pediatric settings. Questionnaires were used to determine parent and student assessment of professional and clinical skills (primary outcome) and parent attitudes towards 3rd year medical students (secondary outcome) at the University of Alberta. Chi Square, Kendall's Tau and Kappa coefficients were calculated to compare parent and student responses in 8 areas: communication, respect, knowledge, listening, history taking, physical examination, supervision, and overall satisfaction. Overall satisfaction with medical student involvement by parents was high: 56.7% of all parents ranked the encounter as 'excellent'. Areas of lesser satisfaction included physician supervision of students. Compared to the parent assessment, students tended to underrate many of their skills, including communication, history taking and physical exam. There was no relationship between parent demographics and their attitude to rating any of the students' skills. Parents were satisfied with medical student involvement in the care of their children. Areas identified for improvement included increased supervision of students in both history taking and physical examination. This is one of the largest studies examining parent attitudes towards pediatric students. The results may enhance undergraduate curriculum development and teaching in pediatric ambulatory clinics and strengthen the ongoing partnership between the community and teaching clinics.


Correlation of observed and model vibrational frequencies for aqueous organic acids: UV resonance Raman spectra and molecular orbital calculations of benzoic, salicylic, and phthalic acids.
The aromatic carboxylic acids benzoic, salicylic and phthalic acid were used to study the interaction of soluble organics compounds with metal cations. To accomplish this, we have developed methods for studying the carboxylic acids using UV resonance Raman (UVRR) combined with molecular orbital density functional theory calculations. The pH values of the acid solutions were based on the pK(a)'s for the different acids to examine the neutral and charged species. Deprotonation of the organic acids was detectable down to 10(-4)M using UVRR (two orders of magnitude lower than previous vibrational spectroscopy studies). Limitations to decreasing the concentration lower using the current UVRR facilities are discussed. Two methods were used to calculate the optimized geometry and frequencies of the acids: explicit and continuum solvation. The frequencies from the experimental spectra were then compared to the theoretical results obtained from the two methods.


Bone characteristics of autosomal dominant hypophosphatemic rickets patients.
Autosomal dominant hypophosphatemic rickets (ADHR) is a rare disease caused by activating mutations in fibroblast growth factor 23 (FGF23) gene. With FGF23 activation, ADHR is a good model to explore the effects of FGF23 on skeletal development and mineralization. However, the bone microarchitecture of ADHR patients is poorly investigated. This study aims to illustrate the bone properties of ADHR patients and clarify the effect of FGF23 on load bearing and non-load bearing bone. Bone microarchitectures of 11 ADHR subjects and sex- and age-matched healthy controls were analyzed by HR-pQCT. The effect of FGF23 mutations on load bearing and non-load bearing bone was explored by comparison of bone microarchitecture in distal radius and distal tibia. The BMD, bone microarchitecture and bone strength were compared between 7 ADHR patients and 7 age- and sex-matched XLH patients. Among 11 subjects with FGF23 mutations, 10 patients presented with obvious symptoms, five of which had received 1-3 years of iron supplement, neutral phosphate, and calcitriol treatments. The symptomatic patients presented with low bone density and fractures in X rays, with decreased Z score of aBMD (L1-L4: -1.3 ± 1.4, femoral neck: -2.1 ± 1.8, total hip: -1.85 ± 1.6). Compared with controls, HR-pQCT analysis of 5 untreated ADHR patients showed increased total area (+61.6 %, p = 0.03) and cortical perimeter (+17.2 %, p = 0.03) in distal radius. No significant differences were found in other parameters in distal radius. In distal tibia, the patients presented obvious defects in cancellous bone, with decreased trabecular vBMD (-62.9 %, p = 0.003), trabecular BV/TV (-48.7 %, p = 0.003) and trabecular number (-42.2 %, p = 0.001). The trabecular separation (+113.3 %, p = 0.007) and trabecular network inhomogeneity (+226.7 %, p = 0.001) were accordingly increased. In addition to another 5 treated patients, the bone microarchitecture changes revealed similar pattern, but the increase of total area and cortical perimeter in distal radius was no longer statistically significant. The non-symptomatic ADHR patient demonstrated slightly decreased total vBMD, trabecular vBMD and trabecular BV/TV in distal tibia. The changing pattern of bone geometry and microarchitecture of ADHR patients were similar to XLH patients but showed less deficit and stronger bone strength. ADHR patients presented increased total area and cortical perimeter in distal radius, and obvious defect in cancellous bone in distal tibia. FGF23 have impairment effect on trabecular bone especially in weight bearing site.


Rapid, precise quantification of large DNA excisions and inversions by ddPCR.
The excision of genomic sequences using paired CRISPR-Cas nucleases is a powerful tool to study gene function, create disease models and holds promise for therapeutic gene editing. However, our understanding of the factors that favor efficient excision is limited by the lack of a rapid, accurate measurement of DNA excision outcomes that is free of amplification bias. Here, we introduce ddXR (droplet digital PCR eXcision Reporter), a method that enables the accurate and sensitive detection of excisions and inversions independent of length. The method can be completed in a few hours without the need for next-generation sequencing. The ddXR method uncovered unexpectedly high rates of large (> 20 kb) excisions and inversions, while also revealing a surprisingly low dependence on linear distance, up to 170 kb. We further modified the method to measure precise repair of excision junctions and allele-specific excision, with important implications for disease modeling and therapeutic gene editing.


[Value of 2-D echocardiography in the detection of stress-induced wall-motion abnormalities in coronary heart disease--a comparison with biplane cineventriculography].
To determine the accuracy of echocardiography in assessment of exercise-induced wall motion abnormalities, the results of stress-echocardiography were compared with exercise-cineventriculography. In 56 consecutive patients biplane cineventriculography at rest and immediately after supine bicycle exercise was performed. Cross-sectional echocardiography was obtained using the apical 2- and 4-chamber view for LV imaging under identical exercise conditions. In 6 of the 56 patients 2-D echo, in 8 patients LV-angio, and in 2 patients both methods were of inadequate quality during exercise. Thus, in 40 patients (34 patients had coronary artery disease) local wall motion of 360 wall segments was analysed. 49 segments (14%) of 24 patients showed exercise-induced ischemic wall motion abnormalities during cineventriculography. Only 24 of these 49 asynergics (49%) were also detected by 2-D-echo. Using cross-sectional echocardiography, ischemia related wall motion abnormalities were best detected laterally and septaly, whereas apical asynergies were identified in 3 of 12 segments only. Thus, the clinical value of exercise 2-D echo as a screening method in patients suspected to have coronary artery disease is limited and restricted to patients with excellent visualization of the left ventricular endocardium.


Dialysis-Chelex method for determination of exchangeable copper in human plasma.
A method based on dialysis in the presence of histidine, and subsequent copper adsorption and preconcentration using Chelex-100 resin, has been developed for selective extraction of exchangeable copper from blood plasma. The method was verified by comparative analysis using two different instrumental techniques, ICP-MS and AAS. The results obtained were in excellent agreement. Accurate quantification of the exchangeable copper and the firmly bound copper was achieved. The method was further validated by analysis of a lyophilised human serum certified reference material.


Purification and characterization of two activities of the intracellular dextransucrase from Leuconostoc mesenteroides NRRL B-1299.
Dextransucrase (sucrose: 1,6-alpha-D-glucan 6-alpha-glucosyltransferase EC 220.127.116.11) activity of Leuconostoc mesenteroides NRRL B-1299 cells has been purified by adsorption on hydroxyapatite, followed by chromatographies on DEAE-cellulose and DEAE-Sephadex. The enzyme activity was readily separated into two principal forms of the enzyme, I and II, by DEAE-cellulose chromatography. Both enzymes I and II were purified to an electrophoretically homogeneous state in which the relative enzyme activities reached 32-and 14-fold of the original specimen, respectively. Molecular weights were 69 000 for the enzyme I and 79 000 for the enzyme II as determined by electrophoresis in sodium dodecyl sulphate-polyacrylamide, and no subunit structure was observed. Enzyme I had an optimum pH at 6.3-6.5 and exhibited a maximal activity at 45 degrees C, while the optimum pH and temperature of enzyme II were pH 5.5-5.9 and 35-40 degrees C. The Km values of enzymes I and II were 10.7 and 250 mM, respectively. The effects of several metal ions, chemical reagents, and addition of various dextrans were also examined. Beside linear alpha-1,6-linkages the polymer synthesized by the enzyme II contained lesser amount of alpha-1,2-and alpha-1,3-linkages, which seems to be a primary characteristic of the B-1299 dextran.


Comparative safety of the sodium glucose co-transporter 2 (SGLT2) inhibitors: a systematic review and meta-analysis.
To estimate the association between the use of sodium glucose co-transporter-2 (SGLT2) inhibitors and postmarket harms as identified by drug regulatory agencies. We conducted a systematic review and meta-analysis of randomised controlled trials (RCT). Six large databases were searched from inception to May 2018. Random effects models were used to estimate pooled relative risks (RRs). SGLT2 inhibitors, compared with placebo or active comparators. Acute kidney injury (AKI), diabetic ketoacidosis (DKA), urinary tract infections (UTI), bone fractures and lower limb amputations. We screened 2418 citations of which 109 were included. Most studies included one of four SGLT2 inhibitors, dapagliflozin, canagliflozin, empagliflozin and ipragliflozin. When compared with placebo, SGLT2 inhibitors were found to be significantly protective against AKI (RR=0.59; 95% CI 0.39 to 0.89; I2=0.0%), while no difference was found for DKA (RR 0.66; 95% CI 0.30 to 1.45, I2=0.0%), UTI (RR 1.02; 95% CI 0.95 to 1.09, I2=0.0%) or bone fracture (RR 0.87; 95% CI 0.69 to 1.09, I2=1.3%). Three studies reported on amputation, with one finding a significant increase risk. No increased risk for either outcome was found when compared with active controls. Subgroup analysis did show an increased risk of UTI with dapagliflozin only (RR 1.21; 95% CI 1.02 to 1.43, I2=0.0%), but no other analysis supported an increased risk of AKI, DKA, UTI or fracture. Current evidence from RCTs does not suggest an increased risk of harm with SGLT2 inhibitors as a class over placebo or active comparators with respect to AKI, DKA, UTI or fracture. However, wide CIs for many comparisons suggest limited precision, and therefore clinically important adverse events cannot be ruled out. Dapagliflozin, appears to independently increase the risk of UTI, although the mechanism for this intraclass variation in risk is unclear. CRD42016038715.


N-methylated diazabicyclo[3.2.2]nonane substituted triterpenoic acids are excellent, hyperbolic and selective inhibitors for butyrylcholinesterase.
Triterpenoic acids (oleanolic, ursolic, betulinic, platanic and glycyrrhetinic acid) were acetylated and coupled with 1,3- or 1,4-diazabicyclo[3.2.2]nonanes to yield amides. Reaction of these amides with methyl iodide at the distal nitrogen of the bicyclic system gave the corresponding quaternary ammonium salts. These compounds were shown to act as excellent inhibitors of the enzyme butyrylcholinesterase (BChE) while being only weak inhibitors for acetylcholinesterase (AChE). Evaluation of the enzyme kinetics revealed these compounds to act as hyperbolic inhibitors for BChE while the results from molecular modeling gave an explanation for their selectivity between AChE and BChE.


Persistent Mullerian duct syndrome with bilateral abdominal testis: surgical approach and review of the literature.
We present two cases of persistent Mullerian duct syndrome (PMDS) with bilateral intra-abdominal testes and review the world's literature with special attention to diagnosis and surgical management. Two consecutive cases of PMDS with bilateral intra-abdominal testes from our institution are presented with detailed descriptions of the presentation, physical examination, laboratory profiles, surgical findings, and treatment undertaken. Follow-up at 1 year postoperatively is included. Bilateral orchiopexy was accomplished in both the cases. In one case this was possible after division of the persistent Mullerian structures in the midline to achieve testicular mobility. In a subsequent case, splitting of the Mullerian complex did not provide adequate mobilization and microvascular autotransplantation was performed with an excellent surgical outcome. Bilateral intra-abdominal testes in the setting of persistent Mullerian duct syndrome are a rare entity and controversy remains as to the ideal surgical treatment. Our two cases represent the first reported examples of open single-stage bilateral orchiopexy with division of the Mullerian complex and preservation of the vas deferens (1-year-old boy) and microvascular autotransplantation (5-year-old boy).


Salmonella on feces, hides and carcasses in beef slaughter facilities in Venezuela.
This study determined Salmonella prevalence at different stages during the slaughtering in three beef slaughter plants (A, B and C) located in the western region of Venezuela (Zulia and Lara states). Each facility was visited three times at monthly intervals, from the months October through December of 2006. Samples were collected from hides (n=80), fecal grabs (n=80) and carcasses (n=80) at the phases of pre-evisceration, after-evisceration and pre-cooler at three sampling sites on the animals (rump, flank and brisket). Salmonella prevalence was higher on hides (36.3%) than on feces (13.8%) (P<0.05). Differences among slaughter plants for overall Salmonella prevalence were observed (P=0.001; A: 3.5%, B: 11.1%, C: 4.4%). From the isolated strains, Salmonella enterica subspecies enterica ser. Saintpaul, Salmonella ser. Javiana and Salmonella ser. Weltevreden were identified. Cattle feces and hides might be considered as important sources of Salmonella for carcass contamination at different slaughter stages. The presence of potentially pathogenic Salmonella serotypes at the slaughtering stages is an evidence of the circulation of this pathogen in the food environment; its presence could increase consumers' risks of infection if proper food handling and preparation techniques are not followed. These data should serve as a baseline for future comparisons in Salmonella prevalence on beef carcasses to be used by the government and industry in order to establish preventive measures and to better address the risks of Salmonella contamination.


[New approaches in dystonia (clinical features, genetic issues and pathophysiology].
Dystonia is defined by the occurrence of abnormal twisting movements and posture, and may be generalized or focal. Many subtypes of dystonias have been described, including monogenic forms (eg. DYT1 and DYT6), secondary dystonias related to metabolic disorders, forms due to brain lesions (including post-anoxic brain injury), and tardive dystonia. Dystonia may also be associated with myoclonus (myoclonus-dystonia, DYT11) or parkinsonism (rapid-onset dystonia parkinsonism DYT1). Paroxystic dystonias are a subgroup of paroxysmal dyskinesias. Diagnosis is based mainly on clinical features, age at onset, outcome, associated neurological signs, and brain MRI. Until recently, basal ganglia dysfunction and alterations of the sensory-motor network with maladaptive cortical plasticity were the core pathophysiological features of dystonia. Cerebellar and cerebello-thalamic anatomic-functional abnormalities may also play a role. The main objectives of this review are to highlight the principal characteristics of dystonias and to provide an overview of known culprit genes and environmental factors. Current pathophysiological knowledge, including anatomic-functional abnormalities, is summarized.


A pediatric case of infliximab-resistant ulcerative colitis successfully treated using vedolizumab.
Pediatric ulcerative colitis is likely to be more severe than adult ulcerative colitis. Failure to thrive should be considered during therapy. A 10-year-old boy was diagnosed with ulcerative colitis based on his clinical presentation and colonoscopy and biopsy results. The administration of 5-aminosalicylic acid and prednisolone resulted in remission ; however, the symptoms reappeared after the discontinuation of prednisolone. Then, infliximab was administered ; however, the patient was resistant to it and appeared to be dependent on prednisolone. Vedolizumab, a monoclonal antibody against ?4?7 integrin, was administered, which resulted in rapid remission. A steady decrease in prednisolone followed, and remission was maintained even after prednisolone discontinuation. Vedolizumab may be effective in pediatric patients with moderate-to-severe refractory ulcerative colitis. Vedolizumab prevents lymphocytes from binding to MAdCAM-1, which is selectively expressed in the gastrointestinal submucosa, leading to the mitigation of the systemic side effects of immunosuppression, such as infections. In Japan, vedolizumab use is not yet approved for use in children, but its effectiveness and safety in children is expected to be investigated in the future. J. Med. Invest. 70 : 294-297, February, 2023.


Analysis of the disease constituent ratio and prognosis of patients undergoing ambulatory surgery in oral and maxillofacial medicine: A monocentric retrospective analysis of 427 patients.
Ambulatory surgery and single-visit surgery are becoming increasingly accepted and practiced. The clinical data of patients undergoing ambulatory surgery were collected, and information on their chief complaint and basic information was specifically included. Follow-up phone calls were conducted 1 and 3 days, 1 and 2 weeks, and 1 month after treatment. Information on their recovery and well-being was collected. A total of 427 patients (males: 224, females: 203, average age: 23.07±11 years) were recruited for this study. A total of 43.55% of the patients chose ambulatory surgery. A total of 62.9% of them selected it for convenience, while 43.55% selected it for pain reduction. The top three diseases treated by ambulatory surgery were impacted teeth (56.7%), jaw cyst (14.75%) and supernumerary teeth (10.07%). Postoperative complications occurred in 248 of the 427 patients, with an incidence rate of 58.08%. The complication that occurred most frequently was postoperative pain (56.44%). Complications frequently occurred on Day 3 after the operation and resolved after 2 weeks. After being diagnosed, ambulatory surgery is an effective mode of treatment for oral and maxillofacial diseases. Oral hygiene, professional postoperative follow-up visits and rigorous anesthesia evaluation are very important for ambulatory surgery for oral and maxillofacial diseases.


Properties and applications of α-galactosidase in agricultural waste processing and secondary agricultural process industries.
Agriculture products form the foundation building blocks of our daily lives. Although they have been claimed to be renewable resources with a low carbon footprint, the agricultural community is constantly challenged to overcome two post-harvest bottlenecks: first, farm bio-waste, a substantial economic and environmental burden to the farming sector, and second, an inefficient agricultural processing sector, plagued by the need for significant energy input to generate the products. Both these sectors require extensive processing technologies that are demanding in their energy requirements and expensive. To address these issues, an enzyme(s)-based green chemistry is available to break down complex structures into bio-degradable compounds that source alternate energy with valuable by-products and co-products. α-Galactosidase is a widespread class of glycoside hydroxylases that hydrolyzes α-galactosyl moieties in simple and complex oligo and polysaccharides, glycolipids, and glycoproteins. As a result of its growing importance, in this review we discuss the source of the enzyme, production and purification systems, and enzyme properties. We also elaborate on the enzyme's potential in agricultural bio-waste management, secondary agricultural industries like sugar refining, soymilk derivatives, food and confectionery, and animal feed processing. Insight into this vital enzyme will provide new avenues for less expensive green chemistry-based secondary agricultural processing and agricultural sustainability. © 2023 Society of Chemical Industry.


Closed-Chamber Anterior Capsulorhexis Under Air Tamponade In White Cataract.
To evaluate the efficacy and safety of a closed-chamber air bubble capsulotomy technique to prevent the Argentinean flag sign (AFS) in patients with white and intumescent cataracts. Sohag University, Faculty of Medicine, Ophthalmology Department, Sohag, Egypt. Prospective interventional noncomparative case series. Eighty-two eyes of 80 patients with white and intumescent cataracts were included. Eyes with any ocular pathology other than cataract or eyes subjected to previous intraocular surgery were excluded. Needle capsulorhexis was performed under a large air bubble tamponade without capsular staining. Efficacy of the technique was assessed intraoperatively by successful completion of capsulorhexis. Safety of the technique was assessed by the occurrence of intraoperative or postoperative complications. Complete capsulorhexis by the closed-chamber air bubble technique was successful in 75 eyes (91.5%), while capsulorhexis extension occurred in seven eyes. In four eyes with extension, the procedure was shifted to the standard technique of CCC (circular curvilinear capsulorhexis), using a viscoelastic device. A shift to extracapsular cataract extraction (ECCE) surgery occurred in the remaining three eyes, as the extension was so far-gone. No intraoperative or postoperative complications were observed. The closed anterior chamber air bubble technique for capsulorhexis is a novel, safe, and effective technique to prevent AFS in white and intumescent cataracts. It is also time-saving and cost-effective, with less likelihood of capsulorhexis extension. The technique is considered a valuable addition to previously described techniques.


Early intervention and long-term outcome with cardiac resynchronization therapy in patients without a history of advanced heart failure symptoms.
MADIT-CRT showed that cardiac resynchronization therapy with a defibrillator (CRT-D) improves long-term outcomes in currently mildly symptomatic heart failure (HF) patients with LBBB regardless of the presence of prior advanced HF symptoms. We aimed to evaluate the long-term benefit of CRT-D in patients who never experienced advanced HF symptoms prior to device implantation. Interaction term analysis was used to compare the clinical and echocardiographic benefit of CRT-D vs. implantable cardioverter defibrillator (ICD)-only therapy during long-term follow-up (median 5.6 years) between LBBB patients with or without a history of advanced HF [defined as NYHA class ≥ III or past hospitalization for worsening HF >3 months prior to enrolment in MADIT-CRT (n = 529 and 752, respectively)]. Multivariable analysis showed that treatment with CRT-D was associated with a significant reduction in the risk of HF or death during long-term follow-up regardless of the presence of prior advanced HF symptoms [hazard ratio 0.53 (P < 0.001) and 0.47 (P < 0.001) in the respective groups of patients with and without prior advanced HF; interaction P for the difference = 0.58]. Echocardiographic response to CRT at 1 year was also similar between the two groups (P > 0.10 for all comparisons). Our findings suggest that treatment with CRT-D is associated with pronounced echocardiographic and long-term clinical benefit in patients with LV dysfunction and LBBB who never experienced advanced HF symptoms. These data further emphasize the benefit of early intervention with CRT in this population.


Chromosomal localization and catalytic properties of the recombinant alpha subunit of human lymphocyte methionine adenosyltransferase.
Human lymphocyte methionine adenosyltransferase (HuLy MAT) consists of heterologous subunits alpha and beta. The cDNA sequence of the alpha subunit of HuLy MAT from Jurkat leukemic T cells was identical to that of the human kidney alpha subunit and highly homologous to the sequence of the extrahepatic MAT from other sources. The 3'-untranslated sequence was found to be highly conserved, suggesting that it may be important in regulating the expression of MAT. The extrahepatic alpha subunit unit of MAT was found to be expressed also in human liver, and no differences were found in the sequence of the alpha subunit from normal and malignant T cells. The sequence of two unspliced introns found in the cDNA clones from the Jurkat library enabled us to isolate genomic clones harboring the human extrahepatic alpha subunit gene and to localize it to the centromere on chromosome arm 2p, an area that corresponds to band 2p11.2. Expression of the alpha subunit cDNA in Escherichia coli yielded two peptides with the immunoreactivity and mobilities of authentic alpha/alpha' subunits from HuLy. The Km of the recombinant alpha subunit was 80 microM, which is 20-fold higher than found for the (alpha alpha')x beta y holoenzyme purified from leukemic lymphocytes and 4-10-fold higher than found for the normal lymphocyte enzyme. The data suggest that the alpha/alpha' subunits mediate the enzyme catalytic activity and that the beta subunit may be a regulatory subunit of extrahepatic MAT.


Notes on two species of Massuria Thorell, 1887 (Arachnida, Araneae, Thomisidae) from China with description of a new species.
Two species assigned to Massuria Thorell, 1887 are reviewed. The female of Massuriabandian Tang & Li, 2010 is described for the first time from Jianfengling National Natural Reserve, Hainan Province, China. The species Diaeasimplex Xu, Han & Li, 2008 is described as a synonym of Massuriabellula Xu, Han & Li, 2008 based on female and male specimens from Guangdong Province, China. Massuriaminsp. nov. described as a new species (female, Fujian Province, China). Detailed illustrations and a distribution map are provided for these three species of Massuria.


Aging of the immune system.
A review of the literature shows that the thymus is the major reason for aging of the immune system. A graph of the mouse thymus mass is linear with age after puberty. However, T cell function is determined by thymic hormone factors and serum thymic factors. The net result is that T cell function is so affected that there is a deficit in T cell responsiveness. For humans a graph of 1-I (I is immunological efficiency) versus age is a straight line on a semi-log plot. The slope of the line is about the same as the incidence of class I cancer incidence vs. age.


Sun exposure and risk of lymphoid neoplasms in Singapore.
Epidemiologic studies have reported an inverse association between sun exposure and non-Hodgkin lymphoma (NHL), but these have been almost exclusively conducted in Western populations residing in temperate locations. We evaluated the association between personal outdoor sun exposure and risk of malignant lymphomas in Singapore. A hospital-based case-control study of 541 incident cases of lymphoid neoplasms and 830 controls were recruited during 2004-2008. Participants were interviewed regarding recreational or occupational outdoor activities during childhood and in adulthood. Basic demographics and potential confounders were also collected. Odds ratios (OR) and 95 % confidence intervals (CI) were calculated using unconditional logistic regression analysis. Compared with individuals who did not have regular sun exposure, a lower risk of NHL was observed for those who reported regular exposure on non-school days during childhood [OR, 0.62; 95 % CI, 0.46-0.83] and non-working days in adulthood [OR, 0.70; 95 % CI, 0.51-0.97]. The protective effect was more evident among women. Our findings support an inverse relationship between intermittent sun exposure and the risk of NHL. These findings are consistent with the growing evidence from various countries, but further studies, especially prospective studies, are needed in Asian populations.


Voluntary alpha control, visually evoked potentials, and peripheral physiological indicants of activation.
To test the hypothesis that voluntary alpha control is mediated in part through self-induced changes in cortical activation level and bodily arousal, changes in several physiological indicants of such activity were systematically examined as a function of the voluntary enhancement and suppression of scalp-recorded alpha activity. Following per-training, 6 subjects attempted to either produce or suppress alpha with eyes closed while receiving continuous anditory feedback of their alpha levels. The results were interpreted as offering tentative support for the hypothesis.


Improving the length-fractionation of DNA during capillary electrophoresis.
The present study develops a path-lengthening strategy for capillary electrophoresis of short double-stranded DNA molecules, in an aqueous solution of neutral polymer (hydroxypropylmethylcellulose). Tests of the dependence of fractionations on pulse times reveal the operation of at least one mechanism in addition to increase in effective path length. Electrophoresis is performed in the following two-stage cycles (cyclic electrophoresis): The first analysis-stage of each cycle is a constant field (forward) capillary electrophoresis. This analysis-stage reveals the length distribution of the shortest DNA molecules not previously analyzed. The second, enhancement-stage of each cycle is zero-integrated field electrophoresis (ZIFE). The enhancement-stage improves the DNA length-fractionation for the next DNA molecules to be analyzed. A slight reverse migration occurs in the enhancement-stage. Increase in both peak separation and peak sharpness contribute to improvement in the length-fractionation of DNA molecules.


[Antiprotease activity of the blood serum of patients with the disseminated form of breast cancer before and after combined treatment].
The paper discusses the results of a study on the blood-serum levels of alpha 1-antitrypsin and alpha 2-macroglobulin in 29 patients with an extensive breast cancer before and within 3 years after a combination therapy including proton "hypophysectomy". A correlation was established between the blood level of alpha 1--antitrypsin and the subclinical stage of generalized metastatic spreading. Consequently, the assay of the level of this inhibitor is suggested as another test for assessment of the efficacy of treatment of such patients and prediction of the results.


The Iranian version of the Premenstrual Symptoms Screening Tool (PSST): a validation study.
The objective of the current study was to translate and test psychometric properties of the Premenstrual Symptoms Screening Tool (PSST) in Iran. Using a standard "forward-backward" procedure, the English version of PSST was translated into Persian. A random sample of university students aged 18 years and over completed the questionnaire in Tehran, Iran. Psychometric properties of the Iranian version of PSST were assessed by performing reliability (internal consistency) and validity analyses [Content Validity Ratio (CVR) and Content Validity Index (CVI)]. In all, 925 female students took part in the study. Of these, 284 (30.7 %) had premenstrual syndrome (PMS) and 119 (12.9 %) had premenstrual dysphoric disorder (PMDD). Reliability of the PSST as measured by internal consistency was found to be satisfactory (Cronbach's alpha coefficient, 0.93). The content validity as assessed by CVR and CVI were desirable (0.7 and 0.8, respectively). The Iranian version of PSST seems to be a reliable and valid measure of detecting PMS and PMDD in Iranian young female populations.


Evaluation of the safety and protection efficacy of an attenuated genotype vii newcastle disease virus strain as a candidate vaccine.
Newcastle disease (ND) is a serious avian infectious disease, causing severe economic loss worldwide. Its prevention depends on comprehensive vaccination scheme against Newcastle disease virus (NDV). However, current vaccine strains are of different genotypes with prevalent circulating strains (genotype VII), with significant genetic distance. Our team previously generated a genotype matched attenuated NDV strain (rmNA-1). In this study, its safety and immunization efficacy were evaluated. Its lentogenic characteristic was stable for 25 generations in embryonated chicken eggs and for six generations in SPF chickens. Overdosed administration did not cause any clinical signs or pathogenic changes in chickens. As to its immunization effect, rmNA-1 stimulated a comparable serum NDV specific antibody level to a LaSota (genotype II) strain based commercial vaccine, and provided full protection against virulent genotype VII strain challenge, with significantly reduced virus shedding period.


Cytogenic and molecular analysis of an aggressive angiomyxoma.
Aggressive angiomyxoma is a rare mesenchymal tumor occurring mainly in the vulvar region extending into the paravaginal and perirectal region. Histologically, this tumor is rich in vascular structures and in collagen fibers and is of myxoid appearance. Cytogenetic and molecular analysis was performed on a case of an aggressive angiomyxoma and revealed clonal karyotypic abnormalities. All 50 metaphases analyzed showed a translocation involving the chromosomal region 12q14-15. Chromosomal aberrations involving the breakpoint region 12q14-15 are frequently seen in a variety of other mesenchymal tumors as uterine leiomyomas, lipomas, hamartomas of the lung, liposarcomas, or hemangiopericytomas. Therefore, this breakpoint region seems to be the most frequent chromosomal abnormality associated with the initiation of human mesenchymal neoplasms. To narrow down the breakpoint region on a molecular level in the cells of the angiomyxoma we performed FISH analysis with different cosmid clones originating from a yeast artificial chromosome and cosmid contig overspanning parts of the region 12q14-15. We were able to narrow down the region to approximately 70-80 kb belonging to an area designated a multiple aberration region, because it also includes the breakpoints of leiomyomas, lipomas, and pleomorphic adenomas with 12q13-15 abnormalities. Our molecular and cytogenic data suggest that angiomyxomas or at least a subset of them molecularly belong to the benign group of mesenchymal tumors showing multiple aberration region involvement.


EMLA cream-induced irritant contact dermatitis.
The Eutectic Mixture of Local Anesthetics (EMLA cream) is a topical anesthetic used for providing pain relief in patients undergoing superficial surgical procedures. Cutaneous side-effects have been reported rarely. We present a case of irritant contact dermatitis induced by EMLA cream in a 6-year-old boy with Wiskott-Aldrich syndrome. Our patient showed clinically a well circumscribed patch corresponding to the site of application of the topical anesthetic. Histopathology showed confluent necrosis of keratinocytes in the upper epidermis, a mixed inflammatory infiltrate with priminent neutrophils in the upper dermis, and focal signs of interface changes including basal cell vacuolization and subepidermal cleft formation. Graft-vs.-host-disease (GVHD), necrolytic migratory erythema, dermatitis enteropathica and pellagra should be considered in the histopathologic differential diagnosis of acute contact dermatitis caused by EMLA.


Laparoscopic surgery for submucosal tumors located at the esophagogastric junction and the prepylorus.
Laparoscopic partial gastric resection is widely accepted as a treatment for gastric submucosal tumors (SMTs). However, SMTs of either end of the stomach are generally managed by subtotal gastrectomies or total gastrectomies. This study was conducted to evaluate surgical techniques for management of SMTs located at the ends of the stomach. Among 63 patients who were diagnosed and underwent laparoscopic surgery for gastric SMTs at Seoul National University Bundang Hospital from May 2003 to May 2007, 11 SMTs located at the ends of the stomach were identified. The clinicopathologic results of these 11 SMTs were analyzed. Laparoscopic partial wedge resections or tumor excisions were successfully performed on all patients except for those who had prepyloric tumors. Six men and five women had SMTs at the ends of the stomach. The patients ranged in age from 21-63 years (mean 43.4 +/- 13.5 years). Of six esophagogastric junctional tumors that showed low, homogeneous contrast enhancement on computed tomography (CT) scans, five were treated by laparoscopic transgastric enucleation and one by tumor-everting resection. One esophagogastric junctional tumor that leaned toward the fundus and showed a 6-cm-diameter endophytic mass with heterogeneous enhancement on CT scan was resected by laparoscopic wedge resection. The mean operation time was 100 min (range 60-210 min). Three laparoscopy-assisted distal gastrectomies and one laparoscopic wedge resection were performed on SMTs located near the prepyloric antrum. There were no intra- or postoperative complications. Duration of postoperative hospital stay ranged from 4-7 days. Laparoscopic local resection is an effective treatment for SMTs located at the esophagogastric junction and can be used instead of a total or proximal gastrectomy. However, gastrectomies should be considered for SMTs located near the pylorus because of the small volume of the lower third of the stomach.


Characteristics of bombesin-induced inhibition of intake of ethanol.
The temporal and neural dependencies of the inhibitory effect of the administration of bombesin tetradecapeptide (BBS) on the intake of ethanol were assessed in the water-deprived rat. Variation of the intraperitoneal (i.p.) injection of neuropeptide--5% ethanol access interval (0-20 min), revealed that suppression induced by bombesin (0.5-4.0 micrograms/kg) was significantly greater and more potent at shorter intervals. The intake of ethanol was less in rats with subdiaphragmatic vagotomies, but bombesin equivalently suppressed the intake. Intracerebroventricular injection of bombesin more potently and completely inhibited the intake of ethanol but bombesin injected intraventricularly, unlike that given intraperitoneally, elicited excessive grooming and scratching behavior. The suppressant effect of bombesin, given intraperitoneally, requires close temporal contiguity of administration and caloric solution access, which is consistent with a satiety action of a neuropeptide. This satiation effect to ethanol of peripherally administered bombesin appears to reflect a non-vagal, extra-ventricular neural action.


Brief report: cognitive performance in autism and Asperger's syndrome: what are the differences?
Autism spectrum disorders include autistic and Asperger's Syndrome (AS), often studied in terms of executive functions (EF), with controversial results. Using Planning Attention Simultaneous Successive theory (PASS; Das et al. in Assessment of cognitive processes: the PASS theory of intelligence. Allyn and Bacon, Boston, MA, 1994), this research compares the cognitive profiles obtained by the Cognitive Assessment System (CAS; Naglieri and Das in Cognitive assessment system. Riverside, Itasca, IL, 1997) of 15 subjects with typical development, 18 with autistic disorder and 20 with AS. Results highlight lower profiles for children with autistic and AS compared with typical development and even lower Planning and Attention processes for the group with autistic disorders than that with Asperger's. Subjects with Asperger's diagnosis do not differ from those with typical development as regards Simultaneous and Successive processes. Results are discussed in the light of current studies about EF.


The cytosolic or the mitochondrial glutathione peroxidase-type tryparedoxin peroxidase is sufficient to protect procyclic Trypanosoma brucei from iron-mediated mitochondrial damage and lysis.
African trypanosomes express three virtually identical glutathione peroxidase (Px)-type enzymes that occur in the cytosol (Px I and II) and mitochondrion (Px III) and detoxify fatty acid-derived hydroperoxides. Selective deletion of the genes revealed that procyclic Trypanosoma brucei lacking either the cytosolic or mitochondrial enzyme proliferate nearly as wild-type parasites, whereas the knockout of the complete genomic locus is lethal. Flow cytometry and immunofluorescence analyses revealed that the Px I-III-deficient parasites lose their mitochondrial membrane potential, which is followed by a loss of the lysosomal signal but not the glycosomal one. Mitochondrial damage and cell lysis are prevented by Trolox, ubiquinone derivatives and the iron chelator deferoxamine, whereas starch-deferoxamine is inefficient. In glucose-rich medium, cell death is attenuated suggesting that oxidants generated by the respiratory chain contribute to the lethal phenotype. Thus, the Px-type peroxidases protect procyclic cells from an iron-mediated oxidative membrane damage that originates at the mitochondrion. This contrasts with the situation in bloodstream cells, where the lysosome is the primarily affected organelle. Strikingly, either the cytosolic or the mitochondrial form of the peroxidases is required and sufficient to protect the mitochondrion and prevent cell lysis.


Effects of sildenafil on myocardial infarct size, microvascular function, and acute ischemic left ventricular dilation.
Adverse cardiac events in patients treated with the phosphodiesterase-5 inhibitor sildenafil for erectile dysfunction raised concerns about its safety in ischemic heart disease. In anesthetized open-chest rabbits, receiving 1.45 mg/kg sildenafil intravenously or saline 30 min prior to ischemia (n=12, each), infarct size (IS, triphenyltetrazolium), the area of no-reflow (ANR, thioflavin S) (% of the risk area, RA, blue dye), and regional myocardial blood flow (RMBF, radioactive microspheres) were measured after 30 min of coronary occlusion and 180 min of reperfusion. Left ventricular hemodynamics and dimensions (echocardiography) were determined in a separate series of animals (n=5, each). Sildenafil significantly lowered arterial blood pressure before occlusion (-17 to -19 mmHg over 30 min), but during ischemia and reperfusion hemodynamics were comparable to controls. IS in treated animals (51+/-4%) did not significantly differ from control animals (47+/-4%). No major arrhythmias or lengthening of QT/QTc occurred. While sildenafil slightly increased RMBF and significantly reduced specific vascular resistance in the RA during reperfusion (51+/-7 versus 73+/-10 mmHg g min/ml, P<0.05), the ANR (46+/-3%) was similar to control animals (44+/-4%). Sildenafil reduced left ventricular dP/dt(max) (P<0.05) and dP/dt(min) (P<0.01) in non-ischemic conditions, and slightly during ischemia, along with a pronounced decrease in ischemic left ventricular end-diastolic pressure (9+/-2 versus 15+/-2 mmHg after saline, P<0.05), but did not attenuate acute ischemic left ventricular dilation. Sildenafil reduced cardiac pre- and afterload, and parameters of left ventricular contractility. Myocardial necrosis and microvascular dysfunction were neither exacerbated nor attenuated.


New insight into the significance of KLF4 PARylation in genome stability, carcinogenesis, and therapy.
KLF4 plays a critical role in determining cell fate responding to various stresses or oncogenic signaling. Here, we demonstrated that KLF4 is tightly regulated by poly(ADP-ribosyl)ation (PARylation). We revealed the subcellular compartmentation for KLF4 is orchestrated by PARP1-mediated PARylation. We identified that PARylation of KLF4 is critical to govern KLF4 transcriptional activity through recruiting KLF4 from soluble nucleus to the chromatin. We mapped molecular motifs on KLF4 and PARP1 that facilitate their interaction and unveiled the pivotal role of the PBZ domain YYR motif (Y430, Y451 and R452) on KLF4 in enabling PARP1-mediated PARylation of KLF4. Disruption of KLF4 PARylation results in failure in DNA damage response. Depletion of KLF4 by RNA interference or interference with PARP1 function by KLF4YYR/AAA (a PARylation-deficient mutant) significantly sensitizes breast cancer cells to PARP inhibitors. We further demonstrated the role of KLF4 in modulating homologous recombination through regulating BRCA1 transcription. Our work points to the synergism between KLF4 and PARP1 in tumorigenesis and cancer therapy, which provides a potential new therapeutic strategy for killing BRCA1-proficient triple-negative breast cancer cells.


Technical problems in living donor transplantation.
Inferior outcomes are generally described with grafts having multiple arteries or renovascular disease. A consecutive series of 261 living donor (LD) transplants performed in the CsA era was classified in three groups with regard to the graft arterial abnormalities and the techniques employed for revascularization. Two hundred eleven recipients had a kidney with one "healthy" renal artery (Group I); 11 patients, multiple arteries, which were reconstructed by various intracorporeal techniques (Group II); 39 patients, one diseased renal artery or multiple arteries, which were reconstructed on the bench (Group III). One- and 3-year graft survivals not censored for death, were 91%, 82%, 100% and 87%, 82%, 100%, respectively, for Group I, Group II, and Group III. An aggressive policy in performing microsurgical bench reconstruction, also for kidneys with one artery that are affected by intrinsic disease, allowed us to obtain a 3-year graft survival of 100% in our more recent consecutive series of 53 LD transplants.


The effects of the Rho-kinase inhibitor Y-27632 on arachidonic acid-, GTPgammaS-, and phorbol ester-induced Ca2+-sensitization of smooth muscle.
The effects of the Rho-kinase inhibitor, Y-27632 [1] on Ca2+-sensitization of force induced by arachidonic acid (AA), phorbol 12,13-dibutyrate (PDBu), GTPgammaS, and by the stable thromboxane analog, 9,11-dideoxy-9alpha,11alpha-methanoepoxy-PGF2alpha (U-46619), were determined in alpha-toxin-permeabilized smooth muscles. Y-27632 relaxed (up to 99%) Ca2+-sensitization by GTPgammaS (10 microM) and U46619 (1 microM), but not by PDBu (20 microM), and reduced GTPgammaS-induced myosin light chain (MLC20) phosphorylation from 28% to 17% (P=0.002). GTPgammaS-induced force sensitization was inhibited by Y-27632 more potently when the inhibitor was added during the plateau of force than prior to stimulation. In alpha-toxin-permeabilized smooth muscle, Y-27632 inhibited AA (50 microM)-induced Ca2+-sensitization of force (by 66 +/- 1.3%) and reduced MLC20 phosphorylation. In contrast, Y-27632 did not relax force Ca2+-sensitized by AA in smooth muscle permeabilized with Triton X-100. We conclude that (i) AA induces Ca2+-sensitization through dual mechanisms, one mediated by Rho-kinase (or a related kinase), and (ii) Rho-kinase is not required for phorbol ester-induced Ca2+-sensitization.


Antihepatotoxic principles of Solanum capsicastrum.
Two new steroidal alkaloids named capsimine and isocapsicastrine were isolated from the root bark of Solanum capsicastrum. Their structures were elucidated by chemical degradation and spectral data as (22R, 25R)-22,26-epiminocholest-5-ene-3 beta, 16 alpha-diol and (22S,25S)-O(3)-beta-D-glucopyranosyl-22,26-epiminocholest-5-ene-3 beta-16 alpha-diol, respectively. Capsicastrine, capsicastrine acetate, isoteinemine acetate, and etioline exhibited strong activity against liver damage induced by CCl4.


Brain-derived neurotrophic factor does not prevent ionizing radiation-induced apoptosis in the developing rat brain.
In an attempt to learn whether brain-derived neurotrophic factor (BDNF) is able to protect neural cells from ionizing radiation-induced apoptosis in the developing brain, BDNF-transfected (F3N-BDNF) and mock-transfected (F3A MT) fibroblasts were grafted into the right germinal periventricular zone of rats at postnatal day 1 (P1). The rats were later irradiated at P3 with a single dose (2 Gy whole body irradiation) of gamma rays. Age-matched non-irradiated and non-grafted irradiated rats at P3 were used as controls. No differences in the number of apoptotic cells were observed between the ipsilateral and contralateral side of either F3N-BDNF- or F3A-MT-grafted irradiated rats Furthermore, counts of apoptotic cells were similar in non-grafted irradiated and grafted irradiated rats. These results show that BDNF has no capacity to prevent ionizing radiation-induced apoptosis in the developing rat brain.


Arterial system configuration and wave reflection.
On the assumption that the arterial segments between branching points are cylindrical, the relationship between total arterial cross-sectional area and the distance from the heart for the entire length of the arterial system is derived. Based on this relationship, a model of the whole arterial system is proposed; it is composed of three tubes of common origin and of unequal lengths and cross-sectional areas. This model may be useful for simulation purposes, with particular reference to pulse wave reflections.


Characterization of a Schistosoma mansoni homologue of the gene encoding the breast basic conserved protein 1/L13 ribosomal protein.
The Schistosoma mansoni gene sequence encoding the breast basic conserved protein 1/ribosomal protein L13 has been isolated from an adult worm cDNA library using the Expressed Sequence Tag strategy. The cDNA codes for a putative protein of 184 amino acids which is approximately 55% identical to other eukaryotic L13 ribosomal proteins. A PCR amplified genomic fragment containing the coding region of the gene was seen to possess only a single large intron interrupting the open reading frame. Studies of gene expression by RT-PCR showed the transcript is expressed in distinct stages of the parasite life cycle. The cDNA was also hybridized with an ordered cosmid library of S. mansoni and the identified cosmids were mapped to chromosomes 3 and W by chromosomal in situ suppression hybridization.


Peripheral CD4+ /CD8+ T cell composition distinct from healthy individuals is shared by ankylosing spondylitis and rheumatoid arthritis.
Ankylosing spondylitis (AS) and rheumatoid arthritis (RA) are chronic inflammatory joint diseases, linking to the alterations of immune cells. We attempted to assess whether the alterations in the composition of CD4+ /CD8+ T cells are different between AS and RA and identify the characteristic cells between male and female patients. The proportions of CD3+ or double positive T cells, 6 CD4+ T subsets and 9 CD8+ T cell subsets were detected by flow cytometry and compared in 30 healthy individuals, 42 AS patients and 45 RA patients. The differentially altered cells were individually analyzed for associations with disease activity parameters. In addition, their proportions were compared between different genders in the 3 groups. The proportions of CD4+ T cells, naive CD4+ T cells and central memory CD4+ T cells were lower in AS patients (P = 0.001, P = 0.002 and P = 0.007, respectively) and RA patients (P = 0.032, P < 0.001 and P = 0.016, respectively), but the proportion of effector memory ones was higher when compared with healthy populations (both P < 0.001), as were the decrease of naive/central memory CD8+ T cells in AS (P = 0.003 and P = 0.016, respectively) and RA (P < 0.001 and P = 0.006, respectively), and the increased tendency of terminally differentiated CD8+ T cells. However, these above-mentioned cells, regulatory T (Treg) cells and CD8+ T cells with different CD127 expressions between AS and RA were similar in proportion. Furthermore, naive CD4+ T cells were positively associated with C-reactive protein (CRP) in AS, whereas CD4+ T cells and terminally differentiated CD8+ T of RA patients were associated with CRP in RA. The gender-related alterations predominantly displayed the overexpressions of Treg cells and naive CD8+ T cells in female patients with AS and RA, respectively. AS patients and RA patients have some similar peripheral CD4+ /CD8+ T cell subsets but are distinct from healthy individuals, which may contribute to disease severity. Females are respectively characterized by the up-regulation of Treg cells and naive CD8+ T cells in AS patients and RA patients. The study offers an in-depth understanding of the role of T cell subsets in the similarities of the disorders and helps us to monitor disease changes and may offer a theoretical basis of developing novel therapies against common targets.


Daily cannabis use is associated with sleep duration differentially across ages.
To assess the relationship between frequency of cannabis use and sleep duration across age in a large US population (235,667 people). Multinomial logistic regression was used to evaluate the association between the frequency of cannabis use and sleep duration using cross sectional data from the 2016-2018 Behavioral Risk Factor Surveillance System. When adjusted for sociodemographic factors, health related variables, and stratified by age we found that young adults (18-44 years) who reported daily-use (≥16 uses a month) had an increased risk ratio (RR [95% CI]) for either short or long sleep (1.22 [1.06-1.40] and 1.52 [1.07-2.16]); midlife adults (45-64 years) who reported daily-use had an increased prevalence of long sleep (1.71 [1.03-2.82]); and older adults (≥65 years) who reported daily-use had an increased prevalence of short sleep (1.61 [1.05-2.49]). Compared to those who reported no cannabis use, individuals who reported daily cannabis use demonstrated a greater prevalence for either short or long sleep duration.


Prevalence of Internet addiction and its associated factors among female students at Jouf University, Saudi Arabia.
Internet addiction is an increasing problem among university students worldwide. The Internet provides numerous educational advantages, but too much Internet use can lead to unfavorable outcomes such as social isolation and poor academic achievement. The objectives of the present study were to assess the prevalence of Internet addiction and its associated factors among female students at Jouf University, Saudi Arabia. A cross-sectional study using a multistage proportionate sampling technique was done. A self-administered questionnaire was distributed to the female students of Jouf University during face-to-face interviews with them. This questionnaire consisted of two parts; the first is a structured one for identifying sociodemographic features, and the second is Young's Internet Addiction Test (YIAT) to assess Internet use among the students. According to the YIAT scale of Internet addiction, 48.6% of the students were scored to be average Internet users. However, 49.5% and 1.9% of the students had moderate and severe addictions, respectively. The vast majority of students (94.6%) preferred home to access the Internet. Communication was the main purpose of using the Internet as it was reported by 47.3% of the students. The majority of the students (79.5%) utilized mobile phones for Internet access while other devices such as a laptop, tablets and desktop were used by 15.4%, 4.3%, and 3.2% of the students, respectively. More than half of the students (54.6%) used the Internet for an average of more than 4 h every day. Also, more than half (51.4%) used it in the evening being the dominant time of using the Internet. Internet addiction was significantly higher among students with high father education, students who sleep 6 h or less, students who utilize the Internet for entertainment purposes, and students who utilize the Internet mainly in the midnight. Nearly half of female students at Jouf University have moderate or severe Internet addiction. The significant predictors of Internet addiction were sleeping hours ≤ 6, midnight as the dominant time for utilizing the Internet and using the Internet for entertainment purposes.


Distributed synchronization of networked drive-response systems: A nonlinear fixed-time protocol.
The distributed synchronization of networked drive-response systems is investigated in this paper. A novel nonlinear protocol is proposed to ensure that the tracking errors converge to zeros in a fixed-time. By comparison with previous synchronization methods, the present method considers more practical conditions and the synchronization time is not dependent of arbitrary initial conditions but can be offline pre-assign according to the task assignment. Finally, the feasibility and validity of the presented protocol have been illustrated by a numerical simulation.


[Distal femoral fractures].
With a prevalence of approximately 0.4%, distal femoral fractures are a rare entity. Nevertheless, due to the high mortality rates, which are comparable to proximal femoral fractures, these fractures are highly important. The clinical symptoms are often quite striking. In combination with preoperative computed tomography (CT) and if necessary, supplemented by CT angiography, the morphological situation can be assessed and the decision making facilitated. There are several different treatment options and the appropriate one should be selected for each individual case. The use of external fixation of the complete knee joint in the sense of damage control surgery, is useful. For definitive treatment retrograde intramedullary nailing and locking plates with angle stable screws are the main options. Modern angle stable and anatomically preformed implants enable surgical treatment using minimally invasive approaches to protect the soft tissues, with a better overall outcome.


Plasticity of spinovestibular projections in response to hemicerebellectomy in newborn rats.
Spinal cord projections to the lateral vestibular nucleus (LVN) are relatively sparse in normal adult black-hooded rats. However, in response to hemicerebellectomy in 2--3-day-old rats an increase in spinovestibular fibers to the ipsilateral lateral vestibular nucleus was observed. This increased input appears to be due to fibers which would normally be destined for the cerebellum but which alternatively project to the functionally related lateral vestibular nucleus. Considering the neuroanatomical relationships between the cerebellum and the vestibular nuclei and in view of the functional compensation frequently reported after cerebellar lesions, the observed remodelling may represent a partial anatomical basis for recovery of function.


Radiographic characteristics of the maxillomandibular complex in neufibromatosis: short communication and literature review.
To report radiographic characteristics of a case of a patient with neurofibromatosis involving the maxillomandibular complex and to point out other dental radiographic characteristics that are found in the scientific literature to help the dentist when faced with a case of neurofibromatosis. Male, 31 years old, referred for radiographic examination. During anamnesis and physical examination, lesions were noted, which were papules and café-au-lait patches larger than 1 cm on the trunk and upper limbs, further to Lisch nodules (brown spots on the iris) and axillary and inguinal ephelides. When investigating the history of the disease, the patient reported the presence of such skin atlterations since birth and mentioned that his mother had been diagnosed with neurofibromatosis. During radiographic evaluation, a significant increase in the caliber of the mandibular canal and mental foramen was observed, bilaterally. Completing the combination of skin characteristics present in the patient, anamnesis data and current history of the disease with interpretation of radiographic alterations, it was possible to arrive at the diagnostic hypothesis of Neurofibromatosis type 1 and correct clinical management. Therefore, it is extremely important for the dentist to be aware of the clinical, physical and radiographic characteristics that this condition presents for the correct management and success of dental treatment.


Transabdominal bowel sonography for the detection of intestinal complications in Crohn's disease.
The course of Crohn's disease is characterised by the occurrence of intestinal complications such as strictures, intra-abdominal fistulas, or abscesses. Standard diagnostic procedures may fail to show these complications, in particular fistulas. To test the value of transabdominal bowel sonography (T) for the detection of intestinal complications in Crohn's disease. T was prospectively performed in 213 patients with Crohn's disease in a university based inflammatory bowel disease referral centre. Thirty three underwent resective bowel surgery and were included in this study. The accuracy of T to detect strictures, intra-abdominal fistulas, or abscesses was compared with surgical and pathological findings. T was able to identify strictures in 22/22 patients and to exclude it in 10/11 patients (100% sensitivity, 91% specificity). Fistulas were correctly identified in 20/23 patients and excluded in 9/10 patients (87% sensitivity, 90% specificity). Intra-abdominal abscesses were correctly detected in 9/9 patients and excluded in 22/24 patients (100% sensitivity, 92% specificity). In experienced hands T is an accurate method for the detection of intestinal complications in Crohn's disease. T is thus recommended as a primary investigative method for evaluation of severe Crohn's disease.


Risk stratification and response to therapy in patients with pulmonary arterial hypertension and comorbidities: A COMPERA analysis.
A diagnosis of idiopathic pulmonary arterial hypertension (IPAH) is frequently made in elderly patients who present with comorbidities, especially hypertension, coronary heart disease, diabetes mellitus, and obesity. It is unknown to what extent the presence of these comorbidities affects the response to PAH therapies and whether risk stratification predicts outcome in patients with comorbidities. We assessed the database of COMPERA, a European pulmonary hypertension registry, to determine changes after initiation of PAH therapy in WHO functional class (FC), 6-minute walking distance (6MWD), brain natriuretic peptide (BNP) or N-terminal fragment of probrain natriuretic peptide (NT-pro-BNP), and mortality risk assessed by a 4-strata model in patients with IPAH and no comorbidities, 1-2 comorbidities and 3-4 comorbidities. The analysis was based on 1,120 IPAH patients (n = 208 [19%] without comorbidities, n = 641 [57%] with 1-2 comorbidities, and n = 271 [24%] with 3-4 comorbidities). Improvements in FC, 6MWD, BNP/NT-pro-BNP, and mortality risk from baseline to first follow-up were significantly larger in patients with no comorbidities than in patients with comorbidities, while they were not significantly different in patients with 1-2 and 3-4 comorbidities. The 4-strata risk tool predicted survival in patients without comorbidities as well as in patients with 1-2 or 3-4 comorbidities. Our data suggest that patients with IPAH and comorbidities benefit from PAH medication with improvements in FC, 6MWD, BNP/NT-pro-BNP, and mortality risk, albeit to a lesser extent than patients without comorbidities. The 4-strata risk tool predicted outcome in patients with IPAH irrespective of the presence of comorbidities.


SARS-CoV-2 specific plasma cells acquire long-lived phenotypes in human bone marrow.
SARS-CoV-2 specific antibody-secreting plasma cells (PC) mediating specific humoral immunity have been identified in the human bone marrow (BM) after COVID-19 or vaccination against SARS-CoV-2. However, it remained unclear whether or not they acquire phenotypes of human memory plasma cells. SARS-CoV-2-specific human bone marrow plasma cells (BMPC) were characterised by tetramer-based, antigen-specific flow cytometry and FluoroSpot assay. SARS-CoV-2 spike-S1-specific PC were detectable in all tested BM samples of previously vaccinated individuals, representing 0.22% of total BMPC. The majority of SARS-CoV-2-specific BMPC expressed IgG and their specificity for the spike S1 protein indicated emergence from a systemic vaccination response. Of note, one-fifth of SARS-CoV-2-specific BMPC showed the phenotype of memory plasma cells, i.e., downregulated CD19 and present or absent CD45 expression. Our data indicate the establishment of phenotypically diverse SARS-CoV-2-specific PC in the human BM after basic mRNA immunization, including the formation of memory phenotypes. These results suggest the induction of durable humoral immunity after basic mRNA vaccination against SARS-CoV-2. The study was supported by funding by the DFG grants TRR130 TP24, ME 3644/8-1, and the Berlin Senate. SR received funding from DFGSFB-1444 C01 Central Service Project.


Efficacy and safety of a "sandwich therapy" based on staged stereotactic radiosurgery and bevacizumab for large brainstem metastases.
Gamma Knife stereotactic radiosurgery (SRS) is an effective therapeutic option for unresectable brainstem metastases. Currently, staged stereotactic radiosurgery (SSRS) has become available for large brainstem metastases(≥ 1 cm3) despite the limitation of peritumoral edema. The authors developed the so-called "sandwich therapy" which integrated 2-stage stereotactic radiosurgery (2-SSRS) with bevacizumab for peritumoral edema reduction and local control of large brainstem metastases. 42 patients with large brainstem metastases ≥1 cm3 who received 2-SSRS simultaneously with bevacizumab were screened from 2019 to 2021 retrospectively. The first SRS margin doses were 13 Gy (range 11-15) and the second SRS margin doses were 12 Gy (range 11-13), one-time vascular endothelial growth factor (VEGF) inhibitor (bevacizumab) of 3.5-5 mg/kg was administrated intravenously the next day after the first SRS. The median interval between the two sessions of SRS was 6 days. Baseline demographics, clinical and radiology imaging follow-ups were recorded to determine symptomatic improvement, peritumoral edema reduction, local control, and disease progression. Median survival was calculated using Kaplan-Meier analysis. Multivariate analysis was performed to identify prognostic factors. The "sandwich therapy" was applied to 42 lesions. Significant reductions of tumor volume (p < 0.05) and peritumoral edema volume (p < 0.01) were achieved at the second SRS in comparison to those at the first SRS. The proportion of favorable Karnofsky performance scale (KPS) (≥80 %) increased significantly at early follow-up time points and reached the highest value of 85.7 %. The median survival time was 9.7 months, the median local control duration was 11.3 months. 8 acute adverse events of CTCAE grade 2 and 3 were observed in 6 patients and resolved with palliative treatment. Tyrosine kinase inhibitor (TKI) treatment was identified as a predictive factor for longer survival. The "sandwich therapy" which integrates 2-SSRS with bevacizumab is a safe and effective option for large brainstem metastases.


Does self-esteem mediate the association between perfectionism and mindfulness among Lebanese university students?
To evaluate the associations between mindfulness, self-esteem and perfectionism in a Lebanese sample of university students, as well as the indirect effect of self-esteem between mindfulness and perfectionism was investigated. This cross-sectional study was carried out between July and September 2021. A total of 363 university students were recruited through convenience sampling through several universities in Lebanon's governorates. An online survey that included a part that collected sociodemographic information, the Freiburg Mindfulness Inventory to assess mindfulness, the Rosenberg Self-Esteem Scale to assess self-esteem and the Big Three Perfectionism Scale to assess perfectionism was completed by participants. Higher self-esteem (Beta = 1.30) was significantly associated with more mindfulness, whereas higher self-critical perfectionism (Beta = - 0.61) was significantly associated with less mindfulness. Self-esteem mediated the association between self-critical and narcissistic perfectionism and mindfulness. More perfectionism was significantly and directly associated with less mindfulness and lower self-esteem, whereas higher self-esteem was significantly associated with more mindfulness. This study provides valuable findings that enable practitioners to effectively identify people needing interventions to improve psychological health through mindfulness, self-esteem and perfectionism. The conclusions that can be deduced from this study can help educational psychologists and counselors guide university students towards effective mindfulness practices that can increase self-esteem levels and balance maladaptive perfectionism that can cause distress and impairment in the social and academic settings.


Late clinical outcome in patients with early ventricular fibrillation after myocardial infarction.
Whether ventricular fibrillation occurring within 48 h after acute myocardial infarction is associated with particular clinical features and poor prognosis, especially in patients with anterior myocardial infarction, is still debated. Therefore, clinical variables and in-hospital and 1 year mortality rates were analyzed in 2,088 patients, aged 18 to 95 years (mean +/- SD 64 +/- 12), admitted to the hospital with acute myocardial infarction between 1979 and mid 1984. One hundred forty-seven patients (7%) had at least one episode of ventricular fibrillation occurring within 48 h of hospital admission. Of these, 25% died during their initial hospitalization compared with 13% of patients without early ventricular fibrillation (p less than 0.001). In greater than 50% of patients with early ventricular fibrillation, the immediate cause of death was left ventricular failure or cardiogenic shock. In contrast, the 1 year mortality rate after hospital discharge was not significantly greater in patients with than in those without early ventricular fibrillation (15 versus 11%, respectively), particularly in the subgroup of patients with anterior myocardial infarction in which the mortality rate tended to be lower in patients with early ventricular fibrillation (8 versus 14%, respectively). Similar mortality results were found when only primary (not associated with left ventricular failure) ventricular fibrillation was analyzed. The left ventricular ejection fraction and the incidence of complex ventricular arrhythmias from 24 h ambulatory electrocardiographic monitoring obtained at hospital discharge were not different in survivors with or without early ventricular fibrillation (0.45 +/- 0.13 versus 0.49 +/- 0.14 and 41 versus 41%, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)


Early milk-feeding regimes in calves exert long-term effects on the development of ovarian granulosa cells.
Nutrition has not only an impact on the general wellbeing of an animal but can also affect reproductive processes. In cattle, feeding regimes can influence the age of puberty onset and alter gonadal development. We analyzed effects of different milk replacer (MR) feeding regimes during rearing on ovarian physiology with specific emphasis on the numbers as well as gene expression characteristics of granulosa cells (GCs) at the age of puberty onset. Two groups of calves received either 10% or 20% of bodyweight MR per day during their first 8 weeks. After weaning, both groups were fed the same mixed ration ad libitum until slaughter at 8 months. Animals of the 20% feeding group had a significantly higher body weight, but the proportion of animals having a corpus luteum at the time of slaughter was not different between groups, suggesting a similar onset of puberty. Calves of the 10% group showed a constant GC count regardless of the number of follicles (r = 0.23) whereas in the 20% group increasing numbers of GCs were detected with a higher follicle count (r = 0.71). As a first effort to find a possible molecular explanation for this unexpected limitation of GC numbers in the 10% group, we comparatively analyzed GC transcriptomes in both diet groups. The mRNA microarray analysis revealed a total of 557 differentially expressed genes comparing both groups (fold change > |1.5| and p < 0.05). OAS1X, MX2 and OAS1Z were among the top downregulated genes in the 20% vs. the 10% group, whereas top upregulated genes comprised BOLA and XCL1. All of these genes are known to be regulated by interferon. Subsequent signaling pathway analysis revealed the involvement of several immune response mechanisms in accordance with a number of interferons as upstream regulators. The results indicate that the plane of MR feeding in early life has an impact on the number and physiology of GCs later in life. This might influence the overall reproductive life initiated by the onset of puberty in cattle. In addition, the observed alterations in GCs of calves fed less MR might be a consequence of interferon regulated immunological pathways.


Thin and ordered hydrogel films deposited through electrospinning technique; a simple and efficient support for organic bilayers.
Thermal behavior of Dipalmitoylphosphatidylcholine (DPPC) bilayers deposited over hydrogel fibers was examined. Thus, membrane stability, water absorption-release, phase transitions and phase transition temperatures were studied through different methods during heating cycles. Hydrogel films were realized using an oligomer mixture (HEMA-PEGDA575/photo-initiator) with adequate viscosity. Then, the fibers were deposited over silicon wafers (hydrophilic substrate) through electrospinning technique using four different voltages: 15, 20, 25 and 30 kV. The films were then exposed to UV light, favoring polymer chain crosslinking and interactions between hydrogel and substrate. For samples deposited at 20 and 25 kV, hierarchical wrinkle folds were observed at surface level, their arrangement distribution depends directly on thickness and associated point defects. DPPC bilayers were then placed over hydrogel scaffold using Langmuir-Blodgett technique. Field emission scanning electron microscopy (FE-SEM) analysis were used to investigate sample surface, micrographies show homogeneous layer formation with chain polymer order/disorder related to applied voltage during hydrogel deposition process, among other parameters. According to the results obtained, it is possible to conclude that the oligomer deposited at 20 kV produce thin homogenous films (~40 nm) with enhanced ability to absorb water and release it in a controlled way during heating cycles. These scaffold properties confer to DPPC membrane thermal stability, which allow an easy detection of phase(s) and phase transitions. Thermal behavior was also studied via Atomic Force Microscopy (roughness analysis). Contact angle measurements corroborate system wettability, supporting the theory that hydrogel thin films act as DPPC membrane enhancers for thermal stability against external stimuli.


Neurocognitive profiles in MSUD school-age patients.
Maple syrup urine disease (MSUD), an inborn error of amino acids catabolism is characterized by accumulation of branched chain amino acids (BCAAs) leucine, isoleucine, valine and their corresponding alpha-ketoacids. Impact on the cognitive development has been reported historically, with developmental delays of varying degree. Currently, earlier diagnosis and improved management allow a better neurodevelopment, without requirement of special education. However, specific impairments can be observed, and so far, results of detailed neurocognitive assessments are not available. The aim of this study was to analyse neurocognitive profiles of French MSUD patients. This was a multicentre retrospective study on MSUD patients who underwent neurocognitive evaluation at primary school age. Twenty-one patients with classical neonatal onset MSUD were included. The patients' mean age at the time of evaluation was 8.7 years. The mean intellectual quotient (IQ) score was in the normal range (95.1 ± 12.6). In a subset of eight patients, a consistent developmental pattern of higher verbal than performance IQ was observed (mean of the difference 25.7 ± 8.7, p < 0.0001). No correlation could be established between this pattern and long-term metabolic balance (BCAA blood levels), or severity of acute metabolic imbalances, or leucine blood levels at diagnosis and time to toxin removal procedure. These data show that some MSUD patients may exhibit an abnormal neurocognitive profile with higher verbal than performance abilities. This might suggest an executive dysfunction disorder that would need to be further investigated by specialized testing. This pattern is important to detect in MSUD, as appropriate neuropsychological treatment strategies should be proposed.


The reliability of specific primary progressive MS criteria in an ethnically diverse population.
Three different diagnostic criteria for primary progressive MS were recently proposed for Caucasian population of Western European region. The objective of the study was to apply these criteria to a series of Brazilian patients with high ethnic diversity background to evaluate reproducibility and reliability. 52 patients classified as form of the disease that is progressive from onset and followed between 2000 and 2006 were included. Thompson, McDonald and Polman criteria were applied based in clinical date and complementary exams. 72% fulfilled all three criteria with moderate agreement (p<0.001). Ten patients fulfilled at least one criterion and four failed to fulfill any of the three criteria. Strong agreement was found between Thompson and McDonald criteria (p<0.001), agreement was moderate between Thompson and Polman criteria (p<0.001) and weak agreement occurred between McDonald and Polman criteria (p=0.042). The main difference between these criteria is the change in the role of CSF, previously a prerequisite for diagnosis. Rigid diagnostic criteria as Thompson have higher specificity, should be used in clinical research protocols, while more flexible criteria as Polman facilitate the diagnosis of PPMS in neurological practice, particularly in initial stages of the disease, because of their potentially higher sensitivity.


Up-Regulation of Neuronal Nitric Oxide Synthase Expression by Cobalt Chloride Through a HIF-1α Mechanism in Neuroblastoma Cells.
Nitric oxide (NO) plays a dual role in response to neural hypoxia. NO is synthesized by three isoforms of nitric oxide synthase (NOS), among which the neuronal NOS (nNOS) is predominant in the nervous system. Hypoxia-inducible factor-1α (HIF-1α) is a transcription factor that is induced under hypoxic conditions, but its correlation with nNOS remains unclear. In the present study, we aimed at clarifying the regulation pattern of the nNOS expression in response to cobalt chloride (CoCl2), a widely used chemical mimic of hypoxia, and the role of HIF-1α in this process in neuroblastoma cells. We found CoCl2 evidently increased the nNOS expression and NO production in human neuroblastoma SK-N-SH cells, but the effect of CoCl2 on NO was partially abrogated by 7-nitroindazole, a selective inhibitor for nNOS. Importantly, we identified a hypoxia response element (HRE) within the nNOS promoter, to which HIF-1α may bind, and CoCl2 greatly enhanced the HIF-1α expression and its binding to the HRE. Meanwhile, we demonstrated that this HRE was functionally important for the activation of the nNOS transcription, and CoCl2 increased the transcriptional activity of the nNOS promoter through this HRE. Taken together, our study shows that CoCl2 may induce the nNOS expression and NO production through a HIF-1α mechanism in neuroblastoma cells, which may provide a potential target for the treatment of neurological hypoxic disorders caused by NO dysregulation.


Age differences in the neural correlates of recollection: transient versus sustained fMRI effects.
Prior functional magnetic resonance imaging findings in young adults indicate that recollection-sensitive neural regions dissociate according to the time courses of their respective recollection effects. Here, we examined whether such dissociations are also evident in older adults. Young and older participants encoded a series of word-image pairs, judging which of the denoted objects was the smaller. At the test, participants judged whether each of a series of test words was old or new. If a word was old, the requirement was to recall the associated image and maintain it over a variable delay period. Older adults demonstrated significantly lower associative memory performance than young adults. Transient recollection effects were identified in the left hippocampus, medial prefrontal cortex, and posterior cingulate, while sustained effects were widespread across left lateral cortex and were also evident in the bilateral striatum. Except for those in the left insula, all effects were age-invariant. These findings suggest that both transient and sustained recollection effects are largely stable across much of the healthy adult life span.


Azotobacter vinelandii mutants that overproduce poly-beta-hydroxybutyrate or alginate.
Azotobacter vinelandii produces two polymers of industrial importance, i.e. alginate and poly-beta-hydroxybutyrate (PHB). Alginate synthesis constitutes a waste of substrate when seeking to optimize PHB production and, conversely, synthesis of PHB is undesirable when optimizing alginate production. In this study we evaluated the effect of a mutation in algA, the gene encoding the enzyme that catalyzes the first step of the alginate biosynthetic pathway in the production of PHB. We also evaluated production of alginate in strain AT6 carrying a phbB mutation that impairs PHB synthesis. The algA mutation prevented alginate production and increased PHB accumulation up to 5-fold, determined in milligrams per milligram of protein. Similarly, the phbB mutation increased alginate production up to 4-fold.