Spectral counting assessment of protein dynamic range in cerebrospinal fluid following depletion with plasma-designed immunoaffinity columns.
In cerebrospinal fluid (CSF), which is a rich source of biomarkers for neurological diseases, identification of biomarkers requires methods that allow reproducible detection of low abundance proteins. It is therefore crucial to decrease dynamic range and improve assessment of protein abundance. We applied LC-MS/MS to compare the performance of two CSF enrichment techniques that immunodeplete either albumin alone (IgYHSA) or 14 high-abundance proteins (IgY14). In order to estimate dynamic range of proteins identified, we measured protein abundance with APEX spectral counting method.Both immunodepletion methods improved the number of low-abundance proteins detected (3-fold for IgYHSA, 4-fold for IgY14). The 10 most abundant proteins following immunodepletion accounted for 41% (IgY14) and 46% (IgYHSA) of CSF protein content, whereas they accounted for 64% in non-depleted samples, thus demonstrating significant enrichment of low-abundance proteins. Defined proteomics experiment metrics showed overall good reproducibility of the two immunodepletion methods and MS analysis. Moreover, offline peptide fractionation in IgYHSA sample allowed a 4-fold increase of proteins identified (520 vs. 131 without fractionation), without hindering reproducibility. The novelty of this study was to show the advantages and drawbacks of these methods side-to-side. Taking into account the improved detection and potential loss of non-target proteins following extensive immunodepletion, it is concluded that both depletion methods combined with spectral counting may be of interest before further fractionation, when searching for CSF biomarkers. According to the reliable identification and quantitation obtained with APEX algorithm, it may be considered as a cheap and quick alternative to study sample proteomic content.


NIRS-IVUS for Differentiating Coronary Plaque Rupture, Erosion, and Calcified Nodule in Acute Myocardial Infarction.
This study sought to investigate the ability of combined near-infrared spectroscopy and intravascular ultrasound (NIRS-IVUS) to differentiate plaque rupture (PR), plaque erosion (PE), or calcified nodule (CN) in acute myocardial infarction (AMI). Most acute coronary syndromes occur from coronary thrombosis based on PR, PE, or CN. In vivo differentiation among PR, PE, and CN is a major challenge for intravascular imaging. The study enrolled 244 patients with AMI who had a de novo culprit lesion in a native coronary artery. The culprit lesions were assessed by both NIRS-IVUS and optical coherence tomography (OCT). Maximum lipid core burden index in 4 mm (maxLCBI4mm) was measured by NIRS. Plaque cavity and convex calcium was detected by IVUS. The OCT diagnosis of PR (n = 175), PE (n = 44), and CN (n = 25) was used as a reference standard. In the development cohort, IVUS-detected plaque cavity showed a high specificity (100%) and intermediate sensitivity (62%) for identifying OCT-PR. IVUS-detected convex calcium showed a high sensitivity (93%) and specificity (100%) for identifying OCT-CN. NIRS-measured maxLCBI4mm was largest in OCT-PR (705 [interquartile range (IQR): 545 to 854]), followed by OCT-CN (355 [IQR: 303 to 478]) and OCT-PE (300 [IQR: 126 to 357]) (p < 0.001). The optimal cutoff value of maxLCBI4mm was 426 for differentiating between OCT-PR and -PE; 328 for differentiating between OCT-PE and -CN; and 579 for differentiating between OCT-PR and -CN. In the validation cohort, the NIRS-IVUS classification algorithm using plaque cavity, convex calcium, and maxLCBI4mm showed a sensitivity and specificity of 97% and 96% for identifying OCT-PR, 93% and 99% for OCT-PE, and 100% and 99% for OCT-CN, respectively. By evaluating plaque cavity, convex calcium, and maxLCBI4mm, NIRS-IVUS can accurately differentiate PR, PE, and CN.


Peripheral arterial disease in older people with intellectual disability in The Netherlands using the ankle-brachial index: results of the HA-ID study.
Older people with an intellectual disability (ID) have been shown to have similar to increased cardiovascular risks as compared to the general population. Peripheral arterial disease (PAD), atherosclerosis distal from the aortic bifurcation, is associated with increased cardiovascular morbidity and mortality. The prevalence of PAD has not been investigated in this population. Therefore, the aim of the present study was to determine the prevalence of PAD in older people with ID in The Netherlands, the rate of prior diagnoses, and correlations with participant characteristics, and to compare the prevalence with PAD in the general Dutch population. 771 people aged 50 years and over participated in ankle-brachial index (ABI) measurement as part of a multi-centre cross-sectional study (HA-ID study). PAD was defined as an ABI<0.9. After excluding those, who met the exclusion criteria, 629 participants remained. PAD was present in 20.7% of the participants and 97% had not been diagnosed before. People with higher age, smokers and people who lived in central settings, walked with support and were more dependent in activities of daily living were more at risk of PAD. Prevalence of PAD is higher than in the general population (17.4% of 562 eligible participants with ID, as compared to 8.1% of 917 Dutch participants of the PANDORA study, a pan-European study into the prevalence of PAD) through all age groups. Because the high prevalence of PAD implies a serious health risk for older people with ID, we recommend that ankle-brachial index measurement is to be routinely performed as part of the cardiovascular risk management in this group.


Gestational trophoblastic neoplasia: an update.
Gestational trophoblastic neoplasia (GTN) includes invasive mole, choriocarcinoma, and placental site trophoblastic tumors. The overall cure rate in treating these tumors currently exceeds 90%. Thorough evaluation and staging allow selection of appropriate therapy that maximizes chances for cure while minimizing toxicity. Nonmetastatic (stage I) and low-risk metastatic (stages II and III, World Health Organization score < 7) GTN can be treated with single-agent chemotherapy, resulting in a survival rate approaching 100%. High-risk metastatic GTN (stage IV, WHO score > or = 7) requires initial multiagent chemotherapy with or without adjuvant radiation and surgery to achieve a survival rate of 80% to 90%.


Clinical pharmacology of tiazofurin (2-beta-D-ribofuranosylthiazole-4-carboxamide, NSC 286193).
A pharmacokinetic study of tiazofurin was carried out in 13 patients treated in a phase I clinical trial of the drug and in 7 patients undergoing surgical resection of brain tumor (in conjunction with studies on penetration of drug into central nervous system tumors). Tiazofurin was found to be rapidly eliminated from the plasma and red blood cell fraction of both groups of patients with kinetics consistent with a two-compartment model of elimination. Operative conditions did not significantly change the pharmacokinetics of tiazofurin in CNS patients. Pharmacokinetics were linear over the dose range 500-2700 mg/m2. The data suggest that there is only a small degree of tissue binding of drug and that the drug is not concentrated by tissues. Uptake into RBC was rapid and elimination from RBC was essentially parallel to drug elimination from plasma. There was no evidence that RBC sequestration of drug contributes to toxicity. Much of the drug was excreted unchanged in the urine, but there was little correlation between creatinine clearance and plasma pharmacokinetics of tiazofurin, suggesting that renal tubular secretion may be a more important method of elimination than is glomerular filtration. Patients with high AUC values and low plasma clearance values were particularly prone to develop toxicity.


Effects of percutaneous coronary arterial thrombectomy during acute myocardial infarction on left ventricular remodeling.
The benefit of primary angioplasty for acute myocardial infarction (AMI) is limited by the no-reflow phenomenon, resulting in chronic left ventricular (LV) remodeling. The aim of this study was to evaluate the impact of thrombectomy with the Rescue percutaneous thrombectomy catheter on LV function after AMI. We performed a retrospective study comparing conventional angioplasty with the combination of angioplasty and thrombectomy using the Rescue catheter. The study population was comprised of 109 consecutive patients with AMI who underwent angioplasty and thrombectomy and 86 controls treated with conventional angioplasty. Baseline clinical and lesion characteristics were similar in the 2 groups. Postprocedural restoration of normal flow (Thrombolysis In Myocardial Infarction grade 3) was more frequent in the thrombectomy group (82% vs 69%, p = 0.03). No differences were observed in cardiac events, including death, reinfarction, and target vessel revascularization (thrombectomy vs controls, 27% vs 33%; p = 0.44) or changes in ejection fraction (p = 0.22) during 6-month follow-up. The incidence of LV remodeling, defined as an increase in LV end-diastolic volume index of >20%, was significantly lower in the thrombectomy group (22% vs 44%; p = 0.01). Multiple logistic regression analysis revealed that thrombectomy with the Rescue catheter contributed significantly to reduction of both no-reflow and LV remodeling. In the setting of primary angioplasty, adjunctive pretreatment with a rescue catheter reduces the no-reflow phenomenon and protects against LV remodeling.


Interleukin-5 expressed by a recombinant virus vector enhances specific mucosal IgA responses in vivo.
Several in vitro studies have shown that murine interleukin-5 (mIL-5) enhances IgA production by activated mucosal B cells. To date, however, there is no evidence that this factor significantly up-regulates mucosal IgA responses in vivo. Here, we show that expression of the gene for mIL-5 in a recombinant vaccinia virus vector markedly increases IgA responses to co-expressed heterologous antigen in the lungs of mice given intranasal inocula of the virus. The elevated local IgA responses to vectors expressing mIL-5 peaked at a fourfold higher level than those elicited by control virus at 14 days after infection and were sustained for at least 4 weeks. Increased IgA responses were abrogated in mice treated with monoclonal antibody against mIL-5 and were not detected in systemic lymphoid tissue. No enhancement of specific IgG levels was found either locally or systemically. Our results indicate that mIL-5 selectively enhances the development of mucosal IgA responses in vivo and suggest that expression of this factor in mucosal vaccine vectors may stimulate local immune reactivity.


Elimination of the allatotropic activity in locusts by microsurgical and immunological methods: evidence for humoral control of the corpora allata, hemolymph proteins, and ovary development.
Transection of the nervous connections (NCC I) between the brain and the corpus cardiacum (group 1) and injection of anti-brain antibodies inhibited vitellogenin synthesis and ovary development in virgin Locusta migratoria females. Removal of the neurosecretory material by these methods changed the titer and the electrophoretic pattern of hemolymph proteins, including vitellogenin. Transection of NCC I at some distance from the brain resulted in bulbs with 30-200 micron diameter which were filled with stainable neurosecretory material (group 2). Oocyte maturation was delayed in group 2 but normal eggs were laid. The same happened if group 1 animals received daily injections of JH-III. It is concluded that a humoral factor is involved in the activation of JH-III biosynthesis. The factor is transported through the NCC I and is released into the hemolymph as allatotropin.


Exploration of the role of Cuproptosis genes and their related long non-coding RNA in clear cell renal cell carcinoma: a comprehensive bioinformatics study.
Clear cell renal cell carcinoma is a common malignant tumor of the urinary system. The mechanism of its occurrence and development is unknown, and there is currently few effective comprehensive predictive markers for prognosis and treatment response. With the discovery of a new cell death process - cuproptosis drew the attention of researchers. We constructed a model for the prediction of clinical prognosis and immunotherapy response through integrative analysis of gene expression datasets from KIRC samples in The Cancer Genome Atlas (TCGA) database. During the course of the study, we found that cuproptosis genes are significantly differentially expressed between clear cell renal cell carcinoma samples and normal samples. Based on this, we put forward the prognostic model for cuproptosis gene related-long non-coding RNA. And through various statistic and external independent cohorts, we proved that the model is accurate and stable, worthy of clinical application and further exploration and validation.


Effect of short-term prostacyclin administration on restenosis after percutaneous transluminal coronary angioplasty.
The effect of short-term prostacyclin (PGI2) administration on the incidence of restenosis after coronary angioplasty was studied in a prospective single-blind randomized trial of 286 patients. Of the 270 patients in whom dilation was successful, 134 received prostacyclin and 136 received placebo. Intracoronary prostacyclin was administered before and after dilation and then intravenously for 48 h. The control group received intracoronary placebo infusions before and after dilation. All patients received aspirin and dipyridamole before and after angioplasty, at least until follow-up angiography. Follow-up angiograms were obtained in 93% of patients in whom angioplasty was successful. Restenosis of one or more lesions was present in 34 patients (27%) who were given prostacyclin compared with 40 patients (32%) in the control group (p = NS). Acute vessel closure and ventricular tachyarrhythmias were more common in the control group than in the patients who received prostacyclin (acute vessel closure occurred in 14 [10.3%] of 136 versus 4 [3.0%] of 134, respectively, p less than 0.01; ventricular tachyarrhythmias occurred in 5 [3.4%] of 147 versus 0 of 139 respectively, p less than 0.05). Short-term administration of prostacyclin did not significantly lower the risk of restenosis after coronary angioplasty.


Pulmonary rehabilitation in an asthma clinic.
For a small proportion of patients with respiratory disorders, the regular medical care provided by general practitioners and chest physicians does not result in control of respiratory symptoms. In the Dutch Asthma Center, Davos, Switzerland, two conditions not available in routine medical settings are instrumental in treating patients with severe respiratory illness. The treatment center is located at 1560 m above sea level in a high altitude climate (low air humidity, low allergen concentration) which helps stabilize respiratory symptoms. Second, an interdisciplinary treatment plan encompasses optimal medical care and psychological interventions (patient education, self-management training). In this treatment facility, we carry out a cohort study on the determinants of the course of respiratory disorders in the adult patients. At admission, at discharge, and at 6-month follow-up, patient are assessed on medical, psychological, and medical outcome variables. At 6-month follow-up, 35% of the patients were readmitted to a hospital for exacerbations of respiratory symptoms. Multivariate analyses point out that both pulmonary function and psychological factors (particularly state anxiety and agoraphobia) predicted admission. Additional data will be presented and the implications for the multidisciplinary treatment will be discussed.


In vivo experimental evidence that the major metabolites accumulating in 3-hydroxy-3-methylglutaryl-CoA lyase deficiency induce oxidative stress in striatum of developing rats: a potential pathophysiological mechanism of striatal damage in this disorder.
3-Hydroxy-3-methylglutaryl-CoA lyase (HL) deficiency is a genetic disorder biochemically characterized by predominant accumulation of 3-hydroxy-3-methylglutaric (HMG) and 3-methylglutaric (MGA) acids in tissues and biological fluids of affected individuals. Clinically, the patients present neurological symptoms and basal ganglia injury, whose pathomechanisms are partially understood. In the present study, we investigated the ex vivo effects of intrastriatal administration of HMG and MGA on important parameters of oxidative stress in striatum of developing rats. Our results demonstrate that HMG and MGA induce lipid and protein oxidative damage. HMG and MGA also increased 2',7'-dichlorofluorescein oxidation, whereas only HMG elicited nitric oxide production, indicating a role for reactive oxygen (HMG and MGA) and nitrogen (HMG) species in these effects. Regarding the enzymatic antioxidant defenses, both organic acids decreased reduced glutathione concentrations and the activities of superoxide dismutase and glutathione reductase and increased glutathione peroxidase activity. HMG also provoked an increase of catalase activity and a diminution of glucose-6-phosphate dehydrogenase activity. We finally observed that antioxidants fully prevented or attenuated HMG-induced alterations of the oxidative stress parameters, further indicating the participation of reactive species in these effects. We also observed that MK-801, a non-competitive antagonist of the N-methyl-D-aspartate (NMDA) receptor, prevented some of these effects, indicating the involvement of the NMDA receptor in HMG effects. The present data provide solid evidence that oxidative stress is induced in vivo by HMG and MGA in rat striatum and it is presumed that this pathomechanism may explain, at least in part, the cerebral alterations observed in HL deficiency.


Does mechanical loading restore ligament biomechanics after injury? A systematic review of studies using animal models.
Mechanical loading is purported to restore ligament biomechanics post-injury. But this is difficult to corroborate in clinical research when key ligament tissue properties (e.g. strength, stiffness), cannot be accurately measured. We reviewed experimental animal models, to evaluate if post-injury loading restores tissue biomechanics more favourably than immobilisation or unloading. Our second objective was to explore if outcomes are moderated by loading parameters (e.g. nature, magnitude, duration, frequency of loading). Electronic and supplemental searches were performed in April 2021 and updated in May 2023. We included controlled trials using injured animal ligament models, where at least one group was subjected to a mechanical loading intervention postinjury. There were no restrictions on the dose, time of initiation, intensity, or nature of the load. Animals with concomitant fractures or tendon injuries were excluded. Prespecified primary and secondary outcomes were force/stress at ligament failure, stiffness, laxity/deformation. The Systematic Review Center for Laboratory animal Experimentation tool was used to assess the risk of bias. There were seven eligible studies; all had a high risk of bias. All studies used surgically induced injury to the medial collateral ligament of the rat or rabbit knee. Three studies recorded large effects in favour of ad libitum loading postinjury (vs. unloading), for force at failure and stiffness at 12-week follow up. However, loaded ligaments had greater laxity at initial recruitment (vs. unloaded) at 6 and 12 weeks postinjury. There were trends from two studies that adding structured exercise intervention (short bouts of daily swimming) to ad libitum activity further enhances ligament behaviour under high loads (force at failure, stiffness). Only one study compared different loading parameters (e.g. type, frequency); reporting that an increase in loading duration (from 5 to 15 min/day) had minimal effect on biomechanical outcomes. There is preliminary evidence that post-injury loading results in stronger, stiffer ligament tissue, but has a negative effect on low load extensibility. Findings are preliminary due to high risk of bias in animal models, and the optimal loading dose for healing ligaments remains unclear.


Self-efficacy beliefs and fear of childbirth in nulliparous women.
To explore how childbirth self-efficacy, i.e. outcome expectancy and efficacy expectancy, was associated with fear of childbirth (FOC) and how efficacy expectancy and FOC, respectively were related to socio-demographic characteristics, mental problems and preference for a caesarean section. In this cross-sectional study, a consecutive sample of 1000 pregnant nulliparous women was sent the Wijma Delivery Expectancy Questionnaire and Childbirth Self-Efficacy Inventory. Statistical analyses were performed on data from 423 women. Outcome expectancy and efficacy expectancy correlated significantly and positively, FOC correlated significantly and negatively with both outcome expectancy and efficacy expectancy. Women with severe FOC (20.8%) had a significantly lower level of education (p = 0.001), and had more often sought help because of mental problems (p = 0.004). They were more likely to have low-efficacy expectancy (p < 0.001) and to prefer a caesarean section instead of a vaginal birth (p < 0.001). Lower efficacy expectancy was associated with higher FOC while preference for a caesarean section was not. Improvement of self-efficacy could be a part of care for women with FOC during pregnancy; however, it would not be enough for fearful women who wish to have a caesarean section.


2022 Malaysian Working Group Consensus Statement on Renal Denervation for management of arterial hypertension.
Hypertension is highly prevalent and a major contributor to cardiovascular mortality and morbidity. In spite of the availability of efficacious, safe and affordable anti-hypertensive drugs, hypertension remains poorly controlled in the majority of hypertensive patients. Various reasons including non-adherence to the anti-hypertensive drugs, account for the poor control. Resistant hypertension is also one of the reasons for poor control of blood pressure (BP). The sympathetic nervous system (SNS) has long been recognized as one of the determinants in the pathophysiology of a raised BP. Overactivity of the SNS is a contributor to sustained arterial hypertension. Renal denervation (RDN) is increasingly recognized as a safe and effective adjunctive therapy to control BP with or without pharmacotherapy. Hence for patients who remain uncontrolled despite all efforts, renal denervation (RDN) is a novel treatment that can potentially improve BP control, hence reducing the major adverse cardiovascular events (MACE). More recent randomized, sham control trials of RDN have shown that RDN produces a sustained lowering of BP. To date, this lowering of BP through RDN is maintained for at least 3 years. Furthermore, this procedure has been found to be safe. Hence this consensus summarises the science behind RDN and the available clinical data to support the use of this therapy. It is hoped that this consensus will offer guidance on the importance of identifying patients who will benefit most from this therapy. A multidisciplinary team approach in the management of the patient undergoing RDN is recommended.


Comparison of weight loss outcomes 1 year after sleeve gastrectomy and Roux-en-Y gastric bypass in patients aged above 50 years.
Safe, effective weight loss with resolution of comorbidities has been convincingly demonstrated with bariatric surgery in the aged obese. They, however, lose less weight than younger individuals. It is not known if degree of weight loss is influenced by the choice of bariatric procedure. The aim of this study was to compare the degree of weight loss between laparoscopic sleeve gastrectomy (LSG) and laparoscopic Roux-en-Y gastric bypass (LRYGB) in patients above the age of 50 years at 1 year after surgery. A retrospective analysis was performed of all patients more than 50 years of age who underwent LSG or LRYGB between February 2012 and July 2013 with at least 1 year of follow-up. Data evaluated at 1 year included age, sex, weight, body mass index (BMI), mean operative time, percentage of weight loss and excess weight loss, resolution/remission of diabetes, morbidity and mortality. Of a total of 86 patients, 54 underwent LSG and 32 underwent LRYGB. The mean percentage of excess weight loss at the end of 1 year was 60.19 ± 17.45 % after LSG and 82.76 ± 34.26 % after LRYGB (P = 0.021). One patient developed a sleeve leak after LSG, and 2 developed iron deficiency anaemia after LRYGB. The remission/improvement in diabetes mellitus and biochemistry was similar. LRYGB may offer better results than LSG in terms of weight loss in patients over 50 years of age.


Antibodies to HIV2 in genital secretions.
In order to study the local immune response to HIV2, three exclusively heterosexual, infected West African patients were selected: one male with full blown AIDS, one female with ARC and one healthy HIV2-carrier female. Sera and genital secretions were obtained and tested for anti-HIV2 IgG and IgA antibodies by Western blotting. In semen and in vaginal secretions, IgG antibodies directed against all viral antigens were detected. In comparison with the IgG response, local IgA antibody response was less intense and heterogenous. Two out of 3 patients had local antibodies of the IgA class directed against the envelope glycoprotein GP105. Such antibodies of both the IgG and IgA classes in genital secretions could play a protective role in heterosexual transmission of HIV2.


Neuraminidase treatment of avian infectious bronchitis coronavirus reveals a hemagglutinating activity that is dependent on sialic acid-containing receptors on erythrocytes.
The interaction of infectious bronchitis virus (IBV) with erythrocytes was analyzed. The binding activity of IBV was not sufficient to agglutinate chicken erythrocytes. However, it acquired hemagglutinating activity after treatment with neuraminidase to remove alpha 2,3-linked N-acetylneuraminic acid from the surface of the virion. Pretreatment of erythrocytes with neuraminidase rendered the cells resistant to agglutination by IBV. Susceptibility to agglutination was restored by resialylation of asialo-erythrocytes to contain alpha 2,3-linked sialic acid. These results indicate that IBV attaches to receptors on erythrocytes, the crucial determinant of which is sialic acid alpha 2,3-linked to galactose. In contrast to other enveloped viruses with such a binding specificity (influenza viruses and paramyxoviruses) IBV lacks a receptor-destroying enzyme.


Review: Histotrophic nutrition and the placental-endometrial dialogue during human early pregnancy.
Development of the placenta must always be in advance of that of the embryo. Evidence from domestic species demonstrates that the placenta is capable of stimulating its own development through a signalling dialogue with the endometrial glands. Placental lactogens produced by the trophoblast lead to increased expression and release of uterine secretions and mitogenic growth factors, including epidermal growth factor, that have a close temporal and spatial relationship with trophoblast proliferation. Here, we review evidence that an equivalent mechanism operates in the human. The same repertoire of receptors is present on the endometrial gland cells, and the epithelial cells have long been known to adopt a hypersecretory phenotype following an implantation. Furthermore, early pregnancy hormones stimulate the secretion of glycodelin-A and osteopontin, two 'uterine milk proteins' that have multiple potential effects at the maternal-placental interface, from organoid cultures derived from endometrial glands. Prolactin appears to be an important stimulant, but unlike in domestic species the human trophoblast does not secrete this hormone. Instead, it is a major product of decidual cells. Hence, complications of pregnancy that have their pathophysiological roots in deficient trophoblast proliferation may be due primarily to problems of decidualisation. Ensuring the endometrium is in an optimal state pre-conceptionally should therefore be a priority for women's health. Trophoblast stemness and proliferative capacity show a sharp decline at the switch from histotrophic to haemotrophic nutrition. This may reflect the increase in oxygen concentration or loss of growth factor support. Either way, there are implications for adaptive growth of the organ.


A simple, universal, efficient PCR-based gene synthesis method: sequential OE-PCR gene synthesis.
Herein we present a simple, universal, efficient gene synthesis method based on sequential overlap extension polymerase chain reactions (OE-PCRs). This method involves four key steps: (i) the design of paired complementary 54-mer oligonucleotides with 18 bp overlaps, (ii) the utilisation of sequential OE-PCR to synthesise full-length genes, (iii) the cloning and sequencing of four positive T-clones of the synthesised genes and (iv) the resynthesis of target genes by OE-PCR with correct templates. Mispriming and secondary structure were found to be the principal obstacles preventing successful gene synthesis and were easily identified and solved in this method. Compensating for the disadvantages of being laborious and time-consuming, this method has many attractive advantages, such as the ability to guarantee successful gene synthesis in most cases and good allowance for Taq polymerase, oligonucleotides, PCR conditions and a high error rate. Thus, this method provides an alternative tool for individual gene synthesis without strict needs of the high-specialised experience.


Joining empirical and modelling approaches to estimate dry deposition of nitrogen in Mediterranean forests.
In Mediterranean areas, dry deposition is a major component of the total atmospheric N input to natural habitats, particularly to forest ecosystems. An innovative approach, combining the empirical inferential method (EIM) for surface deposition of NO3- and NH4+ with stomatal uptake of NH3, HNO3 and NO2 derived from the DO3SE (Deposition of Ozone and Stomatal Exchange) model, was used to estimate total dry deposition of inorganic N air pollutants in four holm oak forests under Mediterranean conditions in Spain. The estimated total deposition varied among the sites and matched the geographical patterns previously found in model estimates: higher deposition was determined at the northern site (28.9 kg N ha-1 year-1) and at the northeastern sites (17.8 and 12.5 kg N ha-1 year-1) than at the central-Spain site (9.4 kg N ha-1 year-1). On average, the estimated dry deposition of atmospheric N represented 77% ± 2% of the total deposition of N, of which surface deposition of gaseous and particulate atmospheric N averaged 10.0 ± 2.9 kg N ha-1 year-1 for the four sites (58% of the total deposition), and stomatal deposition of N gases averaged 3.3 ± 0.8 kg N ha-1 year-1 (19% of the total deposition). Deposition of atmospheric inorganic N was dominated by the surface deposition of oxidized N in all the forests (means of 54% and 42% of the dry and total deposition, respectively). The relative contribution of NO2 to dry deposition averaged from 19% in the peri-urban forests to 11% in the most natural site. During the monitoring period, the empirical critical loads provisionally proposed for ecosystem protection (10-20 kg N ha-1 year-1) was exceeded in three of the four studied forests.


The forgotten future of the phageThe Good Virus Tom Ireland Norton, 2023. 400 pp.
As antibiotic resistance soars, a journalist revisits an often overlooked antimicrobial strategy.


Appearance of new alpha-bungarotoxin-acetylcholine receptors in cultured sympathetic ganglia of chick embryos.
alpha-Bungarotoxin (alpha-BuTX) has been used as a marker for studying the production of alpha-bungarotoxin-acetylcholine receptors (alpha-BuTX-AChRs) in explants of chick embryo sympathetic ganglia cultured in vitro. New alpha-BuTX-AChRs appear rapidly in the explants after blocking of the pre-existent ones with the toxin (40% of the total receptors at 3 h). There is a portion of alpha-BuTX-AChRs in the explants which for a short time is not accessible to the toxin. This portion constitutes the precursor pool of receptors and represents 18% of the total. The precursor pool of receptors supplies the neurons with new receptors for 1-2 h in the absence of protein synthesis. The appearance of new receptors from the precursor pool is an energy-dependent process.


Spinal meningeal melanocytomas: clinical manifestations, radiological and pathological characteristics, and surgical outcomes.
Meningeal melanocytoma is a rare benign tumor, most frequently located in the posterior fossa and spinal canal. The aim of this study was to investigate the clinical manifestations, radiological features, management, and follow-up data of spinal meningeal melanocytomas. We present the clinical data and long-term outcomes from a consecutive surgical series of 16 patients with pathologically diagnosed spinal meningeal melanocytomas. All of the patients underwent surgical resection. Pre and postoperative MRI was performed. Follow-up data and neurological functional assessment is presented and discussed. The mean age at diagnosis was 42.0 years, with a significant male predominance. The primary clinical symptoms were weakness or numbness of the extremities. The appearance of melanocytoma on MRI is typically isointense to hyperintense on T1-weighted images, hypointense on T2-weighted images, and contrast enhancement tends to be remarkable and homogeneous. In most cases, gross total resection is achievable; however, in rare cases with dumbbell-shaped tumors involving the extraspinal region, a staging operation and subtotal resection should be attempted. During a mean follow-up period of 58.1 months, the symptoms were completely relieved in all the patients, and no tumor progression or recurrence was noted. Melanocytic tumors of the central nervous system have a typical appearance on MRI scans, varying with the content and distribution of melanin. However, the differential diagnosis between malignant melanoma and melanocytoma still depends on pathological criteria. Spinal meningeal melanocytoma has a benign course, and it is amenable for gross total resection. The outcome is favorable following complete resection.


Regions of DNA involved in the stringent control of plasmid-encoded rRNA in vivo.
We have examined the transcription of two plasmid-encoded, stable RNAs; a shortened 16S ribosomal RNA and the spacer transfer RNA2Glu from the Escherichia coli rrnB operon. Plasmid deletions were constructed in vitro, in order to examine the DNA regions required for stringent control of rRNA expression in vivo during amino acid starvation. We find that rRNA synthesized from plasmids does exhibit a relA-dependent, stringent response. The DNA sequences required for this regulation do not extend beyond 20 bases downstream of the P1 transcription initiation site. Deletion of P2, the second of the two tandem rRNA promoters, does not weaken the stringent control of transcripts from P1. These results demonstrate that pause sites for RNA polymerase identified in vitro do not play a significant role in the stringent control of rRNA synthesis in vivo and imply that stringent regulation takes place at the level of initiation, rather than elongation, of transcription. Surprisingly, we find that the presence of extra intact rrnB operons (carried by a multicopy plasmid) reduces the magnitude of the stringent response.


SEM studies on acanthocephalan parasite, Echinorhynchus veli infecting the fish Synaptura orientalis (Bl & Sch, 1801).
Echinorhynchus veli (George and Nadakal, 1978), an acanthocephalid worm infesting the estuarine flat fish, Synaptura orientalis, was collected from the Veli lake, Kerala. The parasite was recovered from the intestine of the host fish. The detailed surface morphology was studied with the help of scanning electron microscope. The study revealed a cylindrical, medially swollen proboscis with a flat apex, backward directed hooks, each with smooth surface, broad base, pointed tip and an epidermal elevation at the point of insertion. A pair of sensory pits was seen at the base of the proboscis. The neck was well developed with densely packed epidermal micropores. Paired sensory pits were seen at the base of the neck and a collar between it and the trunk. The epidermis of the trunk has microtriches and micropores. The female genital pore was circular, and terminal in an elevated orifice. In male, the copulatory bursa was directed ventrally, with well-defined rim and several sensory papillae.


A stable cation of a CSi3P five-membered ring with a weakly coordinating chloride anion.
Don't count on counterions: The cyclic five-membered CSi(3)P cation 1 is synthesized in the reaction of benzamidinato-stabilized chlorosilylene and methyl phosphaalkyne. The presence of four π electrons in 1 means it can be considered as a formal, heavier analogue of the cyclopentadienyl cation. Surprisingly the small counteranion (Cl(-)) does not contribute to the ring stability.


Rated age-of-acquisition norms for over 3,200 German words.
Words that have been learned early in life are responded to faster than words that have been acquired later. Subjective ratings of acquisition ages have been successfully employed to study the effect of age of acquisition (AoA). Although a large number of norms exist in many languages, fewer are available for German. Therefore, subjective AoA ratings for 3,259 German words were collected online, including 2,363 nouns and 473 verbs. These words were presented in lists of 140 words, and participants rated the age in years at which they had first learned each word. A split-half correlation testified to a high internal reliability. There were also high correlations with rated AoA values for subsets of the items that had been collected in previous studies, in both German and English. Age and gender were found to influence the ratings very weakly, in that older and male participants tended to give slightly higher age ratings. Education, multilingualism, and frequent usage of languages other than German did not exert an influence on the rating values. These new ratings will extend the currently existing norms available for language and reading research across languages and will provide researchers with a wider choice of word stimuli. The ratings are available expressed in two measurements: age in years, and AoA rated on a 7-point Likert scale.


Behavioral aspects of host recognition by the symbiotic water mite Unionicola formosa (Acarina, Unionicolidae).
The occurrence and specificity of host recognition behavior of adult and nymphal Unionicola formosa and the capability of adult mites to recolonize various mussel species were examined. Adult U. formosa aggregated on excised tissue from their host mussel, Anodonta imbecilis, in preference to that of two other species of mussels. Nymphs also exhibited an aggregation response to host tissue. A radioisotope (51Cr) technique was used to monitor the recolonization behavior of U. formosa. Adult female mites preferentially re-entered A. imbecilis rather than the sympatric mussel A. cataracta. The specificity of this behavior parallels the distribution of this water mite among potential bivalve hosts in the southeastern U.S. Host recognition by U. formosa may contribute to re-establishing contact with a host after accidental separation and probably helps to maintain mite-mussel symbioses. Whether or not larval U. formosa employ similar host recognition behavior while selecting a potential host has not as yet been determined.


A spectrophotometric method for measuring permanganate index (CODMn) by N,N-diethyl-p-phenylenediamine (DPD).
A new spectrophotometric method for measuring permanganate index (chemical oxygen demand using potassium permanganate (KMnO4) as oxidant, CODMn) in water was established. The method was based on the rapid oxidation of N,N-diethyl-p-phenylenediamine (DPD) by residual KMnO4 in digestion solution under neutral pH condition to form the stable pink radical (DPD●+). Only 20 s were enough to form the pink DPD●+. The generated DPD●+ could be quantitatively measured by a visible spectrophotometer at 551 nm. Stoichiometric coefficient of the reaction between KMnO4 and DPD was close to 1:5 (1:5.07). There was a well linear relationship (R2 = 0.999) between the change of the absorbance of DPD●+ at 551 nm and the concentration of CODMn in the range of 0-4.46 mg L-1. Limit of detection of the DPD method was as low as 0.02 mg L-1 CODMn. The DPD method was highly accurate for measuring CODMn in standard solutions with well recovery rates of 99.17%-102.22%, and was well tolerant to the interference of coexistent Cl- and Fe3+. The DPD method was successfully applied for measuring CODMn in real water samples, including surface water, underground water and drinking water. In comparison to the traditional titration method, the proposed DPD method was more convenient to operate, required less samples and digestion reagents (i.e., KMnO4 and H2SO4) and could be employed for online monitor.


Cost Drivers in Carpal Tunnel Release Surgery: An Analysis of 8,717 Patients in New York State.
The annual high volume of carpal tunnel releases (CTRs) has a large financial impact on the health care system. Validating the cost drivers related to CTR in a large, diverse patient population may aid in developing cost reduction strategies to benefit health care systems. Adult patients with carpal tunnel syndrome who underwent CTR were identified in the New York Statewide Planning and Research Cooperative System database from 2016 to 2017. The Statewide Planning and Research Cooperative System is a comprehensive all-payer database that collects all inpatient and outpatient preadjudicated claims in New York. A multivariable mixed model regression with random effects was performed for the facility to assess the variables that contributed significantly to the total charge. The variables included were patient age, sex, anesthesia method, whether the surgery took place in an ambulatory surgery center or a hospital outpatient department, operation time in minutes, primary insurance type, race, ethnicity, Charlson Comorbidity Index, and categories for billed procedure codes. During the period of 2016 to 2017, 8,717 claims were included, with a mean charge per claim of $4,865. General anesthesia was associated with higher charges than local anesthesia. A procedure at a hospital outpatient department was associated with an approximately 48.2% increase in the total charge compared with that at an ambulatory surgery center. A 1-minute increase in the operation time was associated with a 0.3% increase in the total charge. Claims with antiemetics, antihistamines, benzodiazepines, intravenous fluids, narcotic agents, or preoperative antibiotics were associated with higher total charges than claims that did not bill for these. Compared with endoscopic procedures, open procedures had a 44.3% decrease in the total charges. This comprehensive multivariable model has validated that general anesthesia, hospital-based surgery, the use of antibiotics and opioids, longer operative times, and endoscopic CTR significantly increased the cost of surgery. Economic and decision analyses II.


Neovascularisation precedes neural changes in the rat groin skin flap following denervation: an immunohistochemical study.
We have employed immunohistochemical techniques to study neural and vascular changes in rat skin flaps. Following partial or total denervation, flaps were studied at 4, 7 or 12 days using antisera to PGP 9.5 (panneural marker), the neuropeptides CGRP (sensory nerves) and CPON (adrenergic nerves) and an endothelial marker VWF. In partially denervated flaps, moderate increases in PGP-immunoreactive (PGP-IR) and CGRP-IR nerves and a mild increase in CPON-IR nerves in immediate surrounding skin preceded smaller increases in similar nerves around the pedicle. Following total denervation, mild increases in all nerve types at these locations were accompanied by a marked increase of these nerves in distant surrounding skin, 1-2 cm from the suture line. A gradual increase in endothelial cell staining (VWF) of blood vessels was seen in surrounding skin, flap beds and pedicles. Angiogenesis preceded flap reinnervation which occurred initially from surrounding skin, and later from the base, with sensory fibres appearing first.


The management of oral mucous membrane pemphigoid with dapsone and topical corticosteroid.
Mucous membrane pemphigoid (MMP) is a chronic autoimmune subepithelial disease that primarily affects the mucous membranes of patients over the age of 50 years, resulting in mucosal ulceration and subsequent scarring. The purpose of this study was to investigate the efficacy of dapsone in combination with topical corticosteroids to treat the oral lesions of MMP. Twenty-two patients who had been diagnosed with MMP were selected. The oral features were graded according to severity of disease from 1 to 3. Each patient was assigned to one of four groups according to his or her response to therapy. Five patients with mild to moderate disease were treated with topical corticosteroids alone for the duration of the study; 15 patients with moderate to severe disease were treated by topical corticosteroid with dapsone therapy. All of 15 patients had significant benefits. The use of dapsone in combination with topical corticosteroids is one of the most useful methods for the treatment of MMP.


Quantitative orientation measurements in thin lipid films by attenuated total reflection infrared spectroscopy.
Quantitative orientation measurements by attenuated total reflectance (ATR) infrared spectroscopy require the accurate knowledge of the dichroic ratio and of the mean-square electric fields along the three axes of the ATR crystal. In this paper, polarized ATR spectra of single supported bilayers of the phospholipid dimyristoylphosphatidic acid covered by either air or water have been recorded and the dichroic ratio of the bands due to the methylene stretching vibrations has been calculated. The mean-square electric field amplitudes were calculated using three formalisms, namely the Harrick thin film approximation, the two-phase approximation, and the thickness- and absorption-dependent one. The results show that for dry bilayers, the acyl chain tilt angle varies with the formalism used, while no significant variations are observed for the hydrated bilayers. To test the validity of the different formalisms, s- and p-polarized ATR spectra of a 40-A lipid layer were simulated for different acyl chain tilt angles. The results show that the thickness- and absorption-dependent formalism using the mean values of the electric fields over the film thickness gives the most accurate values of acyl chain tilt angle in dry lipid films. However, for lipid monolayers or bilayers, the tilt angle can be determined with an acceptable accuracy using the Harrick thin film approximation. Finally, this study shows clearly that the uncertainty on the determination of the tilt angle comes mostly from the experimental error on the dichroic ratio and from the knowledge of the refractive index.


[Chemical Composition of n-Butanol Fraction from Polygonum amplexicaule var. sinense].
To study chemical composition of n-butanol fraction from Polygonoum amplexicaule var. sinense. TLC,normal-phase silica gel, reveres-phase silica gel, Sephadex-LH and semi-preparative HPLC were used to isolate chemical compositions of n-butanol fraction from Polygonoum amplexicaule var. sinense. Nine compounds were identified as: caffeic acid n-butly ester (1), p-methoxy benzoic acid propyl ester (2),p-E-coumarin quinic acid methyl ester (3),p-Z-coumarin quinic acid methyl ester (4), ethyl ferulate (5), cinchonain I a (6), cinchonain Ib (7), methyl chlorogenate(8), and 6-O-β-D-caffeoylglucose (9). All compounds are isolated from this genus for the first time.


HPV-16 variants' impact on uterine cervical cancer response to radiotherapy: A descriptive pilot study.
Although the large impact of Human papilloma virus (HPV) in cervical cancer is established, its place as a therapeutic target is new and according to the growing literature, could be promising. In the present study, radiosensitivity's difference based on HPV-16 variants is assessed. Variants of Human papilloma virus were identified before the exclusive radiotherapy in patients with cervical cancer. Data were prospectively collected. Fifty-nine patients were screened. Among the 59 screened patients, 34 (57.6%) were identified to be HPV-16 (+), with 13 European and two non-European variants. Of the 34 patients, 15 experienced exclusive radiotherapy. Among them, eight had complete response (seven with European and one with non-European variants), four with European variant had partial response, three with European variant had tumour persistence and one with non-European variant progressed at 3 months. No radiosensitivity difference was established, probably because of the limited population. Non-European variant aggressiveness might be suggested in accordance with the literature, as it was associated with the only tumour progression. Exclusive radiotherapy provides a unique and "pure" model of radioresistance in cervical cancer and could be the missing link between in vitro studies and state of the art chemoradiotherapy studies that probably feature too many parameters to identify radioresistance causes. The present study was a first step, with the future prospects of building a larger cohort study in order to better understand HPV-induced radioresistance and then to be able to propose new made-to-measure treatments.


mcr-1 colistin resistance gene sharing between Escherichia coli from cohabiting dogs and humans, Lisbon, Portugal, 2018 to 2020.
BackgroundThe emergence of colistin resistance is a One Health antimicrobial resistance challenge worldwide. The close contact between companion animals and humans creates opportunities for transmission and dissemination of colistin-resistant bacteria.AimTo detect potential animal reservoirs of colistin-resistant Escherichia coli and investigate the possible sharing of these bacteria between dogs, cats and their cohabiting humans in the community in Lisbon, Portugal.MethodsA prospective longitudinal study was performed from 2018 to 2020. Faecal samples from dogs and cats either healthy or diagnosed with a skin and soft tissue or urinary tract infection, and their cohabiting humans were screened for the presence of colistin-resistant E. coli. All isolates were tested by broth microdilution against colistin and 12 other antimicrobials. Colistin-resistant isolates were screened for 30 resistance genes, including plasmid-mediated colistin resistance genes (mcr-1 to mcr-9), and typed by multilocus sequence typing. Genetic relatedness between animal and human isolates was analysed by whole genome sequencing.ResultsColistin-resistant E. coli strains harbouring the mcr-1 gene were recovered from faecal samples of companion animals (8/102; 7.8%) and humans (4/125; 3.2%). No difference between control and infection group was detected. Indistinguishable multidrug-resistant E. coli ST744 strains harbouring the mcr-1 gene were found in humans and their dogs in two households.ConclusionsThe identification of identical E. coli strains containing the plasmid-mediated mcr-1 gene in companion animals and humans in daily close contact is of concern. These results demonstrate the importance of the animal-human unit as possible disseminators of clinically important resistance genes in the community setting.


Biological activity of leukotriene sulfones on respiratory tissues.
The biological activity of synthetic leukotriene C4, D4 and E4 sulfone has been determined in respiratory smooth muscle in vitro and in vivo. The sulfones of LTC4, LTD4 and LTE4 were potent contractile agonists on indomethacin-treated guinea pig tracheal chains with respective pD2-values of 8.2, 8.0 and 7.9. Contractions were submaximal (75-85% of the cholinergic maximum), slow in onset, prolonged in duration, slowly reversed by washing (compared to acetylcholine or histamine) and were partially reversed by 2 muM FPL-55712. The sulfones of LTC4, LTD4 and LTE4 also contracted indomethacin-treated guinea pig parenchyma (respective pD2's of 7.9 8.2 and 7.8) and rat parenchyma (respective pD2's of 7.1, 7.2 and 7.2) but were inactive on rat trachea (0.01-2.0 muM). When administered intravenously to anaesthetized guinea pigs, the sulfones of LTD4, LTE4 and to a lesser degree LTC4 (respective ED50's - 0.5; 2.0 and 4.6 microgram/kg) elicited dose-dependent increases in inflation pressure which were antagonized by FPL-55712 and indomethacin. Leukotriene C4, D4 and E4 sulfones display a qualitatively similar profile of biological activity to that of their corresponding sulfides.


[Therapy with plasmapheresis and lymphoplasmapheresis combined with immunosuppressive agents in 2 cases of intractable juvenile rheumatoid arthritis].
Two cases of JRA refractory to NSAID' steroids and long-acting drugs were successfully treated with plasmapheresis and combined lympho- and plasma-apheresis respectively. Case 1. A 8 year old female child who had been suffering from systemic JRA for 6 years received a course of three plasmapheresis followed by a 6 month cycle of azathioprine. Clinical and laboratory remission was promptly achieved and is still present after 18 months. Case 2. A 12-year old boy affected by poliarticular JRA received a course of 4 plasmapheresis followed by a 6 month cycle with azathioprine. The opportunity of performing plasma or lymphoplasmapheresis reinforced by immunosuppressive drugs such as azathioprine or methotrexate at low doses in refractory cases of JRA is discussed.


3D printing- creating a blueprint for the future of orthopedics: Current concept review and the road ahead!
The use of 3D printing in Orthopedics is set to transform the way surgeries are planned and executed. The development of X rays and later the CT scan and MRI enabled surgeons to understand the anatomy and condition better and helped plan surgeries on images obtained. 3DGraphy a term used for 3D printed orthopedic patient models and Jigs has gone a step further by providing surgeons with a physical copy of the patient's affected part that can not only be seen but also felt and moved around spatially. Similarly 3D printed Jigs are patient specific devices that are used to ensure optimal screw trajectory and implant placement with minimal exposure. While the use of 3D printed models and Jigs have now become routine, a similar revolution is happening in the field of designing and printing patient specific implants. Metal printing along with enhanced capability to print other biocompatible materials like PEEK and PLA is likely to improve the current implant manufacturing process. On the horizon is another interesting development related to this field - 3D Bio printing. Printing human tissues and organs is considered the final frontier and impressive strides have been made in printing bone graft substitutes and cartilage like material. This paper is an overview of all the current developments and the road ahead in this invigorating field.


Transfer factor therapy in the Wiskott-Aldrich syndrome. Results of long-term follow-up in 32 patients.
Thirty-two patients with the Wiskott-Aldrich syndrome have been treated with transfer factor provided by this laboratory. Apparent clinical benefit was observed in 44 per cent of them. The mean age of the patients who showed clinical benefit was significantly greater than that of the patients who showed no benefit. Conversion of immunologic reactivity correlated with clinical benefit. Thirteen of the patients who received transfer factor are alive, and 17 have died (43 per cent survival). Clinical benefit was correlated with survival. The median survival was greater than five years in the patients who showed clinical benefit, whereas it was 18 months in those who did not show clinical benefit. We conclude that transfer factor caused conversion of immunologic parameters, apparent clinical benefit and prolonged survival in some, but not all, patients with the Wiskott-Aldrich syndrome.


Autoantibodies to insulin are present in sera of patients with autoimmune thyroid disease.
It has been clinically suspected that patients with autoimmune thyroid disease are at an increased risk of developing other autoimmune diseases later in life. To determine the presence and potential importance of a more generalized deregulation of immune response in patients with Grave's disease and Hashimoto's disease, sera from 33 patients with Graves' disease and 16 patients with Hashimoto's disease were screened for the presence of anti-insulin antibodies and anti-insulin-receptor antibodies. An enzyme-linked immunosorbent assay was used to identify the presence of IgG against human insulin. The optical density indicating the presence of IgG against insulin in sera from patients with Graves' disease averaged .172 +/- .024 (mean +/- SE; range .010-.802), compared to the mean normal value of .098 +/- .0009 (range .012-.238) in 33 control subjects. Ten of 33 patients with Graves' disease had values greater than .200, whereas control sera values were less than .200 in all but one case (P less than .005, Graves' sera vs. controls). The sera from patients with Hashimoto's disease had a mean optical density of .110 +/- .016, with 15 of 16 values between .010 and .200. These values were not significantly different from controls with an insulin-binding inhibition assay. Anti-insulin-receptor antibodies were not detected in any of 33 patients with Graves' disease, and cytoplasmic islet cell antibodies were not detected in sera from seven patients with Graves' disease who had insulin-binding antibodies. These data support the hypothesis that the immunologic response in autoimmune thyroid disease may be more heterogeneous and polyclonal than previously believed.


Molecular basis of alpha-thalassemia in Algeria.
An epidemiological molecular study was carried out to evaluate the spectrum and allelic frequency of alpha-thalassemia (alpha-thal) defects in Algeria. A series of 153 randomly selected blood donors was screened for 10 alpha-thal alleles described in the Mediterranean area. In addition, six unrelated cases with hematological and biochemical data suggestive of Hb H disease were investigated. Our data revealed an allele frequency of 4.6%. The presence of alpha(0)-thal determinants (-alpha(20.5) and --MED I) was observed both in Hb H patients and in the randomly collected samples. Overall, the -alpha(3.7) deletion was the most prevalent allele (2.9%), followed by the alpha(Nco I)alpha (HBA2:c.1A>G) allele (0.6%) and by the alpha(Hph I)alpha (HBA2:c.95 + 2_95 + 6delTGAGG), -alpha(20.5), --(MED I) alleles (0.3% each). The -alpha(4.2) deletion was observed in only one Hb H patient. These results outline the heterogeneity of the alpha-thal alleles in Algeria which reflects the anthropological history of the country. Because of their frequency, alpha-thal alleles are probably frequent modulators of prevalent beta-globin gene-related hemoglobinopathies in Algeria.


Pseudomonas phenolilytica sp. nov., a novel phenol-degrading bacterium.
This study describes a bacterium strain RBPA9 isolated from a municipality waste dumping area capable of degrading phenol, proposed as a novel species of Pseudomonas. Cells are Gram-negative, rod shaped, aerobic and motile. The genome is 3.92 Mb, and the G + C content is 64.64%. The overall genome relatedness indices such as in silico DNA-DNA hybridization (isDDH), average nucleotide identity (ANI), and average amino acid identity (AAI) values were below 70% and 95-96%, respectively. Phylogenetic analysis based on genome-wide core genes and 16S rRNA gene sequences revealed that strain RBPA9 clustered with Pseudomonas stutzeri ATCC 17588 T in both the phylogenetic trees. Maximum growth was recorded at 200 mg /L concentration of phenol which was consumed within 24 h. A gene cluster of phenol degradation pathway was detected. The quantitative real-time PCR (RT-PCR) demonstrated the expression of all the genes required in the meta-cleavage pathway of phenol in RBPA9. Our results reveal that strain RBPA9 represents a novel species for which Pseudomonas phenolilytica sp. nov. is proposed. The type strain is RBPA9T (= TBRC 15231 T = NBRC 115284 T).


Cleavage of fibrinogen by proteinases elicits allergic responses through Toll-like receptor 4.
Proteinases and the innate immune receptor Toll-like receptor 4 (TLR4) are essential for expression of allergic inflammation and diseases such as asthma. A mechanism that links these inflammatory mediators is essential for explaining the fundamental basis of allergic disease but has been elusive. Here, we demonstrate that TLR4 is activated by airway proteinase activity to initiate both allergic airway disease and antifungal immunity. These outcomes were induced by proteinase cleavage of the clotting protein fibrinogen, yielding fibrinogen cleavage products that acted as TLR4 ligands on airway epithelial cells and macrophages. Thus, allergic airway inflammation represents an antifungal defensive strategy that is driven by fibrinogen cleavage and TLR4 activation. These findings clarify the molecular basis of allergic disease and suggest new therapeutic strategies.


Effect of intracoronary captopril on coronary blood flow and regional myocardial function in dogs.
To evaluate the cardiac effect of an inhibitor of angiotensin-converting enzyme, the effect of intracoronary (i.c.) captopril on coronary blood flow and regional myocardial function was examined in the anesthetized open-chest dog. Blood flow of the left circumflex coronary artery (LCX), left ventricular pressure (LVP), aortic pressure (AoP) and regional myocardial segment length were measured continuously. Captopril i.v. (0.3 mg/kg) produced an immediate reduction in AoP and an increase in percent shortening of myocardial segments followed by a decrease in coronary vascular resistance and increases in heart rate and LVdP/dt. Reductions in LCX flow induced by i.c. angiotensin were attenuated and i.c. bradykinin-induced increases in LCX flow were augmented after captopril. On the contrary, i.c. infusion of captopril (0.01 mg/min) into the LCX caused no change in hemodynamic variables and myocardial shortening although responses to angiotensin I and bradykinin were markedly modified. These results suggest that captopril may have no direct cardiac effect.


A comparison of two functional tests in quadriplegia: the quadriplegia index of function and the functional independence measure.
Individuals with spinal cord injury are evaluated according to a set of guidelines based on motor, sensory, and functional tests. The resulting scores are used to quantify the extent of neurological injury and functional loss. The purpose of the present study was to compare certain scoring systems using the same group of patients. Twenty-nine subjects with cervical spine cord injury were evaluated by the same examiner using three scales: (1) The American Spinal Cord Injury Association (ASIA) (2) The Quadriplegia Index of Function (QIF) (3) The Functional Independence Measure (FIM) Assessments were made both at admission to, and discharge from, the rehabilitation center. Positive change in motor score is widely used as an indicator of recovery after spinal cord injury. We assessed the relationship of the two functional tests, the FIM and the QIF, to ASIA scores and found strong correlations in both cases. The feeding and dressing categories of QIF showed an even stronger correlation to ASIA motor scores, though the statistical significance was the same for corresponding categories of FIM and QIF. The percent of recovery on ASIA motor scores was significantly correlated only to gain in QIF scores, not FIM. FIM lacks the category of bed activities. Some additions to the FIM may be useful, especially in the feeding and dressing categories, and a category of bed activities could be included as well, in order to improve sensitivity.


Autophagy inhibitors reduce avian-reovirus-mediated apoptosis in cultured cells and in chicken embryos.
Avian reovirus (ARV)-induced apoptosis contributes to the pathogenesis of reovirus in infected chickens. However, methods for effectively reducing ARV-triggered apoptosis remain to be explored. Here, we show that pretreatment with chloroquine (CQ) or E64d plus pepstatin A decreases ARV-mediated apoptosis in chicken DF-1 cells. By acting as autophagy inhibitors, CQ and E64d plus pepstatin A increase microtubule-associated protein 1 light chain 3-II (LC3II) accumulation in ARV-infected cells, which results in decreased ARV protein synthesis and virus yield and thereby contributes to the reduction of apoptosis. Furthermore, ARV-mediated apoptosis in the bursa, heart and intestines of chicken embryos is attenuated by CQ and E64d plus pepstatin A treatment. Importantly, treatment with these autophagy inhibitors increases the survival of infected chicken embryos. Together, our data suggest that pharmacological inhibition of autophagy might represent a novel strategy for reducing ARV-mediated apoptosis.


A new production route of Gadolinium-148 for use in Radioisotope Thermoelectric Generators.
The production of Gd-148 had challenges for available methods in the literature, such as its high cost and low production amount. Therefore, we recommend a new production route of Gd-148 on natural Sm and Eu targets via particle accelerators. For this aim, we calculated and simulated cross-section, activity, the yield of product, and integral yield curves for 21 different nuclear reaction processes under certain conditions. Based on the obtained results, we proposed the radioisotope Gd-148 to use Radioisotope Thermoelectric Generators for deep space and planetary explorations in spacecraft and space-probes as a suitable energy source, instead of Pu-238.


Targeted next generation sequencing of pancreatic solid pseudopapillary neoplasms show mutations in Wnt signaling pathway genes.
Solid pseudopapillary neoplasms of the pancreas are rare neoplasms that have been shown to harbor recurrent somatic pathogenic variants in the beta-catenin gene, CTNNB1. Here, we used targeted next generation sequencing to analyze these tumors for other associated mutations. Six cases of solid pseudopapillary neoplasms were studied. DNA extracted from formalin-fixed paraffin embedded tissue blocks was analyzed using the Ion Torrent platform, with the 50-gene Ampliseq Cancer Hotspot Panel v2 (CHPv2), with further variant validation performed by Sanger sequencing. Four tumors (67%) were confirmed to harbor mutations within CTNNB1, two with c.109T > G p.(Ser37Ala) and two with c.94G > A p.(Asp32Asn). One case showed a frameshift deletion in the Adenomatous Polyposis Coli gene, APC c.3964delG p.(Glu1322Lysfs*93) with a variant allele frequency of 42.6%. Sanger sequencing on non-tumoral tissue confirmed the variant was somatic. The patient with the APC mutation developed metastasis and died. In addition to the four cases harboring CTNNB1 variants, we found a case characterized by poor outcome, showing a rare frameshift deletion in the APC gene. Since the APC product interacts with beta-catenin, APC variants may, in addition to CTNNB1, contribute to the pathogenesis of solid pseudopapillary neoplasms via the Wnt signaling pathway.


The potential biomarkers for the formation and development of intracranial aneurysm.
This study is aimed to understand the pathogenesis of intracranial aneurysm (IA), which has a risk of rupture and is the primary cause of subarachnoid hemorrhage. From Gene Expression Omnibus (GEO) database, GSE75436 was extracted (15 IA tissues and 15 superficial temporal artery tissues). The differentially expressed genes (DEGs) was conducted through limma package, which followed by the enrichment analysis. Combining STRING database, protein-protein interaction (PPI) network was constructed. The modules in PPI network were performed utilizing molecular complex detection (MCODE) algorithm. With Cytoscape software, the transcription factor-miRNA-target regulatory network was constructed. Finally, microarray dataset GSE54083 was downloaded (13 IA tissues and 10 superficial temporal artery tissues) for the verification test. A total of 1332 DEGs were screened in IA tissues compared with superficial temporal artery tissues. Besides, the up-regulated TNF, IL10, IL1B, and CTSS, as well as down-regulated IL6 were included in the top 20 nodes in the PPI networks. Furthermore, in the module A of up-regulated PPI network, TNF, IL10, IL1B, and VCAM1 were interact with each other. In the regulatory network, miR-29A/B/C targeted up-regulated genes. Besides, VCAM1 was implicated in the pathway of leukocyte transendothelial migration. In the verification analysis, between GSE75436 and GSE54083, there were 444 up-regulated and 543 down-regulated co-existence DEGs and 11 co-existence genes involved the Leukocyte transendothelial migration pathway. VCAM1, TNF, CTSS, IL10, IL1B, IL6, and miR-29A/B/C might be the potential biomarkers for the formation and development of IA.


Biochemical and molecular characterization of the novobiocin and rifampicin resistant Aeromonas hydrophila vaccine strain AL09-71N+R compared to its virulent parent strain AL09-71.
To understand the fitness cost of novobiocin- and rifampicin-resistance in an attenuated Aeromonas hydrophiila vaccine strain AL09-71 N+R compared to its virulent parent strain AL09-71, colony size, cell size, cell proliferation rate, chemotactic response, and the ability to invade catfish gill cells of the two strains were compared. Our results revealed that: (1) the cell size and the colony size of AL09-71 N+R was significantly (P<0.05) smaller than that of AL09-71; (2) the proliferation rate of AL09-71 N+R was significantly (P<0.05) slower than that of AL09-71; (3) AL09-71 N+R had a significantly (P<0.05) lower chemotactic response to catfish mucus than that of AL09-71; 4) the ability of AL09-71 N+R to invade catfish gill cells was significantly (P<0.05) lower than that of AL09-71. To understand whether target site mutation might play a role in antibiotic resistance, novobiocin's target site DNA gyrase subunit B gyrB and rifampicin's target site RNA polymerase subunit B rpoB were sequenced from the two strains. Our results revealed the following five mutations: (1) two missense mutations (CGC to ATC resulting in arginine/R to serine/S; TAC to TGC resulting in tyrosine/Y to cysteine/C) between AL09-71 gyrB and AL09-71 N+R gyrB; (2) three missense mutations (GAC to AAC resulting in aspartic acid/D to asparagine/N; CTG to CCG resulting in leucine/L to proline/P; CTG to CCG resulting in leucine/L to proline/P) between AL09-71 rpoB and AL09-71 N+R rpoB. To determine whether any unique DNA sequences were present in AL09-71 but absent in AL09-71 N+R, PCR-select bacterial genome subtractive hybridization was performed. Of 96 clones selected from the subtractive genomic DNA library, 32 sequences were found. None of the 32 sequences was confirmed to be present in AL09-71 but absent in AL09-71 N+R. At the transcription level, 29 of the 32 genes were found to be expressed greater than 10-fold in AL09-71 N+R compared to that in AL09-71.


Electrochemical and Photoelectrochemical Water Oxidation for Hydrogen Peroxide Production.
Hydrogen peroxide (H2 O2 ), as a green fuel and oxidant, has drawn increasing attention in the energy and environmental research. Compared with the traditional anthraquinone process, the electrochemical (EC) and photoelectrochemical (PEC) syntheses of H2 O2 are cost-effective and environmentally friendly. In order to construct membraneless EC/PEC devices for the full H2 O2 synthesis, anodic H2 O2 production by water oxidation, which is less developed than cathodic H2 O2 generation, is highly desirable. Here, we review recent developments for the EC/PEC H2 O2 production by water oxidation, including fundamental aspects, benchmarking activity evaluation, material/catalyst selection, and strategies for increasing selectivity, efficiency, and accumulation. Furthermore, we discuss the challenges and outlook of water oxidation for H2 O2 production, especially device-level development, accumulation and stability, and industrial applications. Our review is intended to stimulate studies further improving EC/PEC H2 O2 production.


A radiometric assay for phenylethanolamine N-methyltransferase and catechol O-methyltransferase in a single tissue sample: application to rat hypothalamic nuclei, pineal gland, and heart.
A simple and highly sensitive method for simultaneous assay of phenylethanolamine N-methyltransferase (PNMT) and catechol O-methyltransferase (COMT) is described. These enzymes are determined in a single tissue homogenate using S-[methyl-3H] adenosyl-L-methionine as methyl donor and sequentially incubating with the substrates phenylethanolamine and epinephrine. The radioactive products of the enzymatic reactions, N-methylphenylethanolamine and metanephrine, are extracted and then separated by thin-layer chromatography. The identity of the reaction products has been established chromatographically and the conditions for both enzymatic reactions in the assay procedure have been defined. Measurement of PNMT activity in the rat pineal gland or in minute fragments of other tissues (e.g., brain nuclei) has not been possible using previously described methods. Activities of PNMT and COMT in the rat pineal gland, various hypothalamic nuclei, and the auricular and ventricular myocardia are herein reported.


Antibodies against Streptococcus mutans and glucosyltransferases in caries-free and caries-active military recruits.
1) Plaque samples from caries-active recruits showed a higher incidence of S. mutans than plaque samples from caries-free recruits. The persistance of the S. mutans serotypes in the individual plaque samples attested to the consistency of S. mutans in human dental plaque. 2) S. mutans serotype d is predominantly present in approximal plaque of caries-active subjects. 3) Serum antibody titers against S. mutans are higher in caries-active individuals. 4) Sera from caries-free or caries-active individuals inhibit the insoluble glucan synthesis by the cellbound GTF to the same extent. 5) It is unlikely that antibodies against S. mutans in parotid saliva or serum play a major role in the protection against dental caries in Dutch military recruits.


Effects of Observational Learning, Overt Activity, and Class Placement on Children's Mediated Associative Learning.
The present study investigated the effects of instructional training on the generation and use of imaginal- or verbal-associative-learning mediators by third-grade children. Instructional training was based on modeling (active practice) or observation of the generation of associative-learning mediators. Ss (N = 40) served both as models (active generators) and observers of mediated associative-learning responses, alternating on successive trials of the task. Verbal-mediation responding resulted in significantly more effective mediator use and recall performance than did imagery or control responding; this effect interacted with class placement. No difference was found between the modeling and observing conditions, indicating that both are effective in promoting mediated associative learning in young children.


Taurocholate increases the biliary output of bilirubin pigments in the rat, due to haemolysis.
In order to investigate the effect of bile salts on the biliary transport of bilirubin, we infused rats with taurocholate at 1-3 mumol/min/kg body weight. When administered in saline, taurocholate induced haemolysis, and a dose-dependent increase in biliary excretion and serum concentration of all bilirubin pigments. The addition of human or rat serum albumin to the infusate, at an albumin:taurocholate molar ratio equal to or above 200, completely prevented haemolysis and the above-mentioned alterations in bilirubin transport, without any change in the bile acid output. In order to mimic a haemolytic condition, an erythrocyte haemolysate was further added to the taurocholate-albumin solution. This again resulted in an increased bilirubin output. We suggest that taurocholate treatment increases bilirubin output due to the increased load of the pigment on the liver, as a result of the bile acid-induced haemolysis.


Studies on the nature and pressure evolution of phase transitions in 1-adamantylamine and 1-adamantanol.
In this paper, several experimental techniques, i.e., differential scanning calorimetry, X-ray diffraction, Fourier transform infrared, Raman, and broadband dielectric spectroscopy were applied to study the nature of the phase transitions in 1-adamantylamine (1-NH2-ADM, C10H17N) and 1-adamantanol (1-OH-ADM, C10H16O). Calorimetric measurements showed one and three endothermic peaks in thermograms for the latter and the former substance, respectively. Indeed, results of spectroscopic investigations indicated that the observed thermal events in 1-NH2-ADM correspond to transitions between various plastic crystal (PC) phases (I, II, III, IV), while the endothermic process in 1-OH-ADM can be assigned to a phase transition between the PC and the ordinary crystal (OC). Especially interesting were the outcomes of dielectric studies carried out both at ambient and high-pressure conditions, during heating and cooling cycles. They showed: i) noticeable changes in the frequency dependencies of the imaginary (ε'') and real (ε') parts of the complex dielectric permittivity that occurred around temperatures of the characteristic endothermic events detected by the calorimetry, and ii) significant fluctuations of ε'' and ε' at pressures attributed to the respective phase transitions. Moreover, the pressure coefficients of the phase transition temperatures were estimated to be approximately equal to 0.2 K/MPa for both compounds. In turn, volume variation (ΔV) at the PC (II)-PC (III) and PC (III)-PC (IV) transition temperatures for 1-NH2-ADM was essentially different than ΔV for the PC-OC transition in 1-OH-ADM.


Stimulus-specific regulation of visual oddball differentiation in posterior parietal cortex.
The frequency at which a stimulus is presented determines how it is interpreted. For example, a repeated image may be of less interest than an image that violates the prior sequence. This process involves integration of sensory information and internal representations of stimulus history, functions carried out in higher-order sensory areas such as the posterior parietal cortex (PPC). Thus far, there are few detailed reports investigating the single-neuron mechanisms for processing of stimulus presentation frequency in PPC. To address this gap in knowledge, we recorded PPC activity using 2-photon calcium imaging and electrophysiology during a visual oddball paradigm. Calcium imaging results reveal differentiation at the level of single neurons for frequent versus rare conditions which varied depending on whether the stimulus was preferred or non-preferred by the recorded neural population. Such differentiation of oddball conditions was mediated primarily by stimulus-independent adaptation in the frequent condition.


Specific expression patterns of epithelial to mesenchymal transition factors in gestational molar disease.
The epithelial to mesenchymal transition, a well-known and re-emerging model in pathology, has not been completely investigated in the field of gestational pathology. This study aims at improving the comprehension of this process in molar disease, even looking for new possible immunohistochemical markers. We have analysed the immunohistochemical expression of Twist1 and Snai2, two of the most important transcription factors involved in epithelial to mesenchymal transition, in formalin-fixed paraffin-embedded samples of 23 spontaneous abortive pregnancies, 22 molar pregnancies (10 partial and 12 complete) and 7 term placentas. Twist1 and Snai2 were highly expressed in stromal villi cells of molar disease. Particularly, Twist1 was highly expressed in complete moles compared to both abortive pregnancies (p < 0.001) and partial moles (p < 0.05). Also Snai2 was more expressed by complete moles, differentiating them from non-molar abortions (p < 0.05). On the basis of the known cadherins and claudins expression in these pathologies, our new findings reinforce the hypothesis of the involvement of epithelial to mesenchymal transition in early molar pregnancies and above all in complete moles. Furthermore, we highlighted that in molar disease not only the trophoblast, but even the villi stromal cells, are involved. Thanks to their specificity, furthermore, these Twist1 and Snai2 could be used as additional immunohistochemical tool in the diagnosis of complete molar disease, with Twist1 as the first choice.


Gender differences in hepatic phylloquinone and menaquinones in the vitamin K-deficient and -supplemented rat.
Gender differences in relation to vitamin K were investigated in the rat. Hepatic phylloquinone and menaquinone (MK-1 to MK-10) concentrations, gamma-carboxyglutamic acid (Gla) excretion, plasma phylloquinone and percent prothrombin were measured in male and female rats on a chow diet (24.5 ng phylloquinone and 8.8 microgram menadione), and on phylloquinone-deficient and -supplemented purified diets (0.38 and 1400 ng phylloquinone/g, respectively). Mean hepatic phylloquinone concentrations varied with dietary intake and ranged from 6.8+/-9.0 pmol/g in the deficient male, to 171. 1+/-56.9 pmol/g in the supplemented female. Menaquinones accounted for a large proportion of total vitamin K in the liver of males and females with MK-4, MK-6, and MK-10 present in highest concentrations. On the chow and supplemented diets, females had significantly higher MK-4, MK-6, and MK-10 concentrations in their livers (P<0.05). On the phylloquinone-deficient diet (-K1), hepatic phylloquinone, MK-4, and to a lesser extent MK-6 (but not MK-10) were significantly reduced (P<0.05). In the phylloquinone-supplemented male and female groups, which did not receive menadione during the experimental period, MK-4 increased above that in the chow groups suggesting synthesis of MK-4 from phylloquinone which was statistically significant in the female (P<0.01). A significant gender difference (P<0.05) was also observed for urinary Gla excretion with less Gla excreted by the females indicating that females may require less dietary phylloquinone than males of the same body weight.


Prognostic significance of neutrophil-lymphocyte ratio and carbohydrate antigen 19-9 in patients with gallbladder carcinoma.
The neutrophil-lymphocyte ratio (NLR) is an immune response-related indicator and it is associated with poor prognosis of various cancers. The carbohydrate antigen19-9 (CA19-9) is a tumor-associated antigen and it has prognostic relevance in gallbladder carcinoma (GBC). We aimed to analyze whether preoperative NLR and serum CA19-9 were associated with outcomes of GBC patients after surgery with curative intent.Between January 2010 and May 2015, 90 resectable GBC patients who underwent curative surgery in our institution were included. All final diagnoses were confirmed by pathologic examination. The demographics, clinical, and histopathology data were analyzed. The Cox regression proportional hazard model and Kaplan-Meier method were used to assess prognostic factors.The cutoff values of 4.33 and 250.90 U/mL were defined as high NLR and high CA19-9, respectively. The univariate analyses showed that TNM stage, lymph node metastasis, the degree of tumor differentiation, margin status, combined hepatectomy, CA19-9, NLR, and PNI were all associated with overall survival (P < .05). According to the multivariable analysis, NLR (hazard ratio (HR) 3.840, 95% confidence interval (95% CI): 2.122-6.947, P < .001), CA19-9 (HR 2.230, 95% CI: 1.297-3.835, P = .004), TNM stage (HR 3.864, 95% CI: 1.819-8.207, P < .001), lymph node metastasis (HR 1.679, 95% CI: 1.005-2.805, P = .048), and margin status (HR 1.873, 95% CI: 1.063-3.300, P = .030) were independent prognostic factors. The median survival time in low NLR and CA19-9 group was better than high NLR and CA19-9 group (P < .05).The preoperative NLR and serum CA19-9 are associated with prognosis of patients with GBC. High NLR and high CA19-9 were predictors of poor long-term outcome among patients with GBC undergoing curative surgery.


The use of monoclonal antibodies in demonstrating the effect of antibody heterogeneity on immune complex size.
The effect of antibody heterogeneity on immune complex size was investigated by using monoclonal antibodies. Five monoclonal antibodies which bind different antigenic determinants of human serum albumin were used to produce immune complexes. When one or two different monoclonal antibodies were used, under antigen-excess, antigen-antibody equivalent, and antibody-excess conditions, hardly any immune complexes larger than mouse IgM were produced. On the other hand, in a representative experiment, when five different monoclonal antibodies were used, immune complexes larger than mouse IgM comprised 21.5% of the total antigen under antigen-antibody equivalent conditions, 18.2% under antibody-excess conditions, and 4.3% under antigen-excess conditions. These results indicate that antibody heterogeneity is one of the important factors determining immune complex size.


Nellore cattle (Bos indicus) and ticks within the Brazilian Pantanal: ecological relationships.
Pantanal is a huge floodplain mostly in Brazil, and its main economic activity is extensive cattle raising, in farms characterized by an extremely wildlife-rich environment. We herein describe tick infestations of cattle and of the natural environment in Pantanal of Nhecolândia in Brazil, at areas with and without cattle during both dry and wet seasons. Environmental sampling resulted in three tick species: Amblyomma sculptum (423 nymphs and 518 adults), Amblyomma parvum (7 nymphs and 129 adults), Amblyomma ovale (3 adults) as well as three clusters and two individuals of Amblyomma sp. larvae. A significantly higher number of adult A. sculptum ticks was found in areas with cattle in the wet season. From 106 examinations of bovines 1710 ticks from three species were collected: Rhipicephalus microplus (55.7% of the total), A. sculptum (38%) and A. parvum (4.1%), as well as 32 Amblyomma sp. larvae. A significant similarity was found between Amblyomma tick fauna from environment and on cattle during both seasons. All A. sculptum females on bovines were flat whereas many of A. parvum females and A. sculptum nymphs were engorging. Although R. microplus was the most abundant tick species on cattle, overall highest tick prevalence on bovines in the dry season was of A. sculptum nymphs. Lack of R. microplus in environmental sampling, relationship between cattle and increase in adult A. sculptum numbers in the environment as well as suitability of bovine for the various tick species are discussed.


Effects of the noradrenaline neurotoxin DSP 4 on monoamine neurons and their transmitter turnover in rat CNS.
Regional effects of DSP 4 on monoamine neurons have been analyzed by chemical assay of endogenous monoamines and their metabolites in rat CNS. The results confirmed that the neurotoxic action of DSP 4 is predominantly on noradrenaline nerve terminal projections originating from locus coeruleus, with the most marked effects on terminal fields localized most distant from the noradrenaline perikarya. DSP 4 treatment (10 days) caused no alteration of the regional DA levels, except in cingulate cortex, where a moderate increase (+40%) was observed, possibly at least partially related to a sprouting of dopamine nerve terminals following the noradrenaline denervation. 5-hydroxytryptamine levels were generally unaltered after DSP 4, except for an about 10-25% reduction in cerebral cortex and hippocampus. There was with time a certain noradrenaline recovery, most likely related to regeneration of noradrenaline nerve terminals, although this process was relatively slow (months). Analysis of catecholamine decline after tyrosine hydroxylase inhibition and metabolite/monoamine ratios, as indices for transmitter utilization rate, indicated an increased noradrenaline turnover in terminals spared by DSP 4, while dopamine turnover appeared to be reduced in many regions (i.a. cerebral cortex, striatum, accumbens, olfactory tubercle and spinal cord), most pronounced in cingulate cortex. The results indicate that noradrenaline neurons have a facilitatory action on dopamine neurons. The DSP 4 treatment did not cause any significant effect on 5-hydroxytryptamine turnover in any of the individual regions analyzed.


Correlation between percentage of brain parenchymal volume and neurocognitive performance in HIV-infected patients.
This study was designed to determine whether neuropsychological function in HIV-infected persons is correlated with loss of brain volume (as measured by percentage of brain parenchymal volume [PBV]). We hypothesized that whole-brain parenchymal volume might correlate with neuropsychologic performance, even before overt clinical dysfunction is apparent. A computer-assisted segmentation technique with thin section MR imaging was used for 15 patients with HIV infection (seven symptomatic, eight asymptomatic) and for five HIV-negative control participants to quantify whole brain and CSF volumes. To determine the degree of brain atrophy, the PBV relative to that of intracranial content was calculated. Neuropsychological performance was assessed by using a standard battery of eight tests (NPZ-8 test battery). HIV-infected patients had significantly lower NPZ-8 scores (t[18] = 2.26, P <.05) and lower PBV (t[18] = 1.79, P <.01) than those of healthy control participants. With the Spearman rank order correlation coefficients, data analyzed for all 20 study participants (15 HIV-infected patients and five noninfected control participants) showed a significant (r = -0.50, P <.05) negative correlation between PBV and NPZ-8 test battery score. In addition, there was a significant negative correlation between subtest score of motor impairment and PBV (r = -0.69, P <.01) and between AIDS dementia complex score (r = -0.64) and PBV (P <.01). These correlations suggest that quantitation of PBV may offer an objective, easily acquired surrogate predictor of neuropsychological impairment and clinically apparent cognitive/motor dysfunction among HIV-infected persons.


Repeat instability: mechanisms of dynamic mutations.
Disease-causing repeat instability is an important and unique form of mutation that is linked to more than 40 neurological, neurodegenerative and neuromuscular disorders. DNA repeat expansion mutations are dynamic and ongoing within tissues and across generations. The patterns of inherited and tissue-specific instability are determined by both gene-specific cis-elements and trans-acting DNA metabolic proteins. Repeat instability probably involves the formation of unusual DNA structures during DNA replication, repair and recombination. Experimental advances towards explaining the mechanisms of repeat instability have broadened our understanding of this mutational process. They have revealed surprising ways in which metabolic pathways can drive or protect from repeat instability.


Noninvasive assessment of autonomic modulation of heart rate variability in the Ts65Dn mouse model of Down syndrome: A proof of principle study.
The Ts65Dn mouse is the most widely used animal model of Down syndrome (DS). Differences in autonomic regulation of heart rate variability (HRV) in individuals with DS have been hypothesized. Pharmacological studies in animal models may help us understand mechanisms underlying observed changes in HRV in people with DS. To investigate the use a new, noninvasive technique to assess cardiac autonomic modulation in Ts65Dn mice under the effect of adrenergic and cholinergic agonists. We recorded electrocardiograms (ECGs) from 12 Ts65Dn and 12 euploid control mice. A 30-min baseline recording was followed by the injection of an adrenergic (isoproterenol [Iso]) or cholinergic (carbachol [CCh]) agonist. Heart rate and HRV were analyzed using a series of methods customized for mice. The ECG apparatus described here allowed us to detect noninvasively long series of heartbeats in freely-moving animals. During baseline conditions, the yield of detectable heartbeats was 3%-27% of the estimated total number of events, which increased to 35%-70% during the 15-min period after either Iso or CCh injections. Ts65Dn mice displayed a robust enhanced Iso-induced negative chronotropic rebound response compared with euploid control mice. We observed a significantly smaller CCh response in Ts65Dn versus control euploid mice in the 6- to 10-min-interval postcarbachol injection. This work showed that the techniques described here are sufficient for this type of study. However, future studies involving the use of more selective pharmacological agents and/or genetic manipulations will be key to advance a mechanistic understanding of cardiac autonomic regulation in DS.


Carcinoma of the bladder - radiotherapy.
Differentiated transitional cell carcinoma responded better to radiotherapy than anaplastic or squamous cell carcinoma. Five year survival was higher in low stage than in high stage carcinoma, and higher in papillary than in solid carcinoma. Tumour control, but also complication rate, increased with higher radiation dose. With the introduction of a two volume irradiation technique, a large tumour dose can be given with a minimum of complications.


Prolactin and growth hormone secretion during long-term incubation of the pituitary pars distalis of mature chum salmon, Oncorhynchus keta.
The effects of hypotonicity of the medium on prolactin (PRL) and growth hormone (GH) release from the pars distalis (PD) of mature chum salmon pituitary were examined during culture for 4 days. Large amounts of PRL and GH were released during the first 6 hr and secretion decreased rapidly thereafter. The amounts of GH released as well as the residual content in PD were 10 times greater than those of PRL. There was no difference in PRL or GH release between the PD incubated in isotonic medium (325 mOsm) and those in hypotonic medium (250 mOsm) even during the first 6 hr. Female PD secreted more PRL than male PD during the first 6 hr, whereas a sex difference was not observed in GH release. Considerable amounts of both PRL and GH remained in the PD at the end of the culture period. The amount of PRL released during 4 days of culture from female PD (37-39%) was greater than that from male PD (23-26%), and the amount of GH released was less in females (28-32%) than that in males (53-54%). Release of PRL and GH appears to be mainly under stimulatory hypothalamic control in the mature chum salmon.


Metabolomics profiling of Polygoni Multiflori Radix and Polygoni Multiflori Radix Preparata extracts using UPLC-Q/TOF-MS.
The side effects caused by Polygoni Multiflori Radix (PMR) and Polygoni Multiflori Radix Praeparata (PMRP) have often appeared globally. There is no research on the changes of endogenous metabolites among PMR- and PMRP-treated rats. The aim of this study was to evaluate the varying metabolomic effects between PMR- and PMRP-treated rats. We tried to discover relevant differences in biomarkers and endogenous metabolic pathways. Hematoxylin and eosin staining and immunohistochemistry staining were performed to find pathological changes. Biochemical indicators were also measured, one-way analysis of variance with Dunnett's multiple comparison test was used for biochemical indicators comparison among various groups. Metabolomics analysis based on ultra-high performance liquid chromatography-quadrupole time of flight mass spectrometry (UPLC-Q/TOF-MS) was performed to find the changes in metabolic biomarkers. Multivariate statistical approaches such as principal component analysis (PCA) and orthogonal partial least square-discriminant analysis (OPLS-DA) were applied to reveal group clustering trend, evaluate and maximize the discrimination between the two groups. MetaboAnalyst 4.0 was performed to find and confirm the pathways. PMR extracts exhibited slight hepatotoxic effects on the liver by increasing aspartate and alanine aminotransferase levels. Twenty-nine metabolites were identified as biomarkers, belonging to five pathways, including alpha-linolenic acid metabolism, taurine and hypotaurine metabolism, glycerophospholipid metabolism, arginine and proline metabolism, and primary bile acid biosynthesis. This study provided a comprehensive description of metabolomic changes between PMR- and PMRP-treated rats. The underlying mechanisms require further research.


A cross-sectional study examining multiple mobility device use and fall status among middle-aged and older adults with multiple sclerosis.
To document the prevalence of multiple mobility device use among adults with multiple sclerosis (MS) (≥ 55 years) and examine the association between falls status (faller/non-faller) and the number of mobility devices used. Cross-sectional data generated through telephone interviews with 353 participants was used for this secondary analysis. Descriptive statistics were used to address the first study purpose. Multiple device use was measured by the number of devices used, which ranged from 0 (never use a cane, walker, manual wheelchair, or power wheelchair/scooter) to 4 (use all four mobility devices at least some of the time). Logistic regression analysis was used to address the second purpose, with fall status used as the dependent variable (non-fallers [<1 per year] versus fallers [≥ 1 per year]). Just under 60% of participants reported the use of at least two mobility devices. For each additional mobility device used, the odds of being a faller increased by 1.47 times (95% CI = 1.14-1.90). Multiple mobility device use was common and the greater number of devices used, the greater the likelihood of being a faller. To prevent falls, this association requires further research to determine directionality.