A novel variant in the FBP1 gene causes fructose-1,6-bisphosphatase deficiency through increased ubiquitination.
Fructose-1,6-bisphosphatase (FBPase) deficiency is an autosomal recessive disorder characterized by impaired gluconeogenesis caused by mutations in the fructose-1,6-bisphosphatase 1 (FBP1) gene. The molecular mechanisms underlying FBPase deficiency caused by FBP1 mutations require investigation. Herein, we report the case of a Chinese boy with FBPase deficiency who presented with hypoglycemia, ketonuria, metabolic acidosis, and repeated episodes of generalized seizures that progressed to epileptic encephalopathy. Whole-exome sequencing revealed compound heterozygous variants, c.761 A > G (H254R) and c.962C > T (S321F), in FBP1. The variants, especially the novel H254R, reduced protein stability and enzymatic activity in patient-derived leukocytes and transfected HepG2 and U251 cells. Mutant FBP1 undergoes enhanced ubiquitination and proteasomal degradation. NEDD4-2 was identified as an E3 ligase for FBP1 ubiquitination in transfected cells and the liver and brain of Nedd4-2 knockout mice. The H254R mutant FBP1 interacted with NEDD4-2 at significantly higher levels than the wild-type control. Our study identified a novel H254R variant of FBP1 underlying FBPase deficiency and elucidated the molecular mechanism underlying the enhanced NEDD4-2-mediated ubiquitination and proteasomal degradation of mutant FBP1.


The effects of low electromagnetic field and lead acetate combination on some hemato-biochemical and immunotoxicological parameters in mice.
The present study was carried out to investigate the potential effects of extremely low-frequency electromagnetic fields (ELF-EMF) and lead acetate on some hemato-biochemical, immune and pathologic variables in mice. A total of 90 female mice were equally divided into six groups. (Gp. 1) kept as control, (Gp. 2) exposed to EMF of 2 millitesla (mT) intensity and 50 Hz frequency (4h/day) for 30 days, (Gps. 3 and 4) were administered lead acetate orally at doses 1 and 5 mg/kg BW, respectively for 30 days. The last 2 groups (5, 6) were exposed to EMF- lead combination for the same period. EMF exposure induced a significant increase in RBCs (p<0.001), WBCs (p<0.01) and platelets (p<0.001) counts, compared to control. However, anemia and leukopenia were recorded with oral administration of Pb acetate. The phagocytosis % and phagocytic index were significantly (p<0.05) increased in mice exposed to EMF for 30 days, but decreased (p<0.01) in the animals given the highest dose of lead. Comparing to unexposed mice, significant variation in biochemical parameters (glucose, enzymes, and protein profiles) were noticed. Combined lead and EMF treatments had antagonizing effect on some previous parameters, whereas mice given the highest dose of lead with EMF aggravated hemato-biochemical and pathological findings. We concluded that combined exposure to ELF-EMF and Pb acetate produced significant changes in the hemato-biochemical and immune parameters which were both real and inconsistent.


Exposure assessment of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) in temporary municipal-waste-incinerator maintenance workers before and after annual maintenance.
Polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) have been found in ambient air around municipal waste incinerators (MWIs), and elevated serum levels in incinerator workers have also been observed in some studies. However, few studies have focused on temporary employees who work intermittently through the annual maintenance and clean-up around different incinerators. The present study aimed to assess the change in serum PCDD/F levels of temporary employees between the beginning of periodic incinerator maintenance and one month the work was completed. Thirty-five volunteer workers, most of them transient and temporary maintenance staff, were recruited from a contractor that provided annual maintenance for four incinerators in this study. Information about each participant was obtained by questionnaire at the beginning of annual maintenance. The questionnaire asked for work history, health status, and diet information. As measured by the PCDD/F levels in blood, a significant increase was observed in workers after a month of maintenance work. The increase was greater in workers who had never done this type of maintenance than in those with previous experience, especially for 2,3,4,6,7,8-HxCDF levels. The data also showed that the laborers and employers need to pay more attention to occupational health issues even for short-term incinerator maintenance workers.


Speciation of magnesium in rat plasma using capillary electrophoresis-inductively coupled plasma-atomic emission spectrometry.
A new method for speciation analysis of magnesium species and quantification of free magnesium concentrations in rat plasma was developed by on-line coupling of CE with inductively coupled plasma-atomic emission spectrometry (ICP-AES). Baseline separation of seven magnesium species was achieved by using a 120 cm (100 microm internal diameter) fused-silica capillary, a 20 kV separation voltage and a solution of 50 mmol/L NaAc-HAc (pH 5.5) as electrolyte buffer. CE-ICP-AES analysis of a rat plasma sample showed the presence of seven magnesium species, one of which was identified as free Mg2+ ion by spiking a Mg2+ standard; the migration time of the Mg2+ peak in the standard and the spiked sample matched with each other. One protein-bound magnesium species in rat plasma is associated with albumin, and the other three species are combined with globulin. The concentration of free magnesium in the plasma was 14.0 mg/L. The other six magnesium species were estimated to be 4-15 mg/L. RSDs of migration time and peak area for the magnesium species from ten replicates were less than 5%. The developed method was also applied to speciation analysis of magnesium species in spiked plasma samples. The recoveries of the free magnesium species in four samples ranged from 95.8 to 103.8%.


Rhizobial symbiosis alleviates polychlorinated biphenyls-induced systematic oxidative stress via brassinosteroids signaling in alfalfa.
The role of symbiotic rhizobia in the alleviation of polychlorinated biphenyl (PCB)-induced phytotoxicity in alfalfa and the brassinosteroid (BR) hormone signaling involved were investigated during phytoremediation. The association between alfalfa and Sinorhizobium meliloti was adopted as a remediation model. Phytotoxicity due to PCB 77 (3,3',4,4'-tetrachlorobiphenyl) exerted adverse impacts on plant performance (biomass accumulation and photosynthesis) and elicited cellular oxidative stress (overproduction of reactive oxygen species, lipid peroxidation, and cell necrosis) which was largely attenuated by pre-inoculation with S. meliloti strain NM. The protective role may have been achieved as a result of strengthening of basic antioxidant defense before stress as evidenced by the augmented activity and gene expression of antioxidative enzymes (peroxidase, glutathione reductase, superoxide dismutase, catalase, and ascorbate peroxidase) of both leaves and roots. In nodulated seedlings peroxidase showed additive increased activity following PCB exposure but the activities of the other four enzymes tended to remain stable after stress. Furthermore, application of strain NM and brassinolide both triggered the accumulation of endogenous BRs and the antioxidant network, while pre-treatment of seedlings with a biosynthetic inhibitor of BRs, brassinazole, abolished the rhizobia-induced activation of detoxification responses towards PCB. These observations indicate that association with S. meliloti NM enhanced the systemic antioxidant defenses of alfalfa to detoxify PCB, at least in part, via BR-dependent signaling pathways. These results contribute to our knowledge of the 'logistic role' played by rhizobia in assisting the phytoremediation of PCB-contaminated soils and suggest an optimum manipulation strategy for bioremediation.


A novel method for intraoperative osseomechanical strength measurements: a biomechanical ex vivo evaluation on proximal femora.
The increasing number of geriatric traumatology cases has intensified the need to reliably and objectively evaluate local bone quality, the latter poses a decisive factor for the choice of an optimal approach to treat osteoporotic fractures. Osteodensitometry imaging techniques are not routinely available in acute operative settings, nor do they provide objective information on local bone properties specifically needed for the prognosis of implant stability. This study sought to verify ex vivo the feasibility and sensitivity of a novel method for the determination of local bone strength in the acute operative setting (intraoperative osseomechanical strength measurement; IOSM) that is based on the principle of material displacement resistance against the force of a rotary indenter. Samples consisted of human femoral heads obtained after total hip replacement. Comparisons were made with results obtained via conventional dual-energy X-ray absorptiometry (DXA) and quantitative computed tomography (qCT). Regression analyses of the results showed a highly significant correlation between the IOSM and the control methods (r = 0.61 and r = 0.56; p < 0.01), indicating that this new approach qualifies as a reliable tool for the intraoperative evaluation of the intrinsic local bone strength. The intraoperative integration of this method may support surgeon on taking proper decisions in terms of optimal surgical approaches and prevention of complications inherent to osteoporotic bone.


Antifungal potential of marine natural products.
Fungal diseases represent an increasing threat to human health worldwide which in some cases might be associated with substantial morbidity and mortality. However, only few antifungal drugs are currently available for the treatment of life-threatening fungal infections. Furthermore, plant diseases caused by fungal pathogens represent a worldwide economic problem for the agriculture industry. The marine environment continues to provide structurally diverse and biologically active secondary metabolites, several of which have inspired the development of new classes of therapeutic agents. Among these secondary metabolites, several compounds with noteworthy antifungal activities have been isolated from marine microorganisms, invertebrates, and algae. During the last fifteen years, around 65% of marine natural products possessing antifungal activities have been isolated from sponges and bacteria. This review gives an overview of natural products from diverse marine organisms that have shown in vitro and/or in vivo potential as antifungal agents, with their mechanism of action whenever applicable. The natural products literature is covered from January 2000 until June 2015, and we are reporting the chemical structures together with their biological activities, as well as the isolation source.


The role of homeobox protein distal-less 3 and its interaction with ETS2 in regulating bovine interferon-tau gene expression-synergistic transcriptional activation with ETS2.
Distal-less 3 (DLX3), a homeodomain transcription factor required for placental development in the mouse, modestly transactivates hCG-alpha subunit gene (hCGA) expression in human choriocarcinoma cells. Because hCG and interferon-tau (IFNT) are expressed in trophectoderm of primates and ruminants, respectively, we have tested the hypothesis that DLX3 regulates the genes for IFNT (IFNT). A bovine IFNT1 promoter (-457 to +66), linked to a luciferase (luc) reporter, was transactivated approximately 20-fold by overexpressing DLX3 in human JAr cells. Elimination of a potential DLX3-binding site (-54 GATAATGAG -46) by either truncation or mutagenesis abolished this effect. A sequence (-59 to -44) encompassing this site bound DLX3 specifically. Coexpression of DLX3 and ETS2, which is known to be a key regulator of IFNT expression, increased reporter activity by more than 250-fold, whereas deletion of the established ETS2 site (-79 to -70) eliminated the ability of DLX3 to transactivate the gene. Conversely, mutation of the DLX3 site significantly reduced the transactivational effects of ETS2. Both DLX3 and ETS2 are coexpressed in JAr cells and in an IFNT-producing, bovine trophoblast cell line, CT-1. The two can be immunoprecipitated together as a complex from CT-1 cells, and RNAi-mediated, partial knockdown of DLX3 expression reduced the production of IFNT by approximately 50+. Together, these results suggest that DLX3 has a central role in controlling IFNT gene expression by associating with ETS2 on the IFNT promoter.


Effects of insulin and sitagliptin on early cardiac dysfunction in diabetic rats.
Cardiac affection is common in diabetic patients. Although insulin exerts a cardioprotective role, it may not be enough to totally prevent this affection. The current study aimed to compare the cardioprotective effect of insulin alone or combined with sitagliptin in a rat model of type 1 diabetes mellitus. Diabetes was induced by a single intraperitoneal injection of streptozotocin (STZ; 60 mg/kg). Diabetic rats were treated with insulin (3 IU), insulin (6 IU), or insulin (3 IU) + sitagliptin (10 mg/kg) for 42 days. Diabetic rats exhibited significant systolic and diastolic cardiac affection with significant elevation of tumor necrosis factor α (TNF-α), interleukin 6 (IL-6) and brain natriuretic peptide (BNP) levels. Treatment with insulin prevented the deterioration of diabetes-induced cardiac condition, an effect that was significantly potentiated by the combined use of sitagliptin. The combined use of sitagliptin and insulin significantly improved the cardioprotective effect of insulin and prevented the early cardiac dysfunction in STZ diabetic rats.


Inflammatory Mechanisms in Hidradenitis Suppurativa.
Hidradenitis suppurativa (HS) is a chronic relapsing disease of follicular occlusion that causes immense clinical and psychosocial morbidity when refractory to treatment. HS is no longer considered a disease of primary infectious etiology, although bacteria play a role. There is increasing evidence that HS is associated with immune dysregulation, based on its clinical association with other immune-mediated disorders, by its response to biologic therapy in the clinical arena, and from molecular research. This article summarizes what is known in relation to the inflammatory pathways in HS.


Substitution of generic warfarin for Coumadin in an HMO setting.
Substitution of generic warfarin for Coumadin presents safety concerns due to warfarin's narrow therapeutic index and because a prior generic formulation was removed from the US market after it was associated with adverse events. To determine whether a health maintenance organization (HMO) can add generic warfarin to its formulary without adversely affecting warfarin management or increasing adverse events. In a prospective, observational study, an HMO that formerly dispensed only Coumadin added a generic warfarin preparation (Barr Laboratories, Pomona, NY) to its formulary. An anticoagulation service (ACS) affiliated with an HMO that was based in St. Louis, MO. The cohort consisted of 182 enrollees in the ACS as of May 1, 1999. At the start of the study, these participants were taking Coumadin; by October 31, 2000, all had switched to Barr warfarin. We collected data 8 months prior to and 10 months after the introduction of generic warfarin for the following endpoints: international normalized ratio (INR) control, frequency of INR monitoring, number of dose changes, and rate of thrombotic and hemorrhagic events. Statistical process control charts were used to differentiate between random variation in the endpoints and changes due to different warfarin formulations, and we used the Wilcoxon signed-rank test to look for a change in any endpoint after patients changed to generic warfarin. No significant differences were found in any endpoint. Substitution of Barr warfarin for Coumadin did not significantly affect INR control, warfarin management, or adverse events. Our findings suggest that HMOs can safely substitute at least 1 generic formulation of warfarin without extra monitoring.


On the Stability of MHD Boundary Layer Flow over a Stretching/Shrinking Wedge.
The steady two dimensional magnetohydrodynamic (MHD) boundary layer flow and heat transfer over a stretching/shrinking permeable wedge is numerically investigated. The partial differential equations governing the flow and heat transfer are transformed into a system of ordinary differential equations using a similarity transformation. These equations are then solved numerically using the boundary value problem solver, bvp4c in Matlab software. It is found that dual solutions exist for a certain range of the shrinking strength. A stability analysis is performed to identify which solution is stable and physically reliable.


Critical Analysis of cGMP Large-Scale Expansion Process in Bioreactors of Human Induced Pluripotent Stem Cells in the Framework of Quality by Design.
Human induced pluripotent stem cells (hiPSCs) are manufactured as advanced therapy medicinal products for tissue replacement applications. With this aim, the feasibility of hiPSC large-scale expansion in existing bioreactor systems under current good manufacturing practices (cGMP) has been tested. Yet, these attempts have lacked a paradigm shift in culture settings and technologies tailored to hiPSCs, which jeopardizes their clinical translation. The best approach for industrial scale-up of high-quality hiPSCs is to design their manufacturing process by following quality-by-design (QbD) principles: a scientific, risk-based framework for process design based on relating product and process attributes to product quality. In this review, we analyzed the hiPSC expansion manufacturing process implementing the QbD approach in the use of bioreactors, stressing the decisive role played by the cell quantity, quality and costs, drawing key QbD concepts directly from the guidelines of the International Council for Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use.


Results of a 2-year randomized, controlled obesity prevention trial: Effects on diet, activity and sleep behaviors in an at-risk young adult population.
Excess weight gain tends to occur in young adulthood. However, research examining effective weight-related interventions for this age group has been limited. As one of seven trials in the EARLY Trials consortium (Early Adult Reduction of weight through LifestYle intervention), the CHOICES Study (Choosing Healthy Options in College Environments and Settings) tested effects of a technology-integrated, young adult weight gain prevention intervention. It was a randomized controlled trial with assessments at baseline (2011) and 4-, 12- and 24-months post-intervention initiation and included 441 participants (ages 18-35) who were students at three Minnesota community colleges. The 24-month intervention included a 1-credit academic course and social networking and support online intervention. This analysis examined effects on 12 secondary behavioral outcomes across three domains: diet (fast food, sugary beverages, breakfast, at-home meal preparation), physical activity/screen time (minutes and energy expenditure in leisure time physical activity, television viewing, leisure time computer use) and sleep (hours of sleep, time required to fall asleep, days not getting enough rest, difficulty staying awake). The intervention resulted in significant reductions in fast food (p=0.007) but increases in difficulty staying awake (p=0.015). There was limited evidence of other behavior changes at 4months (0.05<p<0.1) in the expected direction but differences by treatment condition dissipated over time. Analyses examining summary treatment effects (i.e., modeling effects on all behavioral outcomes simultaneously) indicated significant overall effects (p=0.014), largely driven by 4-month results (p=0.005). Additional research is needed to understand effective obesity prevention among young adults, particularly when addressing multiple weight-related outcomes.


Synergistic effect of progesterone on prostaglandin E modulation of the mitogenic response of human peripheral lymphocytes.
The combined effects of prostaglandins of the E series (PGE) and progesterone (P), both of which are found to increase locally in the secretory endometrium, were studied in mitogen-stimulated cultures of lymphocytes from human peripheral blood. When added separately to lymphocyte cultures, both PGE and P produced a concentration-dependent inhibition of T-cell, but not B-cell, mitogenesis. If added together, these agents caused a much greater inhibition of T-cell mitogenesis, with marked synergy, than that observed with either agent alone. The synergistic inhibition was achieved with endometrial concentrations of PGE and P. The kinetics of PGE- and P-mediated inhibition investigated at various times during lymphocyte activation indicated that both agents affected the early events of T-cell mitogenesis. PGE- and P-inhibition of T-cell mitogenesis were shown to be reversible phenomena by washing the cells treated with these agents. These results suggest that elevated levels of PGE and P in the secretory endometrium, acting synergistically in inhibiting the proliferative response of T cells, may participate in facilitating implantation of histoincompatible fetal tissue in the maternal uterus as non-specific local immunosuppressive factors.


Tie-over bolster dressings may not be necessary to secure small full thickness skin grafts.
The accepted method for securing full thickness skin grafts (FTSG) is with a tie-over bolster dressing, with or without basting sutures. We question the need for tie-over bolster dressings for small FTSGs. We describe our method of FTSG reconstruction and the outcome in a consecutive series. Thirty patients with surgical defects following tumour excision from the face24, scalp1, ear2, and finger3 ranging in diameter from 8 to 45 mm (mean 20 mm) were reconstructed with a FTSG. Interrupted monofilament nylon perimeter sutures only were used, with antibiotic ointment at the wound edge with either a light dressing or no dressing. A series of 30 FTSG secured without a tie-over bolster dressing or basting sutures is described, all with good to excellent long term results. In two cases there was early superficial necrosis of the graft, but ultimately 100% graft take. We have found tie-over bolster dressings and basting sutures to be unnecessary in our series of small FTSG. This technique saves time and material and minimizes handling of the graft.


The acceptance and feasibility of breast cancer screening in the East.
Breast cancer, traditionally a disease prevalent among women in the West, has become an increasing disease burden in the East. In Hong Kong, breast cancer is the number one incident cancer in women. In Asian countries, due to the lower incidence rate, population screening is still controversial. Most screening programs are not population-based, and are either self-financed privately or partially subsidized by the government. Since the first government-subsidized screening program was set up in 1991 in Hong Kong, numerous private and government health centres for women have been set up. A dedicated private Breast Care Centre was set up at the Hong Kong Sanatorium and Hospital (HKSH) in 1999. Over an 8-year period from October 1999 to September 2006, 14,596 women were screened, of whom 11,408 were asymptomatic. This study aims to produce the first report on breast screening experience in the largest cohort of asymptomatic Chinese women in the private sector. An overall malignancy detection rate of 2.3 per 1000 screens and a recall rate of 9.2% were found. Despite culture differences and differences in breast characteristics (denser and smaller breasts), breast screening is feasible and acceptable in the East.


Rapid Classification of Sarcomas Using Methylation Fingerprint: A Pilot Study.
Sarcoma classification is challenging and can lead to treatment delays. Previous studies used DNA aberrations and machine-learning classifiers based on methylation profiles for diagnosis. We aimed to classify sarcomas by analyzing methylation signatures obtained from low-coverage whole-genome sequencing, which also identifies copy-number alterations. DNA was extracted from 23 suspected sarcoma samples and sequenced on an Oxford Nanopore sequencer. The methylation-based classifier, applied in the nanoDx pipeline, was customized using a reference set based on processed Illumina-based methylation data. Classification analysis utilized the Random Forest algorithm and t-distributed stochastic neighbor embedding, while copy-number alterations were detected using a designated R package. Out of the 23 samples encompassing a restricted range of sarcoma types, 20 were successfully sequenced, but two did not contain tumor tissue, according to the pathologist. Among the 18 tumor samples, 14 were classified as reported in the pathology results. Four classifications were discordant with the pathological report, with one compatible and three showing discrepancies. Improving tissue handling, DNA extraction methods, and detecting point mutations and translocations could enhance accuracy. We envision that rapid, accurate, point-of-care sarcoma classification using nanopore sequencing could be achieved through additional validation in a diverse tumor cohort and the integration of methylation-based classification and other DNA aberrations.


Non-peptidic HIV protease inhibitors: C2-symmetry-based design of bis-sulfonamide dihydropyrones.
Potent, non-peptidic, dihydropyrone sulfonamide HIV protease inhibitors have been previously described. Crystallographic analysis of dihydropyrone sulfonamide inhibitor/HIV protease complexes suggested incorporation of a second, C2 symmetry-related sulfonamide group. Selected bis-sulfonamide dihydropyrone analogues display high HIV protease inhibitory activity.


Sarcomas of the head and neck.
With the exception of pediatric RMS, soft tissue sarcomas only rarely arise in the head and neck region. Soft tissue sarcomas include a diverse array of histologic types because of the variety of mesenchymal tissues from which they originate. The combination of infrequent occurrence, varied pathologic features, and the many potential sites of presentation makes these tumors a challenge for the head and neck oncologist and underscore the need for review by a pathologist experienced with soft tissue tumors. Classification schemes that group sarcomas according to grade have been helpful in providing prognostic information. Although local control of the primary tumor is critical to successful treatment of both high- and low-grade lesions, the high rate of distant metastases in high-grade tumors supports the role of combined modality therapy. Compared with other types of head and neck neoplasms, such as squamous cell carcinoma, soft tissue sarcomas have low rates of regional metastases. Surgery generally has been recommended as the primary method of treatment for achieving local control, except in those high-grade tumors arising in sites not amenable to resection. Exceptions to this principle include RMSs of the orbit, paranasal sinuses, and masticator space in children; these are usually treated with radiotherapy and combined multiagent chemotherapy, thereby avoiding the functional and cosmetic impact of surgery. Also, extensive angiosarcomas of the scalp should be treated with multimodality therapy combining surgery and wide-field radiation therapy in an attempt to achieve local control. Adjuvant radiotherapy is generally recommended for high-grade sarcomas, large tumors, close or positive surgical margins, and certain histologic variants. Systemic chemotherapy is recommended for those tumors with a significant risk of distant metastases. Increasingly, neoadjuvant chemotherapy is being used to determine responsiveness to chemotherapy, which can help physicians select patients who may benefit from systemic postoperative therapy. Traditional predictors of treatment failure for soft tissue sarcomas include larger tumor size, high-grade histology, and positive surgical margins. The advent of more advanced reconstructive techniques, including free tissue transfer, has made more aggressive surgical resection of these tumors possible. Nevertheless, a considerable number of ancillary support staff are critical to the patient's postoperative rehabilitation and eventual return to a satisfactory level of function and quality of life. In the future, the discovery of the molecular pathogenesis of specific tumor types, such as the cytogenetic findings in synovial sarcoma, will improve physicians' prognostic abilities and selection of patients who are most likely to benefit from emerging adjuvant therapies.


Surgical rosacea.
Rosacea is a common cutaneous disease. The dermatologic surgeon will frequently encounter rosacea in the surgical patient. To describe observations regarding surgical and wound care techniques that facilitate and improve outcomes in the rosacea patient. We present a compilation of our clinical experience and observations regarding rosacea in the dermatologic surgery. Simple interventions such as disease recognition and treatment, proper wound care and suture selection, as well as patient counseling have proven beneficial in optimizing surgical outcomes in the rosacea patient.


Nuclearly-encoded CBP1 interacts with the 5' end of mitochondrial cytochrome b pre-mRNA.
CBP1 is a nuclearly-encoded protein that is imported into mitochondria and confers stability on the mRNA for cytochrome b. Previous work has shown that CBP1 interacts with the cytochrome b transcript upstream of the coding sequence; a region encompassing some 1,100 nucleotides. The work presented here narrows the site of action of CBP1 to the distal third of this upstream sequence through analysis of mRNA produced from a novel recombinant gene containing segments of the gene for cytochrome b, cob, and the ATP synthase subunit 9 gene, olil. In a wild-type CBP1 strain, the cob-olil-cob gene produces stable, mature mRNA that is translated and contributes a portion of the cytochrome b necessary for optimal growth on non-fermentable medium.


Skin disease is more recalcitrant than muscle disease: A long-term prospective study of 184 children with juvenile dermatomyositis.
Persistent skin manifestations, especially calcinoses, contribute to morbidity in children with juvenile dermatomyositis. To compare the course of skin and muscle involvement and document frequency of calcinosis in juvenile dermatomyositis. Prospective cohort study of 184 untreated children with juvenile dermatomyositis (July 1971 to May 2019) at a single children's hospital. Disease Activity Scores (DASs) were persistently higher for skin versus muscle at all points; clinical inactivity (DAS ≤2) occurred earlier for muscle than skin. Among vascular features for DAS for skin, eyelid margin capillary dilatation was most frequent (54.3%) and persisted longest. Intravenous methylprednisolone reduced DAS for skin more than oral prednisone at 12 months (P = .04). Overall, 16.8% of patients (n = 31) had calcifications, with 4.9% at enrollment. Despite therapy, 25.0% of calcifications recurred and 22.6% failed to resolve; of the latter, 71.4% (n = 5) were present at enrollment. Children with persistent calcifications had longer duration of untreated disease than those whose calcifications resolved (mean 12.5 months) (P < .001). Hydroxychloroquine did not improve DAS for skin (P = .89). DAS does not quantify nailfold capillary dropout. In juvenile dermatomyositis, skin disease presents with greater activity and is more recalcitrant to therapies than muscle disease. Early and aggressive treatment can limit the severity and persistence of calcifications identified later in the disease course.


Expression patterns of odorant-binding proteins in antennae of the moth Manduca sexta.
Monoclonal antibodies (MAbs) were generated to six recombinant proteins (odorant-binding proteins; OBPs) of Manduca sexta. The specificity of each MAb was demonstrated by labeling six immunoblots, each of which contained samples of all six recombinant OBPs. The expression patterns of the six OBPs could be grouped into three classes: (1) one (GOBP1) was expressed in sensilla located throughout each annulus; (2) two (ABPX and ABP2) were expressed in the long sensilla trichoidea bordering a zone that was arranged as an arch on the periphery of each annulus; (3) three (PBP2, PBP3, and GOBP2) were expressed in shorter sensilla occupying a wedge-shaped mid-annular zone of each annulus. In female antennae, sensilla expressing these OBPs were intermixed, and the distinct zonation observed in the male antenna was absent. In males, PBP2 was co-expressed in exactly the same cells of the mid-annular zone as those expressing PBP3 and most of the same cells expressing GOBP2, although its expression overlapped with no or only a few sensilla expressing OBPs of class 1 (GOBP1) or class 2 (ABPX, ABP2). This overlap of expression or lack of overlap between PBP2 and the other OBPs for male antennae was mirrored in female antennae. In view of the restricted spatial expression of OBPs within an annulus and the diversity of possible dimeric combinations of OBPs that arises from the co-expression of multiple OBPs in a given sensillum, OBPs could contribute to the specificity of the olfactory responses of insects.


Genetic Susceptibility to Atrial Fibrillation Is Associated With Atrial Electrical Remodeling and Adverse Post-Ablation Outcome.
This study sought to assess the atrial electrophysiological properties and post-ablation outcomes in patients with atrial fibrillation (AF) with and without the rs2200733 single nucleotide variant. The phenotype associated with chromosome 4q25 of the AF-susceptibility locus remains unknown. In this study, 102 consecutive patients (ages 61 ± 9 years, 64% male) with paroxysmal or persistent AF were prospectively recruited prior to ablation. Patients were genotyped for rs2200733 and high-density left atrial (LA) electroanatomic maps were created using a multipolar catheter during distal coronary sinus (CS) pacing at 600 ms. Voltage, conduction velocity (CV), CV heterogeneity, and fractionated signals of 6 LA segments were determined. Arrhythmia recurrence was assessed by continuous device (51%) and Holter monitoring. Overall, 41 patients (40%) were single nucleotide variant carriers (38 heterozygous, 3 homozygous). A mean of 2,239 ± 852 points per patient were collected. Carriers had relatively increased CV heterogeneity (45.7 ± 7.5% vs. 35.9 ± 2.3%; p < 0.001), complex signals (9.4 ± 2.9% vs 6.0 ± 1.2%; p = 0.008), regional LA slowing, or conduction block (31.7 ± 8.2% vs. 17.9 ± 1.9%; p = 0.013) particularly in the posterior and lateral walls. There were no differences in CV, voltage, atrial refractoriness, or sinus node function. At follow-up (median: 27 months; range 19 to 31 months), carriers had lower arrhythmia-free survival (51% vs. 80%; p = 0.003). On multivariable analysis, carrier status was independently associated with CV heterogeneity (p = 0.001), complex signals (p = 0.002), and arrhythmia recurrence (p = 0.019). These data provide the first evidence that the rs2200733-tagged haplotype alters LA electrical remodeling and is a determinant of long-term outcome following AF ablation. The molecular mechanisms underpinning these changes warrant further investigation.


Primary aortoenteric fistula: pitfalls in the diagnosis of a rare condition.
Primary aortoenteric fistula is such a rare condition that even specialist vascular surgeons may not encounter one in their lifetime practice. Any hope of survival depends upon prompt diagnosis and surgical treatment. We present our experience of one such case, particularly the difficulties encountered in confirming the diagnosis, and go on to review the literature on the subject. Both our experience and that reported by previous authors would suggest that specialized investigations such as computed tomography, contrast angiography, radionuclide scintigraphy, and gastrointestinal endoscopy have only a limited contribution to make in confirming the diagnosis of primary aortoenteric fistula. The diagnosis should be considered in any patient with gastrointestinal blood loss and an abdominal aortic aneurysm. Specialized investigations should not be relied upon to confirm or refute the diagnosis. Any prospect of survival is dependant upon prompt diagnosis and surgical intervention. The latter is only achieved by a high index of clinical suspicion.


An atom economic synthesis and AChE inhibitory activity of novel dispiro 7-aryltetrahydro-1H-pyrrolo[1,2-c][1,3]thiazole and 4-aryloctahydroindolizine N-methylpiperidin-4-one hybrid heterocycles.
The 1,3-dipolar cycloaddition of azomethine ylides generated in situ from acenaphthenequinone and α-amino acids viz. 1,3-thiazolone-4-carboxylic acid and piperidine-2-carboxylic acid to a series of 1-methyl-3,5-bis[(E)-arylmethylidene]tetrahydro-4(1H)-pyridinones afforded novel spiro[5.2″]acenaphthene-1″-onespiro[6.3']-5'-arylmethylidene-1'-methylpiperidin-4'-one-7-aryltetrahydro-1H-pyrrolo[1,2-c][1,3]thiazoles and spiro[2.2″]acenaphthene-1″-onespiro[3.3']-5'-arylmethylidene-1'-methylpiperidin-4'-one-4-aryloctahydroindolizines respectively in quantitative yields. These compounds were evaluated for their AChE inhibitory activity and compound 3c was found to be the most potent with IC50 1.86 μmol/L.


Comparing aflatoxin contamination in chilies from Punjab, Pakistan produced in summer and winter.
A comparison was made of total aflatoxins (AFs) in 43 samples of chilies collected during winter and 42 in summer to determine the effect of season on contamination. The samples were analyzed by HPLC with fluorescence detection. The limits of detection and quantification for AFB1 and AFG1 were 0.05 μg/kg and 0.50 μg/kg, whilst for AFG2 and AFB2 they were 0.10 μg/kg and 0.60 μg/kg. In the winter samples, AFs were detected in 18 (72%) whole and 14 (60%) ground chilies, with concentration ranges 0.00-52.30 μg/kg and 0.00-74.60 μg/kg respectively. In the summer samples, 17 (64%) whole and 12 (76%) ground chilies were contaminated with AFs at concentrations 0.00-61.50 μg/kg and 0.00-95.90 μg/kg respectively. The percentage of samples greater than the European Union statutory limit for AFB1 and total AF for whole chilies were 48 and 36%, compared with ground chili values of 50 and 45%, respectively, in the winter season. In the summer season, the samples greater than the European Union limit for AFB1 and total AF in whole chilies were 52 and 38%, compared with values of 54 and 49% in ground chilies respectively. AF contamination was found to be higher in summer chili samples and hence winter chilies may provide a better quality product with respect to AF contamination. The ability to undertake this analysis in Pakistan will enhance greatly the ability to improve chili production in that country, as described herein.


Correction: Tickner, Z.J.; Farzan, M. Riboswitches for Controlled Expression of Therapeutic Transgenes Delivered by Adeno-Associated Viral Vectors. Pharmaceuticals 2021, 14, 554.
In the original article, Renzl, C [...].


MMP-3 -1171 5A/6A promoter polymorphism and cancer susceptibility: an updated meta-analysis and trial sequential analysis.
Purpose: Previous studies of MMP-3 -1171 5A/6A in cancers have produced inconclusive outcomes. This updated meta-analysis was performed to clarify the link between this variant and cancer. Methods: Databases including PubMed, Google Scholar, EMBASE and Cochrane were searched for data collection. The associations were calculated by odds ratios with 95% CIs. Results: 63 eligible studies with 14,252 cases and 15,176 controls were included. The codominant 2, codominant 3, dominant, recessive and allele models were found to be significantly associated with 1.28-, 1.13-, 1.13-, 1.19- and 1.13-fold enhanced overall risk of cancer, respectively. Stratification analysis revealed a 1.28-times enhanced risk of esophageal cancer (codominant 1), 1.29- and 1.26-fold (codominant 3) and 1.18- and 1.28-fold (recessive model) enhanced risk in colorectal and gastrointestinal cancers, respectively, 1.30-, 1.35- and 1.22-times in codominant model 1, dominant and allele models for breast cancer, 1.56-fold (codominant 2) for gynecological cancer and 2.40-times in codominant model 2 for hepatocellular cancer. Conclusion: This meta-analysis suggests a significant association between the MMP-3 -1171 5A/6A variant and cancer. This meta-analysis was registered at INPLASY (registration number: INPLASY202280049).


Comorbidity and KPS are independent prognostic factors in stage I non-small-cell lung cancer.
To determine the prognostic role of comorbidity in Stage I non-small-cell lung cancer (NSCLC) treated with surgery or radiotherapy (RT). One hundred sixty-three patients with clinical Stage I NSCLC were analyzed for overall survival (OS) and comorbidity. One hundred thirteen patients underwent surgery (surgical group) and 50 patients received definitive radiotherapy (RT group). Ninety-six percent of the surgical group had lobectomy or pneumonectomy, and negative margins were achieved in 96% of the patients. The median dose to the tumor for the RT group was 61.2 Gy (range 30.8-77.4). The Cumulative Illness Rating Scale for Geriatrics (CIRS-G) and the Charlson scale were used to rate comorbidity. Karnofsky performance scores (KPS) were available in 42 patients; the rest of the scores were determined retrospectively by two physicians independently, with 97% agreement. The OS was 44% for the surgical group and 5% for the RT group at 5 years. Noncancer-related mortality was observed in 31% and 62% of the surgical and RT patients, respectively. On univariate analysis, performed on all patients (n = 163), squamous cell histologic type (p <0.001), clinical Stage T2 (p = 0.062), tumor size >4 cm (p = 0.065), >40 pack-year tobacco use (p <0.001), presence of a CIRS-G score of 4 (extremely severe, CIRS-G4: [+]) (p <0.001), severity index of >2 (p <0.001), Charlson score >2 (p = 0.004), KPS <70 (p <0.001), and treatment with RT (p <0.001) were associated with a statistically significant inferior OS. Multivariate analysis with histologic features, clinical T stage, age, tobacco use, KPS, comorbidity [CIRS-G(4)] and treatment group on all patients showed that squamous cell histology, >40 pack-year tobacco use, KPS <70, and presence of CIRS-G(4) were independently associated with an inferior OS. Treatment modality, T stage, and age did not have any statistically significant effect on OS. Statistically significant differences were found between the surgical and RT groups in Charlson score (p = 0.001), CIRS-G total score (p = 0.004), severity index (p = 0.006), CIRS-G4(+) (p <0.001), KPS (p <0.001), amount of tobacco use (p = 0.002), clinical tumor size (p <0.001), clinical T stage (p = 0.01), forced expiratory volume in 1 s (p = 0.001), and age (p = 0.008), in favor of the surgical group. The presence of significant comorbidity and KPS of <70 are both important prognostic factors, but were found to be independent of each other in Stage I NSCLC. Therefore, comorbidity and KPS assessment are recommended when analyzing the prognostic effects of tumor or treatment-related factors on OS.


Dynamic nanopore long-read sequencing analysis of HIV-1 splicing events during the early steps of infection.
Alternative splicing is a key step in Human Immunodeficiency Virus type 1 (HIV-1) replication that is tightly regulated both temporally and spatially. More than 50 different transcripts can be generated from a single HIV-1 unspliced pre-messenger RNA (pre-mRNA) and a balanced proportion of unspliced and spliced transcripts is critical for the production of infectious virions. Understanding the mechanisms involved in the regulation of viral RNA is therefore of potential therapeutic interest. However, monitoring the regulation of alternative splicing events at a transcriptome-wide level during cell infection is challenging. Here we used the long-read cDNA sequencing developed by Oxford Nanopore Technologies (ONT) to explore in a quantitative manner the complexity of the HIV-1 transcriptome regulation in infected primary CD4+ T cells. ONT reads mapping to the viral genome proved sufficiently long to span all possible splice junctions, even distant ones, and to be assigned to a total of 150 exon combinations. Fifty-three viral RNA isoforms, including 14 new ones were further considered for quantification. Relative levels of viral RNAs determined by ONT sequencing showed a high degree of reproducibility, compared favourably to those produced in previous reports and highly correlated with quantitative PCR (qPCR) data. To get further insights into alternative splicing regulation, we then compiled quantifications of splice site (SS) usage and transcript levels to build "splice trees", a quantitative representation of the cascade of events leading to the different viral isoforms. This approach allowed visualizing the complete rewiring of SS usages upon perturbation of SS D2 and its impact on viral isoform levels. Furthermore, we produced the first dynamic picture of the cascade of events occurring between 12 and 24 h of viral infection. In particular, our data highlighted the importance of non-coding exons in viral RNA transcriptome regulation. ONT sequencing is a convenient and reliable strategy that enabled us to grasp the dynamic of the early splicing events modulating the viral RNA landscape in HIV-1 infected cells.


Transient isolated ocular motor abnormality related to perilesional edema of an acute medullary microbleed: A case report and review of the literatures.
We report a case of transient isolated gaze-evoked nystagmus with ocular lateropulsion in a patient with an acute medullary microbleed which was detected by brain magnetic resonance imaging. Considering the correlation between the neural structures involved by the lesion and the ocular motor symptoms of this patient, we suggest that the perilesional edema of the acute medullary microbleed is responsible for this transient ocular motor abnormality.


Endoscopic ultrasonography-fine needle aspiration of solid pancreatic masses: Do we need the fourth pass? A prospective study.
Endoscopic ultrasonography-guided fine-needle aspiration (EUS-FNA) is important for the differential diagnosis of solid pancreatic lesions. Sample adequacy is related to the number of needle passes, and European guidelines recommend three to four needle passes with a standard EUS-FNA needle. We aimed to evaluate the optimal number of passes with standard EUS-FNA needles in solid pancreatic lesions. Patients with solid pancreatic masses without cystic component >20% on computed tomography scan, and without biliary metallic stents, or coagulation problems were included prospectively. Standard 22G needles were used (maximum four passes); each sample was paraffin-embedded and analyzed separately. Final diagnosis was established by EUS-FNA, repeat EUS-FNA, surgery, or follow-up. Sixty-one of 65 patients were included. The final diagnoses were adenocarcinoma (n = 44, 72%), neuroendocrine tumor (NET) (n = 10, 16%), metastasis (n = 1, 4%) and nonmalignant lesion (n = 6, 10%). Immunohistochemical staining was possible in 17 cases. The diagnosis was established by the first pass in 62% of cases (n = 38), by the second in 15% (n = 9), by the third in 15% (n = 9), and by the fourth in 3% (n = 2). The diagnostic accuracy for all four passes compared to the first three passes was 95% vs 92% (P = .5). The contribution of the fourth pass was not different between adenocarcinoma and NET (2% vs 10%, respectively; P = .667). Three passes with standard EUS-FNA was optimal for a specific diagnosis of solid pancreatic masses, regardless of the histological type of the lesion.


Effects of poppy seed meal on egg production and hatching results in quail (Coturnix coturnix japonica).
This study was carried out to determine the effects of poppy seed meal (PSM) on the egg production and hatching of quail. Two experiments were undertaken. In the first experiment, 240 quail, reared under the same conditions, were divided into four groups of 60 birds each (which were, in turn, subdivided into four subgroups of 15 each for experimental replicates). The first, second, and third experimental groups were fed ratios containing 5%, 15%, and 25% PSM, respectively, over 12 weeks. The control group received no PSM. Feed and water were supplied ad libitum and artificial light was provided for 16 h a day. During the first experiment, egg production, egg weight, and feed conversion ratio (FCR) were measured. In the second experiment, 240 female and 48 male quail were divided into four groups, as in the preceding experiment, and the birds were housed in cages at a ratio of one male to five females. The diets and their presentation to the quails were the same as in the preceding experiment. The results of the two experiments showed that the addition of PSM to the feed ratios increased egg production, feed consumption, and feed conversion per kg of eggs and per dozen eggs. However, PSM addition at all percentages significantly reduced (P<0.001) hatchability, mainly because of increased infertility and embryonic deaths.


Reciprocal transrepression between FOXF2 and FOXQ1 controls basal-like breast cancer aggressiveness.
FOXF2 and FOXQ1, forkhead box transcription factor superfamily members, are encoded by neighboring genes located on human chromosome 6p25.3 and play opposite roles in epithelial-mesenchymal transition (EMT) and metastasis in basal-like breast cancer (BLBC). However, the relationship between FOXF2 and FOXQ1 in cancer remains unknown. Here, we found mutual transcriptional repression between FOXF2 and FOXQ1, and the reciprocal negative feedback loop controlled EMT, aggressiveness, and chemoresistance in BLBC cells. We further demonstrated that FOXF2 recruited nuclear receptor corepressor 1 and histone deacetylase 3 to the FOXQ1 promoter to inhibit its transcription in BLBC cells, but FOXQ1 did not exert such an effect on FOXF2. Our findings reveal novel mechanisms underlying the determination of BLBC aggressiveness and the transrepressive function of FOXF2 in a basal-like cell subtype-specific manner. Therefore, blocking the vicious cycle of the abnormal reciprocal feedback loop between FOXF2 and FOXQ1 to induce cell differentiation and restore tissue homeostasis is a promising strategy for the treatment of aggressive BLBC.-Kang, L.-J., Yu, Z.-H., Cai, J., He, R., Lu, J.-T., Hou, C., Wang, Q.-S., Li, X.-Q., Zhang, R., Feng, Y.-M. Reciprocal transrepression between FOXF2 and FOXQ1 controls basal-like breast cancer aggressiveness.


The Importance of Mean Neutrophil Volume (MNV) Values in Differential Diagnosis of Bacterial Sepsis.
Bloodstream infections are a significant cause of morbidity and mortality in hospitalized patients. Blood cultures and other laboratory tests are used for diagnosis. Among these tests, the mean neutrophil volume (MNV) value is reported as a potential indicator that supports the diagnosis of sepsis. Our study identified the MNV values of patients via microorganisms cultivated from blood cultures and examined the role of these MNV values in the early diagnosis of bloodstream infections. Our study surveyed retrospectively 148 adult patient blood culture samples that had been sent to our laboratory. BACTEC 9050 (Becton Dickinson, USA) and BACTEC FX 40 (Becton Dickinson, USA) devices were used in the blood culture isolation procedures. The average MNV value was found to be 159.0 (+11.3) in patients whose sepsis originated from Gram-negative bacteria, and the average MNV value was measured as 152.4 (+14.5) among patients whose sepsis originated from Gram-positive bacteria. When comparing groups of patients having Gram-negative bacteria and patients having Gram-positive bacteria, a statistically significant difference (p = 0.041) in the MNV values was observed. The MNV value was found to be statistically significant in discrimination of Gram-negative and Gram-positive sepsis. Considering these findings, measuring the MNV values can help initiate proper antibiotic treatment more quickly, and we think that this will help lower the mortality rate. However, these findings should be supported with further studies.


Prolonged expiratory duration with elevated lung volume in newborn infants.
End-expiratory lung volume increased 17.5 +/- 5.4 ml (mean +/- SD) in full term infants and 7.7 +/- 2.1 ml in premature infants when 5--7 cm H2O continuous negative pressure (CNEG) was applied around the thorax. In the full term infants, respiratory rate decreased from 52--43 min-1 (P less than 0.001), mean inspiratory duration (ti) was unchanged, and mean duration of expiration (te) increased from 0.62 +/- 0.14 (SE)-0.84 +/- 0.22 sec (P less than 0.001) after application of CNEG. Te of occluded efforts on CNEG was also prolonged (P less than 0.005), although less than te of spontaneous breaths on CNEG (P less than 0.005). Te increased in four of eight premature infants when CNEG was applied. We conclude that phasic vagal feedback regulates Ti and te is controlled by tonic vagal activity. Premature infants may have an optimum functional residual capacity (FRC) which can be achieved with continuous distending pressure, resulting in regularization of respiration.


High AMY1 copy number protects against obesity in Portuguese young adults.
The highly polymorphic copy number variation (CNV) in the salivary amylase gene (AMY1) has been associated with obesity in different populations. However, some authors have failed to reproduce these findings. To investigate the association between AMY1 CNV and obesity in young adults of Portuguese origin. This study evaluated AMY1 gene copy number (CN) in 262 individuals: 155 females and 107 males, aged 18-34 years-old (mean age = 21.08). The number of AMY1 copies was estimated in a QX100 droplet digital PCR (ddPCR) system (Bio-Rad Laboratories, Hercules, CA). Defining a case group with obese and overweight individuals, logistic regression did not show a significant association between AMY1 CNV and risk of overweight/obesity in the whole population (p = 0.489). However, after testing case-control data in the sub-set of samples above the third quartile (CN ≥10), a significant association was found between lower AMY1 copy number and risk of obesity (OR = 0.532; p = 0.034), even when adjusted for age and sex (OR = 0.527; p = 0.039). In concordance, all participants with >10 AMY1 copies were normal weight controls (n = 20) or overweight (n = 6). The results suggest that high AMY1 gene copy number protects against obesity in Portuguese young adults.


Smooth muscle metaplasia and innervation in interstitium of endometriotic lesions related to pain.
To investigate the pathogenesis of endometriotic pain. Retrospective nonrandomized immunohistochemical study. A university hospital, Department of Gynecology. Twenty human endometriotic specimens were selected from different lesions including ovarian endometrioma, peritoneal lesion, and deep infiltrating lesion. Premenopausal women with histologically diagnosed endometriosis were selected (mean age 39 years; range, 25-53 years). The chief complaint was dysmenorrhea, dyschezia, and dyspareunia. A rat endometriosis model was induced in 10 SLC-Sprague-Dawley rats (8 weeks old) by surgical autotransplantation of the uterus. Immunohistochemical staining of endometriotic specimens for alpha-smooth muscle actin (ASMA), neural cell adhesion molecule (NCAM), and nerve growth factor (NGF) expression. Comparison of the immunoreactive staining of ASMA, NCAM, and NGF expression in human endometriosis and a rat endometriosis model. Morphological analysis revealed thick interstitium in both human and rat endometriotic lesions. The major components of fibrotic interstitium are smooth muscle cells, stained by anti-ASMA antibody, and nerve cells, stained by anti-NCAM antibody. Inflammatory cells are also present (e.g., macrophages and lymphocytes) as revealed by anti-NGF antibody staining. These results suggest that the contraction of smooth muscle cells and the hyperalgia derived from innervation in the interstitial area is related to pain in endometriosis.


NAPSB as a predictive marker for prognosis and therapy associated with an immuno-hot tumor microenvironment in hepatocellular carcinoma.
Napsin B Aspartic Peptidase, Pseudogene (NAPSB) was associated with CD4 + T cell infiltration in pancreatic ductal adenocarcinoma. However, the biological role of NAPSB in hepatocellular carcinoma (HCC) remains to be determined. The expression of NAPSB in HCC as well as its clinicopathological association were analyzed using data from several public datasets. qRT-PCR was used to verify the relative expression of NAPSB in patients with HCC using the Zhongnan cohort. Kaplan-Meier analyses, and univariate and multivariate Cox regression were conducted to determine the prognosis value of NAPSB on patients with HCC. Then enrichment analyses were performed to identify the possible biological functions of NAPSB. Subsequently, the immunological characteristics of NAPSB in the HCC tumor microenvironment (TME) were demonstrated comprehensively. The role of NAPSB in predicting hot tumors and its impact on immunotherapy and chemotherapy responses was also analyzed by bioinformatics methods. NAPSB was downregulated in patients with HCC and high NAPSB expression showed an improved survival outcome. Enrichment analyses showed that NAPSB was related to immune activation. NAPSB was positively correlated with immunomodulators, tumor-infiltrating immune cells, T cell inflamed score and cancer-immunity cycle, and highly expressed in immuno-hot tumors. High expression of NAPSB was sensitive to immunotherapy and chemotherapy, possibly due to its association with pyroptosis, apoptosis and necrosis. NAPSB was correlated with an immuno-hot and inflamed TME, and tumor cell death. It can be utilized as a promising predictive marker for prognosis and therapy in HCC.


Evaluation of sustainable susceptibility to Plasmodium vivax infection among colonized Anopheles darlingi and Anopheles deaneorum.
The colonization of mosquitoes susceptible to Plasmodium vivax via direct membrane feeding assay (DMFA) has the potential to significantly advance our knowledge of P. vivax biology, vector-parasite interaction and transmission-blocking vaccine research. Anopheles darlingi and Anopheles deaneorum are important vectors of malaria in the Western Brazilian Amazon. Since 2018, well-established colonies of these species have been maintained in order to mass produce mosquitoes destined for P. vivax infection. Plasmodium susceptibility was confirmed when the colonies were established, but susceptibility needs to be maintained for these colonies to remain good models for pathogen transmission. Thus, the susceptibility was assessed of colonized mosquitoes to P. vivax isolates circulating in the Western Amazon. Laboratory-reared mosquitoes from F10-F25 generations were fed on P. vivax blood isolates via DMFA. Susceptibility was determined by prevalence and intensity of infection as represented by oocyst load seven days after blood feeding, and sporozoite load 14 days after blood feeding. The effect of infection on mosquito survival was evaluated from initial blood feeding until sporogonic development and survival rates were compared between mosquitoes fed on infected and uninfected blood. Correlation was calculated between gametocytaemia and prevalence/intensity of infection, and between oocyst and sporozoite load. Significant differences were found in prevalence and intensity of infection between species. Anopheles darlingi showed a higher proportion of infected mosquitoes and higher oocyst and sporozoite intensity than An. deaneorum. Survival analysis showed that An. deaneorum survival decreased drastically until 14 days post infection (dpi). Plasmodium vivax infection decreased survival in both species relative to uninfected mosquitoes. No correlation was observed between gametocytaemia and prevalence/intensity of infection, but oocyst and sporozoite load had a moderate to strong correlation. Colonized An. darlingi make excellent subjects for modelling pathogen transmission. On the other hand, An. deaneorum could serve as a model for immunity studies due the low susceptibility under current colonized conditions. In the application of DMFA, gametocyte density is not a reliable parameter for predicting mosquito infection by P. vivax, but oocyst intensity should be used to schedule sporozoite experiments.


The tumor biopsy remains essential for breast cancer diagnosis and characterization. Indeed, the treatment is decided according to histological subtype, and according to the presence of targetable molecular alterations. Notably, the presence of hormone receptors, ERBB2 hyperexpression or the existence of PIK3CA or ESR1 mutations are among the alterations commonly investigated. But these biological characteristics are determined only partially by tumor biopsy, due to tumor heterogeneity or tumor plasticity that happens spontaneously or under treatment. Liquid biopsy, and in particular circulating tumor DNA and circulating tumor cells, is a non-invasive method to identify and characterize the presence of cancer in the blood. The aim of this review is to determine the value of liquid biopsy to enhance or replace the data provided by a tumor biopsy.


Mutation of Haemophilus influenzae transforming DNA in vitro with near-ultraviolet radiation: action spectrum.
Mutations were produced in purified transforming DNA from Haemophilus influenzae by near-UV radiation and were assayed as mutants among cells transformed with irradiated DNA. The maximum efficiency of mutation induction was at around 334 nm, and the efficiency dropped off steeply at lower and higher wavelengths. The difference between the action spectrum for mutation and that for the oxygen-independent inactivation of transforming DNA, which had a shoulder at 365 nm, indicates that there are different lesions involved in the inactivating and mutagenic effects of near-UV. The presence of histidine during irradiation enhanced the mutagenic effect at 334 and 365 nm, although it protected against inactivation at 365 nm. The effective near-UV wavelengths for in vitro mutation are to some extent the same as the effective wavelengths for mutation in vivo reported previously. These findings indicate that mutations are produced in vivo by near-UV with DNA as the primary target molecule rather than by a secondary non-photochemical reaction between DNA and some other cell component.


A note on modelling the dynamics of budding yeast populations using branching processes.
A multitype branching process is proposed as a model for the behaviour of populations of the budding yeast Saccharomyces Cerevisiae. Using the idea of branching processes counted by random characteristics, we are able to obtain explicit expressions describing different aspects of the asymptotic composition of such populations. The main purpose of this note is to show that the branching process approach is an alternative to deterministic population models based on differential equation methods.


Development and characterization of monoclonal antibodies reactive with paraquat.
A set of three anti-paraquat monoclonal antibodies(MoAbs), named APM-1, APM-2 and APM-3, has been isolated. In order to evaluate the ability of these MoAbs to recognize various kinds of bipyridyl herbicides and similar congeners of paraquat, a competition enzyme-linked immunosorbent assay (ELISA) using avidin-biotin complex (ABC) was developed. All three antibodies strongly recognized paraquat and slightly did the other analogs. These three MoAbs are therefore advantageous to a toxicological study of paraquat and of its localization in tissues.


The YJR127C/ZMS1 gene product is involved in glycerol-based respiratory growth of the yeast Saccharomyces cerevisiae.
A putative yeast mitochondrial upstream activating sequence (UAS) was used in a one-hybrid screening procedure that identified the YJR127C ORF on chromosome X. This gene was previously designated ZMS1 and is listed as a transcription factor on the SGD website. Real time RT-PCR assays showed that expression of YJR127C/ZMS1 was glucose-repressible, and a deletion mutant for the gene showed a growth defect on glycerol-based but not on glucose- or ethanol-based medium. Real time RT-PCR analyses identified severely attenuated transcript levels from GUT1 and GUT2 to be the source of that growth defect, the products of GUT1 and GUT2 are required for glycerol utilization. mRNA levels from a large group of mitochondria- and respiration-related nuclear genes also were shown to be attenuated in the deletion mutant. Importantly, transcript levels from the mitochondrial OLI1 gene, which has an associated organellar UAS, were attenuated in the DeltaYJR127C mutant during glycerol-based growth, but those from COX3 (OXI2), which lacks an associated mitochondrial UAS, were not. Transcriptome analysis of the glycerol-grown deletion mutant showed that genes in several metabolic and other categories are affected by loss of this gene product, including protein transport, signal transduction, and others. Thus, the product of YJR127C/ZMS1 is involved in transcriptional control for genes in both cellular genetic compartments, many of which specify products required for glycerol-based growth, respiration, and other functions.


[Petromastoid clearance using plastic surgery in the surveillance of large cavities. Re-suture of the ear canal (author's transl)].
The lesions met with in chronic otitis media and especially in cholesteatoma, can be extremely widespread and, after petromastoid clearance there is a large resulting cavity which is impossible to survey by the usual route of the ear canal. A plastic surgical technique on the ear canal, is described which allows an easy surveillance without esthetic damage. It consists of a resection of a part of the cartilage of the ear canal with conservation of the skin of the inner and outer ear canal. This skin is used to re-suture the piece of cartilage which has been sectioned.


Supplementation of ram's semen extender with Mito-TEMPO II: Quality evaluation and flow cytometry study of post-thawed spermatozoa.
Cryopreservation is an effective method to spread qualified ram spermatozoa for reproductive goals in different farms, but cryopreservation's shocks reduce sperm quality. This study investigated the efficacy of the new mitochondria-targeted antioxidant Mito-TEMPO on post-thawed quality of spermatozoa in sheep. Collected samples were divided into five groups and after dilution, received different doses of Mito-TEMPO (0, 0.5, 5, 50 and 500 µM), and frozen. Thawed sperm motility parameters, malondialdehyde content, membrane functionality, abnormal morphology, mitochondria activity, acrosome integrity, DNA fragmentation, ROS concentration, viability and apoptotic-like changes, were evaluated. According to the results, Mito-TEMPO (5 and 50 μM) improved (p ≤ 0.05) motility parameters, average path velocity, membrane functionality, mitochondria activity and viability compared with the other groups. Moreover, apoptotic-like changes, lipid peroxidation and ROS concentration were lower (p ≤ 0.05) in groups received 5 and 50 μM Mito-TEMPO. Mito-TEMPO showed no effect (p > 0.05) on sperm acrosome integrity, morphology and DNA fragmentation. In conclusion, Mito-TEMPO as a targeted antioxidant could be an efficient cryo-additive to enhance quality parameters of post-thawed ram semen.


Epidemiology and risk factors of infections after solid organ transplantation.
Infection remains a significant complication after solid organ transplantation (SOT). The incidence of various pathogens varies widely depending on the presence of specific factors, according to which patients can be classified into different risk categories that may merit tailored prophylaxis strategies. Both the endogenous origin of microorganisms (previous colonization or latent infection) and new acquisition (primary infection from donor or environment) should be considered. Bacterial infections predominate in patients with complex hospital stays or anatomical alterations. Viral infections, caused both by opportunistic (CMV, EBV, BKV, etc.) and common viruses (influenza, respiratory virus, VVZ, etc.), are of great importance, and may contribute to chronic rejection. Fungal infections are uncommon nowadays, but cause high mortality and deserve prophylaxis for a subset of patients. Parasitic infections are a clear threat, mainly in transplanted patients or those travelling to endemic areas. Physicians attending SOT recipients should be aware of these risk factors, which include specific host characteristics, type of transplantation, microorganism and immunosuppressive policy.


Analysis of omeprazole, midazolam and hydroxy-metabolites in plasma using liquid chromatography coupled to tandem mass spectrometry.
A method has been developed and validated for the quantitation of midazolam, alphahydroxy-midazolam, omeprazole, and hydroxyomeprazole from one 250 microL sample of human plasma using high performance liquid chromatography coupled to tandem mass spectrometry. The method was validated for a daily working range of 0.400-100 ng/mL, with limits of detection between 2 and 15 pg/mL. The inter-assay variation was less than 15% for all analytes at four control concentrations and the samples were stable for three freeze-thaw cycles under the analysis conditions and 24 h in the post-preparative analysis matrix. This method was used to analyze samples in support of clinical studies probing the activity of the cytochrome P-450 enzyme system.


The Role of Telehealth During the COVID-19 Pandemic Across the Interdisciplinary Cancer Team: Implications for Practice.
This literature review aims to explore the role of telehealth during the COVID-19 pandemic across the interdisciplinary cancer care team. Electronic databases including CINAHL, MEDLINE, PsychINFO, Scopus, and gray literature were searched using Google Scholar up until September 2020. Although the safe and effective delivery of cancer care via telehealth requires education and training for health care professionals and patients, telehealth has provided a timely solution to the barriers caused by the COVID-19 pandemic on the delivery of interdisciplinary cancer services. Globally, evidence has shown that telehealth in cancer care can leverage an innovative response during the COVID-19 pandemic but may provide a long-lasting solution to enable patients to be treated appropriately in their home environment. Telehealth reduces the travel burden on patients for consultation, affords a timely solution to discuss distressing side effects, initiate interventions, and enable possible treatment additions and/or changes. Global public health disasters pose significant and unique challenges to the provision of necessary services for people affected by cancer. Oncology nurses can provide a central contribution in the delivery of telehealth through transformational leadership across all domains and settings in cancer care. Oncology nurses provide the "hub of cancer care" safely embedded in the interdisciplinary team. Telehealth provides a solution to the current global health crisis but could also benefit the future provision of services and broad reach clinical trials.


Dihydroartemisinin downregulates vascular endothelial growth factor expression and induces apoptosis in chronic myeloid leukemia K562 cells.
Dihydroartemisinin (DHA), a more water-soluble active metabolite of artemisinin derivatives, is safe and the most effective antimalarial analog of artemisinin. In the present investigation, we assessed the effect of DHA on vascular endothelial growth factor (VEGF) expression and apoptosis in chronic myeloid leukemia (CML) K562 cells. The results demonstrated that in addition to its antiproliferation effect on CML cells, DHA was also found to induce K562 cells apoptosis. The percentage of apoptotic cells was increased to 6.9 and 15.8% after being treated with 5 and 10 micromol/l DHA for 48 h, respectively (P<0.001). In order to analyze the effect of DHA on VEGF expression in K562 cells, we assessed the level of VEGF expression by western blot; detected the form of VEGF mRNA by RT-PCR and examined the level of VEGF secreted in conditioned media (CM) by ELISA assay. All these experiments suggested that DHA could inhibit the VEGF expression and secretion effectively in K562 cells, even at a lower concentration (2 micromol/l, P<0.05). Moreover, we further assessed the stimulating angiogenic activity of CM from K562 cells on CAM model. The angiogenic activity was decreased in response to the CM from K562 cells pretreated with DHA in a dose-dependent manner. Taken together, these results from our study together with its known low toxicity make it possible that DHA might present potential antileukemia effect as a treatment for CML therapy, or as an adjunct to standard chemotherapeutic regimens.


Weak shoulder and arm sparing signs in amyotrophic lateral sclerosis.
Various signs of selective involvement have been reported in amyotrophic lateral sclerosis (ALS). In this study, we describe two new signs, "weak shoulder" and "arm sparing" signs. Subjects were retrospectively identified from our electrodiagnosis database. Medical Research Council scores of relevant muscles were evaluated. Weak shoulder was defined as the deltoid (Del) muscle being weaker than the biceps brachii (BB)/triceps brachii (TB) muscles; that is, Del was weaker than either or both of the muscles and no stronger than either. Arm sparing was defined as both Del and the first dorsal interosseous (FDI) being weaker than BB/TB. Sensitivities of these signs were compared with other signs of selective involvement. The specificities of these signs were investigated in patients with cervical spondylotic amyotrophy (CSA) and multifocal motor neuropathy (MMN). We reviewed 130 patients with ALS, 64 patients with CSA, and 16 patients with MMN. The weak shoulder and the arm sparing signs were observed in 73% and 55% of patients with ALS, 44% and 2% of patients with CSA (93% and 0% of patients with proximal CSA), respectively, and no patients with MMN. The sensitivity of the weak shoulder was higher than with conventional signs, whereas that of the arm sparing sign showed no difference. The weak shoulder sign was highly sensitive in ALS, and was specific when compared with MMN. The arm sparing sign was highly specific for ALS. These two new signs are promising as clinical clues in the diagnosis of ALS.


Cochlear nonlinearity and gain control as determinants of the response of primary auditory neurons to harmonic complexes.
Earlier papers reported a calculational "model" (Greenwood, 1985, 1986a, b, c), composed of an initial compressive nonlinearity accompanied by a nonlinear gain control and "rectification", to demonstrate in principle certain "two-tone suppression" effects seen in primary fiber response. This paper presents simulations of primary fiber response to more complex stimuli. When harmonic stimuli of Horst et al. (1985a, b; 1986a, b) are used, the calculated spectra closely agree with the empirical spectra they report, from fibers with both low or high spontaneous activity. Thus, the simulated nonlinear motion of the cochlear partition reflects a sufficient degree of compressive distortion and an appropriate form to calculate spectra closely matching the empirical spectra in respect to the pattern of intermodulation products and the main "enhancement" and cancellation effects. Comparison of spectra of full and truncated waveforms assess added effects of "rectification". The gain control, as for simpler stimuli, leads to appropriate growth and saturation of Fourier coefficients and average firing rate. The implication that the measured transverse motion of the cochlear partition could provide sufficient distortion to account for the main features of the histograms has a two-part corollary: that the mechanism which produces severe overall saturation does not add much compressive distortion to the waveform passed radially to IHC and primary fibers, and that the IHC is kept operating chiefly in an approximately linear part of its range, by a prior gain control that is, perforce, cochlear and mechanical. Further consequences, for IHC synapses and fiber types, stem from the sufficiency of negatively biased waveform centerlines in generating simulated period histograms and spectra which match those of at least some primary fibers characterized by low spontaneous activity.


Commentary: How useful is 'burden of disease' to set public health priorities for infectious diseases?
For many infectious diseases, a low burden of disease does not equate to reduced potential public health importance. Many zoonotic infectious diseases have the potential for human-to-human transmission with potentially devastating consequences as currently seen with Ebola. Policymakers should not be lulled into thinking that the best use of resources is to allocate them only to the most obvious current problems.


Current trends in pharmaceutical treatment of dry eye disease: A review.
Dry eye disease (DED), keratoconjunctivitis sicca or dysfunctional tear syndrome, is the most prevalent ophthalmic disease which affects a substantial segment of people worldwide with increasing frequency. It is considered a multifactorial disease of the ocular surface and tear film, characterized by a variation of signs and symptoms. The symptoms range from mild to severe itching, burning, irritation, eye fatigue, and ocular inflammation that may lead to potential damage to the cornea, conjunctiva and even vision loss. Correspondingly, depending on the different manifestations and pathophysiology, the treatment must be tailored specifically to each patient by targeting the specific mechanisms implicated in their disease. Currently, there are several medical products and techniques available or under investigation for the treatment of DED. The present article focused on the pathophysiology of DED, the new diagnostic approach and the recently developed drug delivery systems or devices reducing the progress of the disease and treating the causes.


Gravity Observations and Apparent Density Changes before the 2017 Jiuzhaigou Ms7.0 Earthquake and Their Precursory Significance.
An Ms7.0 earthquake struck Jiuzhaigou (China) on 8 August 2017. The epicenter was in the eastern margin of the Tibetan Plateau, an area covered by a dense time-varying gravity observation network. Data from seven repeated high-precision hybrid gravity surveys (2014-2017) allowed the microGal-level time-varying gravity signal to be obtained at a resolution better than 75 km using the modified Bayesian gravity adjustment method. The "equivalent source" model inversion method in spherical coordinates was adopted to obtain the near-crust apparent density variations before the earthquake. A major gravity change occurred from the southwest to the northeast of the eastern Tibetan Plateau approximately 2 years before the earthquake, and a substantial gravity gradient zone was consistent with the tectonic trend that gradually appeared within the focal area of the Jiuzhaigou earthquake during 2015-2016. Factors that might cause such regional gravitational changes (e.g., vertical crustal deformation and variations in near-surface water distributions) were studied. The results suggest that gravity effects contributed by these known factors were insufficient to produce gravity changes as big as those observed, which might be related to the process of fluid material redistribution in the crust. Regional change of the gravity field has precursory significance for high-risk earthquake areas and it could be used as a candidate precursor for annual medium-term earthquake prediction.


Sulfate radical induced degradation of Methyl Violet azo dye with CuFe layered doubled hydroxide as heterogeneous photoactivator of persulfate.
Persulfate (PS)-based advanced oxidation processes have aroused considerable attentions due to their higher efficiency and wider adaptability to the degradation of bio-recalcitrant organic contaminants. In this study, Cu-Fe layered doubled hydroxide (CuFe-LDH) was employed to degrade Methyl Violet (MV) through heterogeneous photo-activation of PS under visible-light irradiation. The reaction kinetics, degradation mechanism, catalyst stability were investigated in detail. Under the conditions of CuFe-LDH (3:1) dosage 0.2 g/L, PS concentration 0.2 g/L and without initial pH adjustment, 20 mg/L MV was almost completely degraded within 18 min. Electron Spin Resonance (ESR) test and radical quenching experiment indicated that sulfate radicals (SO4-) were the dominant reactive oxidants for the MV decolorization, while hydroxyl radicals (OH) were also involved. The CuFe-LDH/PS/Vis system was applicable at wide range of pH level (3-9). However, extreme pH level would lead to the reduction or transformation of SO4-. The catalyst CuFe-LDH exhibited excellent stability and maintained relatively high catalytic activity to PS even after four recycles. Mechanism study revealed that the redox cycle of Fe3+/Fe2+ and Cu2+/Cu3+ assisted by visible-light irradiation accounted for the enhanced generation of radicals in CuFe-LDH/PS/Vis system, resulting in the improved degradation of organic contaminants. Overall, the CuFe-LDH/PS/Vis process could be a promising approach for the removal of refractory organic pollutants in wastewater.


Stem cell treatment and cerebral palsy: Systemic review and meta-analysis.
Perinatal complications may result in life-long morbidities, among which cerebral palsy (CP) is the most severe motor disability. Once developed, CP is a non-progressive disease with a prevalence of 1-2 per 1000 live births in developed countries. It demands an extensive and multidisciplinary care. Therefore, it is a challenge for our health system and a burden for patients and their families. Recently, stem cell therapy emerged as a promising treatment option and raised hope in patients and their families. The aim is to evaluate the efficacy and safety of stem cell treatment in children with CP using a systematic review and meta-analysis. We performed a systematic literature search on PubMed and EMBASE to find randomized controlled clinical trials (RCT) investigating the effect of stem cell transplantation in children with CP. After the review, we performed a random-effects meta-analysis focusing on the change in gross motor function, which was quantified using the gross motor function measure. We calculated the pooled standardized mean differences of the 6- and/or 12-mo-outcome by the method of Cohen. We quantified the heterogeneity using the I-squared measure. We identified a total of 8 RCT for a qualitative review. From the initially selected trials, 5 met the criteria and were included in the meta-analysis. Patients' population ranged from 0.5 up to 35 years (n = 282). We detected a significant improvement in the gross motor function with a pooled standard mean difference of 0.95 (95% confidence interval: 0.13-1.76) favoring the stem cell group and a high heterogeneity (I 2 = 90.1%). Serious adverse events were rare and equally distributed among both intervention and control groups. Stem cell therapy for CP compared with symptomatic standard care only, shows a significant positive effect on the gross motor function, although the magnitude of the improvement is limited. Short-term safety is present and further high-quality RCTs are needed.


[Intraerythrocyte levels of glucose-6-phosphate dehydrogenase in patients with essential arterial hypertension and persons with normal blood pressure. Modifications after nifedipine therapy].
We evaluate the levels of intra-erythrocytes glucose-6-phosphate-dehydrogenase (G6PD) in a group of 50 patients with essential blood hypertension (EBH) (214.76 +/- 38.31 mU/ml) and in a group of 50 healthy persons (130.11 +/- 49.50 mU/ml) (p minor of 0.0005). At the same time, we have analysed the intraerythrocyte levels of G6PD of 34 healthy sons and daughters of hypertensive parents, detecting high levels in 13 of them. We observed a significant decrease of G6PD after the therapy with nifedipine in 20 hypertensive patients (p less than 0.0005). We conclude that measurement of G6PD enzyme activity can prove to be a good marker of essential hypertension. This change occurs at an early stage and is noticeable in high risk persons with family predisposition. It is possible to modify levels by working on cation changes through membrane cells by calcium-antagonist.


Draft Genome Sequences of Antibiotic-Resistant Serratia sp. Strains Isolated from Raw Sewage and Sediment Sludge in Georgia, USA.
Sewage has been established as a prime matrix for monitoring the emergence and dissemination of etiologic agents and antibiotic resistance determinants in a population. Here, we report the draft genomes and the acquired resistance genes of 11 antibiotic-resistant Serratia sp. isolates that were detected in samples from wastewater treatment plants.


Evaluation of baits for oral vaccination of European wild boar piglets.
The objective of this study was to develop and evaluate new baits for the oral delivery of vaccine preparations to 2-4 month-old wild boar piglets. Baits were prepared using a matrix composed of wild boar feed, wheat flour, paraffin, sacarose and cinnamon-truffle powder attractant with polyethylene capsules dipped into the matrix to introduce vaccine formulation. Physical stability studies demonstrated that baits were stable for at least three days at temperatures as high as 42 degrees C. Recombinant Escherichia coli expressing the membrane-displayed BM95-MSP1a fusion protein were used to test bacterial viability in the baits and the antibody response in orally immunized wild boar. The E. coli viability was not significantly affected after bait incubation at 25 and 37 degrees C for 96h. Bait acceptance studies using artificial feeders in the field showed that baits were accepted by 2-3month-old animals, the preferred age for vaccination. Orally immunized wild boar piglets excreted recombinant E. coli in the feces and developed antibody titers to recombinant BM95-MSP1a protein, thus confirming that vaccine composition was released and reached the wild boar gastrointestinal track. The results of these experiments support the use of these baits for oral delivery of vaccine formulations to 2-4month-old wild boar piglets.


Heavy metal accumulation (lead and cadmium) and ion exchange in three species of Sphagnaceae : II. Chemical equilibrium of ion exchange and the selectivity of single ions.
Chemical equilibrium of ion exchange reactions of Sphagnum mosses was investigated for four different combinations of ions. These combinations were: Pb/Ca; Cd/Ca; Pb/K; Cd/K. Based on the law of mass action one can find mathematical descriptions of ion exchange reactions which permit one to calculate a state of equilibrium. As a measure of ion selectivity a rational coefficient of selectivity can be used. For culture experiments three different species of Sphagnum originating from a bog ("Hochmoor") in southern Bavaria were used. With these species coefficients of selectivity of the mentioned heavy metals were determined in solutions of different total ion content. Furthermore for each test solution of fixed total concentration the portions of the ions were altered. The Sphagnum material shows the behaviour of a relative ideal ion exchanger, whose states of equilibrium can be well described by the mathematical equations. The coefficients of selectivity are independent of Sphagnum species. In all cases they reflect the preferred binding of lead to the exchanger. Cadmium and calcium are adsorbed in approximately equivalent amounts. Representative limit isotherms permit an estimation of the errors caused by variation of the rational coefficients of selectivity. The applicability of the determined interactions and regularities on heavy metal monitoring programs is discussed.


The value of computed tomography scans in the diagnosis and management following invasive arterial investigation.
We report a patient who was known to have an abdominal aortic aneurysm and was being assessed for endovascular repair by calibrated angiogram, who developed back pain and cardiovascular collapse, where a computed tomography scan proved essential in establishing the correct diagnosis - bleed from an accessory renal artery branch.


Influence of Brugia malayi life stages and BmAFII fraction on experimental Leishmania donovani infection in hamsters.
The influence of live Brugia malayi parasites and a Sephadex G-200 fraction of the adult parasite extract (BmAFII) on the progression of Leishmania donovani infection was studied. Inbred hamsters were first infected with B. malayi infective 3rd stage larvae (L3), adult worms or microfilariae (mf), and then with L. donovani amastigotes (Ld), or vice versa or received both the infections simultaneously; a group of animals were first immunized with BmAFII and then infected with Ld. L. donovani parasite burden was determined between 17 and 19 days post amastigote challenge (p.a.c.) and, in case of immunized animals, between 32 and 35 days p.a.c also. Nitric oxide (NO) release from peritoneal macrophages and cellular proliferative responses of lymphnode cells were assessed in BmAFII-immunized animals given leishmania infection or no infection. Leishmanial parasite burden was significantly reduced in animals exposed to filarial L3 before amastigote inoculation and in animals given filarial adult worms after or together with amastigotes. Prior immunization of leishmania-infected animals with BmAFII also reduced the leishmanial parasite burden (17-19 days p.a.c.: >90%; 32-35 days p.a.c.: 60%). These animals showed upregulation of NO release and cellular proliferative responses to promastigote antigen or BmAFII stimulation in vitro. The findings show, for the first time, that B. malayi L3/adult worms or immunization with BmAFII inhibits progression of L. donovani infection in hamsters and this is associated with upregulation of NO and lymphocyte proliferative responses indicating that Th1 response might be responsible for this.


The male urethral sphincter complex revisited: an anatomical concept and its physiological correlate.
The anatomy of the male urethral sphincter has not been stable since it was first described more than 150 years ago. Although 18th and 19th century historical descriptions of the urethral sphincter are most accurate and comprehensive, modern textbooks lack details and include inaccuracies and misleading illustrations. This is an attempt to achieve a revised concept of the male urethral sphincter complex. A thorough review of the English literature in the last 100 years, and of pertinent Germinal publications and textbooks of the 19th and 20th centuries was done. Also, we reviewed urodynamic findings in male patients in whom the urethral sphincters had been expectedly damaged in the proximal or distal part by surgery during the last 20 years. The current concept of urethral sphincter anatomy does not differ much from that described and illustrated in the 19th century. The disagreement between the historical and recent descriptions is primarily concerned with the cranial extension of the skeletal muscle component and the caudal extension of the smooth muscle component in the urethral wall. The male urethral sphincter complex is composed of an inner lissosphincter of smooth muscle and an outer rhabdosphincter of skeletal muscle. It extends in the form of a cylinder around the urethra from the vesical orifice to the perineal membrane. While the rhabdosphincter is most marked around the membranous urethra and becomes gradually less distinct toward the bladder, the lissosphincter has its main part at the vesical orifice and is thinner in its further course in the urethra. The lissosphincter is primarily concerned with the function of continence at rest. On the other hand, the rhabdosphincter has a dual genitourinary function, namely active continence during stress conditions and antegrade semen propulsion.


Ectopic pregnancy with use of progestin-only injectables and contraceptive implants: a systematic review.
Use of contraception lowers a woman's risk of experiencing an ectopic pregnancy. In the case of method failure, however, progestin-only contraceptives may be more likely to result in ectopic pregnancies than some other methods such as combined hormonal and barrier contraceptives. To describe ectopic pregnancy risk associated with use of implants and progestin-only injectable contraceptives through a systematic review of published studies. We searched electronic databases for articles in any language published through May 2015 describing studies of progestin-only injectables and implants. We also searched bibliographies and review articles for additional studies. Studies that reported any pregnancies were included in the review. Independent data extraction was performed by two authors based on predefined data fields, and where possible, we calculated the proportion of pregnancies that were ectopic and the ectopic pregnancy incidence rate per 1000 woman-years. Fifty-three studies of implants and 28 studies of injectables were identified; 79% reported pregnancy location. The proportion of ectopic pregnancy ranged from 0 to 100% with an incidence of 0-2.9 per 1000 woman-years in studies of marketed levonorgestrel implants. Studies of etonogestrel implants and the injectables, depot-medroxyprogesterone acetate and norethisterone enanthate, reported few ectopic pregnancies. Progestin-only contraceptive implants and injectables protect against ectopic pregnancy by being highly effective in preventing pregnancy overall; however, the absolute risk of ectopic pregnancy varies by type of progestin. Risk of ectopic pregnancy should not be a deterrent for use or provision of these methods.


Differential changes of autonomic nervous system function with age in man.
To assess the relationship between aging and autonomic nervous system function, cardiovascular and pupillary autonomic nervous system reflexes were measured in subgroups of 103 normal male subjects ranging in age from 19 to 82 years (mean age = 39 years). Both the plasma norepinephrine level, a measure of cardiovascular sympathetic nervous system activity, and the mean arterial blood pressure increased with age (r = 0.68 and 0.67, respectively, both p less than 0.001). In contrast, the plasma epinephrine level, a measure of adrenomedullary sympathetic nervous system activity, was unrelated to age (r = 0.08, p = NS). Respiratory variation of heart rate during beta-adrenergic blockade, an index of cardiac parasympathetic nervous system activity, was reduced in older subjects (r = -0.54, p less than 0.001). Thus, there was evidence of an age-related increase of cardiovascular sympathetic nervous system activity and a reduction of cardiac parasympathetic nervous system activity. These findings are consistent with the hypothesis that there is sympathetic nervous system and parasympathetic nervous system compensation of cardiovascular function in response to an age-related decrease in baroreceptor sensitivity. However, dark-adapted pupil size during parasympathetic nervous system blockade, an index of iris sympathetic nervous system activity, declined with age (r = -0.81, p less than 0.001). The latency time for the pupillary response to a light stimulus, an index of iris parasympathetic nervous system activity, was prolonged in older subjects (r = 0.58, p less than 0.001). Thus, both sympathetic nervous system and parasympathetic nervous system inputs to the iris were diminished in older subjects, findings consistent with the generalized decrease of peripheral somatic nerve function that has been reported with aging in man. It is concluded that autonomic nervous system function also declines with aging, but that other age-related changes such as a decline of baroreceptor sensitivity may lead to compensatory autonomic nervous system response, which could mask underlying functional defects.


Direct needle insufflation for pneumoretroperitoneum: anatomic confirmation and clinical experience.
The feasibility and safety of direct needle insufflation to create pneumoretroperitoneum was assessed by an imaging study and clinical experience. A total of 10 patients without previous retroperitoneal surgery or diseases received computed tomography scans of the retroperitoneum 2 cm above the iliac crest. Distances between quadratus lumborum and colon (Q-C distance) were measured in the supine and lateral positions. Changes of Q-C distance were calculated when the patient was changed from the supine to the lateral position. Operative charts on 38 retroperitoneoscopic procedures were collected prospectively to assess complications related to direct needle insufflation, which was performed by inserting a 14 G Veress needle blindly along the posterior axillary line 2 cm above the iliac crest. Q-C distance increased from 8.7 to 27.3 mm (left side) and 4.6 to 18.1 mm (right side) when the patient was changed from the supine to the lateral position, both P values < 0.05. An average distance of 23 mm between colon and quadratus lumborum was found when patients were lying laterally. The misplacement of a Veress needle was encountered in 1 patient, in which a prefascia insufflation resulted in conversion of the endoscopic procedure. Needle puncture caused no visceral or great vessel injury. Significant anterior movement of the colon was found when patients were changed from the supine to the lateral position. It provided a window for inserting the Veress needle blindly into the retroperitoneum. The high success rate (97%) and low complication rate of direct needle insufflation were found in actual clinical applications. We considered needle insufflation a safe and effective method of establishing a pneumoretroperitoneum for any retroperitoneoscopic procedure.