Internet-based guided self-help comprehensive behavioral intervention for tics (ICBIT) for youth with tic disorders: a feasibility and effectiveness study with 6 month-follow-up.
Practice guidelines endorse comprehensive behavioral intervention for tics (CBIT) as first-line treatment for tic disorders (TD) in youth. Nevertheless, CBIT is rarely available due to various barriers. This study evaluated the feasibility and potential effectiveness of an Internet-based, self-help CBIT program (ICBIT) guided by parents with minimal therapist support delivered via telepsychotherapy. Forty-one youths, aged 7-18 years, were randomly assigned to receive either ICBIT (n = 25) or a wait-list (WL) condition (n = 16) in a crossover design. ICBIT was feasible to implement and at post-treatment, 64% of the participants have improved significantly. Results demonstrated clinically meaningful reductions in tic severity and improved youth global impairment and functioning. Gains were maintained over a 6-month follow-up period. The effect size for the primary outcome measure (Yale Global Tic Severity Scale) ranged between large effect size (Cohen"s d = 0.91) at post-intervention to very large effect size (Cohen's d = 2.25) 6 months after the end of the acute intervention. These were comparable to face-to-face delivery treatment trials for TD. Participants rated the intervention as highly acceptable and satisfactory. Youth receiving ICBIT experienced improvement in self-esteem and comorbidity. Finally, during the COVID-19 pandemic, the ICBIT program enabled the delivery of the intervention consecutively without interruption. The results observed provide preliminary evidence of the feasibility and effectiveness of this innovative modality to assist youth with TD and remove various barriers to treatment, including those during a public crisis, such as the COVID-19 pandemic. Larger studies with an active control group are warranted.Trial registration URL: http://clinicaltrials.gov, ClinicalTrials.gov Identifier: NCT04087616.


Inflammatory tissue reaction in rabbit bowel injected with Crohn's homogenates.
Homogenates from the terminal ileum of a patient with Crohn's disease with granulomas were prepared as snap-frozen or fresh and were injected into the ascending colonic walls of New Zealand white rabbits. Control animals were injected with 1 per cent bovine serum albumin alone. The rabbit bowel was examined after 1 year, and lesions were noted in each of the rabbits injected with Crohn's disease homogenate, irrespective of the type of tissue preparation. The observed lesions were diffuse and occurred both at the injection site and in the terminal ileum. These changes were not noted in the control group. This work confirms earlier results in the same animal model and suggests that either fresh or snap-frozen homogenates will produce the intestinal lesion but that bovine albumin alone will not.


Analgesic efficacy and quadriceps strength of adductor canal block versus femoral nerve block following total knee arthroplasty.
Femoral nerve blocks (FNBs) provide effective analgesia after total knee arthroplasty, but have been associated with quadriceps weakness. Adductor canal block (ACB) is a promising alternative option that delivers a primarily sensory blockade. The aim of this study was to determine whether ACB provides superior quadriceps strength and similar pain control than FNB. A systematic search of PubMed, the Cochrane Library, EMBASE and Web of Science was conducted without publication data or language restriction. Comparative studies comparing ACB with FNB were included. Two authors independently assessed data extraction and quality of the studies. Nine studies with 639 patients were identified. Results of meta-analysis showed that compared with FNB, ACB preserved quadriceps muscle strength better than FNB [MD24h = 1.14, 95 % CI (0.38, 1.91), p < 0.01; MD48h = 0.40, 95 % CI (0.16, 0.64), p < 0.01], while there were no significant differences in pain score during rest at 24 h [SMD = - 0.04, 95 % CI (-0.17, 0.26); n.s] or 48 h [SMD = - 0.10, 95 % CI (-0.27, 0.08); n.s], pain score during activity at 24 h [SMD = 0.13, 95 % CI (-0.36, 0.62); n.s] or 48 h [SMD = - 0.13, 95 % CI (-0.37, 0.12); n.s], opioid consumption at 24 h [SMD = - 0.01, 95 % CI (-1.68, 1.66); n.s] or 48 h [SMD = - 0.92, 95 % CI (-6.86, 5.01); n.s], length of hospital stay [MD = 0.05, 95 % CI (-0.91, 1.00); n.s] and nausea or vomiting [RR = 1.17, 95 % CI (0.62, 2.20); n.s] between ACB and FNB. ACB preserved the strength of quadriceps more than FNB and achieves similar analgesic effects in post-operative pain. Level III.


Social behavior of butterfly and surgeonfishes on coral reefs: Some mirror experiments.
Chaetodontids and acanthurids showed behavior that suggested the existence of dominance systems. There was no sign that the poster colors of chaetodontids serve as territorial signals, and there were interspecific differences in their responses to the mirror. Aggressive butterfly fishes usually butted with the spines of the forward part of the dorsal fin; surgeonfishes used their scalpels in attacks against their images.


Multiple Sclerosis Severity Score (MSSS) improves the accuracy of individualized prediction in MS.
The MSBase prediction model of treatment response leverages multiple demographic and clinical characteristics to estimate hazards of relapses, confirmed disability accumulation (CDA), and confirmed disability improvement (CDI). The model did not include Multiple Sclerosis Severity Score (MSSS), a disease duration-adjusted ranked score of disability. To incorporate MSSS into the MSBase prediction model and compare model accuracy with and without MSSS. The associations between MSSS and relapse, CDA, and CDI were evaluated with marginal proportional hazards models adjusted for three principal components representative of patients' demographic and clinical characteristics. The model fit with and without MSSS was assessed with penalized r2 and Harrell C. A total of 5866 MS patients were started on disease-modifying therapy during prospective follow-up (age 38.4 ± 10.6 years; 72% female; disease duration 8.5 ± 7.7 years). Including MSSS into the model improved the accuracy of individual prediction of relapses by 31%, of CDA by 23%, and of CDI by 24% (Harrell C) and increased the amount of variance explained for relapses by 49%, for CDI by 11%, and for CDA by 10% as compared with the original model. Addition of a single, readily available metric, MSSS, to the comprehensive MSBase prediction model considerably improved the individual accuracy of prognostics in MS.


Optimum scratch assay condition to evaluate connective tissue growth factor expression for anti-scar therapy.
To evaluate a potential anti-scar therapy, we first need to have a reliable in vitro wound model to understand dermal fibroblast response upon cell injury and how cytokine levels are changed upon different wound heal phases. An in vitro wound model with different scratch assay conditions on primary human foreskin fibroblast monolayer cultures was prepared and cytokine levels and growth properties were evaluated with the aim of determining optimum injury conditions and observation time. Morphological characteristics of differently scratched fibroblasts from 0 to 36 h post injury (1 line, 2 lines and 3 lines) were investigated. The expression of connective tissue growth factor, CTGF, which is a key mediator in hyper-tropic scarring, and relative intensity of CTGF as a function of time were determined by western blot and gelatin Zymography. After injury (1 line), CTGF level was increased more than 2-fold within 1 h and continuously increased up to 3-fold at 6 h and was leveled down to reach normal value at 36 h, at which cell migration was complete. In more serious injury (2 lines), higher expression of CTGF was observed. The down regulation of CTGF expression after CTGF siRNA/lipofectamine transfection in control, 1 line and 2 lines scratch conditions were 40%, 75% and 55%, respectively. As a model anti-CTGF based therapy, CTGF siRNA with different ratios of linear polyethyleneimine (PEI) complexes (1:1, 1:5, 1:10, 1:20 and 1:30) were prepared and down-regulation efficacy of CTGF was evaluated with our optimized scratch assay, which is 1 line injury at 6 h post injury observation time. As the cationic linear PEI ratio increased, the down regulation efficacy was increased from 20% (1:20) to 55% (1:30). As CTGF level was increased to the highest at 6 h and leveled down afterwards, CTGF level at 6 h could provide the most sensitive response upon CTGF siRNA transfection. The scratch assay in the present study can be employed as a useful experimental tool to differentiate between anti-scar therapies for their down regulation efficacy of CTGF.


Cinnamyl Chalcone Based AIE Fluorescent Probes for Sensitive Detection of Hydrazine and its Application in Living Cells.
Widely utilized in the chemical industry and agriculture, hydrazine is easily absorbed by living things and can cause physical harm when in touch for an extended period of time. As a result, a novel cinnamaldehyde chalcone C5 was produced by Friedel Crafts process and aldol condensation reaction. Triphenylamine was used as the raw material for hydrazine determination in both reactions. Chalcone C5 exhibits significant AIE behavior in a mixed mixture of ethanol and water in addition to having great selectivity and a low detection limit (0.119 nm) for hydrazine. The solvent effect test revealed a linear relationship between the Stokes shift of C5 in the solvent and the rise in solvent orientation polarization. It is important to note that C5 is not harmful to MCF-7 cells, mouse kidney cells, or pig kidney cells. Furthermore, research on cell imaging has demonstrated that probe C5 may be utilized to image the fluorescence of hydrazine in active MCF-7 cells.


The impact of fabric conditioning products and lint filter pore size on airborne microfiber pollution arising from tumble drying.
Vented tumble dryers release moist warm air from the drying process to the external environment, usually through pipework linking the appliance to a vent in an exterior wall. Although such dryers contain a lint filter to remove fibers from this air stream, recent reports suggest that this process is incomplete, leading to microfibers being released in the ducted warm air and subsequently polluting the external environment. Microfiber release from wash loads comprising 10 100% cotton and 10 100% polyester T-shirts (total load mass ratio 48% cotton, 52% polyester) was measured at different stages of the washing and drying process to compare the quantities of fibers released 'down the drain', collected in the dryer lint filter, and released to air from the tumble dryer. Testing under both European and North American washing conditions found that the quantities of microfibers released to air during tumble drying were significant and comparable to levels released 'down the drain' during washing. Use of conventional rinse-added liquid fabric conditioner increased microfiber accumulation on the dryer lint filter, with reduced release from the dryer exhaust observed at the highest fabric conditioner dose tested (21.6% and 14.2% reduction under North American and European conditions, respectively). Conventional liquid fabric conditioner did not significantly impact microfiber release from the washing machine, in line with previous studies. A fabric conditioner specially designed for anti-wrinkle performance reduced microfiber release from the dryer exhaust at all levels tested (by 17.6-35.6%, depending on dose), apparently by increasing the efficiency of microfiber accumulation in the lint filter. Tumble dryer sheets were also found to cause a reduction in microfiber release from the dryer exhaust (by 14.1-34.9%, depending on the dose/product), likely driven by collection of liberated fibers on the sheet during the drying process. The use of both antiwrinkle liquid fabric conditioner and dryer sheet enabled a 44.9% reduction in microfiber emissions from the dryer exhaust. In all studies, the fiber mass collected on the lint filter or emitted from the dryer exhaust was richer in cotton fibers (range 83.4-96.3% on the lint filter, 93.0-99.8% from the dryer exhaust) than the wash load composition (48% cotton). Moreover, fibers collected by the lint filter contained a higher proportion of polyester than emissions from the dryer exhaust (range 3.7-16.6% on the lint filter, 0.2-7.0% from the dryer exhaust). There is significant variation in the porosity of lint filters among installed vented tumble dryers. Single-variable testing of the impact of lint filter design concluded that reducing screen pore size significantly reduces airborne microfiber release during tumble drying; a reduction in lint filter pore size from 0.2 mm2 to 0.04 mm2 reduced release by 34.8%. As some lint filters have pore sizes of around 1 mm2, there is enormous scope to reduce microfiber release from dryers though improved lint filter design. However, it is suggested that a step-change in appliance design away from vented dryers to only fully-sealed condenser dryers might be necessary to eliminate the contribution of tumble drying to airborne microfiber pollution.


Update on vena cava filters.
Inferior vena cava (IVC) filter placement has increased dramatically over the past two decades. Filters are indicated to prevent pulmonary embolism in patients with venous thromboembolism (VTE) and a contraindication to anticoagulation or a complication of anticoagulation. Some of this increased use is the result of expanding relative indications for filter placement, including placement for primary prophylaxis. The US Food and Drug Administration has approved 11 filters for permanent deployment, two of which--the Günther-Tulip (Cook Medical, Bloomington, IN) and the OptEase (Cordis Endovascular, Miami Lakes, FL)--are optionally retrievable. Once anticoagulation is deemed safe, all patients should be fully anticoagulated to prevent propagation and recurrent thromboembolism. Complications related to IVC filters include procedure-related issues, device complications, and secondary VTE. Therefore, the decision regarding filter placement and/or retrieval must be individualized.


MedChemTrain e-School 2020: Event Highlights and Insights into Virtual Symposium Organization.
Virtual events are flourishing with the world lockdown due to the COVID-19 pandemic. As a result of the cancelation or postponement of scheduled physical meetings, a revolution in medicinal chemistry scientific meetings occurred, leading to an increase in new strategies to share science. One example are online events, namely e-schools or webinars. Taking this into consideration, we decided to promote the MedChemTrain e-School 2020, a virtual event aiming to bring together the scientific community and share some updates in the medicinal chemistry field. After organizing this free event, with more than 1.4 thousand participants worldwide, we decided to share some insights about the logistics behind organizing a virtual symposium to help scientists with this new challenge in science communication.


Staff Knowledge, Awareness, Perceptions, and Beliefs About Infection Prevention in Pediatric Long-term Care Facilities.
The burden of healthcare-associated infection worldwide is considerable, and there is a need to improve surveillance and infection control practices such as hand hygiene. The aims of this study were to explore direct care providers' knowledge about infection prevention and hand hygiene, their attitudes regarding their own and others' hand hygiene practices, and their ideas and advice for improving infection prevention efforts. This exploratory study included interviews with direct care providers in three pediatric long-term care facilities. Two trained nurse interviewers conducted semistructured interviews using an interview guide with open-ended questions. Two other nurse researchers independently transcribed the audio recordings and conducted a thematic analysis using a strategy adapted from the systematic text condensation approach. From 31 interviews, four major thematic categories with subthemes emerged from the analysis: (a) hand hygiene products; (b) knowledge, awareness, perceptions, and beliefs; (c) barriers to infection prevention practices; and (d) suggested improvements. There was confusion regarding hand hygiene recommendations, use of soap or sanitizer, and isolation precaution policies. There was a robust "us" and "them" mentality between professionals. One essential driver of staff behavior change is having expectations that are meaningful to staff, and many staff members stated that they wanted more in-person staff meetings with education and hands-on, practical advice. Workflow patterns and/or the physical environment need to be carefully evaluated to identify systems and methods to minimize cross-contamination. Further studies need to evaluate if personal sized containers of hand sanitizer (e.g., for the pocket, attached to a belt or lanyard) would facilitate improvement of hand hygiene in these facilities.


Prefrontal responses to drug cues: a neurocognitive analysis.
The construct of craving has been central to addiction research for more than 50 years. Only recently have investigators begun to apply functional neuroimaging techniques to the study of drug cue reactivity, and a small but growing number of studies implicate a distributed system of brain regions in the pathogenesis of craving. The internal consistency of this burgeoning literature has thus far been disappointing, however, leaving open the question of which brain regions contribute to craving. Here we review neuroimaging studies of cue-elicited craving in the context of a framework drawn from behavioral research indicating that perceived drug use opportunity significantly affects responses to the presentation of drug cues. Using this framework provides a way to reconcile discrepant findings among brain-imaging studies of cue-elicited craving.


Genomic structure of the Xenopus laevis liver transcription factor LFB1.
Liver factor B1 [LFB1, also called hepatocyte nuclear factor 1 (HNF1)] is a tissue-specific vertebrate transcription factor that is present in the liver, intestine, stomach and kidney. The LFB1 protein contains an unusual homeobox that is characterized by an insertion of 21 amino acids (aa) not found in any other homeodomain protein. We have isolated and characterized the genomic sequences encoding the LFB1 of Xenopus laevis. By comparing the genomic sequences with the cDNA clones, we could identify nine exons. In general, the position of the introns is identical to the one previously found in the rat. However, the C-terminal activation domain of LFB1 contains, in each species, an exon that is split in two in the other species. The homeobox of the X. laevis LFB1 contains an intron at exactly the position where the 21 aa typical for LFB1 are inserted. This is in agreement with the structure found in the rat gene and supports the notion that the LFB1 homeobox evolved separately from the other genes encoding homeodomain proteins.


Cannabinoid receptor-2 stimulation suppresses neuroinflammation by regulating microglial M1/M2 polarization through the cAMP/PKA pathway in an experimental GMH rat model.
Excessive inflammatory responses are involved in secondary brain injury during germinal matrix hemorrhage (GMH). The process of microglial polarization to the pro-inflammatory M1 or anti-inflammatory M2 phenotypes is considered to occur in a major immunomodulatory manner during brain inflammation. We previously found that cannabinoid receptor-2 (CB2R) stimulation attenuated microglial accumulation and brain injury following experimental GMH. However, whether CB2R has effects on microglial polarization after GMH remains unclear. Herein, we investigated the effects of CB2R stimulation on neuroinflammation after experimental GMH and the potential mechanisms that mediate M1/M2 microglial phenotype regulation. The results indicated that during the GMH acute phase, microglia primarily polarized to the M1 phenotype and induced an overwhelming release of pro-inflammatory cytokines. However, JWH133, a selective CB2R agonist, significantly prevented the pro-inflammatory cytokine release while promoting an M1 to M2 phenotype transformation in microglia, resulting in an increased anti-inflammatory cytokine release. Moreover, in thrombin-induced rat primary microglial cells, JWH133 reduced the pro-inflammatory cytokine levels and M1 phenotype by enhancing the acquisition of the M2 phenotype. Additionally, JWH133 facilitated synthesis of cyclic AMP (cAMP) and its downstream effectors, phosphorylated cAMP-dependent protein kinase (p-PKA) and exchange protein activated by cyclic-AMP 1 (Epac1). The promoting effects of JWH133 on M2 polarization were attenuated with a specific PKA inhibitor but not with an Epac inhibitor, indicating that the cAMP/PKA signaling pathway was involved in the JWH133 effects. This is the first study to propose that promotion of microglial M2 polarization through the cAMP/PKA pathway participates in the CB2R-mediated anti-inflammatory effects after GMH induction. The results will help to further understand the mechanisms that underlie neuroprotection by CB2R in GMH and promote clinical translational research for CB2R agonists.


Late sequelae of perioperative myocardial infarction.
The late suquelae of myocardial injury occurring at the time of direct myocardial revascularization are unknown. Fifty of 500 consecutive patients undergoing aortocoronary bypass grafting developed both electrocardiographic and enzymatic evidence of myocardial injury. They were matched with 50 patients of similar age, sex, history of previous infarction, severity of angina, degree of coronary arteriosclerosis, and level of ventricular function as determined by preoperative angiographic studies. The conduct of the operation was identical in each group except for prolongation of total cross-clamp time in those patients with myocardial injury. The total number of vessels grafted, the conduit used, and the operative mean graft flow were similar. Results of treadmill stress testing at 24 to 36 months were not significantly different between groups. Angina status, long-term survival, graft patency, and ventricular function were not adversely affected by intraoperative myocardial injury. However, postoperative ventricular function and stress test performance were related to graft patency.


Central nervous system side effects are less important after iv regional anesthesia with ropivacaine 0.2% compared to lidocaine 0.5% in volunteers.
Following release of a double tourniquet for intravenous regional anesthesia (IVRA), ropivacaine was shown to have a longer duration of action and less central nervous system (CNS) side effects than lidocaine. This study examines the correlation of CNS side effects to plasma levels of lidocaine 0.5% and ropivacaine 0.2% when injected intravenously for IVRA. In a double-blind, cross-over study, ten volunteers received IVRA with 40 mL ropivacaine 0.2% or lidocaine 0.5% at least four days apart. Both cuffs of a double-cuff tourniquet remained inflated until they could no longer be tolerated. The incidence, duration and intensity of CNS side effects were recorded at three, ten, and 30 min after tourniquet release and correlated with simultaneous venous blood samples. There was a lower incidence of CNS side effects with ropivacaine (6/10 volunteers) when compared to lidocaine (10/10 volunteers). There was also less duration of these side effects (mean +/- SD, 5.1 +/- 5.2 min vs 11.7 +/- 6.7 min). Measured total plasma levels were highest at ten minutes with ropivacaine 0.2% (1.2 +/- 0.3 microg.mL(-1)) and at three minutes with lidocaine 0.5% (1.7 +/- 0.6 microg.mL(-1)). Peak CNS symptoms correlated with measured venous plasma levels for lidocaine, but occurred earlier with ropivacaine. We observed a lower incidence of CNS side effects with ropivacaine as compared to lidocaine. Although ropivacaine's greater lipid solubility should, theoretically, lead to more CNS side effects, this was, likely, offset by slower release from tissues and lesser percentage of unbound (free) drug.


How Personality, Emotions and Situational Characteristics Affect Learning from Social Interactions in the Workplace.
The present study examines the effects of social interactions' situational characteristics, emotions, and personality on self-perceived learning from social interactions at work based on diary and survey data. The sample comprises 43 German vocational education and training (VET) trainees in various apprenticeship programs. During the diary period of ten working days, the participants were instructed to record five typical social interactions at work every day. Quantitative data of 1,328 social interactions were analyzed by means of multilevel analysis. Regarding social interactions' characteristics, the analysis revealed the baseline level of instrumentality, an interruption of the social interaction, its instrumentality and questions asked by the trainee during the interaction as positive predictors of self-perceived learning. A trainee's higher speech proportion, however, was a negative predictor. Regarding state emotions, the emotional experiences of bored and motivated were identified as significant positive predictors of learning from social interactions at work. Emotions' baseline level as well as personality traits had no significant influence. The results indicate that social interactions' situational characteristics have the biggest influence on self-perceived learning from social interactions.


[Azotemia in rabbits and rodents - an update].
An impaired elimination of urinary excreted substances and an increase of these substances in the blood (azotemia) can also occur in rabbits and rodents. In addition to renal diseases, prerenal and postrenal conditions can induce azotemia. A systematic evaluation of patients with azotemia is needed to find a diagnosis and to initiate an effective treatment. The article provides an overview of the diagnosis and therapy of common causes of azotemia in rabbits and rodents.


Effects of Ergosterol, Isolated from Scleroderma Polyrhizum Pers., on Lipopolysaccharide-Induced Inflammatory Responses in Acute Lung Injury.
The present study aimed to investigate the protective role of ergosterol, isolated from Scleroderma polyrhizum Pers., in lipopolysaccharide (LPS)-induced acute lung injury (ALI). ALI was induced in mice by LPS (0.5 mg/kg), and ergosterol (25 and 50 mg/kg) was administrated orally 1 h prior to LPS administration. Ergosterol pretreatment at doses of 25 and 50 mg/kg decreased LPS-induced lung histopathological changes, lung wet-to-dry weight ratio. In addition, pretreatment with ergosterol inhibited inflammatory cells and proinflammatory cytokines including tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6). Furthermore, we demonstrated that ergosterol blocked the activation of nuclear factor-kappaB (NF-κB), cyclooxygenase (COX)-2, and nitric oxide synthase (iNOS) pathways. The results presented here suggest that the protective mechanism of ergosterol may be attributed partly to the inhibition of NF-κB, COX-2, and iNOS pathways.


Globular body production, their anatomy, DNase gel analysis and NDP kinase activity in root tips of Poncirus trifoliata L.
Green globular bodies were developed from Poncirus trifoliata L. root tip explants as a response to addition in the substrate of different growth regulators. From the globular bodies, shoots initiated and grew. Median section of the globular bodies reveals that they are composed of parenchyma cells and originate from the pericycle. The activity of DNases during shoot formation from globular bodies was influenced by the type and concentration of plant growth regulators that were added in the nutrient substrate. Peptide bands formation was also influenced by the increase of BA concentration. Consequently, BA, NAA and IAA combination influenced 5'-triphosphonucleosides (NTPs) appearance and activity in the presence of metal. Peptide bands resulted from the electrophoretic analysis of endogenous protein phosphorylation, proved to be catalytic subunits of NDP kinases, as they all phosphorylate diphosphonucleosides. The enzymes DNases and NDP kinases could be used as a scientific tool for the study of shoot formation from P. trifoliata L. green globular bodies.


Patchiness and compensatory growth in a fungus-Collembola system.
The compensatory growth potential of a grazed fungal biomass was mathematically expressed as a function of patchiness in its distribution and demonstrated in an experiment using the fungivorous collembolan Onychiurus armatus and the soil fungi Verticillium bulbillosum and Penicillium spinulosum. The model addresses the regrowth potential in relation to patch fragmentation, travelling time and consumption rate of the collembolan and the mean relative growth rate of the fungus. It suggests that the mean relative growth rate required for regrowth decreases with patch fragmentation and increases with the mean growth rate of the fungus. The experiments were performed with a system of soil-filled vials provided with fungi and collembolans. The size of the vials and the length of the tubes connecting them were varied to give different patch sizes and travelling times. The respiratory activity of fungi after grazing increased as a unit of mycelium was distributed into smaller connected vials. The slow growing species V. bulbillosum showed a greater but delayed response to grazing in comparison with the fast growing P. spinulosum. An increased travelling time delayed the growth response in both species.


Stable expressions of nine genes within human oocytes and live birth by in vitro fertilisation.
The development of new methods to cryopreserve human ovarian cortex tissues without damage is needed for the improvement of quality of life of female cancer patients. Here, we show that stable expressions of nine genes within human oocytes by using a supercooling procedure are necessary in order to produce human mature eggs and experience live births with high safety profiles by in vitro fertilisation. Our method will be helpful in order to preserve the fertility of female cancer patients or healthy women.


Patients' perspective of disease and medication adherence for type 2 diabetes in an urban area in Bangladesh: a qualitative study.
Patients' perspective of diabetes and adherence to its prescribed medications is a significant predictor of glycemic control and overall management of the disease. However, there is a paucity of such information in Bangladesh. This study aimed to explore patients' perspective of diabetes, their experience of taking oral hypoglycemic medications and explore factors that contribute to medication adherence in patients with type 2 diabetes in Bangladesh. We conducted in-depth face-to-face interviews with 12 type 2 diabetes patients attending a tertiary hospital in Dhaka city between February and March, 2014. Participants were purposively sampled representing different age groups, education levels, years since diagnosis with diabetes, and glycemic status, to achieve maximum variation sampling. All interviews were conducted using a topic guide and were audio-recorded, transcribed verbatim, checked for errors, coded and analyzed by means of a qualitative content analysis framework. The data analysis generated rich information on the participants' knowledge and perception on diabetes, its causes, self-management, medication use, adverse effects of medication use, medication adherence, and impact of diabetes, Although most of the participants demonstrated substantive knowledge on diabetes and its consequences, they also reported numerous misconceptions about the disease. Knowledge on diabetes medication, their appropriate use and side effects was rather poor. Respondents also reported non-compliance to dietary and physical activity advice by their physicians and concerns on diabetes diabetes-induced psychological stress. High cost of medications, concerns over medication side effects and forgetfulness was noted as factors for non-adherence to medication. Participants' knowledge and perception on diabetes are key factors determining their adherence to medications and, thereby, diabetes management. Healthcare providers should explore to better understand patients' perspective on diabetes, medication beliefs, identify psychological stress and provide more effective health education interventions to enhance medication adherence.


Effects of dipeptidyl peptidase-4 inhibitors on beta-cell function and insulin resistance in type 2 diabetes: meta-analysis of randomized controlled trials.
Dipeptidyl peptidase-4 (DPP-4) inhibitors are a novel family of glucose-lowering agents. Accumulating evidence suggests that DPP-4 inhibitors preserve pancreatic beta-cell function, but results in previous studies have been inconsistent. We assessed the effects of DPP-4 inhibitors on the homoeostasis model assessment beta-cell function (HOMA-B) or insulin resistance (HOMA-IR) index in patients with type 2 diabetes through a systematic review and meta-analysis of randomized controlled trials (RCTs). Relevant articles were identified from PubMed, Embase, and Cochrane Library databases up to December 27, 2016. We calculated weighted mean differences (WMDs) and 95% confidence intervals (CIs) in each included trial and pooled the data using a random-effects model. Fifty-two trials were included in the present analysis. Compared with placebo control, DPP-4 inhibitors as monotherapy significantly improved HOMA-B (WMD 9.15; 95% CI 7.48, 10.81). Similarly, DPP-4 inhibitors as add-on therapy in combination with other drugs showed significant improvement in HOMA-B (WMD 9.04; 95% CI 5.72, 12.37). However, we found no significant improvement in HOMA-IR following treatment with DPP-4 inhibitors as mono-therapy or as add-on therapy. In conclusion, DPP-4 inhibitors as monotherapy or as add-on therapy significantly improved beta-cell function but had no significant effect on insulin resistance in type 2 diabetes.


Changes in myelinated nerve fibers and skeletal muscle of rats exposed to high doses of permethrin.
Neurological signs and segmental demyelination in a cervical nerve were observed in rats treated orally with permethrin (300 mg/kg/day) for 5 days. Inflammatory and degenerative signals were recorded in the diaphragm muscle. These effects were more intense with the trade grade than with the technical grade product. The possible influence of the percentage of cis:trans isomers on the intensity of the observed effects is discussed.


Substrate-Specific Reduction of Tetrazolium Salts by Isolated Mitochondria, Tissues, and Leukocytes.
Tetrazolium salts are commonly used in cytochemical and biochemical studies as indicators of metabolic activity of cells. Formazans, formed by reduction of tetrazolium salts, behave as pseudo-solutions during initial incubation, which allows monitoring their optical density throughout incubation. The criteria and conditions for measuring oxidative activity of mitochondria and dehydrogenase activity in reduction of nitroblue tetrazolium (NBT) and methyl thiazolyl tetrazolium (MTT) in suspensions of isolated mitochondria, tissue homogenates, and leukocytes were investigated in this work. We found that the reduction of these two acceptors depended on the oxidized substrate - NBT was reduced more readily during succinate oxidation, while MTT - during oxidation of NAD-dependent substrates. Reduction of both acceptors was more sensitive to dehydrogenase inhibitors that to respiratory chain inhibitors. The reduction of NBT in isolated mitochondria, in leukocytes in the presence of digitonin, and in liver and kidney homogenates was completely blocked by succinate dehydrogenase inhibitors - malonate and TTFA. Based on these criteria, activation of succinate oxidation was revealed from the increase in malonate-sensitive fraction of the reduced NBT under physiological stress. The effect of progesterone and its synthetic analogs on oxidation of NAD-dependent substrates by mitochondria was investigated using MTT. Both acceptors are also reduced by superoxide anion; the impact of this reaction is negligible or completely absent under physiological conditions, but can become detectable on generation of superoxide induced by inhibitors of individual enzyme complexes or in the case of mitochondrial dysfunction. The results indicate that the recording of optical density of reduced NBT and MTT is a highly sensitive method for evaluation of metabolic activity of mitochondria applicable for different incubation conditions, it offers certain advantages in comparison with other methods (simultaneous incubation of a large set of probes in spectral cuvettes or plates); moreover, it allows determination of activity of separate redox-dependent enzymes when selective inhibitors are available.


A workflow from untargeted LC-MS profiling to targeted natural product isolation.
Liquid chromatography-mass spectrometry (LC-MS or HPLC-MS) is an extremely sensitive analytical technique that enables the detection of metabolites with a vast range of chemistries and molecular masses. Extracts from any biological starting material are first fractionated chromatographically on a stationary phase suitable for the retention of the target molecules. The eluent is then transferred directly to the ionization source for MS detection. There is a vast range of chromatographic separation methods and MS configurations. This chapter describes a method for the detection of a broad range of metabolites using reversed phase (C18) LC-MS as well as a method for the isolation of targeted metabolites of interest.


Capillary Isoelectric Focusing: Mass Spectrometry Method for the Separation and Online Characterization of Monoclonal Antibody Charge Variants at Intact and Subunit Levels.
Monoclonal antibodies (mAbs) are one of the most widely used types of protein therapeutics. Charge variants are important quality attributes for evaluating developability, activity, and safety for mAb therapeutics. Here, we report a novel online capillary isoelectric focusing-mass spectrometry (CIEF-MS) method for mAb charge variant analysis using an electrokinetically pumped sheath-flow nanospray ion source on a time-of-flight (TOF) MS with a pressure-assisted chemical mobilization. Key factors that enable online CIEF-MS include effective capillary electrophoresis-MS (CE-MS) interface with enhanced sensitivity, utilization of MS-friendly electrolytes, beneficial effects of glycerol that reduces non-CIEF electrophoretic mobility and limits band broadening, appropriate ampholyte type and concentration selection for balanced separation resolution and MS detection sensitivity, optimized sheath liquid composition to realize high-resolution CIEF separation and effective MS electrospray ionization, as well as judiciously selected CIEF running parameters. The fundamental premise of CIEF has been verified by the linear correlation between isoelectric point (pI) values and migration time using a mixture of pI markers. By achieving high separation resolutions that are similar as those obtained from imaged CIEF (iCIEF), this method successfully provides highly sensitive MS identification for intact mAb charge variants. Furthermore, a middle-up sample treatment workflow can be adopted to provide in-depth charge variant analysis at subunit level for mAbs with complex charge heterogeneity. The mAb subunit CIEF-MS reveals the source of charge variant with enhanced resolution on both CIEF separation and MS spectra. This novel CIEF-MS method is a valuable tool with distinct advantage for objective and accurate assessment of charge heterogeneity of protein therapeutics.


Quinones contained in wastewater as redox mediators for the synergistic removal of azo dye in microbial fuel cells.
The present paper aimed to investigate the roles of quinones contained in wastewater and the enhanced effects on microbial fuel cells (MFCs) under different redox conditions. The feasibility of using wastewater rich in quinones to act as co-substrate and redox mediators (RMs) library to strengthen the synergistic removal of azo dye in MFCs was evaluated. The results demonstrated that quinones achieved enhanced effects on electricity generation and COD removal of MFC better at higher current intensity. The addition of pure quinone decreased electron transfer resistance (Rct) of MFCs from 4.76 Ω to 2.13 Ω under 1000 Ω resistance and 1.16 Ω-0.75 Ω under 50 Ω resistance. Meanwhile, higher coulombic efficiency was achieved. Compared with sodium acetate, using quinone-rich traditional Chinese medicine (TCM) wastewater as the co-substrate enhanced the synergistic removal of reactive red 2 (RR2) in MFCs from 79.58% to 92.45% during 24 h. RR2 was also degraded more thoroughly due to the accelerated electron transfer process mediated by RMs. Microbial community analysis demonstrated that the presence of quinone in TCM wastewater can enrich different exoelectrogens under varied redox conditions and thus influenced the enhanced effects on MFC. Metagenomic functional prediction results further indicated that the abundance of functional genes involved in carbohydrate metabolism, membrane transport metabolism, biofilm formation, and stress tolerance increased significantly in presence of RMs. Redundancy analyses revealed that RMs addition was the more important factor driving the variation of the microorganism community. This study revealed the potential effect of quinones as redox mediators on the bioelectrochemical system for pollutants removal.


Subcutaneous blood flow in early male pattern baldness.
The subcutaneous blood flow (SBF) was measured by the 133Xe washout method in the scalp of 14 patients with early male pattern baldness. Control experiments were performed in 14 normal haired men matched for age. The SBF in the scalp of the normal individuals was about 10 times higher than previously reported SBF values in other anatomical regions. In patients with early male pattern baldness, SBF was 2.6 times lower than the values found in the normal individuals (13.7 +/- 9.6 vs 35.7 +/- 10.5 ml/100 g/min-1). This difference was statistically significant (p much less than 0.001). A reduced nutritive blood flow to the hair follicles might be a significant event in the pathogenesis of early male pattern baldness.


Effect of estrogen and tamoxifen on the expression pattern of AP-1 factors in MCF-7 cells: role of c-Jun, c-Fos, and Fra-1 in cell cycle regulation.
The activated transcription factor ERα plays an important role in the breast development and progression of cancer. In a non-classical pathway ER interacts with other transcription factors AP-1, NFkB, SP1, etc. AP-1 transcription factors control rapid responses of mammalian cells to stimuli that impact proliferation, differentiation, and transformation. AP-1 factors are leucine zipper proteins belonging to members of the Jun family (c-Jun, JunB, and JunD) and Fos family (c-Fos, FosB, Fra-1, and Fra-2) proteins. Although AP-1 factors are well characterized, not much is known about the expression pattern of the AP-1 factors in breast cancer cells. Hence to determine which AP-1 factors are expressed and regulated by estrogen, we used human breast cancer MCF-7 cells as in vitro model system. The MCF-7 cells were treated with or without estradiol-17β (E2) or antiestrogen tamoxifen (TMX) and the cell proliferation and viability was assessed by MTT assay. The expression of different AP-1 factors was analyzed by semi-quantitative RT-PCR. The cells treated with E2 found to increase the cell proliferation by more than 35 % and TMX an antiestrogen decreased by 29 % compared to control. The E2 found to induce the expression of c-Jun, Fra-1, and c-Fos, while TMX decreased the expression. In addition TMX also decreased the mRNA levels of Jun-D and Fra-2. These results suggest that the AP-1 factors c-Jun, c-Fos, and Fra-1 may be involved in the proliferation and transformation of MCF-7 cells. E2 also found to induce cyclin D1 and cyclin E1 mRNA transcripts of cell cycle regulators while TMX significantly decreased compared to control. Further E2 induced the anti-apoptotic Bcl-2 and TMX decreased mRNA transcripts. The data presented here support the E2-ERα-mediated MCF-7 cell proliferation and confirms the role of AP-1 factors in cell cycle regulation.


Basic dye adsorption onto an agro-based waste material--sesame hull (Sesamum indicum L.).
The aim of this project was to establish an economical and environmentally benign biotechnology for removing methylene blue (MB) from wastewater. The adsorption process of MB onto abandoned sesame hull (Sesamum indicum L.) (SH) was investigated in a batch system. The results showed that a wide range of pH (3.54-10.50) was favorable for the adsorption of MB onto SH. The Langmuir model displayed the best fit for the isothermal data. The exothermic adsorption process fits a pseudo-second-order kinetic model. The maximum monolayer adsorption capacity (359.88 mg g(-1)) was higher than most previously investigated low-cost bioadsorbents (e.g., peanut hull, wheat straw, etc.). This study indicated that sesame hull is a promising, unconventional, affordable and environmentally friendly bio-measure that is easily deployed for removing high levels of MB from wastewater.


A non-randomized controlled stepped wedge trial to evaluate the effectiveness of a multi-level mammography intervention in improving appointment adherence in underserved women.
Considerable racial and socio-economic disparities exist in breast cancer. In spite of the existence of numerous evidence-based interventions (EBIs) aimed at reducing breast cancer screening barriers among the underserved, there is a lack of uptake or sub-optimal uptake of EBIs in community and clinical settings. This study evaluates a theoretically based, systematically designed implementation strategy to support adoption and implementation of a patient navigation-based intervention, called Peace of Mind Program (PMP), aimed at improving breast cancer screening among underserved women. The PMP will be offered to federally qualified health centers and charity clinics in the Greater Houston area using a non-randomized stepped wedge design. Due to practical constraints of implementing and adopting in the real-world, randomization of start times and blinding will not be used. Any potential confounding or bias will be controlled in the analysis. Outcomes such as appointment adherence, patient referral to diagnostics, time to diagnostic referral, patient referral to treatment, time to treatment referral, and budget impact of the intervention will be assessed. Assessment of constructs from the consolidated framework for implementation research (CFIR) will be assessed during implementation and at the end of the study (sustainment) from each participating clinic. Data will be analyzed using descriptive statistics (chi-square tests) and generalized estimating equations (GEE). While parallel group randomized controlled trials (RCT) are considered the gold standard for evaluating EBI efficacy, withholding an effective EBI in practice can be both unethical and/or impractical. The stepped wedge design addresses this issue by enabling all clinics to eventually receive the EBI during the study and allowing each clinic to serve as its own control, while maintaining strong internal validity. We expect that the PMP will prove to be a feasible and successful strategy for reducing appointment no-shows in underserved women. NCT02296177.


Lack of teratogenicity of aluminum hydroxide in mice.
The embryotoxic and teratogenic potential of aluminum hydroxide, a therapeutic drug used as an antacid and phosphate binder, was investigated in Swiss mice. Mated female mice were given by gavage daily doses of 0, 66.5, 133 or 266 mg/kg of A1 (OH)3 on gestation days 6 through 15 and killed on gestation day 18. Females were evaluated for body weight gain, food consumption, appearance and behavior, survival rates, and reproduction data. No significant effects attributable to A1(OH)3 were noted in comparison of maternal body weight and food consumption values, appearance and behavior. No treatment-related changes were recorded in the number of total implants, resorptions, the number of live and dead fetuses, fetal size parameters or fetal sex distribution data. Gross external, soft tissue and skeletal examination of the A1-treated fetuses did not reveal differences at any dose in comparison with the controls. Thus, no evidence of maternal toxicity, embryo/fetal toxicity or teratogenicity was observed with A1(OH)3 in mice.


Influence of the nutritional status on facial morphology in young Japanese women.
Evidence regarding the possible influence of nutritional status on the facial morphology has thus far been insufficient. We examined whether or not the physical body compositions and dietary behaviors were correlated with any morphological characteristics of the face. One hundred and fifteen young Japanese women participated. Variables representing the dietary behaviors were extracted from self-reported survey data, and corresponding three-dimensional (3D) facial images and body compositions were examined. Multivariate analyses identified significant relationships between the nutritional status and facial topography (p < 0.05). The clustering method revealed the existence of three dietary condition patterns ("balanced diet", "high-calorie-diet" with obesity tendency, and "imbalanced low-calorie-diet" with sarcopenic obesity tendency). Among these three patterns, a round face (increased facial width; analysis of variance [ANOVA], p < 0.05) was observed in the high-calorie-diet pattern, while the imbalanced low-calorie-diet pattern showed a more masculine face (increased face height, decreased eye height, increased non-allometric sexual shape differences; ANOVA, p < 0.05), thus suggesting the possibility of sex-hormonal influences. In summary, the body composition and dietary behaviors were found to influence the facial morphology, and potential biological influences were discussed.


Anterior cervical discectomy and fusion with zero-profile versus stand-alone cages for two-level cervical spondylosis: A retrospective cohort study.
To assess the mid-long-term clinical and radiological outcomes of zero-profile (ZP) compared with stand-alone (ST) cages for two-level anterior cervical discectomy and fusion (ACDF). We included 77 patients (39 women and 38 men) who underwent two-level ACDF between May 5, 2016, and May 5, 2020, and who were followed up for at least 1 year. The subjects were divided into the ST (n = 38) and ZP (n = 39) group. For the evaluation of functional status, Japanese Orthopedic Association (JOA), Neck Disability Index (NDI), and Visual Analogue Scale (VAS) scores were used. Additionally, radiological outcomes and procedure complications were observed at final follow-up. Both groups had excellent clinical outcomes at the final follow-up. There were no significant intergroup (ZP vs. ST) differences in the fusion rate (91.02% vs. 90.79%, P > 0.05) and postoperative dysphagia (15.4% vs. 2.6%, P = 0.108). However, the disc height at the final follow-up in the ZP group was higher than that in the ST group (6.86 ± 0.84 vs. 6.17 ± 1.03, P = 0.002). The ZP group accomplished a lower loss of cervical lordosis (18.46 ± 4.78 vs. 16.55 ± 4.36, P = 0.071), but without reaching statistical significance. ACDF with either ZP or ST cages turns out to be a dependable strategy for two-level ACDF in terms of clinical results. However, compared with the ST, the ZP cage may achieve a significantly lower loss of disc height.


[The impact of ultrasound in urology].
Ultrasound is of great importance in the diagnosis of acute and chronic diseases in urology, such as kidney colic, testicular torsion, low-grade kidney trauma or for follow-up of vesicoureteral reflux, evaluation of infertility, measurement of residual urinary volume and the detection of cancer. An ultrasound examination is time and cost-effective without exposure to ionizing radiation and is routinely performed by practitioners as well as in the clinical daily routine. With technical innovations, such as contrast-enhanced ultrasound or real time elastography, it would for instance be possible to extend the application field of ultrasound. However, in some fields of investigation ultrasound still lacks accuracy and despite its many advantages the validity of ultrasound findings sometimes has to be verified with computed tomography (CT) or magnetic resonance imaging (MRI).


Acetabular revision in total hip arthroplasty with tantalum augmentation and lyophilized bovine xenograft.
To evaluate a mixed technique of acetabular reconstruction, which uses trabecular metal in the form of tantalum augments associated with lyophilized bovine xenograft. Fifteen patients were evaluated prospectively, who underwent acetabular reconstruction with impacted lyophilized bovine xenograft associated with the use of tantalum augments. The main outcome was the failure of the tantalum-bone interface. The population had a mean age of 58.33 years ± 14.27; the majority was female, 80%. Of the total subjects, 66.7% were operated for failure in primary arthroplasty. The mean follow-up time was 45.2 months ± 11.39. The failure rate of the method in the period and population studied was 6.7%. An extremely high index (93.3%) of success was observed in an average time of 45.2 months of follow-up. Data were comparable to current literature, demonstrating that the technique employed and proposed is adequate for hip reconstruction in young patients.


Effects of oxidized and glycated LDL on gene expression in human retinal capillary pericytes.
Modified (oxidized and/or glycated) low-density lipoproteins (LDLs) have been implicated in retinal pericyte loss, one of the major pathologic features of early-stage diabetic retinopathy. To delineate underlying molecular mechanisms, the present study was designed to explore the global effects of modified LDL on pericyte gene expression. Quiescent human retinal pericytes were exposed to native LDL (N-LDL), glycated LDL (G-LDL), and heavily oxidized-glycated LDL (HOG-LDL) for 24 hours, and gene expression was evaluated by DNA microarray analysis. Several of the gene responses were checked, and in each case confirmed by reverse-transcription real-time PCR. HOG-LDL induced a gene expression pattern markedly distinct from that of N-LDL or G-LDL, whereas G-LDL elicited gene expression similar to that of N-LDL. A comparison of responses to HOG-LDL versus N-LDL revealed 60 genes with expression that varied by > or =1.7-fold. The HOG-LDL-responsive genes included members of functional pathways, such as fatty acid, eicosanoid, and cholesterol metabolism; fibrinolytic regulation; cell growth and proliferation; cell stress responses; the kinin system; and angiogenesis. HOG-LDL elicits gene expression in retinal pericytes that may contribute to pericyte loss and other retinal abnormalities in diabetic retinopathy. Observed proapoptotic and proangiogenic responses to HOG-LDL may be of particular importance in this regard. The genes identified through these studies provide potential therapeutic targets for the prevention and treatment of diabetic retinopathy.


Acute myeloid leukemia associated with FGFR1 abnormalities.
Hematologic malignancies associated with FGFR1 abnormalities present in heterogeneous forms, including myeloproliferative neoplasm, acute myeloid leukemia (AML), T- or B-lineage lymphoblastic leukemia/lymphoma, and even mixed phenotype acute leukemia. In the present report, we describe three new cases of AML associated with FGFR1 abnormalities: AML with minimal differentiation with 45,XY,-7,t(8;13)(p11.2;q12), acute myelomonocytic leukemia with eosinophilia with 48,XY,t(8;9)(p11.2;q33),+19,+21, and AML with minimal differentiation with 46,XX,add(8)(p11.2). FGFR1 abnormalities were confirmed by fluorescence in situ hybridization. We reviewed the records of 19 patients reported from Asian countries, and found that approximately 40 % of cases manifested as acute leukemia associated with myeloid lineage, and 47 % were not accompanied with eosinophilia. These findings highlight the need for detection of FGFR1 abnormalities, not only in myeloproliferative disorder, but also in AML patients even without eosinophilia. The prognosis for this group of neoplasms is poor, and there is no recognized effective targeted treatment. Two patients, including our case, who received allogeneic hematopoietic stem cell transplantation (allo-HSCT) survived. Accumulation of long-term follow-up data can help determine the allo-HSCT protocol or the need for new therapeutic trials to improve the survival rate of patients with FGFR1 abnormalities.


A novel multi-kinase inhibitor pazopanib suppresses growth of synovial sarcoma cells through inhibition of the PI3K-AKT pathway.
Synovial sarcoma is an aggressive soft tissue sarcoma with only a modest response to conventional cytotoxic agents. In the present study, we evaluated the potential antitumor effects of a novel anti-angiogenesis agent, pazopanib, against synovial sarcoma cells. We found that pazopanib directly inhibited the growth of synovial sarcoma cells by inducing G1 arrest. Multiplex analyses revealed that the PI3K-AKT pathway was highly suppressed in pazopanib-sensitive synovial sarcoma cells. Furthermore, administration of pazopanib highly suppressed the tumor growth in a xenograft model. Taken together, these results suggest pazopanib as a possible agent against synovial sarcoma and may warrant further clinical studies.


Linear quantification of a streptavidin-alkaline phosphatase probe for enzyme-linked immuno mass spectrometric assay.
The alkaline phosphatase-streptavidin (AP-SA) probe released adenosine (∼267.2 Da) from the substrate adenosine monophosphate (AMP), where a signal may be detected from as little as 0.5 μl of a 0.1-pg/ml dilution of the probe (2.6 × 10(-22) mol). The signal from the AP-SA probe was linear from 1 to 50 pg/ml by monitoring adenosine release at 268 m/z (M + H) with liquid chromatography, electrospray ionization, and quadrupole mass spectrometry (LC-ESI-MS). The safe limit of detection and quantification of the AP-SA probe was approximately 0.5 pg/well or 5 pg/ml. Enzyme-linked immuno mass spectrometric assay (ELIMSA) using the AP-SA probe provided a linear signal response for prostate-specific antigen (PSA) against external standards from 1 to 500 pg/ml. The ELIMSA showed a safe limit of detection and quantification at 5 pg PSA/well or 50 pg/ml (false positive detection rate P ≤ 0.01). Female samples of 100 μl plasma/well were read against standards and blanks made in normal female plasma, and the lowest sample quantified was approximately 9.8 pg/well or 98 pg/ml. Here ELIMSA was applied to measure PSA in plasma from female, normal male, prostatectomy patient, and cancer patient samples that showed significant differences by analysis of variance (ANOVA).


Pathological risk factors and predictive endoscopic factors for lymph node metastasis of T1 colorectal cancer: a single-center study of 846 lesions.
Determining the depth of invasion of early stage colorectal cancer has been emphasized as a means of improving endoscopic diagnostic accuracy. Recent studies have focused on other pathological risk factors for lymph node metastasis (LNM). We investigated the significance of depth of invasion and predictive properties of other endoscopic findings. We retrospectively investigated 846 patients with submucosal invasive (T1) colorectal cancer who received an accurate pathological diagnosis and were treated between January 2005 and December 2016. Pathological risk factors associated with LNM were reviewed. We divided patients into groups: low-risk T1 colorectal cancer (LRC; no risk factors) and high-risk T1 colorectal cancer (HRC; exhibiting lymphovascular invasion, tumor budding grade of 2/3, and/or poor differentiation) and studied predictive endoscopic factors for HRC. Significant risk factors for LNM in multivariate analysis were lymphovascular invasion [odds ratio (OR) 8.09; 95% confidence interval (CI) 3.84-17.1], tumor budding (OR 1.89; 95% CI 1.09-3.29), and histological differentiation (OR 2.09; 95% CI 1.12-3.89). The LNM-positive rate with only deep submucosal invasion was 1.6%. Significant predictive factors for HRC in multivariate analysis identified rectal tumor location (OR 1.92; 95% CI 1.35 -2.72, depression (OR 2.73; 95% CI 1.96 -3.80), protuberance within the depression (OR 2.58; 95% CI 1.39- 4.78), expansiveness (OR 2.39; 95% CI 1.27- 4.50), and loss of mucosal patterns (OR 1.90; 95% CI 1.20 -3.01) as significant factors. Rectal tumor location, depression, protuberance within the depression, expansiveness, and loss of mucosal patterns could be predictive factors for HRC.


Issues in the use of complementary/alternative medical therapies for low back pain.
Unfortunately, current conventional medical treatments for low back pain are not managing the problem effectively. Standard approaches to treatment, particularly surgery, may even be exacerbating the problem and causing additional pain and suffering. In this context, it is understandable that LBP patients often go outside of the conventional medical system seeking other treatment options. Conventional medical physicians who treat low back pain patients, therefore, need to become more aware of the fact that their patients may be using complementary/alternative medical therapies.


An improved procedure for bile acid extraction and purification and tissue distribution in the rat.
Two bile acid extraction procedures were compared using endogenously radiolabeled tissues and feces. The method of Setchell et al. (J. Lipid Res. 24, 1085-1100, 1983) resulted in essential complete extraction, whereas that of Manes and Schneider (J. Lipid Res. 6, 376-377, 1971) gave recoveries between 56-82%. The time requirement for the method of Setchell et al. could be drastically reduced with no loss in extraction efficiency. Using extracts from endogenously labeled material, a purification procedure using C18 solid-phase extraction cartridges was developed that recovers greater than 90% of bile acids. The distribution of bile acids within the intestinal tract and liver of the rat was determined.


Recent Advances in Resting-State Electroencephalography Biomarkers for Autism Spectrum Disorder-A Review of Methodological and Clinical Challenges.
Electroencephalography (EEG) has been used for almost a century to identify seizure-related disorders in humans, typically through expert interpretation of multichannel recordings. Attempts have been made to quantify EEG through frequency analyses and graphic representations. These "traditional" quantitative EEG analysis methods were limited in their ability to analyze complex and multivariate data and have not been generally accepted in clinical settings. There has been growing interest in identification of novel EEG biomarkers to detect early risk of autism spectrum disorder, to identify clinically meaningful subgroups, and to monitor targeted intervention strategies. Most studies to date have, however, used quantitative EEG approaches, and little is known about the emerging multivariate analytical methods or the robustness of candidate biomarkers in the context of the variability of autism spectrum disorder. Here, we present a targeted review of methodological and clinical challenges in the search for novel resting-state EEG biomarkers for autism spectrum disorder. Three primary novel methodologies are discussed: (1) modified multiscale entropy, (2) coherence analysis, and (3) recurrence quantification analysis. Results suggest that these methods may be able to classify resting-state EEG as "autism spectrum disorder" or "typically developing", but many signal processing questions remain unanswered. We suggest that the move to novel EEG analysis methods is akin to the progress in neuroimaging from visual inspection, through region-of-interest analysis, to whole-brain computational analysis. Novel resting-state EEG biomarkers will have to evaluate a range of potential demographic, clinical, and technical confounders including age, gender, intellectual ability, comorbidity, and medication, before these approaches can be translated into the clinical setting.


Pharmacogenomics in depression and antidepressants.
Genetic factors are believed to play a major role in the variation of treatment response and the incidence of adverse effects to medication. The aim of pharmacogenetics is to elucidate this variability according to hereditary differences. Considering current hypotheses for the mechanisms of action of antidepressants, most investigations to date have concentrated on mutations in genes coding either for the pathways in the serotonergic and noradrenergic systems or for drug-metabolizing enzymes. Recent studies shifted the emphasis on the main mechanism of drug action from changes in neurotransmitter concentration or receptor function toward long-lasting adaptive processes within the neurons. Although the results are controversial, many studies support the hypothesis that psychopharmacogenetics will help predict an individual's drug response, while minimizing the side effects. The inclusion of functional genomics, which investigates the complex gene and/or protein expression in response to a given drug, may lead to the development of novel and safer drugs.


Gas exchange abnormalities in asthma.
There has been considerable interest over the last years in gas exchange abnormalities that occur in patients with bronchial asthma, particularly during an acute attack. These are essentially characterized by moderate to severe hypoxemia and hypocapnia caused by considerable ventilation-perfusion (VA/Q) mismatch. Using the multiple inert gas elimination technique, the baseline pattern of VA/Q distribution reveals a bimodal bloodflow distribution of VA/Q ratios but no shunt. Inhaled salbutamol seems to be an efficient and safe therapeutic approach, owing to the lack of deleterious effect on VA/Q inequality. Following intravenous salbutamol there is development of further VA/Q mismatch for the same degree of bronchodilation, the resulting deleterious effect on PaO2 being offset by the improvement in mixed venous PO2 due to increased cardiac output.


[Mapping of cloned fragments of the macrophage-stimulating factor receptor gene (FMS) from a non-amplified library of human leukocyte genes].
For investigation of FMS gene polymorphism and mutations that reveal functionally meaning in leukemia and myelodysplastic disorders the overlapping recombinants lambda-clones inserted by FMS gene fragments have been obtained from human leukocyte genomic library in the EMBL 3A phage by using oligonucleotide prode (27 nucleotides) based on 12 exon of the FMS gene. 15 DNA probes were prepared by subcloning the lambda-clones obtained in the pBSKS+ plasmid. The probes obtained allow to analyse extracellular, transmembrane and tyrosine kinase regions of the FMS gene independently.


Viral precursor polyproteins: keys of regulation from replication to maturation.
Many viruses use a replication strategy involving the translation of a large polyprotein, which is cleaved by viral and/or cellular proteases. Several of these viruses severely impact human health around the globe, including HIV, HCV, Dengue virus, and West Nile virus. This method of genome organization has many benefits to the virus such as condensation of genetic material, as well as temporal and spatial regulation of protein activity depending on polyprotein cleavage state. The study of polyprotein precursors is necessary to fully understand viral infection, and identify possible new drug targets; however, few atomic structures are currently available. Presented here are structures of four recent polyprotein precursors from viruses with a positive sense RNA genome.


Safety of High-Dose Unfractionated Heparin for Prophylaxis of Venous Thromboembolism in Hospitalized Obese Patients.
Optimal dosing of unfractionated heparin (UFH) for thromboprophylaxis in the obese patient population is uncertain because of their high-risk, prothrombotic state and a complexity of pharmacokinetic considerations. Literature on the appropriateness of the use of a higher dose UFH regimen remains unclear and inconsistent. To evaluate the safety of the use of 7500 units every 8 hours (high-dose) of subcutaneous UFH compared with the use of 5000 units every 8 hours (standard-dose) of subcutaneous UFH for thromboprophylaxis in obese patients (defined as BMI ≥30 kg/m2). In a retrospective cohort study, 326 adult patients were included, with a BMI ≥30 kg/m2, who were admitted to a large, urban academic medical center between September 1, 2015, and September 1, 2018. Patients received either high-dose or standard-dose UFH for at least 48 hours. The primary end point was the incidence rate of bleeding events, defined as a ≥2-g/dL fall in hemoglobin level or receipt of transfusion of 2 or more units of packed red blood cells (pRBCs) from the start of the UFH order. The incidence rate of bleeding was significantly higher in those who received high-dose UFH (43%) compared with those who received standard-dose UFH (29%; P = 0.008). No significant difference was found between venous thromboembolism event rates. High-dose UFH was associated with an increased bleeding event rate compared with standard-dose UFH in patients with a BMI ≥30 kg/m2. This raises safety concerns about the appropriateness of utilizing this regimen in this population.


USP53 plays an antitumor role in hepatocellular carcinoma through deubiquitination of cytochrome c.
Despite of advances in treatment options, hepatocellular carcinoma (HCC) remains nearly incurable and has been recognized as the third leading cause of cancer-related deaths worldwide. As a deubiquitinating enzyme, the antitumor effect of ubiquitin-specific peptidase 53 (USP53) has been demonstrated on few malignancies. In this study, we investigated the potential antitumor role of USP53 in HCC. The results showed that USP53 was downregulated in HCC tissues as well as in HCC cell lines using both in silico data as well as patient samples. Furthermore, the ectopic expression of USP53 inhibited the proliferation, migration and invasion, and induced the apoptosis of HCC cells. Co-immunoprecipitation (CO-IP) assay and mass spectrometry (MS) combined with the gene set enrichment analysis (GSEA) identified cytochrome c (CYCS) as an interacting partner of USP53. USP53 overexpression increased the stability of CYCS in HCC cells following cycloheximide treatment. Finally, the overexpression of CYCS compensated for the decreased apoptotic rates in cells with USP53 knocked down, suggesting that USP53 induced the apoptosis in HCC cells through the deubiquitination of CYCS. To summarize, we identified USP53 as a tumor suppressor as well as a therapeutic target in HCC, providing novel insights into its pivotal role in cell apoptosis.


Mode of binding of cytochrome b5 to phospholipid bilayers in lamellar structure.
X-ray diffraction studies were made on the multilamellar systems produced by incubation of phospholipid bilayers and the membrane protein, cytochrome b5, or non-membrane proteins (albumin, ovalbumin and beta-lactoglobulin A) at pH 8.1 in aqueous 5 mM CaCl2 solutions. Detergent-extracted cytochrome b5 (soluble aggregate) forms two types of lamellar phase with dipalmitoyl phosphatidylcholine bilayers, depending upon the incubation temperature. One type, which has a repeat distance of 114 A, is formed above 34 degrees C, where the binding of cytochrome b5 to the bilayers is hydrophobic. The other type, with a repeat distance of 153 A, is formed below 34 degrees C, where the binding is electrostatic. It is also suggested that cytochrome b5 is monomeric in the former phase but remains aggregated in the latter phase. When dimyristoyl phosphatidylcholine is used, the boundary temperature for the two types shifts to 12 degrees C. These boundary temperatures coincide with the thermal pretransition points of hydrated dipalmitoyl phosphatidylcholine and dimyristoyl phosphatidylcholine, respectively. Trypsin-treated cytochrome b5 (monomeric) and the three non-membrane proteins exhibit only binding of the electrostatic type to the bilayers, indipendently of the incubation temperature. The observed repeat distances suggest that in these cases two layers of protein molecules are incorporated between the bilayers.


The relationship between community pharmacists' social distance from and their confidence in interacting with patients with depression in Japan.
Background Prejudice and stigma against patients with depression may lead to adverse clinical consequences, such as delayed initiation and premature termination of therapy. However, available survey results on pharmacists' awareness showed a lack of confidence in dealing with patients with depression. Until recently, few studies addressed the relationship between pharmacists' perceptions of their social distance from and confidence in interacting with patients with depression. Objectives To determine pharmacists' social distance from patients with depression, and investigate the factors that influence and are influenced by it. Methods The target population of this survey study included 161 pharmacists employed at a drug chain who had participated in the Effects of Community Pharmacists' Medication Assistance for the Patients Taking Antidepressant study. The questionnaire included six questions on social distance, which were adopted from a previous nation-wide survey of mental illness stigma in Japan. Other questions related to the respondents' attributes, personal experience (living with depression, abusive behaviors inflicted by patients with depression), and confidence in interacting with patients with depression. Results Valid responses were obtained from 77 study participants. The mean total social distance score was 12.49. Greater total social distance scores were associated with greater difficulty in relationship building, information provision, and comprehension of condition. Pharmacists' personal experience of living with depression decreased their social distance from patients with depression, whereas their experience of being verbally or physically abused by patients with depression increased their social distance. Conclusion This study showed that community pharmacists in Japan had less social distance from patients with depression than the general population. A significant negative correlation was observed between community pharmacists' social distance from and their confidence in interacting with patients with depression.


Photodynamic diagnosis-assisted transurethral resection using oral 5-aminolevulinic acid decreases residual cancer and improves recurrence-free survival in patients with non-muscle-invasive bladder cancer.
5-aminolevulinic acid is a protoporphyrin IX precursor used for photodynamic diagnosis. We aimed to clarify the therapeutic benefits of orally administered 5-aminolevulinic acid for photodynamic diagnosis in transurethral resection for non-muscle-invasive bladder cancer in a real-world setting. From January 2009 to December 2020, patients with non-muscle-invasive bladder cancer who have undergone visually complete resection by initial transurethral surgery were included. Histopathologically confirmed residual and intravesical recurrent bladder cancer was retrospectively investigated and compared with transurethral resection using photodynamic diagnosis and conventional white light. One hundred and fifteen patients who underwent photodynamic diagnosis-transurethral resection were compared with 346 patients who underwent white light-transurethral resection. The second transurethral resection revealed that the proportion of residual T1 and Ta bladder cancer was lower in the photodynamic diagnosis group than in the white light group (10.3% vs. 33.8%, P = 0.001). Additionally, the proportion of intravesical recurrence was lower in the photodynamic diagnosis group than in the white light group (9.6% vs. 41.9%, P < 0.001). Using 5-aminolevulinic acid-photodynamic diagnosis during transurethral resection of bladder tumor improved the cumulative intravesical recurrence-free survival (log-rank test: P < 0.001). Furthermore, multivariate analyses indicated that not using 5-aminolevulinic acid-photodynamic diagnosis was an independent predictive factor for residual bladder cancer (odds ratio, 6.16; 95% confidence interval, 2.10-18.0; P < 0.001) and intravesical recurrence (hazard ratio, 2.01; 95% confidence interval, 1.05-3.83, P = 0.034). Introducing 5-aminolevulinic acid-photodynamic diagnosis into transurethral resection reduces residual bladder cancer and improves the cumulative intravesical recurrence-free survival in patients with non-muscle-invasive bladder cancer, indicating its possible therapeutic benefits.


Awareness of diabetes risks is associated with healthy lifestyle behavior in diabetes free American adults: evidence from a nationally representative sample.
The purpose of this study was to examine whether diabetes free healthy non-Hispanic Whites (NHW), non-Hispanic Blacks (NHB), and Mexican-Americans (MA) who are told of their diabetes risk were more likely to adopt healthy lifestyle behavior defined as current weight control, physical activity and reduced fat/calories intake than those who were not told that they were at increased risk. A nationally representative data (n=5073) from the 2007-2008 U.S. National Health and Nutrition Examination Surveys were used for this investigation. Odds ratio from multiple logistic regression analysis was used to determine whether diabetes free NHW, NHB, and MA who are told of their increased diabetes risk were more likely than those who are not told of their diabetes risk to adopt healthy lifestyle behavior. Being told of increased diabetes risk was associated with increased adoption of healthy lifestyle behaviors as indicated by odds ratio of 2.38 (95% CI=1.34-4.05) in NHW, 2.46 (95% CI=1.20-5.05) in NHB and 2.27 (95% CI=1.32-3.89) in MA who have no diabetes, after adjusting for age, sex, race/ethnicity, hypertension, education, household income and total cholesterol. Awareness of increased risk for diabetes is associated with implementing healthy lifestyle behaviors in diabetes free healthy American adults. Population-based programs designed to assess and communicate diabetes risk may be helpful in preventing or delaying the onset of type 2 diabetes. Programs designed along racial/ethnic line may be needed to reduce racial/ethnic differences in rates of type 2 diabetes.


[Peroxidase isoenzymes in cress seedlings (Lepidium sativum L.) and their inhibition by silybin, silydianin and silychristin].
Seedlings of Lepidium sativum L. contain 4 peroxidase isoenzymes. Their de-novo-synthesis is significantly depressed in the presence of 3.10(-3) moles/1 silybin, silydianin and silychristin in the incubation medium respectively.


High-efficiency production of bioactive recombinant human fibroblast growth factor 18 in Escherichia coli and its effects on hair follicle growth.
Using fusion tags, expression of recombinant human fibroblast growth factor 18 (rhFGF18) in mammalian cells and Escherichia coli has been extensively used for fundamental research and clinical applications, including chondrogenesis and osteogenesis, hair growth, and neuroprotection. However, high-level rhFGF18 expression is difficult and the products are often not homogeneous. Furthermore, fusion-tagged protein has higher immunogenicity and lower bioactivity, and the removal of the fused tag is expensive. To overcome the limitations of fusion-tagged expression of protein and to prepare soluble highly bioactive rhFGF18, we have developed a rapid and efficient expression strategy. Optimized hFGF18 gene was amplified by polymerase chain reaction and cloned into pET22b and pET3c vectors, then transformed into E. coli strains Origima (DE3) and BL21 (DE3)PlysS. The best combination of plasmid and host strain was selected, and only Origima (DE3)/pET3c-rhFGF18 was screened for high-level expressed rhFGF18. Under optimal conditions in a 30-L fermentor, the average bacterial yield and expression level of rhFGF18 of three batches were more than 652 g and 30 % respectively, after treatment with 1 mM isopropyl-thio-β-galactopyranoside for 10 h at 25 °C. The target protein was purified by CM Sepharose FF and heparin affinity chromatography. The purity of rhFGF18 was shown by HPLC to be higher than 95 %, and the yield was 155 mg/L. In vitro MTT assays demonstrated that the purified rhFGF18 could stimulate significant proliferation of NIH3T3 cells, and animal experiments showed that rhFGF18 could effectively regulate hair growth. In conclusion, this may be a better method of producing rhFGF18 to meet the increasing demand in its pharmacological application.


Vestibular hypersensitivity to sound in superior canal dehiscence: large evoked responses in the legs produce little postural sway.
Patients with superior canal dehiscence (SCD) typically have enhanced sound-evoked vestibular reflexes, such as vestibulo-collic and vestibulo-ocular reflexes. We wished to investigate whether sound-evoked lower limb EMG responses and postural sway are also enhanced in this condition. Eight patients with CT confirmed SCD (11 affected ears) and 8 age-matched normal controls participated. Three sound-evoked responses were measured; vestibulo-collic reflexes (i.e. vestibular-evoked myogenic potentials, VEMPs), lower limb vestibulo-spinal reflexes and body sway (centre of pressure in mm). Sound stimuli were 500 Hz air-conducted tone bursts of varying lengths (VEMPs: 2 ms; vestibulo-spinal: 20 ms; sway: 1s and 200 ms) set at fixed levels above each subject's VEMP threshold. SCD patients had very large VEMP and vestibulo-spinal responses following high intensity stimulation, but at the matched intensity of 15 dB above threshold amplitudes were similar in both SCD patients and controls. The amplitude of both responses increased linearly with increasing stimulus intensity in both groups. Large ( approximately 20mm), stereotyped sway responses were present in only one (atypical) patient with high intensity stimulation. Small ( approximately 2mm) sway responses were present in the remaining patients, and began immediately following the vestibulo-spinal responses. Despite the presence of large vestibular reflexes, there is usually very little body sway in response to loud sounds in SCD patients. Large short-latency vestibulo-spinal reflexes in SCD do not necessarily evoke large sway responses.


Characterization of Prepractice Injury Prevention Exercises of High School Athletic Teams.
Static and dynamic exercises are performed before activity to decrease injury risk and increase performance. Although evidence supports using dynamic over static stretching and performing Fédération Internationale de Football Association (FIFA) 11+ as a dynamic prepractice routine, we do not know the frequency at which these exercises are utilized in high school populations. We hypothesize that there is a wide variety of preparticipation exercises performed by high school athletes, and that few high school teams perform FIFA 11+ as an injury prevention program in its entirety. Observational study. Level 4. High school prepractice routines were observed for 185 teams (football, soccer, basketball, and lacrosse) over 1 season. The percentages of team warm-up routines that included components of FIFA 11+ were calculated, and the chi-square test was used to compare sex, sport, and level of competition. Of a total 644 warm-up observations, 450 (69.9%) included only non-FIFA 11+ exercises, 56 (8.7%) included at least 1 FIFA 11+ exercise, and 38 (5.9%) included only jogging; 69 (10.6%) consisted only of sport-specific activities. The type of warm-up differed significantly between males and females ( P = 0.002), sports ( P < 0.001), and level of competition ( P < 0.001). Static stretching and athletes stretching on their own were observed in 14% and 15% of all observations. No team performed the FIFA 11+ injury prevention routine in its entirety. The type of warm-up differed by sex, sport, and level of competition. Static stretching was performed more frequently than anticipated, and an entire FIFA 11+ warm-up was never performed. We need to identify the exercises that decrease injury and increase performance and better inform the athletic population about the risks and benefits of static and dynamic warm-up programs.


Synthesis and structure activity relationships of carbamimidoylcarbamate derivatives as novel vascular adhesion protein-1 inhibitors.
Vascular adhesion protein-1 (VAP-1) is a promising therapeutic target for the treatment of diabetic nephropathy. Here, we conducted structural optimization of the glycine amide derivative 1, which we previously reported as a novel VAP-1 inhibitor, to improve stability in dog and monkey plasma, and aqueous solubility. By chemical modification of the right part in the glycine amide derivative, we identified the carbamimidoylcarbamate derivative 20c, which showed stability in dog and monkey plasma while maintaining VAP-1 inhibitory activity. We also found that conversion of the pyrimidine ring in 20c into saturated rings was effective for improving aqueous solubility. This led to the identification of 28a and 35 as moderate VAP-1 inhibitors with excellent aqueous solubility. Further optimization led to the identification of 2-fluoro-3-{3-[(6-methylpyridin-3-yl)oxy]azetidin-1-yl}benzyl carbamimidoylcarbamate (40b), which showed similar human VAP-1 inhibitory activity to 1 with improved aqueous solubility. 40b showed more potent ex vivo efficacy than 1, with rat plasma VAP-1 inhibitory activity of 92% at 1h after oral administration at 0.3mg/kg. In our pharmacokinetic study, 40b showed good oral bioavailability in rats, dogs, and monkeys, which may be due to its improved stability in dog and monkey plasma.


Polypeptide synthesis in sea urchin embryogenesis: an examination with synthetic polyribonucleotides.
After fertilization of the sea urchin egg the rate of protein synthesis by a cell-free ribosomal system increases markedly. This increase can be attributed to newly synthesized messenger RNA, since (i) the rate of polypeptide synthesis elicited by synthetic messenger polyribonucleotides changes only slightly after fertilization, and (ii) the enzymes for the formation of amino acyl transfer RNA's of phenylalanine and leucine and the polymerization of polypeptide are in excess in the unfertilized egg. Polypeptide synthesis has been characterized in development to the gastrula stage.


Performance comparison of parameter estimation techniques for the quantitation of local cerebral blood flow by dynamic positron computed tomography.
Local CBF (LCBF) can be quantitated from positron computed tomographic (PCT) data and physiologically based mathematical models by several general methods. Those using a dynamic sequence of PCT scans allow the simultaneous estimation of both LCBF and p, the indicator's tissue-blood partition coefficient. This article presents a comparison of three rapid estimation techniques for use with inert diffusible radioindicators and serial PCT, each of which is based on the original Kety model. One method, developed in our laboratory, involves minimizing the mean squared discrepancy between measured data and model predictions, whereas the other two methods, recently reported in the literature, are weighted integration techniques that involve multiplying the measured data by time-dependent weighting functions. Simulation studies of noise propagation and other sources of error were performed under a variety of simulated conditions. Functional images of LCBF and p were calculated using each method for both phantom and human subject data. Errors can differ by as much as a factor of 2-3 between methods, with each having its own unique advantages and disadvantages.


Retrograde transneuronal transfer of the C-fragment of tetanus toxin.
The transport of the C-fragment of tetanus toxin after intramuscular injection was studied with immunohistochemical techniques. Brainstems and spinal cords of mice were examined after injections of C-fragment into the tongue or forearm. Labelled motorneurons were observed within 6 h. By two days after injection, the bulk of detectable C-fragment had passed out of motorneurons into the surrounding neuropil. C-fragment, like native tetanus toxin, appears to be transferred from motorneurons to presynaptic structures to an extent not observed with other proteins. It is useful in the study of retrograde transneuronal transfer since it lacks the toxicity of tetanus toxin.


Stabbing headache in patients with autoimmune disorders.
Stabbing headache is a relatively rare type of chronic "primary" headache with distinctive features with respect to more common forms of paroxysmal headache, such as cluster headache and trigeminal neuralgia. Drug treatment is empirical because of the lack of knowledge on the pathophysiology of stabbing headache. We examined 26 patients recruited over 10 years, who met the diagnostic criteria for stabbing headache. Interestingly, more than half of these patients had autoimmune disorders, including multiple sclerosis, Sjögren's disease, Systemic Lupus Erythematosus, Behçet's disease, autoimmune vasculitis, and antiphospholipid antibody syndrome. We speculate that stabbing headache may develop as a result of neuroinflammation and, at least in some cases, may be an epiphenomenon of focal demyelinating lesions of the upper or lower brain stem.


Conventional vs. daylight photodynamic therapy for patients with actinic keratosis on face and scalp: 12-month follow-up results of a randomized, intra-individual comparative analysis.
Daylight PDT (DLPDT) is a new PDT procedure. Several trials demonstrate that DLPDT achieves similar response rates with conventional PDT (CPDT) in the treatment of non-hyperkeratotic actinic keratoses (AKs) in a nearly painless way. It seems that DLPDT represents a more convenient and equally effective treatment modality. Data on long-term efficacy of DLPDT are limited. To compare short- and long-term efficacy, safety and tolerability of DLPDT with that of CPDT in face and scalp AKs. The study, an intra-individual right-left comparison study, was conducted in three centres in North, Center and South Greece. Eligible patients received either DLPDT or CPDT randomly allocated to alternate sides of face or scalp. Patients were evaluated at baseline, 3 and 12 months after treatment. Assessments included lesion response at 3 and 12 months, PDT-associated pain during PDT session, local skin reactions 3 days after treatment as well as patients' preference 3 months after treatment. A total of 46 patients completed the study. Three months after treatment, the overall lesion complete response rate was 78% for DLPDT and 80.6% for CPDT. At the 12-month follow-up, response rate decreased to 71.8% and 73.7% for DLPDT and CPDT accordingly. Regarding response based on lesion grade, response rates obtained in grade-I lesions were higher with DLPDT, while treatment with CPDT resulted to higher rates of cured grade-II lesions at both follow-up visits. Results were not supported by statistical significance. DLPDT was associated with significantly lower pain and reduced severity of local skin reactions. Patients' preference favoured DLPDT. Our study demonstrated that DLPDT is similar to CPDT in terms of long-term efficacy and recurrence rates in the treatment of face and scalp AKs. DLPDT demonstrated a better tolerability profile as it was associated with lower pain and less severe adverse events.


Minilaparoscopy-assisted natural orifice total colectomy: technical report of a minilaparoscopy-assisted transrectal resection.
Experimental experience and the technological evolution of minimally invasive surgical devices have allowed initial reports describing the clinical applicability of natural orifice translumenal endoscopic surgery (NOTES). Colorectal resections are an interesting target for the NOTES platform. Theoretically, the transrectal approach could overcome the proposed limitations of transvaginal access, increasing NOTES clinical applicability. Hybrid procedures such as minilaparoscopy-assisted natural orifice surgery (MA-NOS) are the safe progression to pure NOTES. This report describes the first clinical case of a transrectal MA-NOS total colectomy. The patient was a 36-year-old man with severe ulcerative colitis (UC) who experienced failure of immunosuppressive therapy. The standard steps of laparoscopic total colectomy were respected, with basic triangulation maintained throughout the case. A transrectal endoscopic device was used for optic assistance, colon dissection, ileum section, and specimen retrieval. Transrectal MA-NOS total colectomy was assisted by three laparoscopic ports: a 12-mm port used as the terminal ileostomy site, a 2-mm needle epigastric port, and a 5-mm umbilical port used as a drain site at the final intervention. No intraoperative complications occurred. The total operative time was 240 min. Oral intake was initiated on postoperative day 2. Because of UC rectal activity, a course with azathioprine was completed, and the patient was discharged receiving 1 g of rectal mesalazine for maintenance. The final pathology demonstrated pancolonic inflammatory bowel disease in the form of UC with severe activity. Transrectal MA-NOS total colectomy was feasible and safe in the reported case. Improvement in NOTES instrumentation and selective clinical applications are mandatory before clinical trials.


Parental urinary biomarkers of preconception exposure to bisphenol A and phthalates in relation to birth outcomes.
Bisphenol A (BPA) and phthalates are ubiquitous non-persistent endocrine disrupting chemicals whose relation with infant birth size is not clearly understood. We examined associations between maternal and paternal preconception urinary concentrations of total BPA and 14 phthalate metabolites and birth size for 233 infants. Multiple linear regression models were used to estimate parental quartiles of BPA and phthalates in relation to birth weight, length, head circumference, and ponderal index with separate models run for each parent adjusting for age, smoking, body mass index, education, alcohol, parity, and creatinine. Models also included an interaction term for each chemical and infant sex and were further adjusted to include the other partner's chemical concentrations. In maternal models adjusted for partner's exposure and covariates, reductions in birth weight (range: 178-215 g; p < 0.05) were observed for the 2nd quartile of maternal monomethyl phthalate, mono-[(2-carboxymethyl) hexyl] phthalate and mono-n-octyl phthalate when compared with the 1st quartiles. The 3rd quartile of monoethylhexyl phthalate (mEHP) was also associated with a 200.16 g (95 % CI: -386.90, -13.42) reduction. Similar reductions in birth weight were observed for the 2(nd) quartile of paternal mEHP (β = -191.93 g; 95 % CI: -381.61, -2.25). Additionally, select maternal urinary metabolites were associated with decreased head circumference, birth length and gestational age. However, paternal concentrations were generally associated with increased birth length and gestational age. We observed some suggestion that preconception maternal and paternal urinary concentration of BPA and specific phthalate metabolites may be associated with smaller birth size and increased gestational age, though the findings appeared to be parent and chemical specific.


The Regulation of HBV Transcription and Replication.
Hepatitis B virus (HBV) is a major human pathogen lacking a reliable curative therapy. Current therapeutics target the viral reverse transcriptase/DNA polymerase to inhibit viral replication but generally fail to resolve chronic HBV infections. Due to the limited coding potential of the HBV genome, alternative approaches for the treatment of chronic infections are desperately needed. An alternative approach to the development of antiviral therapeutics is to target cellular gene products that are critical to the viral life cycle. As transcription of the viral genome is an essential step in the viral life cycle, the selective inhibition of viral RNA synthesis is a possible approach for the development of additional therapeutic modalities that might be used in combination with currently available therapies. To address this possibility, a molecular understanding of the relationship between viral transcription and replication is required. The first step is to identify the transcription factors that are the most critical in controlling the levels of HBV RNA synthesis and to determine their in vivo role in viral biosynthesis. Mapping studies in cell culture utilizing reporter gene constructs permitted the identification of both ubiquitous and liver-enriched transcription factors capable of modulating transcription from the four HBV promoters. However, it was challenging to determine their relative importance for viral biosynthesis in the available human hepatoma replication systems. This technical limitation was addressed, in part, by the development of non-hepatoma HBV replication systems where viral biosynthesis was dependent on complementation with exogenously expressed transcription factors. These systems revealed the importance of specific nuclear receptors and hepatocyte nuclear factor 3 (HNF3)/forkhead box A (FoxA) transcription factors for HBV biosynthesis. Furthermore, using the HBV transgenic mouse model of chronic viral infection, the importance of various nuclear receptors and FoxA isoforms could be established in vivo. The availability of this combination of systems now permits a rational approach toward the development of selective host transcription factor inhibitors. This might permit the development of a new class of therapeutics to aid in the treatment and resolution of chronic HBV infections, which currently affects approximately 1 in 30 individuals worldwide and kills up to a million people annually.