The triple variable index combines information generated over time from common monitoring variables to identify patients expressing distinct patterns of intraoperative physiology.
Mean arterial pressure (MAP), bispectral index (BIS), and minimum alveolar concentration (MAC) represent valuable, yet dynamic intraoperative monitoring variables. They provide information related to poor outcomes when considered together, however their collective behavior across time has not been characterized. We have developed the Triple Variable Index (TVI), a composite variable representing the sum of z-scores from MAP, BIS, and MAC values that occur together during surgery. We generated a TVI expression profile, defined as the sequential TVI values expressed across time, for each surgery where concurrent MAP, BIS, and MAC monitoring occurred in an adult patient (≥18 years) at the University of Pittsburgh Medical Center between January and July 2014 (n = 5296). Patterns of TVI expression were identified using k-means clustering and compared across numerous patient, procedure, and outcome characteristics. TVI and the triple low state were compared as prediction models for 30-day postoperative mortality. The median frequency MAP, BIS, and MAC were recorded was one measurement every 3, 5, and 5 min. Three expression patterns were identified: elevated, mixed, and depressed. The elevated pattern displayed the highest average MAP, BIS, and MAC values (86.5 mmHg, 45.3, and 0.98, respectively), while the depressed pattern displayed the lowest values (76.6 mmHg, 38.0, 0.66). Patterns (elevated, mixed, depressed) were distinct across the following characteristics: average patient age (52, 53, 54 years), American Society of Anesthesiologists Physical Status 4 (6.7, 16.1, 27.3%) and 5 (0.1, 0.6, 1.6%) categories, cardiac (2.2, 6.5, 16.1%) and emergent (5.8, 10.5, 12.8%) surgery, cardiopulmonary bypass use (0.3, 2.6, 9.8%), intraoperative medication administration including etomidate (3.0, 7.3, 12.6%), hydromorphone (47.6, 26.3, 25.2%), ketamine (11.2, 4.6, 3.0%), dexmedetomidine (18.4, 16.6, 13.6%), phenylephrine (74.0, 74.8, 83.0), epinephrine (2.0, 6.0, 18.0%), norepinephrine (2.4, 7.5, 21.2%), vasopressin (3.4, 7.6, 21.0%), succinylcholine (74.0, 69.0, 61.9%), intraoperative hypotension (28.8, 33.0, 52.3%) and the triple low state (9.4, 30.3, 80.0%) exposure, and 30-day postoperative mortality (0.8, 2.7, 5.6%). TVI was a better predictor of patients that died or survived in the 30 days following surgery compared to cumulative triple low state exposure (AUC 0.68 versus 0.62, p < 0.05). Surgeries that share similar patterns of TVI expression display distinct patient, procedure, and outcome characteristics.


Acral melanoma detection using a convolutional neural network for dermoscopy images.
Acral melanoma is the most common type of melanoma in Asians, and usually results in a poor prognosis due to late diagnosis. We applied a convolutional neural network to dermoscopy images of acral melanoma and benign nevi on the hands and feet and evaluated its usefulness for the early diagnosis of these conditions. A total of 724 dermoscopy images comprising acral melanoma (350 images from 81 patients) and benign nevi (374 images from 194 patients), and confirmed by histopathological examination, were analyzed in this study. To perform the 2-fold cross validation, we split them into two mutually exclusive subsets: half of the total image dataset was selected for training and the rest for testing, and we calculated the accuracy of diagnosis comparing it with the dermatologist's and non-expert's evaluation. The accuracy (percentage of true positive and true negative from all images) of the convolutional neural network was 83.51% and 80.23%, which was higher than the non-expert's evaluation (67.84%, 62.71%) and close to that of the expert (81.08%, 81.64%). Moreover, the convolutional neural network showed area-under-the-curve values like 0.8, 0.84 and Youden's index like 0.6795, 0.6073, which were similar score with the expert. Although further data analysis is necessary to improve their accuracy, convolutional neural networks would be helpful to detect acral melanoma from dermoscopy images of the hands and feet.


[Quality assurance in cytopathology: aspects of the cytohistological correlation].
The cytohistologic diagnosis concordance of the cervix-uterine lesions was evaluated and the discordances are discussed. To evaluate the level of cytohistological diagnosis agreement in the Pathology Division of the Adolfo Lutz Institute and enhance parameters of quality assurance system developed in this institution. The authors retrospectively evaluated 157 pairs of cytologyhistology consecutive cases from the files the Adolfo Lutz Institute (Division of Pathology). Agreement was found in 119 cases (75.8%); in the remaining cases cytologic diagnosis were higher than histology in 11 lesions (7.0%) and lower in 27 (17.2%). We also observed that in 5 cases previously diagnosed as inflammatory, 2 of them were reclassified as CIN 1; after revision, the others remained as inflammatory even though they had a histological diagnosis of CIN. These results showed the role of cytohistological correlation to the quality assurance system of cytological diagnosis and also stressed the importance of taking the cervical scrapes with accurate care to avoid diagnostic discrepancies.


Community asthma clinics: 1993 survey of primary care by the National Asthma Task Force.
To establish a baseline of work done in primary care asthma clinics in the United Kingdom and to assess the degree of clinical delegation to nurses and the appropriateness of their training. Prospective questionnaire survey of asthma care in general practices and a subsidiary survey of all family health services authorities (FHSAs) of the number of asthma clinics in their area. All 14,251 general practices in the United Kingdom and 117 FHSAs or health boards (Scotland and Northern Ireland). Questionnaires were returned by 4327 (30.4%) general practices, 54% being completed by practice nurses and 22% by general practitioners; in 24% profession was not stated. In all, 77.2% (3339/4327) of respondents ran an asthma clinic. 60 FHSAs state the number of asthma clinics at the time of the general practice survey (total 3653 clinics); within responding FHSAs 1702 (46.6%) practices running an asthma clinic replied to the general practice survey. Clinics exclusive for patients with asthma mostly occurred in practices with five or more general practitioners (70.2%), compared with single-handed practices (31.7%). The average number of asthma clinics run per practice was five a month; the average duration was 2 hours and 20 minutes. 1131 (48.8%) nurses ran clinics by themselves, 1180 (47.9%) with the doctor, and 39 (1.7%) had no medical input. Comprehensive questioning occurred other than for nasal (872, 26.1%) or oesophageal (335, 10.0%) symptoms and use of aspirin and non-steroidal drugs (1161, 33.4%). Growth in children was measured by only a third of respondents. Of the 1131 nurses who ran clinics alone, 251 (22.2%) did so without formal training entailing assessment. Asthma clinics are now common in general practice and much of their work is done by nurses, a significant minority of whom may not have had sufficient training. As this survey is probably biased toward the more asthma aware practices, greater deficiencies in training and standards may exist in other practices. Further evaluation of the effectiveness of asthma clinics is needed.


Plastic debris in the Mediterranean Sea: Types, occurrence and distribution along Adriatic shorelines.
Small plastic debris in sediments from five beaches were investigated to evaluate their occurrence and abundance in the Northern Adriatic coast for the first time. Plastic debris extracted from sediments were counted, weighted and identified by Fourier-transform infrared spectroscopy (FT-IR). A total of 1345 items of debris (13.491g) were recorded, with a mean density of 12.1 items kg-1 d.w. and 0.12gkg-1 d.w. Fragments were the most frequent type of small plastics debris detected. In terms of abundance, microplastics (<5mm) accounted for 61% of debris, showing their wide distribution on Adriatic coasts, even far-away from densely populated areas. The majority of the polymers found were polyolefins: there were greater quantities of polyethylene and polypropylene compared to other types of plastic. Primary microplastics accounted for only 5.6% of the total plastic debris. There were greater quantities of microplastics at sites subjected to stronger riverine runoff. The results will provide useful background information for further investigations to understand the sink and sources of this emergent and priority contaminant.


Isolation and characterization of eight polymorphic microsatellite loci in the endangered Hawaiian tree snail, Achatinella sowerbyana.
Achatinella is a genus of highly endangered native Hawaiian tree snails. Eight polymorphic microsatellite markers were isolated for one species, Achatinella sowerbyana, to be used in assessing remaining genetic diversity and gene flow in the often small and isolated populations. All loci amplified consistently in every individual tested. The number of alleles per locus in a sample of 40 individuals varied from three to 13, and the observed heterozygosity ranged from 0.308 to 0.846. These markers will be used to assist in the development of conservation and management plans.


Oxaliplatin resistance induced by ERCC1 up-regulation is abrogated by siRNA-mediated gene silencing in human colorectal cancer cells.
Oxaliplatin is used to treat patients with colorectal cancer (CRC); however, half the patients fail to benefit. The excision repair cross-complementing group-1 (ERCC1) gene was studied and it was hypothesized that its inducible expression contributes to cellular resistance. Thirty CRC cell lines were treated with oxaliplatin and sensitivity was determined by apoptosis. Four sensitive and resistant cell lines were analyzed for oxaliplatin effect on ERCC1 expression and two resistant cell lines were subjected to siRNA-mediated gene silencing. There was no correlation of basal ERCC1 mRNA expression with response to oxaliplatin. ERCC1 mRNA was induced at 24, 48, and 72 hours (71-264%, p<0.05) and ERCC1 protein at 48 hours (123-521%, p<0.05) post-oxaliplatin treatment in resistant cells only. siRNA-mediated silencing of ERCC1 sensitized the CRC cells to oxaliplatin-induced apoptosis, and increased cleaved PARP. ERCC1 gene expression is inducible, contributes to oxaliplatin resistance, and is reversible by targeted suppression of ERCC1, identifying ERCC1 as a potential target for drug development.


MR imaging of sports injuries of the hip.
While MR imaging studies of the hip have been directed routinely toward the diagnosis of atraumatic osteonecrosis (ON), interest has increased in the use of MR imaging to diagnose other disorders of the hip and surrounding soft tissues, especially in athletic injuries. In addition to reviewing MR imaging technical considerations and image appearance, this article discusses applications of MR imaging to diagnosing pelvic and hip fractures, muscle contusion and strain, tendon injuries, acetabular labral tears, bursitis, and osteitis pubis and chronic symphyseal injury.


Zika: what we know and don't know.
Since the initial reports of a link between Zika and microcephaly, researchers across the world began working toward understanding the virus. In a short amount of time, Zika has become a household name, prompting worldwide concern. The virus is spread rapidly by mosquito bites. We currently do not have a vaccine for Zika. But with the recent findings, vaccine companies are mobilizing their resources to expedite efforts to shave years off the typical decade-long process of vaccine development.


A novel fluorescence ratiometric method confirms the low solvent viscosity of the cytoplasm.
Two homologous indocyanine dyes, Cy3.18 and Cy5.18, can be used as a ratio pair for fluorometric determination of solvent viscosity. Succinimidyl ester derivatives of these dyes can be attached to inert carrier macromolecules, such as Ficoll 70, for measurement of intracellular or intravesicular solvent viscosity. When the viscosity of the solvent was varied by various methods, the fluorescence intensity ratio (Cy3/Cy5) in a mixture of Cy3.18-Ficoll 70 (Cy3F70) and Cy5.18-Ficoll 70 (Cy5F70) in solution was found to be solely a function of solvent viscosity and was insensitive to other solvent parameters such as dielectric constant, temperature, and the ability of the solvent to form hydrogen bonds. Most important, it was insensitive to the presence of large macromolecules, such as proteins, which increase the shear viscosity but have little effect on solvent viscosity. Following microinjection into the cytoplasm of living tissue culture cells, no binding of Cy3F70 or Cy5F70 to intracellular components was detected by fluorescence recovery after photobleaching. Fluorescence intensity ratio imaging of Cy3F70 and Cy5F70 in non-motile interphase CV1 and PtK1 cells showed that the solvent viscosity of cytoplasm was not significantly different from water and showed no spatial variation.


Survival patterns among lymphoma patients with a family history of lymphoma.
Genetic factors are important in the etiology and pathogenesis of chronic lymphocytic leukemia (CLL), Hodgkin's lymphoma (HL), and non-Hodgkin's lymphoma (NHL). Only a few small studies have assessed clinical characteristics and prognosis for familial patients, with inconsistent findings. Using population-based registries from Sweden and Denmark, 7,749 patients with CLL, 7,476 patients with HL, and 25,801 patients with NHL with linkable first-degree relatives were identified. Kaplan-Meier curves were constructed to compare survival in patients with lymphoma with and without a family history of lymphoma. The risk of dying was assessed using adjusted Cox proportional hazard models. We found 85 patients with CLL (1.10%), 95 patients with HL (1.28%), and 206 patients with NHL (0.80%) with a family history of any lymphoma. Five-year mortality was similar for patients with CLL (hazard ratio [HR], 1.28; 95% CI, 0.95 to 1.72), HL (HR, 0.78; 95% CI, 0.49 to 1.25), and NHL (HR, 0.91; 95% CI, 0.74 to 1.12) versus without a family history of any lymphoma. Mortality was also similar for patients with versus without a family history of the same lymphoma. T-cell/anaplastic lymphoma patients with a family history of NHL had poorer outcome 5-years after diagnosis (HR, 5.38; 95% CI, 1.65 to 17.52). Results were similar for 10 years of follow-up. With the exception of T-cell/anaplastic lymphoma, survival patterns for patients with CLL, HL, and NHL with a family history of lymphoma were similar to those for sporadic patients, suggesting that most familial lymphomas do not have an altered clinical course. Our findings provide no evidence to modify therapeutic strategies for patients with CLL, HL, or NHL based solely on family history.


Enhancement of drug-induced apoptosis by antisense oligodeoxynucleotides targeted against Mdm2 and p21WAF1/CIP1.
The p53 tumor suppressor gene plays an important role in DNA damage-induced apoptosis and, in general, inactivation of p53 contributes to poor response to chemotherapy. Apoptotic activity of p53 may be negatively modulated by expression of its downstream mediators, including Mdm2 and p21WAF1/CIP1. Consequently, these cellular pathways also represent potential targets for cancer therapy. This study investigated the effect of antisense oligodeoxynucleotides (ODNs), targeted against Mdm2 and p21WAF1/CIP1 on drug-mediated cell killing. Exposure of U2-OS osteosarcoma cells to DNA damaging agents, cisplatin or mitomycin C, caused upregulated expression of Mdm2 and p21WAF1/CIP1. Transient transfection of cells with antisense ODNs to Mdm2 mRNA inhibited Mdm2 protein expression and markedly enhanced apoptotic cell death induced by these drugs. Moreover, when p21WAF1/CIP1 expression was blocked by antisense transfection, drug-mediated cell killing was further accelerated. These results suggest that the enhanced expression of Mdm2 and p21WAF1/CIP1 may inhibit p53-mediated apoptosis and render cells resistant to the effects of DNA damaging agents. Consequently, antisense ODNs targeted against Mdm2 and p21WAF1/CIP1 could be employed in a potential therapeutic strategy sensitizing tumor cells to certain antineoplastic agents.


Muscle Fatigue Sensor Based on Ti3 C2 Tx MXene Hydrogel.
MXene-based hydrogels have received significant attention due to several promising properties that distinguish them from conventional hydrogels. In this study, it is shown that both strain and pH level can be exploited to tune the electronic and ionic transport in MXene-based hydrogel (M-hydrogel), which consists of MXene (Ti3 C2 Tx )-polyacrylic acid/polyvinyl alcohol hydrogel. In particular, the strain applied to the M-hydrogel changes MXene sheet orientation which leads to modulation of ionic transport within the M-hydrogel, due to strain-induced orientation of the surface charge-guided ionic pathway. Simultaneously, the reorientation of MXene sheets under the axial strain increases the electronic resistance of the M-hydrogel due to the loss of the percolative network of conductive MXene sheets during the stretching process. The iontronic characteristics of the M-hydrogel can thus be tuned by strain and pH, which allows using the M-hydrogel as a muscle fatigue sensor during exercise. A fully functional M-hydrogel is developed for real-time measurement of muscle fatigue during exercise and coupled it to a smartphone to provide a portable or wearable digital readout. This concept can be extended to other fields that require accurate analysis of constantly changing physical and chemical conditions, such as physiological changes in the human body.


Care providers' troubled conscience related to an implementation of a time management system in residential care for older people-a participatory action research study.
Care providers in residential care for older people often refer to time shortage, a problem that may generate troubled conscience. The aim of the study was to describe a PAR process to assist care providers in municipal residential care for older people to constructively deal with their troubled conscience related to an implemented time management system. This intervention study was carried out with 14 care providers and their manager in residential care for older people with the support of participatory action research (PAR). The recorded PAR sessions were transcribed and compiled with inspiration from content analysis. The participants were given oral and written information and gave their written informed consent. The PAR process was found to empower the participants to form their own structure of the practical professional planning, adapted to the residents needs and to their daily work. In this process, participants used their troubled conscience as a driving force and as an asset. Instead of launching change without any deeper information, it is important to carefully prepare, involve and inform those who are going to execute the change. Meeting places should be arranged wherein care providers have the opportunity to share and reflect on challenging situations that can generate troubled conscience, especially when comprehensive changes in their work are going to be implemented.


DNA-binding transcription factor NF-1A negatively regulates JC virus multiplication.
JC virus (JCV) DNA replication occurs in the nuclei of infected cells. The level of JCV genome expression depends on nucleotide sequences in the viral regulatory region and their interaction with host-cell nuclear transcription factors. Our previous studies showed a higher level of NF-1X in JCV-permissive cells compared with the other members of the NF-1 family, NF-1A, B and C, which suggests that NF-1X plays a positive role in JCV multiplication. It remained unclear whether a reduction in the level of NF-1A, which is expressed abundantly in JCV-non-permissive cell types, leads to an increase in JCV multiplication. In this study, we show that downregulation of NF-1A expression in JCV-non-susceptible progenitor and HeLa cells results in a reversion to susceptibility for JCV multiplication. These data demonstrate that a higher level of NF-1A protein in JCV-non-permissive cell types, compared with the level of NF-1X, may be acting as a negative regulator at the JCV promoter to control JCV multiplication.


Plasmacytoma with Amyloidosis in a Nutria A Case Report.
A fatal neoplastic disease in a nutria is described. The salient features were proliferation of plasma cells and amyloidosis. A comparison is made with the lesions of viral plasmacytosis of mink and multiple myeloma of man.


Selective COX-2 Inhibitors: A Review of Their Structure-Activity Relationships.
Non-steroidal anti-inflammatory drugs (NSAIDs) are the competitive inhibitors of cyclooxygenase (COX), the enzyme which mediates the bioconversion of arachidonic acid to inflammatory prostaglandins (PGs). Their use is associated with the side effects such as gastrointestinal and renal toxicity. The therapeutic anti-inflammatory action of NSAIDs is produced by the inhibition of COX-2, while the undesired side effects arise from inhibition of COX-1 activity. Thus, it was though that more selective COX-2 inhibitors would have reduced side effects. Based upon a number of selective COX-2 inhibitors (rofecoxib, celecoxib, valdecoxibetc.) were developed as safer NSAIDs with improved gastric safety profile. However, the recent market removal of some COXIBs such as rofecoxib due to its adverse cardiovascular side effects clearly encourages the researchers to explore and evaluate alternative templates with COX-2 inhibitory activity. Recognition of new avenues for selective COX-2 inhibitors in cancer chemotherapy and neurological diseases such as Parkinson and Alzheimer's diseases still continues to attract investigations on the development of COX-2 inhibitors. This review highlights the various structural classes of selective COX-2 inhibitors with special emphasis on their structure-activity relationships.


Assessment of polystyrene nano plastics effect on human salivary α-amylase structural alteration: Insights from an in vitro and in-silico study.
The study found that the enzyme activity of human salivary α-amylase (α-AHS) was competitively inhibited by nanoplastic polystyrene (PS-NPs), with a half-inhibitory concentration (IC50) of 92 μg/mL, while the maximum reaction rate (Vmax) remained unchanged at 909 μg/mL•min. An increase in the concentration of PS-NPs led to a quenching of α-AHS's fluorescence with a slight red shift, indicating a static mechanism. The binding constant (Ka) and quenching constant (Kq) were calculated to be 2.92 × 1011 M-1 and 1.078 × 1019 M-1• S-1 respectively, with a hill coefficient (n) close to one and an apparent binding equilibrium constant (KA) of 1.54 × 1011 M-1. Molecular docking results suggested that the interaction between α-AHS and PS-NPs involved π-anion interactions between the active site Asp197, Asp300 residues, and van der Waals force interactions affecting the Tyr, Trp, and other residues. Fourier transform infrared (FT-IR) and circular dichroism (CD) analyses revealed conformational changes in α-AHS, including a loss of secondary structure α-helix and β-sheet. The study concludes that the interaction between α-AHS and PS-NPs leads to structural and functional changes in α-AHS, potentially impacting human health. This research provides a foundation for further toxicological analysis of MPs/NPs in the human digestive system.


[Quality of service provided to heart surgery patients of the Unified Health System-SUS].
To evaluate the service quality provided to heart surgery patients during their hospital stay, identifying the patient's expectations and perceptions. To associate service quality with: gender, age and the use of extracorporeal circulation. We studied 82 elective heart surgery patients (52.4% females and 47.6% males), operated by midsternal thoracotomy, age: 31 to 83 years (60.4 +/- 13.2 years); period: March to September 2006. Service quality was evaluated in two instances: the expectations at pre-operative and the perceptions of the service received on the 6th post-operative; through the application of the modified SERVQUAL scale (SERVQUAL-Card). The result was obtained by the difference of the sum of the scores on perception minus those of the expectations, and through statistical analysis. The SERVQUAL-Card scale was statistically validated, showing adequate level of internal consistency. We found a higher frequency of myocardial revascularization 55 (67.0%); first heart surgery 72 (87.8%) and the use of ECC 69 (84.1%). We noticed high mean values for expectations and perceptions with significant results (P<0.05). We observed a significant relationship between the quality of service with: gender, in empathy (P= 0.04) and age, in reliability (P = 0.02). There was no significant association between ECC and quality of service. Service quality was satisfactory. The patient demonstrated a high expectation to hospital medical service. Women present a higher perception of quality in empathy and younger people in reliability. The use of ECC is not related to service quality in this sample. The data obtained in this study suggest that the quality of this health service can be monitored through the periodical application of the SERVQUAL scale.


Barriers and enablers for midwives to use the Nursing and Midwifery Board of Australia's Endorsement for scheduled medicines for midwives.
In 2010, the Nursing and Midwifery Board of Australia introduced a new registration standard: Endorsement for scheduled medicines for midwives. The endorsement enables midwives to provide women with Medicare-rebatable care, prescribe relevant medications, and order relevant Medicare-rebatable diagnostics. Translating endorsement education into clinical midwifery practice has been slow, indicating the presence of barriers affecting midwives' ability to use this standard, despite it increasing their scope for service provision. To discover the mechanisms affecting midwives' ability to work to full scope of practice after completing a programme of study leading to endorsement. An observational (non-experimental) design was used. Midwives who had completed an education programme leading to endorsement were invited to complete a survey. Descriptive statistics were used to analyse the quantitative questions and content analysis was conducted on the qualitative data. Results indicated that barriers - such as the limitations of Medicare provisions for endorsed midwives and a general lack of support for the role - restrict endorsed midwives' ability to provide quality maternity services. Having some form of support for the role may act as an enabler, in addition to midwives having personal determination and confidence in their ability to use the endorsement. Recommendations to strengthen the endorsed midwife's role include facilitating endorsement use in the public sector, relaxing Medicare Benefit Schedule and Pharmaceutical Benefit Scheme restrictions, raising awareness of the role and scope, and improving midwives' pre-endorsement preparation. This study highlights the need for an all-of-system approach to support and develop the endorsed midwife's role.


Novel Applications of FIB-SEM-Based ToF-SIMS in Atom Probe Tomography Workflows.
Atom probe tomography (APT) is used to quantify atomic-scale elemental and isotopic compositional variations within a very small volume of material (typically <0.01 µm3). The small analytical volume ideally contains specific compositional or microstructural targets that can be placed within the context of the previously characterized surface in order to facilitate a correct interpretation of APT data. In this regard, careful targeting and preparation are paramount to ensure that the desired target, which is often smaller than 100 nm, is optimally located within the APT specimen. Needle-shaped specimens required for atom probe analysis are commonly prepared using a focused ion beam scanning electron microscope (FIB-SEM). Here, we utilize FIB-SEM-based time-of-flight secondary ion mass spectrometry (ToF-SIMS) to illustrate a novel approach to targeting <100 nm compositional and isotopic variations that can be used for targeting regions of interest for subsequent lift-out and APT analysis. We present a new method for high-spatial resolution targeting of small features that involves using FIB-SEM-based electron deposition of platinum "buttons" prior to standard lift-out and sharpening procedures for atom probe specimen manufacture. In combination, FIB-ToF-SIMS analysis and application of the "button" method ensure that even the smallest APT targets can be successfully captured in extracted needles.


Antibodies against human oxidized low-density lipoprotein (LDL) as markers for human plasma modified lipoproteins.
Recently there has been much interest in the significance and role of antibodies to oxidized LDL (oxLDL) in the plasma of humans and animals with evolving atherosclerosis. The focus in our study was directed on the significance of these antibodies for the pathogenesis and diagnosis of atherosclerosis. Antibodies to human in vitro oxidized LDL were generated following the method of Kohler & Milstein and used to detect oxidized LDL in human plasma. Several antibodies to oxLDL were selected for high reactivity against circulating modified LDL. The relationship between antibodies against oxLDL and circulating oxLDL level in human plasma was examined in normal and hyperlipidemic patients. The slight inverse relationship observed between anti-oxLDL antibodies and oxLDL levels in normal patients suggests that anti-oxLDL antibodies have a role in the clearance of oxLDL from circulation. These antibodies can be considered as markers of the pathogenic determinants of atherosclerosis, such as enhanced lipid oxidation, pro-inflammatory stage, and impaired vasodilatation.


Self-assembled linear bundles of single wall carbon nanotubes and their alignment and deposition as a film in a dc field.
A one-step process of solubilization of single wall carbon nanotubes (SWCNT) in an organic solvent has enabled us to polarize them asymmetrically in a dc electric field. Quaternary ammonium ion-capped SWCNTs readily suspend in organic solvents; under the influence of a dc electric field, they assemble as stretched bundles anchored on the positive electrode. At low dc applied field (approximately 40 V), all of the SWCNTs from the suspension are deposited on the electrode, thus providing a simple methodology to design robust SWCNT films. At higher applied voltages (>100 V), the SWCNT bundles stretch out into the solution and orient themselves perpendicular to the electrode surface. The alignment of these bundles is responsive to the ON-OFF cycles of the applied electric field. The possibility of modulating the alignment of SWCNT in an electric field opens new ways to achieve electrical contacts in nano- to micro-devices.


Diabetes Care in Venezuela.
The incidence of type 2 diabetes (T2D) and its economic burden have increased in Venezuela, posing difficult challenges in a country already in great turmoil. The aim of this study was to review the prevalence, causes, prevention, management, health policies, and challenges for successful management of diabetes and its complications in Venezuela. A comprehensive literature review spanning 1960 to 2015 was performed. Literature not indexed also was reviewed. The weighted prevalence of diabetes and prediabetes was estimated from published regional and subnational population-based studies. Diabetes care strategies were analyzed. In Venezuela, the weighted prevalence of diabetes was 7.7% and prediabetes was 11.2%. Diabetes was the fifth leading cause of death (7.1%) in 2012 with the mortality rate increasing 7% per year from 1990 to 2012. In 2012, cardiovascular disease and diabetes together were the leading cause of disability-adjusted life years.T2D drivers are genetic, epigenetic, and lifestyle, including unhealthy dietary patterns and physical inactivity. Obesity, insulin resistance, and metabolic syndrome are present at lower cutoffs for body mass index, homeostatic model assessment, and visceral or ectopic fat, respectively. Institutional programs for early detection and/or prevention of T2D have not been established. Most patients with diabetes (∼87%) are cared for in public facilities in a fragmented health system. Local clinical practice guidelines are available, but implementation is suboptimal and supporting information is limited. Strategies to improve diabetes care in Venezuela include enhancing resources, reducing costs, improving education, implementing screening (using Latin America Finnish Diabetes Risk Score), promoting diabetes care units, avoiding insulin levels as diagnostic tool, correct use of oral glucose tolerance testing and metformin as first-line T2D treatment, and reducing health system fragmentation. Use of the Venezuelan adaptation of the transcultural Diabetes Nutrition Algorithm for lifestyle recommendations and the Latin American Diabetes Association guidelines for pharmacologic interventions can assist primary care physicians in diabetes management.


Secretin induces rapid increases in inositol trisphosphate, cytosolic Ca2+ and diacylglycerol as well as cyclic AMP in rat pancreatic acini.
Previous studies have shown that the dose-response relationship for secretin-stimulated cyclic AMP accumulation is different from that for secretin-stimulated enzyme secretion in the rat exocrine pancreas. Here we show that secretin concentrations of 10(-10) M and higher stimulated a rise in cyclic AMP levels, with maximum effect on cyclic AMP accumulation being achieved already with 10(-8) M-secretin. However, at this concentration of secretin, enzyme secretion rates were approximately half-maximal. Unexpectedly, at concentrations of secretin greater than 10(-8) M there was evidence suggestive of phosphatidylinositol bisphosphate hydrolysis with rapid increases in inositol trisphosphate, cytosolic free calcium and diacylglycerol content of rat pancreatic acini. Furthermore, there was a dose-response relationship among secretin concentration (in the range 10(-8) M-2 X 10(-6) M), increases in inositol trisphosphate and increases in cytosolic free calcium ([Ca2+]i). Contrary to what has been previously believed, these results clearly indicate that in rat pancreatic acini secretin not only stimulates cyclic AMP accumulation but also raises inositol trisphosphate, [Ca2+]i and diacylglycerol. Thus, two second messenger systems may play a role in the regulation of secretin-induced amylase release.


Improving the Food Environment in Hospitals and Senior Meal Programs.
Most Americans consume more than the recommended daily level of sodium, and tools are needed to assess and improve food practices related to sodium. We describe how the Sodium Practices Assessment Tool (SPAT) was developed and used in 19 hospitals and senior meal facilities in upstate New York. Initial results identified opportunities for improvement in food preparation, presentation, and purchasing practices to reduce sodium consumption. Pre-post comparison results showed significant increases in the use of herbs, spices, unsalted butter, fruits and vegetables, and in the availability of lower-sodium foods. Food service sites can use SPAT to assess sodium practices, inform development of action plans, and measure change over time.


Formaldehyde Release From Baby Wipes: Analysis Using the Chromotropic Acid Method.
Formaldehyde is a common preservative and strong sensitizer. The aim of the study was to evaluate the release of formaldehyde from baby/toddler wet wipes using the chromotropic acid method (CAM). An online search of best-selling baby wipes was conducted. None declared formaldehyde or formaldehyde-releasing preservatives. Standard CAM procedures were used: a 1 × 1-in square of fresh wipe was placed in a bottle with an open vial of 4 mg/1 mL of chromotropic acid and sulfuric acid solution, sealed, and stored for 48 hours. Formalin and water served as controls. A blinded investigator graded color change (negative, indeterminate, mild, moderate, or strong). For quality control, 20% of all samples as well as all positives were retested. Fifty-one popular and highly reviewed baby and toddler wet wipe products were tested using CAM. Twelve wipes (24%) released formaldehyde (8 mild, 4 moderate/strong). Chromotropic acid method testing of 9 wipes (18%) was indeterminate and 30 (59%) were negative. Almost one quarter of baby/toddler wet wipes released formaldehyde when evaluated with CAM. Patients and clinicians should be aware of this potentially undeclared source of this common allergen.


Population receptivity to tobacco advertising/promotions and exposure to anti-tobacco media: effect of Master Settlement Agreement in California: 1992-2002.
Tobacco marketing contributes to adolescent smoking initiation, and the Master Settlement Agreement (MSA), therefore, included relevant restrictions. We analyzed data from large population surveys of Californians, conducted in 1992 (11,905 adults, ages 18 years and older), 1993 (5,531 adolescents, ages 12 to 17 years), and 1996 (6,252 adolescents, 18,616 adults) before the MSA, and in 1999 (6,090 adolescents, 14,729 adults) and 2002 (5,857 adolescents, 20,525 adults) following its implementation. Camel lost favorite-advertisement popularity after 1996, and between 1999 and 2002, there were large increases in the percentages declining to name a favorite advertisement. Ownership of tobacco promotional items declined from its peak in 1996. Furthermore, in 2002, close to 90% of adolescents and young adults reported seeing anti-tobacco messages on television in the past month, significantly higher than 1996. These trends indicate less receptivity to tobacco advertising and promotions following the MSA but leave room for additional restrictions to further reduce receptivity.


Diet culture and obesity in northern Africa.
The etiology of obesity in North Africa is not well understood and few studies shed any light on its development among women. This study compiles what is known about the prevalence of obesity and its determinants in Morocco and Tunisia. Results from the authors' two surveys on nutrition-related disease among reproductive-age women (sample size: 2800) and their children (1200 children under 5 y and 500 adolescents) were combined with data from four national income and expenditure surveys (dating from 1980) to assess obesity trends and development in Morocco and Tunisia. Overall levels of obesity, identified by body mass index (BMI) > or = 30 kg/m(2), were 12.2% in Morocco and 14.4% in Tunisia. Obesity is significantly higher among women than among men in both countries (22.7% vs. 6.7% in Tunisia and 18% vs. 5.7% in Morocco) and prevalence among women has tripled over the past 20 y. Half of all women are overweight or obese (BMI > 25) with 50.9% in Tunisia and 51.3% in Morocco. Overweight increases with age and seems to take hold in adolescence, particularly among girls. In Tunisia, 9.1% of adolescent girls are at risk for being overweight (BMI/age > or = 85th percentile). Prevalence of overweight and obesity are greater for women in urban areas and with lower education levels. Obese women in both countries take in significantly more calories and macronutrients than normal-weight women. The percentage contribution to calories from fat, protein and carbohydrates seems to be within normal limits, whereas fat intake is high (31%) in Tunisia and carbohydrate intake (65-67%) is high in Morocco. These are alarming trends for public health professionals and policy makers in countries still grappling with the public health effects of malnutrition and micronutrient deficiencies. Health institutions in these countries have an enormous challenge to change cultural norms that do not recognize obesity, to prevent significant damage to the public's health from obesity.


Respiratory syncytial virus infection in tropical and developing countries.
Little is known about the epidemiology of respiratory syncytial virus (RSV) infection in tropical and developing countries; the data currently available have been reviewed. In most studies, RSV was found to be the predominant viral cause of acute lower respiratory tract infections (ALRI) in childhood, being responsible for 27-96% of hospitalised cases (mean 65%) in which a virus was found. RSV infection is seasonal in most countries; outbreaks occur most frequently in the cold season in areas with temperate and Mediterranean climates and in the wet season in tropical countries with seasonal rainfall. The situation on islands and in areas of the inner tropics with perennial high rainfall is less clear-cut. The age group mainly affected by RSV in developing countries is children under 6 months of age (mean 39% of hospital patients with RSV). RSV-ALRI is slightly more common in boys than in girls. Very little information is available about the mortality of children infected with RSV, the frequency of bacterial co-infection, or the incidence of further wheezing after RSV. Further studies on RSV should address these questions in more detail. RSV is an important pathogen ill young children in tropical and developing countries and a frequent cause of hospital admission. Prevention of RSV infection by vaccination would have a significant impact on the incidence of ALRI in children in developing countries.


Motorcycle racer with unilateral forearm flexor and extensor chronic exertional compartment syndrome.
We discuss a case of a 26-year-old man, a motorcycle racer, who presented with progressive pain, weakness and swelling of his right forearm and loss of power in his index finger, experienced during motor racing. Chronic exertional compartment syndrome (CECS) of both flexor and extensor compartments of his forearm was diagnosed by dynamic intracompartmental muscle pressure measurements. After fasciotomies, all symptoms were resolved and the patient was able to improve on his preinjury racing skills, without any limitations. A literature review and a surgical 'how-to' for correct release of the extensor and deep flexor compartments of the forearm are provided.


Indications that the Cgamma2 homology region is not a regular domain.
The Cgamma2 homology region of rabbit IgG does not behave like a domain. Thus, there is no trans-interaction between the two Cgamma2 regions; instead there is an unusual cis-interaction between Cgamma2 and Cgamma3 regions. The observations were made on the plasmin digestion products Facb (IgG minus the Cgamma3 region) and pFc' (Cgamma3 region), which did not dissociate under neutral conditions but dissociated in 3M guanidine solution (that is, cis-interaction between Cgamma2 and Cgamma3). The Facb fragment split into subunits with equal molecular weights under neutral conditions on partial reduction and alkylation (that is, lack of trans-interaction between the two Cgamma2 in the molecule).


Antenatal diagnosis of Pena-Shokeir syndrome (type I) with ultrasonography and magnetic resonance imaging.
A case of Pena-Shokeir syndrome type I was diagnosed prenatally with ultrasonography and magnetic resonance imaging (MRI) in a woman with a possible previous occurrence. Initial ultrasonographic examination at 18.5 weeks' gestation demonstrated an unusual appearance of the fetal spine in an otherwise unremarkable fetus. However, subsequent sonographic examinations at 26 and 28.5 weeks demonstrated polyhydramnios and multiple skeletal, brain, and facial abnormalities. Magnetic resonance imaging, performed to further evaluate the fetal brain, confirmed the sonographic findings. However, MRI was not useful in further differentiating the diagnosis. A 1024-g, premature male fetus was delivered at 30 weeks' gestation and died within 30 minutes of delivery. The fetus had multiple congenital anomalies consistent with Pena-Shokeir syndrome type I.


How Many Operative Performance Ratings Does a Pediatric Surgery Fellow Need to Be Deemed Practice Ready?
Identifying the number of cases required for a fellow to achieve competence has been challenging. Workplace-based assessment (WBA) systems make collecting performance data practical and create the opportunity to translate WBA ratings into probabilistic statements about a fellow's likelihood of performing to a given standard on a subsequent assessment opportunity. We compared data from two pediatric surgery training programs that used the performance rating scale from the Society for Improving Medical Professional Learning (SIMPL). We used a Bayesian generalized linear mixed effects model to examine the relationship past and future performance for three procedures: Laparoscopic Inguinal Hernia Repair, Laparoscopic Gastrostomy Tube Placement, and Pyloromyotomy. For site one, 26 faculty assessed 9 fellows on 16 procedures yielding 1094 ratings, of which 778 (71%) earned practice-ready ratings. For site two, 25 faculty rated 3 fellows on 4 unique procedures yielding 234 ratings of which 151 (65%) were deemed practice-ready. We identified similar model-based future performance expectations, with prior practice-ready ratings having a similar average effect across both sites (Site one, B = 0.25; Site two, B = 0.25). Similar prior practice-ready ratings were needed for Laparoscopic G-Tube Placement (Site one = 13; Site two = 14), while greater differences were observed for Laparoscopic Inguinal Hernia Repair (Site one = 10; Site two = 15) and Pyloromyotomy (Site one = 10; Site two = 15). Our approach to modeling operative performance data is effective at determining future practice readiness of pediatric surgery fellows across multiple faculty and fellow groups. This method could be used to establish minimum case number requirements. Original manuscript, Study of Diagnostic Test. II.


Hot-Tip Scanning Electrochemical Microscopy: Theory and Experiments Under Positive and Negative Feedback Conditions.
Hot-tip scanning electrochemical microscopy (HT-SECM) is a novel surface characterization technique utilizing an alternating current (ac) polarized disk microelectrode as a probe. A high-frequency (∼100 MHz) ac waveform applied between the tip and a counter electrode causes the resistive heating of the surrounding electrolyte solution that leads also to the electrothermal fluid flow (ETF). The effects of the temperature and the convection driven by the ETF result in the increased rate of mass transfer of the redox species. In this paper, HT-SECM was studied in positive and negative feedback modes, for which approach curves and cyclic voltammograms were recorded. The experimental data showed that the use of a hot tip leads to a more pronounced feedback compared to that at room temperature. Numerical simulations performed in COMSOL Multiphysics supported the experimental findings. Additional analytical approximations were developed that could be used to predict the faradaic response in HT-SECM experiments. Finally, a possible contribution to the current from the Soret effect was studied theoretically. A good understanding of HT-SECM was achieved, both experimentally and theoretically, suggesting that this methodology could be applied to investigate electrode kinetics under the conditions of elevated temperature and increased rate of mass transfer.


Altered expression level of Sigma1 receptor gene in human colorectal cancer.
Nonopioid Sigma1 receptor (Sig1R) influences numerous metabolism functions including regulation of ion channels, reaction on stress and response to growth signals. Due to this influence, Sigma1 receptor ligands show anti-proliferative and cytotoxic action on tumor cells. Additionally its increased level is observed in some types of tumors. Colorectal cancer is one of the most common cancers worldwide and its clinical development is well described. The aim of the study was evaluation of Sigma1 receptor mRNA expression level in human colorectal cancer and colorectal cancer liver metastases at different stages of tumor development. The mRNA was isolated from 30 patients: 18 with colorectal cancer (CRC) and 12 with colorectal cancer liver metastases (CRCLM). The cDNA of Sig1R gene was amplified by polymerase chain reaction using specific primers. The level of Sig1R mRNA expression was determined by measurement of optical density. Sig1R expression level was increased in CRC and CRCLM. The highest level of Sig1R mRNA was observed in UICC stage III. We also showed significant interactions of UICC stage and tumor localization with Sig1R expression level. There were no interactions between UICC stage and age of patients, although we observed significantly decreased level of Sig1R mRNA in older patients. Clinical advancement stage, localization of tumor and age of patients seems to be an important factors influencing Sigma1 receptor expression level. It is probably due to double nature of Sig1R action - in certain conditions it could act pro- or antiapoptotic. This action might depend on Sig1R activity resulting from its expression level.


Purification and characterization of human brain prolyl endopeptidase.
Prolyl endopeptidase (EC 18.104.22.168) was purified from human brain by a series of column-chromatographic steps using DEAE-cellulose DE-52, hydroxyapatite, phenyl-Sepharose, Sephacryl S-200 and f.p.l.c. (Mono Q). The enzyme was purified by a factor of 943 and was homogeneous in a SDS/polyacrylamide gel as judged by Coomassie Blue staining. The Mr estimated by SDS/PAGE is 79,600, and under native conditions on Sephacryl S-200 it is 85,600. Therefore the enzyme exists as a monomer. With benzyloxycarbonylglycylproline p-nitroanilide as substrate, the optimum pH of the enzyme is 6.8, and with the substrate concentration between 0.059 mM and 0.37 mM the Km is 9.0 x 10(-4) M. The pI of the enzyme is 4.75. The enzyme is classified as a serine proteinase, as it is strongly inhibited by di-isopropyl fluorophosphate. However, other serine proteinase inhibitors do not inhibit the enzyme significantly, suggesting that the active site of prolyl endopeptidase differs from that of classical serine proteinases such as trypsin. Polyclonal antibodies were raised against purified human brain prolyl endopeptidase in rabbits. Western-blot analysis, enzyme-inhibition assays, antibody binding and immunoprecipitation experiments indicated that the polyclonal antibodies are both specific and inhibitory to the enzyme activity.


Akkermansia muciniphila Enhances the Antitumor Effect of Cisplatin in Lewis Lung Cancer Mice.
Recently, intestinal flora plays a vital role in the occurrence and development of tumors and there is link between cancer immunotherapy and Akkermansia muciniphila (Akk). However, the therapeutic efficacy of Akk in lung cancer remained unclear. Hence, our study is aimed at investigating the antitumor effects of cisplatin (CDDP) combined with Akk on lung cancer. Using the murine lung cancer model by subcutaneously inoculating Lewis lung cancer model, 50 mice were divided into five groups: normal, model, CDDP, CDDP+Akk, and CDDP+antibiotics. After treatment within 5 weeks, compared with the model group, the administered group improved the changes of tumor pathomorphology. Compared with the CDDP group, CDDP combining with Akk slowed down the growth of tumor volume, downregulated the levels of ki-67, p53, and factor-associated suicide (Fas) ligand proteins and upregulated Fas proteins, increased the levels of interferon-γ, interleukin-6, and tumor necrosis factor-α, and suppressed the expression of CD4+CD25+Foxp3+ Treg in mouse peripheral blood and spleen. In addition, transcriptome analysis indicated that Akk combining with CDDP increased obviously the levels of IFI27l2 and IGFBP7 and was related to those pathways including the cytokine-cytokine receptor interaction, Th17 cell differentiation, FOXO, JAK-STAT, and PI3K-Akt signaling pathways. These results suggested that the therapeutic efficacy of the combined treatment of Akk and CDDP was superior to the only CDDP treatment, which could enhance immune regulation and would be a promising strategy for the treatment of lung cancer.


Broadband X-ray ptychography using multi-wavelength algorithm.
Ptychography is a rapidly developing scanning microscopy which is able to view the internal structures of samples at a high resolution beyond the illumination size. The achieved spatial resolution is theoretically dose-limited. A broadband source can provide much higher flux compared with a monochromatic source; however, it conflicts with the necessary coherence requirements of this coherent diffraction imaging technique. In this paper, a multi-wavelength reconstruction algorithm has been developed to deal with the broad bandwidth in ptychography. Compared with the latest development of mixed-state reconstruction approach, this multi-wavelength approach is more accurate in the physical model, and also considers the spot size variation as a function of energy due to the chromatic focusing optics. Therefore, this method has been proved in both simulation and experiment to significantly improve the reconstruction when the source bandwidth, illumination size and scan step size increase. It is worth mentioning that the accurate and detailed information of the energy spectrum for the incident beam is not required in advance for the proposed method. Further, we combine multi-wavelength and mixed-state approaches to jointly solve temporal and spatial partial coherence in ptychography so that it can handle various disadvantageous experimental effects. The significant relaxation in coherence requirements by our approaches allows the use of high-flux broadband X-ray sources for high-efficient and high-resolution ptychographic imaging.


Short communication: Latin-style fresh cheese enhances lactic acid bacteria survival but not Listeria monocytogenes resistance under in vitro simulated gastrointestinal conditions.
Different studies in humans have provided evidence about the health benefits of probiotics. However, most probiotic strains do not maintain good viability in the harsh conditions of the gastrointestinal tract (GIT). In the present study, Latin-style fresh cheese produced with potential probiotic bacteria was tested to evaluate this cheese type as a food carrier for the delivery of viable microorganisms after exposure to simulated GIT conditions. The resistance of 28 lactic acid bacteria (LAB) strains and Listeria monocytogenes upon exposure to acidic conditions (pH 2.5) and bile and pancreatic enzymes (0.3% of bile salts and 0.1% of pancreatin) was evaluated in vitro. When compared with fresh cultures, fresh cheese greatly improved LAB survival to simulated GIT conditions, as no loss of viability was observed in either acidic conditions (pH 2.5) or bile salts and pancreatin environment over a 3-h period. In opposition, L. monocytogenes did not survive after 1h under acidic conditions. These data demonstrated that Latin-style fresh cheese could play an important role in probiotic protection against gastrointestinal juices, enhancing delivery within the gut and thereby maximizing potential health benefits of LAB.


Review article: the role of rebamipide in the management of inflammatory disease of the gastrointestinal tract.
Rebamipide stimulates the generation of endogenous prostaglandins in the gastric mucosa and is reported to accelerate ulcer healing. This review discusses whether rebamipide can prevent Helicobacter pylori infection, reduce inflammation, accelerate healing after eradication, promote ulcer healing, and prevent progression of preneoplastic lesions. Furthermore, we evaluate its usefulness in other inflammatory conditions of the gastrointestinal tract. We conclude that rebamipide is an important candidate for long-term suppression of gastro-intestinal inflammation, particularly if reducing the complications of H. pylori infection without eradicating the organism becomes accepted. If its ability to accelerate mucosal normalization is confirmed, rebamipide could be added to eradication regimens. Little information exists on whether such therapy could help limit the development of pre-neoplastic lesions. In light of the dearth of effective drugs to control inflammation in idiopathic inflammatory bowel disease, the potential of any promising new and safe compound deserves to be fully explored. The next step is to devise a targeted plan of translational research, so that results from the bench may be used to design rigorously controlled international clinical trials.


Effect of black soybean extract on the suppression of the proliferation of human AGS gastric cancer cells via the induction of apoptosis.
Black soybean is known to have a health-promoting effect because of its high content of polyphenolic compounds and antioxidant activities. The objective of the present study was to investigate the chemopreventive effects of black soybean extract against human AGS gastric cancer cells and its possible mechanism in inducing apoptosis. Black soybean extract was obtained by extracting black soybean with acidified aqueous acetone, and its phytochemical constituents, as determined by HPLC-DAD methods, were demonstrated to contain various phenolics. The black soybean extract inhibited AGS cell growth in a dose-dependent manner with an IC(50) of 3.69 mg/mL as measured by the MTT assay. This growth inhibition effect was further confirmed by the CFDA-SE assay. Flow cytometry analysis showed that black soybean extract dose-dependently induced apoptosis of AGS cells. Moreover, the involvement of black soybean extract in inducing apoptosis was confirmed by the expression of Bax, Bcl-2, caspase-3, and PARP. The results of the present study indicated that black soybean extract could be used as an apoptosis inducer in AGS cells and a natural chemopreventive agent in the treatment of human gastric cancer.


Coronavirus Disease 2019 (COVID-19) in Solid Organ Transplant Recipients: A Case-Control Study.
BACKGROUND It is unclear whether solid organ transplant (SOT) patients have more severe coronavirus disease 2019 (COVID-19) and worse outcome than the general population. MATERIAL AND METHODS We conducted a case-control study on 32 SOT recipients and 84 non-SOT controls matched for age and sex admitted for confirmed COVID-19. The primary endpoint was in-hospital all-cause mortality rate. Secondary endpoints included severe acute respiratory distress syndrome (ARDS), use of high-flow oxygen therapy, and length of hospital stay. RESULTS The median (IQR) Charlson comorbidity index (CCI) at admission was significantly higher in SOT recipients (6 (3-8) vs 3 (2-4); P<0.01). Fever was less frequent in SOT recipients (78% vs 94%, P=0.01). SOT recipients had a higher median SaO2/FiO2 at admission (452 [443-462] vs 443 [419-452], P<0.01) and reached the worst SaO2/FiO2 value later during hospitalization 15 (10-21) vs 11 (9-14) days, P=0.01). Both groups had a similar severe ARDS rate during hospitalization (33% vs 28%) (p=0.59). There were no significant differences during hospitalization in terms of highest level of respiratory support needed, or length of hospital stay: 8.5 (5.5-21) vs 11.5 (6.5-16.5) days; P=0.34) in SOT recipients when compared to controls. In-hospital all-cause mortality rates were significantly higher in SOT recipients (21.9% vs 4.7%, P<0.01; OR 1.08; 95% CI 0.10-10.98), but among patients who died, median CCI was similar between groups (8 [6-8] vs 7 [6-8]). CONCLUSIONS In our experience, hospitalized SOT recipients for COVID-19 had higher in-hospital mortality compared to non-SOT patients, probably due to the greater number of underlying comorbidities, and not directly related to chronic immunosuppression.


Differential regulation of the slow and rapid components of guinea-pig cardiac delayed rectifier K+ channels by hypoxia.
The aim of this study was to examine the effects of acute hypoxia on the slow (I(Ks)) and rapid (I(Kr)) components of the native delayed rectifier K+ channel in the absence and presence of the beta-adrenergic receptor agonist isoproterenol (isoprenaline; Iso) using the whole-cell configuration of the patch-clamp technique. Hypoxia reversibly inhibited basal I(Ks). The effect could be mimicked by exposing the cells to the thiol-specific reducing agent dithiothreitol (DTT) and attenuated upon exposure of cells to the membrane-impermeant thiol-specific oxidizing compound 5,5'-dithio-bis[2-nitrobenzoic acid] (DTNB). In the presence of hypoxia, the K(0.5) for activation of I(Ks) by Iso was significantly decreased from 18.3 to 1.9 nm. DTT mimicked the effect of hypoxia on the sensitivity of I(Ks) to Iso while DTNB had no effect. Hypoxia increased the sensitivity of I(Ks) to histamine and forskolin suggesting that the effect of hypoxia is not occurring at the beta-adrenergic receptor. The increase in sensitivity of I(Ks) to Iso could be attenuated with addition of PKCbeta peptide to the pipette solution. While hypoxia and DTT inhibited basal I(Ks) they were without effect on I(Kr.) In addition, Iso did not appear to alter the magnitude of I(Kr) in the absence or presence of hypoxia. These data suggest that hypoxia regulates native I(Ks) through two distinct mechanisms: direct inhibition of basal I(Ks) and an increase in sensitivity to Iso that occurs downstream from the beta-adrenergic receptor. Both mechanisms appear to involve redox modification of thiol groups. In contrast native I(Kr) does not appear to be regulated by Iso, hypoxia or redox state.


Precision Intravenous Immunoglobulin Dosing and Clinical Outcomes: A Retrospective Chart Review.
Intravenous immunoglobulin (IVIg) is used for treatment of acute neurologic conditions such as Guillain-Barre syndrome, chronic inflammatory demyelinating polyradiculoneuropathy relapse, and myasthenia gravis exacerbation. Precision dosing (adjusted or ideal body weight) is proposed to conserve IVIg. There have been no published studies comparing clinical outcomes in traditional dosing (actual body weight) with precision dosing. In 2014, our institution began dosing patients with precision dosing. This decision was largely performed by administration rather than physicians' preference. We sought to analyze our retrospective data to understand the change in dosing methods with neurologic outcomes. We performed a retrospective review of all patients hospitalized at a single center who received IVIg for myasthenia gravis, Guillain-Barre syndrome, and chronic inflammatory demyelinating polyradiculoneuropathy from January 2010 to October 2017. We collected baseline information and clinical outcomes including mortality, readmission, need for second rescue treatment, length of stay, discharge disposition, treatment-related adverse events, and modified research council posttreatment sum score. Length of stay was significantly shorter with precision dosing. There was no statistically significant difference in discharge disposition, readmission, rescue treatment, or modified research council posttreatment sum score with precision dosing. Precision dosing did not adversely affect short-term neurologic outcomes.


Enhanced stretch-induced myogenic tone in the basilar artery of spontaneously hypertensive rats.
We investigated if the magnitude of myogenic tone in the basilar artery of SHR differs from that in WKY and, if so, whether RhoA- or PKC-dependent mechanisms were involved. Myogenic tone was developed in response to stretch. Stretch-induced myogenic contraction was significantly greater in the SHR than WKY in the presence of external Ca(2+). However, in the absence of external Ca(2+), stretch did not evoke a myogenic tone. The [Ca(2+)](i)-induced contraction was larger in SHR than WKY and the [Ca(2+)](i)-force curve was significantly shifted to the left in SHR compared to WKY. Y-27632 significantly inhibited stretch-induced myogenic tone, but the inhibitory effect was larger in the SHR than WKY. However, PKC inhibitors had no significant effect on the myogenic tone. RhoA and PKCepsilon were expressed at higher levels in the SHR compared to the WKY. RhoA and PKCalpha translocated from the cytosol to the cell membrane in response to stretch in both animals, but PKCepsilon was translocated only in SHR. Our results strongly suggest that stretch-induced myogenic tone is enhanced in SHR, and the activation of RhoA/Rho kinase plays an important role in the enhanced myogenic tone in SHR.


Syntheses, Characterization and Crystal Structures of Schiff Base Zinc(II) Complexes with Antibacterial Activity.
Two polynuclear zinc(II) complexes, [Zn2(L1)2(OH2)2] (1) and [ZnL2(μ1,1-N3)]n (2), where L1 is the dianionic form of N,N'-bis(3,5-difluoro-2-hydroxybenzylidene)-1,3-diaminopropane, and L2 is the monoanionic form of 2-[(2-dimethylaminoethylimino)methyl]-4,6-difluorophenol, have been prepared and structurally characterized by elemental analyses, IR and UV-Vis spectroscopy, as well as single-crystal X-ray diffraction. In complex 1, the Zn atom is in octahedral coordination, with the donor atoms of the Schiff base ligands L1 and one water O atom. In complex 2, the Zn atom is in trigonal-bipyramidal coordination, with the three donor atoms of the Schiff base ligand L2 and two azido N atoms. The complexes have strong antibacterial activity against B. subtilis and S. aureus.


Precise and broad scope genome editing based on high-specificity Cas9 nickases.
RNA-guided nucleases (RGNs) based on CRISPR systems permit installing short and large edits within eukaryotic genomes. However, precise genome editing is often hindered due to nuclease off-target activities and the multiple-copy character of the vast majority of chromosomal sequences. Dual nicking RGNs and high-specificity RGNs both exhibit low off-target activities. Here, we report that high-specificity Cas9 nucleases are convertible into nicking Cas9D10A variants whose precision is superior to that of the commonly used Cas9D10A nickase. Dual nicking RGNs based on a selected group of these Cas9D10A variants can yield gene knockouts and gene knock-ins at frequencies similar to or higher than those achieved by their conventional counterparts. Moreover, high-specificity dual nicking RGNs are capable of distinguishing highly similar sequences by 'tiptoeing' over pre-existing single base-pair polymorphisms. Finally, high-specificity RNA-guided nicking complexes generally preserve genomic integrity, as demonstrated by unbiased genome-wide high-throughput sequencing assays. Thus, in addition to substantially enlarging the Cas9 nickase toolkit, we demonstrate the feasibility in expanding the range and precision of DNA knockout and knock-in procedures. The herein introduced tools and multi-tier high-specificity genome editing strategies might be particularly beneficial whenever predictability and/or safety of genetic manipulations are paramount.


Treating traumatic injuries of the diaphragm.
Traumatic diaphragmatic injury (DI) is a unique clinical entity that is usually occult and can easily be missed. Their delayed presentation can be due to the delayed rupture of the diaphragm or delayed detection of diaphragmatic rupture, making the accurate diagnosis of DI challenging to the trauma surgeons. An emergency laparotomy and thorough exploration followed by the repair of the defect is the gold standard for the management of these cases. We report a case of blunt DI in an elderly gentleman and present a comprehensive overview for the management of traumatic injuries of the diaphragm.


Tetracycline-induced steatosis in primary canine hepatocyte cultures.
Primary hepatocyte cultures prepared from male beagle dog liver were used to determine susceptibility of the canine liver to tetracycline-induced steatosis. The effects of the drug on mitochondrial lipid metabolism and intracellular triglyceride accumulation were monitored at the same time that steatosis was detected by light microscopy and quantitated using lipid-specific stains. Exposure of primary canine hepatocyte cultures to tetracycline for 24-48 h resulted in concentration-dependent, significant increases in the Oil Red O-stained lipid inclusions. Microscopic examination of the total stained areas suggested that increases over control levels were due primarily to the increase in the size of the lipid inclusions rather than in the number. Biochemical analyses for triglyceride content and histological staining with Nile red, another neutral lipid-specific dye, confirmed a specific increase in intracellular triglyceride following a 24-h exposure to noncytotoxic levels of tetracycline beta-oxidation studies based on the oxidation of [14C]palmitic acid or [14C]palmitoyl carnitine demonstrated a concentration-dependent inhibition of mitochondrial but not peroxisomal beta-oxidation in hepatocytes after a 24-h exposure to tetracycline. In vitro incubation of tetracycline with mitochondria isolated from dog liver showed similar concentration-dependent inhibition. This study clearly indicates that the canine hepatocyte is susceptible to tetracycline-induced steatosis. Triglyceride accumulation was concomitant with the inhibition of mitochondrial lipid metabolism, indicating that this is a primary mechanism leading to steatosis in dog hepatocytes following tetracycline exposure.


Structural basis for the antibody neutralization of herpes simplex virus.
Glycoprotein D (gD) of herpes simplex virus (HSV) binds to a host cell surface receptor, which is required to trigger membrane fusion for virion entry into the host cell. gD has become a validated anti-HSV target for therapeutic antibody development. The highly inhibitory human monoclonal antibody E317 (mAb E317) was previously raised against HSV gD for viral neutralization. To understand the structural basis of antibody neutralization, crystals of the gD ectodomain bound to the E317 Fab domain were obtained. The structure of the complex reveals that E317 interacts with gD mainly through the heavy chain, which covers a large area for epitope recognition on gD, with a flexible N-terminal and C-terminal conformation. The epitope core structure maps to the external surface of gD, corresponding to the binding sites of two receptors, herpesvirus entry mediator (HVEM) and nectin-1, which mediate HSV infection. E317 directly recognizes the gD-nectin-1 interface and occludes the HVEM contact site of gD to block its binding to either receptor. The binding of E317 to gD also prohibits the formation of the N-terminal hairpin of gD for HVEM recognition. The major E317-binding site on gD overlaps with either the nectin-1-binding residues or the neutralizing antigenic sites identified thus far (Tyr38, Asp215, Arg222 and Phe223). The epitopes of gD for E317 binding are highly conserved between two types of human herpesvirus (HSV-1 and HSV-2). This study enables the virus-neutralizing epitopes to be correlated with the receptor-binding regions. The results further strengthen the previously demonstrated therapeutic and diagnostic potential of the E317 antibody.


Pharmacokinetic overview of oral second-generation H1 antihistamines.
Specific H1 antihistamines have become the standard of treatment for relief of symptoms associated with seasonal allergic rhinitis. First-generation antihistamines are small lipophilic molecules that are associated with numerous adverse events largely because of their propensity to cross the blood-brain barrier and their cholinergic activity. Second-generation antihistamines, being more lipophobic, offer the advantages of a lack of CNS and cholinergic effects such as sedation and dry mouth, which are commonly seen in first-generation antihistamines. Their longer duration of action also enables a more patient-friendly dosing regimen which increases patient compliance. This paper reviews the pharmacokinetic properties of these second-generation agents and is intended to provide comparisons that help explain differences in dosing profiles and drug interactions for members of this class of drugs. With the announced withdrawal of terfenadine from the U.S. market in early 1997, 4 second-generation antihistamines are currently widely available: astemizole, loratadine, cetirizine, and fexofenadine. Terfenadine and astemizole both produce significant cardiac QT interval prolongation that may progress to a rare but fatal cardiac ventricular tachycardia known as torsades de pointes. While only terfenadine has been withdrawn due to its adverse effects profile, significant warnings were recently issued for astemizole. The pharmacokinetic profiles of loratadine and cetirizine are reflective of the advantages of these agents as non-cardiotoxic antihistamines. With respect to the newest agent fexofenadine, the major metabolite of terfenadine, published reports are minimal, but its pharmacokinetics differs from that of terfenadine.


Recurrent splice-site mutation in MBTPS2 underlying IFAP syndrome with Olmsted syndrome-like features in a Chinese patient.
Mutations in MBTPS2 have been reported to cause a broad phenotypic spectrum of X-linked genodermatoses, including IFAP (ichthyosis follicularis; atrichia and photophobia) syndrome (OMIM 308205) with or without BRESHECK (brain anomalies, retardation of mentality and growth, ectodermal dysplasia, skeletal malformations, Hirschsprung disease, ear deformity and deafness, eye hypoplasia, cleft palate, cryptorchidism, and kidney dysplasia/hypoplasia) syndrome, keratosis follicularis spinulosa decalvans (KFSD; OMIM 308800) and an X-linked form of Olmsted syndrome. We report a recurrent intronic mutation in MBTPS2 (c.671-9T>G) in a Chinese patient with the typical triad of IFAP syndrome (i.e. ichthyosis, atrichia and photophobia), along with pachyonychia, palmoplantar and periorificial keratoderma, which were reminiscent of Olmsted syndrome. Interestingly, this mutation was previously reported in two cases of IFAP without keratoderma, which suggests clinical heterogeneicity of the same mutation in MBTPS2. The concomitance of Olmsted syndrome-like features in this patient with IFAP may challenge the existence of the X-linked form of Olmsted syndrome as an independent condition.


NLRC4 Inflammasome Is an Important Regulator of Interleukin-18 Levels in Patients With Acute Coronary Syndromes: Genome-Wide Association Study in the PLATelet inhibition and patient Outcomes Trial (PLATO).
Interleukin 18 (IL-18) promotes atherosclerotic plaque formation and is increased in patients with acute coronary syndromes. However the relative contribution of genetic variants to the IL-18 levels has not been fully determined. Baseline plasma IL-18 levels were measured in 16 633 patients with acute coronary syndrome, of whom 9340 had genetic data that passed genotype quality control. A 2-stage genome-wide association study was performed, followed by combined analyses using >10 million genotyped or imputed genetic markers. Single nucleotide polymorphisms at 3 loci (IL18, NLRC4, and MROH6) were identified (P<3.15×10(-8)) in the discovery cohort (n=3777) and replicated in the remaining patients (n=5563). In the pooled data (discovery+replication cohort), 7 independent associations, in 5 chromosomal regions, were associated with IL-18 levels (minimum P=6.99×10(-72)). Six single nucleotide polymorphisms are located in predicted promoter regions of which one disrupts a transcription factor binding site. One single nucleotide polymorphism in NLRC4 is a rare missense variant, predicted to be deleterious to the protein. Altogether, the identified genetic variants explained 8% of the total variation in IL-18 levels in the cohort. Our results show that genetic variants play an important role in determining IL-18 levels in patients with acute coronary syndrome and we have identified genetic variants located in the IL-18 gene (IL18) or close to genes that are involved in procaspase-1 activation (NLRC4 and CARD16, CARD17, and CARD18). These associations also highlight the importance of the NLRC4 inflammasome for IL-18 production in acute coronary syndrome patients.


Ceftriaxone rescues hippocampal neurons from excitotoxicity and enhances memory retrieval in chronic hypobaric hypoxia.
Exposure to high altitude is known to cause impairment in cognitive functions in sojourners. The molecular events leading to this behavioral manifestation, however, still remain an enigma. The present study aims at exploring the nature of memory impairment occurring on chronic exposure to hypobaric hypoxia and the possible role of glutamate in mediating it. Increased ionotropic receptor stimulation by glutamate under hypobaric hypoxic conditions could lead to calcium mediated excitotoxic cell death resulting in impaired cognitive functions. Since glutamate is cleared from the synapse by the Glial Glutamate Transporter, upregulation of the transporter can be a good strategy in preventing excitotoxic cell death. Considering previous reports on upregulation of the expression of Glial Glutamate Transporter on ceftriaxone administration, the therapeutic potential of ceftriaxone in ameliorating hypobaric hypoxia induced memory impairment was investigated in male Sprague Dawley rats. Exposure to hypobaric hypoxia equivalent to an altitude of 7600 m for 14 days lead to oxidative stress, chromatin condensation and neuronal degeneration in the hippocampus. This was accompanied by delayed memory retrieval as evident from increased latency and pathlength in Morris Water Maze. Administration of ceftriaxone at a dose of 200 mg/kg for 7 days and 14 days during the exposure on the other hand improved the performance of rats in the water maze along with decreased oxidative stress and enhanced neuronal survival when compared to hypoxic group without drug administration. An increased expression of Glial Glutamate Transporter was also observed following drug administration indicating faster clearance of glutamate from the synapse. The present study not only brings to light the effect of longer duration of exposure to hypobaric hypoxia on the memory functions, but also indicates the pivotal role played by glutamate in mediating excitotoxic neuronal degeneration at high altitude. The therapeutic potential of ceftriaxone in providing neuroprotection in excitotoxic conditions by increasing Glial Glutamate Transporter expression and thereby enhancing glutamate uptake from the synapse has also been explored.


Scaling up health intervention: is planning in Nigeria becoming evidence based?
With scaling up effective health interventions towards achieving the health-related Millennium Development Goals high on the policy agendas of many developing nations, the costs and as well as benefits of these health interventions are extremely vital in resource poor settings such as Nigeria. Despite the body of evidence on the significance of economic analysis, the integration of economic analysis into planning and decision making is limited in developing countries. This paper commented on the use of a cost analysis study for planning in Nigeria and need to institutionalize such practice.


Ureteral stent displacement associated with deep massage.
A 51 year-old woman with a history of ureteral stenosis and calculi noted recurrence of severe left flank pain while undergoing a deep body massage using the Rolfing method. Displacement of her left ureteral double J stent was noted in the emergency department. The pain and associated incontinence resolved with restoration of the stent to its original position. Practitioners should be aware of this potential complication related to forceful massage pressures.


Energy-transfer quantum dynamics of HeH+ with He atoms: Rotationally inelastic cross sections and rate coefficients.
Two different ab initio potential energy surfaces are employed to investigate the efficiency of the rotational excitation channels for the polar molecular ion HeH+ interacting with He atoms. We further use them to investigate the quantum dynamics of both the proton-exchange reaction and the purely rotational inelastic collisions over a broad range of temperatures. In current modeling studies, this cation is considered to be one of the possible cooling sources under early universe conditions after the recombination era and has recently been found to exist in the interstellar medium. The results from the present calculations are able to show the large efficiency of the state-changing channels involving rotational states of this cation. In fact, we find them to be similar in size and behavior to the inelastic and reaction rate coefficients obtained in previous studies, where H atoms were employed as projectiles. The same rotational excitation processes, occurring when free electrons are the collision partners of this cation, are also compared with the present findings. The relative importance of the reactive, proton-exchange channel and the purely inelastic channels is also analyzed and discussed. The rotational de-excitation processes are also investigated for the cooling kinetics of the present cation under cold trap conditions with He as the buffer gas. The implications of the present results for setting up more comprehensive numerical models to describe the chemical evolution networks in different environments are briefly discussed.


Recombinant anticoagulant factors for adjunctive treatment of sepsis.
Inflammation and coagulation play a pivotal role in the pathogenesis of sepsis. Increasing evidence points to an extensive cross-talk between these two systems, whereby inflammation not only leads to activation of coagulation, but coagulation also considerably affects inflammatory activity. The intricate relationship between inflammation and coagulation has major consequences for the pathogenesis of microvascular failure and subsequent multiple organ failure, as a result of severe infection and the associated systemic inflammatory response. Molecular pathways that contribute to inflammation-induced activation of coagulation have been precisely identified. Important factors include endothelial-bound anticoagulant mechanisms, such as the antithrombin system, the (activated) protein C/thrombomodulin system, and tissue factor pathway inhibitor, which are all impaired during sepsis. Restoration of these anticoagulant pathways is currently evaluated in several clinical studies. Production of these physiological anticoagulants by recombinant technology greatly facilitates this adjunctive treatment strategy.


Sex-Dependent Mechanisms of Chronic Pain: A Focus on Microglia and P2X4R.
For over two decades, purinergic signaling in microglia has persisted in the spotlight as a major pathomechanism of chronic pain. Of the many purinoreceptors, the P2X4R of the ionotropic family, has a well-described causal role underlying chronic neuropathic pain. This review will briefly examine microglial P2X4R signaling in the spinal cord as it relates to chronic pain through a historical lens, followed by a more in-depth examination of recent work, which has revealed major sex differences. We also discuss the generalizability of sex differences in microglial and P2X4R signaling in other pain conditions as well as in nonspinal regions. Finally, we speculate on remaining gaps in the literature as well as what can be done to address them with the ultimate goal of using our collective knowledge to treat chronic pain effectively and in both sexes. SIGNIFICANCE STATEMENT: Effective treatments are lacking for chronic pain sufferers, and this may be explained by the vast sex differences underlying chronic pain mechanisms. In this minireview, we focus on the roles of microglia and P2X4R in chronic pain, with specific attention to the circumstances under which these pathomechanisms differ between males and females. By delineating the ways in which pain occurs differently between the sexes, we can start developing successful therapies for all.


How reliable are data for the ecotoxicity of trivalent chromium to Daphnia magna?
Risk assessments from the European Union and the World Health Organization report values for acute and chronic toxicity of Cr(III) to Daphnia magna in the range of 0.6 mg/L to 111 mg/L and 0.047 mg/L to 3.4 mg/L, respectively. To understand whether factors other than the use of different test media and data reporting contribute to this variability, the authors tested the acute (48-h) and chronic (21-d) toxicities of Cr(III) to D. magna according to Organisation for Economic Co-operation and Development (OECD) methods. Filterable (0.45-µm) chromium concentrations were measured at 0 h, 6 h, 24 h, and 48 h, the latter value corresponding to the total duration of the acute tests and to the interval between medium renewals in chronic tests. In highly alkaline media (4.9 meq/L), Cr concentrations decreased rapidly below the analytical detection limit, and no toxicity was observed. In less alkaline media (approximately 0.8 meq/L), the decrease in filterable Cr concentrations was inversely proportional to the quantity of added Cr(III). The authors concluded that existing data likely underestimate the ecotoxicity of Cr(III) to D. magna. A reliable assessment of the hazard of Cr(III) to D. magna must consider that exposure concentrations can decrease markedly from the beginning to the end of a test and that medium alkalinity strongly influences the outcome of laboratory toxicity tests.


Projections of total hip replacement in Sweden from 2013 to 2030.
The continuously increasing demand for joint replacement surgery in the past decades imposes higher constraints on the budgets of hospitals and healthcare providers. We undertook an analysis of historical trends in total hip replacement performed in Sweden between 1968 and 2012 in order to provide projections of future demand. We obtained data on total hip replacements registered every year and on the evolution of the Swedish population between 1968 and 2012. We assumed the existence of a maximum incidence. So we adopted a regression framework that assumes the existence of an upper limit of total hip replacement incidence. We found that the incidence of total hip replacement will continue to increase until a projected upper incidence level of about 400 total hip replacements per 10(5) Swedish residents aged 40 years and older will be reached around the year 2107. In 2020, the estimated incidence of total hip replacement will be 341 (95% prediction interval (PI): 302-375) and in 2030 it will be 358 (PI: 317-396). Using official forecasted population growth data, about 18,000 operations would be expected to be performed in 2020 and 20,000 would be expected to be performed in 2030. Growing incidence, population growth, and increasing life expectancy will probably result in increased demand for hip replacement surgery. Our findings could serve as a basis for decision making.


Is the Extracellular Impedance High and Non-resistive in Cerebral Cortex?
A recent commentary to Biophysical Journal criticized a previous study published in the same journal by Gomes et al. in 2016, and an alternative interpretation of the measurements was proposed. We reply here to these criticisms and provide some additional clarification, in particular, about a possible misinterpretation of the electrical circuit corresponding to these experiments. We suggest that, indeed, the extracellular impedance in cerebral cortex could be high and non-resistive, and we propose further experiments to settle this issue.


Effects of partial-body cryotherapy (- 110°C) on muscle recovery between high-intensity exercise bouts.
The aim of this study was to evaluate the effects of a single partial-body cryotherapy bout between training sessions on strength recovery. 12 young men (23.9±5.9 years) were randomly exposed to 2 different conditions separated by 7 days: 1) Partial-body cryotherapy (subjects were exposed to 3 min of partial-body cryotherapy at - 110 °C between 2 high-intensity training sessions); 2) Control (subjects were not exposed to partial-body cryotherapy between 2 high-intensity training sessions). Subjects were exposed to partial-body cryotherapy after the first training session. The 2 knee extension high-intensity training sessions were separated by a 40-min rest interval. Knee extension training consisted of 6 sets of 10 repetitions at 60°.s(-1) for concentric actions and 6 sets of 10 at 180.s(-1) for eccentric actions. The decrease in eccentric peak torque and total work was significantly (p<0.05) less after partial-body cryotherapy (5.6 and 2%, respectively) when compared to control (16 and 11.6%, respectively). However, the decrease in concentric peak torque and total work was not different (p>0.05) between partial-body cryotherapy (9.4 and 6.5%, respectively) and control (7.5 and 5.2%, respectively). These results indicate that the use of partial-body cryotherapy between-training sessions can enhance eccentric muscle performance recovery.


Can emotions precipitate seizures--a review of the question.
Controversy over the emotional precipitation of seizures has existed for centuries. Large clinical studies have generally shown a significant proportion of seizures to be triggered by emotions. Laboratory studies have been reported which describe seizure activity on the electroencephalogram during periods of experimentally induced stress. However, very few pf these studies have utilized control groups and blind evaluators. The numerous reports of the emotional precipitation of seizures indicate that the phenomenon may be genuine and that further research is warranted. Research on emotional interventions such as biofeedback techniques and psychotherapy is the next step, and early work on these interventions is promising.


Tibial and peroneal artery anastomosis.
A technique of distal tibial and peroneal artery bypass vein grafting is presented. The principles are those of limited arterial dissection and minimal handling of the involved artery. A technique of suture placement that is applicable for all end-to-side anastomoses is described.


Computational Analysis of Muscular Dystrophy Sub-types Using A Novel Integrative Scheme.
To construct biologically interpretable gene sets for muscular dystrophy (MD) sub-type classification, we propose a novel computational scheme to integrate protein-protein interaction (PPI) network, functional gene set information, and mRNA profiling data. The workflow of the proposed scheme includes the following three major steps: firstly, we apply an affinity propagation clustering (APC) approach to identify gene sub-networks associated with each MD sub-type, in which a new distance metric is proposed for APC to combine PPI network information and gene-gene co-expression relationship; secondly, we further incorporate functional gene set knowledge, which complements the physical PPI information, into our scheme for biomarker identification; finally, based on the constructed sub-networks and gene set features, we apply multi-class support vector machines (MSVMs) for MD sub-type classification, with which to highlight the biomarkers contributing to sub-type prediction. The experimental results show that our scheme can help identify sub-networks and gene sets that are more relevant to MD than those constructed by other conventional approaches. Moreover, our integrative strategy improves the prediction accuracy substantially, especially for those 'hard-to-classify' sub-types.


Characterization of the secondary structure of calmodulin in complex with a calmodulin-binding domain peptide.
The interaction between calcium-saturated chicken calmodulin and a peptide corresponding to the calmodulin-binding domain of the chicken smooth muscle myosin light chain kinase has been studied by multinuclear and multidimensional nuclear magnetic resonance methods. Extensive 1H and 15N resonance assignments of calmodulin in the complex have been obtained from the analysis of two- and three-dimensional nuclear magnetic resonance spectra. The assignment of calmodulin in the complex was facilitated by the use of selective labeling of the protein with alpha-15N-labeled valine, alanine, lysine, leucine, and glycine. These provided reference points during the main-chain-directed analysis of three-dimensional spectra of complexes prepared with uniformly 15N-labeled calmodulin. The pattern of nuclear Overhauser effects (NOE) seen among main-chain amide NH, C alpha H, and C beta H hydrogens indicates that the secondary structure of the globular domains of calmodulin in the complex closely corresponds to that observed in the calcium-saturated state of the protein in the absence of bound peptide. However, the backbone conformation of residues 76-84 adopts an extended chain conformation upon binding of the peptide in contrast to its helical conformation in the absence of peptide. A sufficient number of NOEs between the globular domains of calmodulin and the bound peptide have been found to indicate that the N- and C-terminal regions of the peptide interact with the C- and N-terminal domains of calmodulin, respectively. The significance of these results are discussed in terms of recently proposed models for the structure of calmodulin-peptide complexes.


[Experimental models of lysosomal phase reactivity in blood leukocytes exposed to low doses of potassium cyanide].
Cytochemical analysis of acid phosphatase was used to evaluate lysosomal membranes stability under oral intake of potassium cyanide by rats over one month in daily doses of 1.30 mg/kg (1/10 LD50) and 0.65 mg/kg (1/20 LD50). The authors demonstrated phase-related dose-dependent changes in the lysosomal state, and the main response feature was associated with functional activation that usually followed the membrane alteration.


Excess sphingomyelin disturbs ATG9A trafficking and autophagosome closure.
Sphingomyelin is an essential cellular lipid that traffics between plasma membrane and intracellular organelles until directed to lysosomes for SMPD1 (sphingomyelin phosphodiesterase 1)-mediated degradation. Inactivating mutations in the SMPD1 gene result in Niemann-Pick diseases type A and B characterized by sphingomyelin accumulation and severely disturbed tissue homeostasis. Here, we report that sphingomyelin overload disturbs the maturation and closure of autophagic membranes. Niemann-Pick type A patient fibroblasts and SMPD1-depleted cancer cells accumulate elongated and unclosed autophagic membranes as well as abnormally swollen autophagosomes in the absence of normal autophagosomes and autolysosomes. The immature autophagic membranes are rich in WIPI2, ATG16L1 and MAP1LC3B but display reduced association with ATG9A. Contrary to its normal trafficking between plasma membrane, intracellular organelles and autophagic membranes, ATG9A concentrates in transferrin receptor-positive juxtanuclear recycling endosomes in SMPD1-deficient cells. Supporting a causative role for ATG9A mistrafficking in the autophagy defect observed in SMPD1-deficient cells, ectopic ATG9A effectively reverts this phenotype. Exogenous C12-sphingomyelin induces a similar juxtanuclear accumulation of ATG9A and subsequent defect in the maturation of autophagic membranes in healthy cells while the main sphingomyelin metabolite, ceramide, fails to revert the autophagy defective phenotype in SMPD1-deficient cells. Juxtanuclear accumulation of ATG9A and defective autophagy are also evident in tissues of smpd1-deficient mice with a subsequent inability to cope with kidney ischemia-reperfusion stress. These data reveal sphingomyelin as an important regulator of ATG9A trafficking and maturation of early autophagic membranes.


Generation of recombinant human monoclonal antibodies to rotavirus from single antigen-specific B cells selected with fluorescent virus-like particles.
Technical difficulties have severely limited the yield of methods for the generation of human antiviral monoclonal antibodies (Mabs) in the past. We describe here a novel method for the efficient development of human Mabs against viruses. Rotavirus (RV) is a major cause of gastroenteritis in infants and adults worldwide. We generated fluorescent virus-like particles (VLPs) to identify and physically sort single RV-specific B cells from healthy adult blood donors, or RV-infected infants or adults. We expanded the sorted single B cells in culture, tested for RV-specific antibody secretion, and cloned and sequenced the antibody heavy and light chain variable region (VH and VL) genes. The percentage of wells that produced antibodies after sorting and expanding RV-specific adult B cell clones was high at 23%. The overall efficiency of RV-specific antibody gene recovery after the isolation, confirmation, and cloning of RV-specific VH segments was 1.3% of sorted cells in adults. RV-specific variable gene segments also were obtained from acutely infected infants, although infant B cells did not proliferate and differentiate in culture as well as adult B cells. We expressed recombinant Fabs incorporating the VH and VL genes from RV-specific B cell clones using a new modified bacterial Fab expression vector that we describe. Finally, we demonstrated binding of purified Fabs to RV proteins by immunofluorescence and ELISA. This method for the generation of recombinant human Mabs to RV from single antigen-specific B cell clones selected with fluorescent VLPs could be used to generate human Mabs to many other viruses whose proteins can self-assemble into VLPs.


Pseudocarbynes: Charge-Stabilized Carbon Chains.
Carbyne is the long-sought linear allotrope of carbon. Despite many reports of solid carbyne, the evidence is unconvincing. A recent report of supposed carbyne shows gold clusters in transmission electron microscopy (TEM) images. In order to determine the effects of such clusters, we performed ab initio calculations of uncapped and capped linear carbon chains and their complexes with gold clusters. The results indicate that gold dramatically alters the electron densities of the C≡C bonds. The resulting charge-stabilization of the carbon chains leads to pseudocarbynes. These findings are corroborated in calculations of the structures of crystals containing isolated carbon chains and those intercalated with gold clusters. Calculated Raman spectra of these pseudocarbynes with gold clusters are in better agreement with experiment than calculated spectra of isolated carbon chains. The current work opens the way toward the design and development of a new class of metal-intercalated carbon compounds.