Leprosy and AIDS: a review of the literature and speculations on the impact of CD4+ lymphocyte depletion on immunity to Mycobacterium leprae.
The potent effects of HIV infection on the human immune system, the complexity of the host-parasite interaction in leprosy, and the paucity of current information on the natural history of co-infected patients makes this area a fertile ground for clinical and immunologic investigation. Several studies have now validated the prediction that there exists a large cohort of patients, particularly in Africa, who are concurrently infected with HIV and M. leprae. Sparse but tantalizing evidence suggests that infection with HIV may increase the incidence of leprosy among individuals with subclinical infection with M. leprae, either through shortening the incubation period or by increasing disease penetrance. Similarly, active mycobacterial disease may accelerate the course of HIV disease, as has been postulated to occur during concurrent infections with certain other viral and bacterial pathogens in HIV-positive patients. A subtle and complex interplay between HIV and leprosy may thus result which will impact the observed epidemiology of both illnesses in regions where both are prevalent. Possible effects of the HIV epidemic on leprosy control programs have been outlined by the World Health Organization and in an editorial by Turk and Rees. The published experience provides few guidelines for the clinical care of co-infected patients. The initial response to conventional therapeutic regimens appears to be excellent, but no follow-up data have been included. The possible absence of ENL in these patients would simplify care for multibacillary disease, if this observation is confirmed in larger field studies.(ABSTRACT TRUNCATED AT 250 WORDS)


Comparative Proteomic Analysis of Proteins Involved in Bioenergetics Pathways Associated with Human Sperm Motility.
Sperm motility is the most important parameter involved in the fertilization process and it is strictly required for reproductive success. Although sperm movements are essential for the physiologic fertilization process, the data, deriving from studies focused on the research of altered cell pathways involved in asthenozoospermia, offer only limited information about the molecular mechanism underlying sperm motility. The aim of this study was to identify proteins involved in human sperm motility deficiency by using label-free mass-spectrometry liquid chromatography (LC-MS/MS). For this purpose, we selected sperm samples with three different classes of progressive motility: low, medium (asthenozoospermic samples) and high (normozoospermic samples). We found that several differential expressed proteins in asthenozoospermic samples were related to energetic metabolism, suggesting an interesting link between bioenergetics pathways and the regulation of sperm motility, necessary for the flagellum movement. Therefore, our results provide strong evidence that mass spectrometry-based proteomics represents an integrated approach to detect novel biochemical markers of sperm motility and quality with diagnostic relevance for male infertility and unravel the molecular etiology of idiopathic cases.


RNAi-mediated tocopherol deficiency impairs photoassimilate export in transgenic potato plants.
Tocopherols (vitamin E) are lipophilic antioxidants presumed to play a key role in protecting chloroplast membranes and the photosynthetic apparatus from photooxidative damage. Additional nonantioxidant functions of tocopherols have been proposed after the recent finding that the Suc export defective1 maize (Zea mays) mutant (sxd1) carries a defect in tocopherol cyclase (TC) and thus is devoid of tocopherols. However, the corresponding vitamin E deficient1 Arabidopsis mutant (vte1) lacks a phenotype analogous to sxd1, suggesting differences in tocopherol function between C4 and C3 plants. Therefore, in this study, the potato (Solanum tuberosum) ortholog of SXD1 was isolated and functionally characterized. StSXD1 encoded a protein with high TC activity in vitro, and chloroplastic localization was demonstrated by transient expression of green fluorescent protein-tagged fusion constructs. RNAi-mediated silencing of StSXD1 in transgenic potato plants resulted in the disruption of TC activity and severe tocopherol deficiency similar to the orthologous sxd1 and vte1 mutants. The nearly complete absence of tocopherols caused a characteristic photoassimilate export-defective phenotype comparable to sxd1, which appeared to be a consequence of vascular-specific callose deposition observed in source leaves. CO2 assimilation rates and photosynthetic gene expression were decreased in source leaves in close correlation with excess sugar accumulation, suggesting a carbohydrate-mediated feedback inhibition rather than a direct impact of tocopherol deficiency on photosynthetic capacity. This conclusion is further supported by an increased photosynthetic capacity of young leaves regardless of decreased tocopherol levels. Our data provide evidence that tocopherol deficiency leads to impaired photoassimilate export from source leaves in both monocot and dicot plant species and suggest significant differences among C3 plants in response to tocopherol reduction.


Evaluation of GeneXpert MTB/RIF for detection of pulmonary tuberculosis at peripheral tuberculosis clinics.
Tuberculosis is one of the most common infectious diseases in China, while delayed patient finding obstructed disease control, especially for smear-negative patients. The current study was undertaken to evaluate the diagnostic accuracy of GeneXpert MTB/RIF compared with conventional methods in the detection of pulmonary tuberculosis patients. A total of 295 spot sputum samples from confirmed pulmonary tuberculosis patients were evaluated from September 2014 to June 2015. Each sample was examined by acid-fast bacillus smear microscopy, culture and GeneXpert MTB/RIF. The sputum culture on Löwenstein-Jensen (L-J) was considered as the gold-standard. After testing by smear, 68.81% (203/295) was negative and 31.19% (92/295) was positive. As the gold-standard, L-J culture detected 37.97% (112/295) positive of all specimens, while the positivity for GeneXpert MTB/RIF was 46.44% (137/295). Compared with L-J culture, the combined sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) for GeneXpert MTB/RIF were 94.64%, 82.97%, 77.37% and 96.18% respectively. For smear-negative specimens, the sensitivity, specificity, PPV and NPV for GeneXpert MTB/RIF were 96.00%, 83.05%, 44.44% and 99.32%; while for smear-positive specimens, the corresponding accuracy values were 94.25%, 80.00%, 98.80% and 44.44%. The findings of study indicated that GeneXpert MTB/RIF assay demonstrated a high sensitivity in detecting Mycobacterium tuberculosis compared to smear method and a high NPV among smear negative patients.


Autoregulation of blood flow: Vessel diameter changes in response to different temperatures.
Autoregulation of blood flow is a marvelous phenomenon balancing blood supply and tissue demand. Although many chemically-based explanations for this phenomenon have been proposed and some of them are commonly used today, biomechanical aspects of this phenomenon was neglected. The biomechanical aspect provides insights to us to model vessel diameter changes more precisely and comprehensively. One important aspect of autoregulation phenomenon is temperature changes of the tissue resulted from tissue metabolism. We hypothesize that temperature changes can affect the mechanical properties of the vessel wall leading to vessel diameter changes during autoregulation. Mechanical modeling of vessel diameter changes can also be useful to explain other phenomena in which the vessel diameter changes in response to temperature alterations. Through the mechanical modeling of any vessel, the analysis of temperature-induced changes in vessel diameter can be done more precisely.


Cloning, expression, and characterization of 5-aminolevulinic acid synthase from Rhodopseudomonas palustris KUGB306.
The hemA gene encoding 5-aminolevulinic acid synthase (ALAS) was cloned from the genomic DNA of photosynthetic bacterium Rhodopseudomonas palustris KUGB306. The deduced protein (ALAS) of this gene contained 409 amino acids. The hemA gene was subcloned into an expression vector pGEX-KG and the encoded protein was overexpressed as a fusion protein with glutathione-S-transferase (GST) in Escherichia coli BL21. The recombinant ALAS was purified and isolated free of the fusion partner (GST) by affinity purification on glutathione-Sepharose 4B resin and cleavage of the purified fusion protein by thrombin protease. The optimum pH and temperature of the recombinant ALAS was found to be at pH 7.5-8.0 and 35-40 degrees C, respectively. The Km value of the enzyme was 2.01 mM for glycine and 49.55 microM for succinyl-CoA. The enzyme activity was strongly inhibited by Pb2+, Fe2+, Co2+, Cu2+, and Zn2+ at 1 mM, but slightly affected by Mg2+ and K+. The recombinant ALAS required pyridoxal 5'-phosphate (PLP) as a cofactor for catalysis. Removal of this cofactor led to complete loss of the activity. Ultraviolet-visible spectroscopy with the ALAS suggested the presence of an aldimine linkage between the enzyme and PLP.


An open, randomized, comparative study of oral fluconazole, itraconazole and terbinafine therapy in onychomycosis.
In this open, randomized and comparative study, the safety and efficacy of systemic fluconazole, itraconazole and terbinafine was investigated in 50 patients with distal subungual toenail onychomycosis diagnosed clinically and mycologically. The patients with positive mycology and also the patients with positive microscopy and negative culture were investigated. The treatment duration was 3 months, and the follow-up period was 6 months. Patients were randomly assigned: 16 patients received 150 mg fluconazole once weekly, 18 patients received 200 mg itraconazole twice daily with meals during the first week of each month, and 16 patients received 250 mg/day terbinafine during the treatment period. In a clinical evaluation, at the endpoint of the follow-up period, the clinical cure rates were 81.3% (13/16) in the terbinafine group, 77.8% (14/18) in the itraconazole group, and 37.5% (6/16) in the fluconazole group. The mycological cure rates were 75% (12/16), 61.1% (11/18) and 31.2% (5/16), respectively. The overall assessment rates were 62.5% (10/16), 61.1% (11/18) and 31.2% (5/16), respectively. Statistically significant intra-group reductions from baseline symptom severity values were seen at the endpoint of treatment and at the endpoint of the follow-up period for all three treatment groups in onycholysis, subungual hyperkeratosis, affected-area percentage score and total score parameters (p < 0.001). At the endpoint of the follow-up period, statistically significant differences between the treatment groups were seen in clinical, mycological and overall assessment (p < 0.05). However, while no statistically significant difference between the terbinafine and itraconazole groups was seen, there was a clinical and statistical difference between the other groups and the fluconazole group. Treatment was not stopped for side effects such as mild gastrointestinal and central nervous system symptoms. These effects were noted in four patients in the fluconazole group (25%), five patients in the itraconazole group (27.8%), and three patients in the terbinafine group (18.75%). The clinical laboratory data on all three drug groups did not show any statistically or clinically significant intra-group changes from baseline values at the endpoint (p > 0.05). This comparative study of systemic fluconazole, itraconazole and terbinafine showed that all three drugs were effective and safe in the treatment of onychomycosis. However, fluconazole, at these doses and treatment durations, was the least effective. With regard to cost-effectiveness, side effects and the cure rates, terbinafine could be the drug of choice in the short-term treatment of toenail onychomycosis.


A novel human papillomavirus and host DNA methylation score and detection of cervical adenocarcinoma.
The widespread introduction of Pap testing in the 1960s was followed by substantial reductions in the incidence of cervical squamous cell cancer (SCC). However, the incidence of cervical adenocarcinoma (ADC) did not decrease, likely because of low Pap test sensitivity for ADC and adenocarcinoma in situ (AIS). This study assessed a novel human papillomavirus (HPV) and host DNA Methylation Score for AIS and ADC screening. We measured methylation levels at CpG sites in the L2/L1 open reading frames of HPV16, HPV18, and HPV45-as well as 2 human loci, DCC and HS3ST2. Specifically, we tested exfoliated cervicovaginal cells from women in the HPV Persistence and Progression (PaP) cohort who were positive for 1 of HPV16, 18, or 45, including: 1) 176 with AIS/ADC, 2) 353 with cervical intraepithelial neoplasia-3 (CIN3) or SCC, and 3) controls who either cleared (HPV-Clearers; n = 579) or had persistent HPV16, 18, or 45 infection (HPV-Persisters; n = 292). CpG site-specific methylation percentages were measured using our reported next-generation methods. The Methylation Score was the average methylation percentage across all 35 CpG sites tested. Each individual CpG site had higher methylation percentages in exfoliated cervicovaginal cells collected from patients with AIS/ADC, and as well as those with CIN3/SCC, relative to either control group (weakest P = .004). The Methylation Score for AIS/ADC had a sensitivity of 74% and specificity of 89%. The multivariate odds ratio (OR) between the Methylation Score (4th vs 1st quartile) for AIS/ADC was ORq4-q1 = 49.01 (PBenjamini-Hochberg = 4.64E-12), using HPV-Clearers as controls. CIN3/SCC had similar, albeit weaker, associations with the Methylation Score. HPV16/18/45-infected women with Methylation Scores in the highest quartile had very high odds of AIS/ADC, suggesting they may warrant careful histologic evaluation of the cervical transition zone (eg, conization or loop electrosurgical excision procedure [LEEP]).


Nursing informatics: a universal nursing language.
Knowledge is necessary to use information to make decisions in nursing situations. Nursing data must be grouped in a logical way so the interrelationship between the classification systems and specific concrete phenomena in individual nursing situations is useful. The processing and management of information for decision making is the focus in nursing and health care. The development and use of language that represents the domain of nursing is an essential dimension in an integrated and coordinated health care delivery system. The design and acceptance of a universal nursing language system is critical to collect appropriate data to document nursing's role, functions, and effectiveness in health care by the year 2000. This is an urgent message for nurses to be involved in the technology and nursing care documentation to demonstrate quality care that is cost effective. In the next century, nurses must speak a common nursing language that has the same meaning for all nurses.


[Application of a new catheter to be determining cardiac output through cor sinistrum with thermodilution].
To introduce a new device and catheter to be determining cardiac output through cor sinistrum with thermodilution. Own control was introduced in eight dogs. A pulmonary thermodilution catheter was used through the cor dextrum of the dogs; The new-design catheter was used through artery, and there were two situs to fix:its ahead in left atrium and the ahead near to aortic root. Determine cardiac output (CO) under four circumstances: controlled respiration, apnoea, in controlled hypotension, after controlled hypotension. There was no significant difference among the parameters (CO and cardiac index) of all circumstances. The CO determined by the pulmonary thermodilution catheter and those by the new-design catheter in two situs were positive correlation, and the gamma were 0.986, 0.989; likewise, the cardiac index (CI) was positive correlation, and the gamma were 0.983, 0.985. The CO and CI by the new-design catheter between two situs were positive, and the gamma were 0.992, 0.988. The parameters by the new-design catheter and those by the pulmonary thermodilution catheter were concordant. In comparison with the pulmonary thermodilution catheter, the new-design catheter had simple device and could be easily operated.


Bilateral and unilateral vertical ground reaction forces and leg asymmetries in soccer players.
The purposes of this study were to assess unilateral and bilateral vertical jump performance characteristics, and to compare the vertical ground reaction force characteristics of the impulse and landing phase of a vertical jump between the dominant and non-dominant leg in soccer players. The sample consisted of 20 male soccer players (22.80 ± 2.71 years, 1.88 ± 0.06 m, 76.47 ± 8.80 kg) who competed in the third division of the Spanish football league. Vertical jump performance was determined by testing the impulse and landing phase of a bilateral vertical jump, dominant leg vertical jump and non-dominant leg vertical jump. Significant differences (p < 0.05) between dominant and non-dominant legs were found in counter movement jump (CMJ) flight time (LA = -2.38%, d = 0.33), CMJ flight height (LA = -4.55%, d = 0.33) and CMJ speed take-off (LA = -2.91%, d = 0.42). No significant differences were found between the dominant and non-dominant leg in the F1 and F2 magnitudes during the landing phase, the time from the first contact of the foot with the ground to the production of F1, the time from the second contact of the foot with the ground to the production of F2, and the time to stabilization of the landing phase. Although differences were found between the dominant and non-dominant leg in the impulse phase of the jump, no significant differences were found between dominant and non-dominant legs in the landing phase of vertical jump variables.


Stress management in highway maintenance workers.
The efficacy of a work-based stress management training program was assessed in highway maintenance workers. Volunteers received training in electromyogram (EMG) biofeedback (n = 17) or muscle relaxation (n = 12) or served as wait-list controls (n = 9). Daily one-hour training sessions were conducted at the workplace for two consecutive workweeks. The biofeedback group showed significant posttraining decreases in forehead EMG levels compared with controls, while decreases found in the muscle relaxation group were not significant. All groups reported significant increases in quality of sleep and in feeling refreshed at work and decreases in subjective tension levels. A three-month follow-up study revealed regression of EMG levels toward baseline in all groups although the percentages of EMG reductions at follow-up were larger for the trained groups than for controls. All groups showed significant improvement on measures of anxiety, somatic complaints, sleep behavior, job satisfaction, and alcohol use. The results support other recent studies indicating the usefulness of work-site stress management programs, although the specificity of training effects and the durability of physiological effects over time remain questionable.


Composition and structure of helminth communities in eels Anguilla anguilla from Italian coastal lagoons.
The composition and diversity of total and intestinal helminth component and infracommunities were determined in eels Anguilla anguilla from four shallow coastal lagoons near Rome. The lagoons differed principally in respect of their salinity. Only Lake Burano still received an input of freshwater, and both freshwater and marine helminth species were found in eels. In the other three lagoons all helminth species were marine or euryhaline and communities were dominated by digeneans. At component community level these three lagoons were far more similar to each other than to Lake Burano, but this distinction almost disappeared at infracommunity level. Species richness and diversity declined with increasing salinity at component community level but not at infracommunity level. Despite the changes and differences in helminth community composition, intestinal helminth infracommunity structure was very similar in all four lagoons and to that in eels from freshwater localities in Britain. The findings thus provide further and unexpected support for the view that some factor(s) other than supply side ones must be operating to produce the fundamental structural similarity in helminth communities in eels.


Natural history and risk stratification of patients undergoing non-invasive ventilation in a non-ICU setting for severe COPD exacerbations.
Non-invasive ventilation (NIV) delivered in an intensive care unit (ICU) has become the cornerstone in the treatment of patients with severe chronic obstructive pulmonary disease (COPD) exacerbations. A trend towards managing these patients in non-ICU setting has emerged in recent years, although out-of-hospital survival by this approach and how to prognosticate it is unknown. We aimed to investigate these issues. We consecutively recruited 100 patients (49 males; median age 82 years) who received NIV treatment for acute respiratory failure due to COPD exacerbation in non-ICU medical wards of our hospital, between November 2008 and July 2012. We assessed survival (both in-hospital and out-of-hospital) of all these patients, and analyzed baseline parameters in a Cox proportional hazards model to develop a prognostic score. The median survival in the study population was 383 days (240-980). Overall survival rates were 71.0, 65.3, and 52.7 % at 1, 3, and 12 months, respectively. Age >85 years, a history of heart disorders and a neutrophil count ≥10 × 10(9) were associated with higher mortality at Cox's analysis (χ (2) = 35.766, p = 0.0001), and were used to build a prognostic score (NC85). The presence of two or more factors determined the deepest drop in survival (when NC85 ≥2, mortality at 1, 3, and 12 was 60.7, 70.4, and 77.2 %, respectively, while when NC85 = 0 were 4.0, 4.0, and 14.0 %). A simple model, based on three variables (age, neutrophil count and history of heart disease), accurately predicts survival of COPD patients receiving NIV in a non-ICU setting.


Novel chromene derivatives as TNF-alpha inhibitors.
A novel series of chromene-based TNF-alpha inhibitors is described. These chromene derivatives inhibit bacterial lipopolysaccharide (LPS) stimulated production of TNF-alpha from human peripheral blood mononuclear cells (PBMC). Additionally, these compounds inhibit NF-kB mediated transcription activation.


Frontline: Epitope recognition on the myelin/oligodendrocyte glycoprotein differentially influences disease phenotype and antibody effector functions in autoimmune demyelination.
Preliminary observations of humoral immunity against the myelin oligodendrocyte glycoprotein (MOG) in experimental allergic encephalomyelitis (EAE) and human multiple sclerosis (MS) suggest that a subset of anti-MOG autoantibodies directed against conformational epitopes is of pathogenic predominance. Here, we provide proof that in marmoset EAE, autoantibodies reactive against conformational epitopes of MOG are not only responsible for aggravating demyelination, but also an essential factor for disease dissemination in space within the central nervous system, a hallmark for typical forms of human MS. In terms of effector mechanisms, IgG deposition and complement activation occur exclusively in association with presence of these conformational antibodies, while microglial/macrophage activation appears to be a common immunopathological finding regardless of the fine determinant specificity of anti-MOG antibodies. These findings highlight for the first time the complex heterogeneity of function and pathogenicity in the polyclonal anti-MOG antibody repertoire of outbred species. Because the linear and conformational antibody determinants of MOG are shared between marmosets and humans, these results are directly relevant to understanding effector mechanisms of organ damage in MS.


MiR-98 Protects Nucleus Pulposus Cells against Apoptosis by Targeting TRAIL in Cervical Intervertebral Disc Degeneration.
The excessive apoptosis of nucleus pulposus (NP) cells is a major risk factor in the progress of cervical intervertebral disc degeneration (IVDD). In this study, we investigated the impact of miR-98 on apoptosis of NP cells and the potential molecular mechanisms. Lipopolysaccharide (LPS) was used to establish an NP cell IVDD model. The sponging effect of miR-98 on TRAIL 3'UTR was predicted by ENCORI and assessed by the dual-luciferase reporter gene system. The expression levels of miR-98, TRAIL, and TRAIL pathway-related genes were tested by qRT-PCR, Western blot, and immunofluorescence analysis. Cell apoptosis was analyzed by Hoechst 33258 staining and flow cytometry. Cell viability was analyzed by MTT assay. It was found that the expression level of miR-98 was downregulated, while the level of TRAIL was upregulated in IVDD tissues, and their levels were negatively and positively associated with the clinical MRI grade, respectively. The LPS treatment resulted in a significant decrease of the miR-98 expression level and an increase of the TRAIL expression level in NP cells. miR-98 reduced NP cell apoptosis under LPS treatment in vitro. miR-98 directly targeted TRAIL. Moreover, the mRNA and protein levels of DR5, FADD, cleaved caspase8, cleaved caspase3, and cleaved PARP were downregulated by miR-98 overexpression. Overexpression of TRAIL partially reversed the suppressive roles of miR-98 on cell apoptosis and activation of the TRAIL pathway. We concluded that miR-98 inhibited apoptosis of NP cells by inactivating the TRAIL pathway via targeting TRAIL in IVDD NP cells. These results indicated that miR-98 might be a therapeutic target for IVDD.


The optimal threshold for removing noise from speech is similar across normal and impaired hearing-a time-frequency masking study.
Hearing-impaired listeners' intolerance to background noise during speech perception is well known. The current study employed speech materials free of ceiling effects to reveal the optimal trade-off between rejecting noise and retaining speech during time-frequency masking. This relative criterion value (-7 dB) was found to hold across noise types that differ in acoustic spectro-temporal complexity. It was also found that listeners with hearing impairment and those with normal hearing performed optimally at this same value, suggesting no true noise intolerance once time-frequency units containing speech are extracted.


Accurate Noninvasive Arterial Assessment of the Wounded Lower Limb: A Clinical Challenge for Wound Practitioners.
Arterial investigations are an essential part of lower extremity wound assessment. The results of these investigations assist the wound clinician to determine the etiology of the wound, predict healing capacity, and inform further management. There are a number of noninvasive testing methods available to practitioners, all with varying levels of reliability and accuracy. Clinical wound assessment guidelines give varied recommendations when it comes to lower limb vascular assessment in the presence of a wound. This leaves clinicians with little guidance on how to choose the most appropriate test, and uncertainty remains about which tests provide the most accurate information in different patient-specific contexts. Conditions such as advanced age, diabetes, and renal disease are known to affect the accuracy of some commonly used lower limb arterial assessment methods, and alternate testing methods should be considered in these cases. This seminal review discusses the reliability and accuracy of lower limb vascular assessment methods used to guide lower limb arterial assessment in the presence of wounds.


Creation of experimental urethral obstruction in utero: evaluation of fetal renal function.
Management of obstructive uropathy diagnosed in utero would be markedly enhanced by the availability of a simple, safe and quantitative fetal renal function test to predict postnatal renal function. In order to allow experimental evaluation of such a parameter, we adjusted a model of complete urethral obstruction with urachal ligation in 30 fetal lambs at 79 or 99 days of gestation. The method described allows obstruction in male and female fetuses as early as 79 days of gestation, with an overall high survival rate (control: 12/14; obstructed: 23/29), although lower (6/12) when obstruction is performed early (79 days) during gestation. Consequences of obstruction were examined on the 121st day of gestation. Severe hydronephrosis, ureteral and calyceal dilatation, with or without ascites and pulmonary hypoplasia were observed in all fetuses; creatinine clearance determined in utero was decreased in both groups with obstruction (early and late) vs control group: 1.15 +/- 0.5, NS, and 0.58 +/- 0.4, p < 0.01 vs 1.61 +/- 0.8 ml/min/kg respectively. In both obstructed groups, fetuses with ascites displayed lower plasma creatinine concentration and higher creatinine clearance values when compared to fetuses without ascites. In conclusion, the experimental model of obstructive uropathy described appears efficient and easily reproducible, allowing therefore the evaluation of a predictive parameter of postnatal renal function. Our preliminary results suggest that renal fetal function is more dependent on the degree of obstruction than on the term of its creation.


Insulin depot formation in subcutaneoue tissue.
The size and geometry of an insulin depot that is formed during subcutaneous administration by an insulin pump is evaluated. A novel method is used to visualize accurately the depot formation for small volumes of insulin (of the order of 10-100 µl) at a given point in time. Conventional visualization methods such as magnetic resonance imaging are unable to provide such accurate measurements because of their coarse imaging resolution and long measurement time. The described method consists of subcutaneously infusing dyed insulin into porcine tissue and subsequently shock freezing it with liquid nitrogen. The frozen sample is then sliced into thin layers using a cryomicrotome. A digital image of each layer is taken and then processed with proprietary software, which identifies the dyed areas on each layer and reconstructs a three-dimensional model of the insulin depot with a planar resolution of 30 × 30 µm(2) and a depth resolution of 100 µm. Since this process is not viable for living organisms, porcine tissue was used immediately following slaughter of the animal. To date, it is most often assumed that the insulin depot takes the shape of a sphere around the tip of the cannula (e.g., 50 µl insulin equates to a spherical radius of 2.3 mm). However, in practice, such a depot form is never observed. Instead, the insulin depot initially spreads laterally (i.e., parallel) to the skin surface and in the collagen matrix that binds the adipose cells together. The depot outreach increases with larger infused volumes, e.g., maximum outreach measured at 5.0/5.7/7.1 mm (quartiles, n = 17) for 50 µl of infused insulin. Beyond a given infused volume (approximately 100 µl), the insulin also starts to spread perpendicular to the skin surface. It is concluded that formation of the insulin depot depends on the opening of channels at the boundaries between adipose cells. Hence the insulin follows a path of least resistance and depot formation is determined by the local structure of the subcutaneous tissue.


An outcomes assessment of a hybrid-PBL course in treatment planning.
As part of an ongoing process of curriculum development, a new course was developed at the University of British Columbia (UBC) to address problems such as lack of student enjoyment of the learning process, poor student preparedness for clinical treatment planning, and underdeveloped critical thinking skills in students beginning their clinical experience. The new course utilized a hybrid problem-based learning (PBL) format that provided students with an overview in lecture format while encouraging active learning in small group tutorials and seminars. Half the second-year class was randomly selected to participate in the new course, while the other half received the standard lectures on the subject. An outcomes assessment examined whether the aforementioned problems had been addressed in the new course. Course participants completed a post-course evaluation, and all students completed a self-assessment of their preparedness and progress in treatment planning and diagnostic. Clinical instructors, who were blind to the identity of the new course participants, were asked to independently assess each student using the same criteria. Results indicate that students who participated in the hybrid-PBL course enjoyed the learning process and later rated themselves as being better prepared and improving more in the areas of treatment planning and diagnostic records than their counterparts, although the latter results were not routinely statistically significant (p < 0.1). Instructor evaluations supported these differences between the two groups.


Rivaroxaban, an oral, direct factor Xa inhibitor: a new option for thromboprophylaxis.
Patients undergoing major orthopedic surgery, including total hip arthroplasty (THA) and total knee arthroplasty (TKA), are at high risk for developing venous thromboembolism (VTE). Although largely a preventable complication, VTE develops in a significant proportion of patients, highlighting the need for improved methods of VTE prevention. Current thromboprophylactic options are limited by unpredictable pharmacokinetics and pharmacodynamics (vitamin K antagonists), parenteral/subcutaneous administration (heparin and low-molecular-weight heparins), complicated dosing, and increased risk of bleeding.Rivaroxaban is an oral, direct Factor Xa inhibitor that has recently received marketing authorization in the United States for prophylaxis of deep vein thrombosis in patients undergoing hip or knee replacement surgery. The clinical pharmacology of rivaroxaban supports a convenient, oral, once-daily dosing regimen without the need for routine coagulation monitoring after THA or TKA. A comprehensive phase II and III study program supports its safety and efficacy for VTE prevention after THA or TKA. Phase III results have demonstrated the superior efficacy of rivaroxaban regimens compared with enoxaparin regimens, with similar rates of major bleeding. This article provides an overview of the phase II and III results that support the use of this agent for the prevention of VTE after elective total hip or knee replacement.


A Carboxylate to Amide Substitution That Switches Protein Folds.
Metamorphic proteins are biomolecules prone to adopting alternative conformations. Because of this feature, they represent ideal systems to investigate the general rules allowing primary structure to dictate protein topology. A comparative molecular dynamics study was performed on the denatured states of two proteins, sharing nearly identical amino-acid sequences (88 %) but different topologies, namely an all-α-helical bundle protein named GA 88 and an α+β-protein named GB 88. The analysis allowed successful design of and experimental validation of a site-directed mutant that promotes, at least in part, the switch in folding from GB 88 to GA 88. The mutated position, in which a glutamic acid was replaced by a glutamine, does not make any intramolecular interactions in the native state of GA 88, such that its stabilization can be explained by considering the effects on the denatured state. The results represent a direct demonstration of the role of the denatured state in sculpting native structure.


[TREATMENT OUTCOME OF OBSTRUCTIVE PYELONEPHRITIS DUE TO URETERAL CALCULI WITH DISSEMINATED INTRAVASCULAR COAGULOPATHY].
Because obstructive pyelonephritis secondary to ureteral stones can easily cause sepsis and concomitant disseminated intravascular coagulation (DIC), it is a potentially lethal disease. However, the optimal treatment for such severe patients has yet to be established. In this study, we aimed at clarifying the effectiveness of emergent drainage and DIC treatments for patients with septic DIC due to obstructive pyelonephritis. In additon, we also evaluated the impact of recombinant human thrombomodullin (rTM) for severe patients with DIC. From September 2006 to May 2013, 31 patients with obstructive pyelonephritis secondary to ureteral stones who met the acute DIC criteria from the Japanese Association of Acute Medicine were treated at our institution. All patients received emergent drainage of urinary tract and anti-DIC treatment, as well as administration of antibiotics and adequate volume infusion. To evaluate the impact of rTM, patients received rTM were compared with those managed by other DIC therapeutic agents. The mean patients' age was 73 years old, and 27 patients (87.1%) were in a state of septic shock. All patients, except for one patient (3.2%) who died 6 days after drainage, could recover from sepsis and comcomitant DIC. Interestingly, thrombocyte count, creatinine, and SOFA Score in rTM group were recovered faster than those in no rTM group (p = 0.017, 0.0038, and 0.0006, respectively). These results indicate that most patients with DIC caused by obstractive pyelonephritis can be successfully managed by emergency drainage and anti-DIC treatment. In addition, rTM may be effective for the treatment of such severe patients by improving organ failure associated with disordered coagulation.


Comparison of Positive/Inconclusive Xpert Xpress SARS-CoV-2 with the Standard M nCoV Real-Time Detection.
COVID-19, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), can be diagnosed using rapid real-time PCR, real-time reverse transcription PCR (rRT-PCR), or rapid antigen testing. Among these, rRT-PCR is considered the gold standard assay. The Xpert Xpress SARS-CoV-2 assay is a rapid real-time PCR test, approved by the Korean Disease Control and Prevention Agency in 2020. The overall concordance and positive concordance rates of the Xpert assay with the STANDARD M nCoV Real-Time Detection kit were determined. All samples with positive or inconclusive Xpert test results from July 2021 to February 2023 that underwent confirmatory testing using the reference rRT-PCR assay were included in the analysis. Samples from 224 patients (93 men and 131 women) with a median age of 59 years (range 15 - 90 years) were included. Of 212 samples that tested positive using Xpert, 112 (52.8%) were true positves and 100 (47.2%) were false positives on rRT-PCR testing. The overall concordance and positive concordance rates were 52.8% (112/212) and 54.5% (112/224), respectively. In the Xpert positive group, the samples had a lower Ct value for the E gene than the N2 gene. The Ct values for the E and N2 genes were significantly lower in the positive group than in the inconclusive group. Positive or inconclusive Xpert results should be confirmed by the gold standard rRT-PCR for early control of this disease. Furthermore, Korea's policy should be reconsidered given the high false-positive rate of the rapid real-time PCR Xpert Xpress SARS-CoV-2 assay.


Pulmonary endothelial cells exhibit sexual dimorphism in their response to hyperoxia.
Abnormal pulmonary vascular development is a critical factor in the pathogenesis of bronchopulmonary dysplasia (BPD). Despite the well-established sex-specific differences in the incidence of BPD, the molecular mechanism(s) behind these are not completely understood. Exposure to a high concentration of oxygen (hyperoxia) contributes to BPD and creates a profibrotic environment in the lung. Our objective was to elucidate the sex-specific differences in neonatal human pulmonary microvascular endothelial cells (HPMECs) in normoxic and hyperoxic conditions, including the propensity for endothelial-to-mesenchymal transition. HPMECs (18- to 24-wk gestation donors, 6 male donors and 5 female donors) were subjected to hyperoxia (95% O2 and 5% CO2) or normoxia (air and 5% CO2) up to 72 h. We assessed cell migration and angiogenesis at baseline. Cell proliferation, viability, and expression of endothelial (CD31) and fibroblast markers (α-smooth muscle actin) were measured upon exposure to hyperoxia. Female HPMECs had significantly higher cell migration when assessed by the wound healing assay (40.99 ± 4.4%) compared with male HPMECs (14.76 ± 3.7%) and showed greater sprouting (1710 ± 962 μm in female cells vs. 789 ± 324 in male cells) compared with male endothelial cells in normoxia. Hyperoxia exposure decreased cell viability (by 9.8% at 48 h and 11.7% at 72 h) and proliferation (by 26.7% at 72 h) markedly in male HPMECs, whereas viability was sustained in female endothelial cells. There was greater expression of α-smooth muscle actin (2.5-fold) and decreased expression (5-fold) of CD31 in male HPMECs upon exposure to hyperoxia. The results indicate that cellular sex affects response in HPMECs in normoxia and hyperoxia. NEW & NOTEWORTHY Cellular sex affects response in human neonatal pulmonary microvascular endothelial cells in normoxia and hyperoxia. Under normoxic conditions, female human neonatal pulmonary microvascular endothelial cells display greater migration and angiogenic sprouting compared with male endothelial cells. Compared with female endothelial cells, hyperoxia exposure decreased cell viability and proliferation and increased α-smooth muscle actin and decreased CD31 expression in male endothelial cells, indicating an increased endothelial-mesenchymal transition.


[M-VAC chemotherapy for advanced urothelial cancer--side effects and their management].
Since the M-VAC (methotrexate, vinblastine, doxorubicin, cisplatin) regimen was reported by Sternberg in 1985, it has been widely accepted for the treatment of metastatic transitional cell carcinoma. This regimen has a significantly high response rate, but bone marrow suppression and gastrointestinal (GI) symptoms are inevitable. To complete this M-VAC regimen, preventive therapy for side effects is necessary. From November 1986 to March 1993, a total of 72 patients were admitted and received M-VAC therapy at our hospital. All of them had metastatic or invasive transitional cell carcinoma and they received a total of 163 complete courses of M-VAC therapy. We examined the side effects of this M-VAC regimen, and evaluated the effectiveness of colony-stimulating factor for prevention of granulocytopenia or granisetron for prevention of GI symptoms. Twenty-three patients (39 courses) were given recombinant colony-stimulating factor. This cytokine prevented the nadir of neutropenia and shortened the period to reach the nadir and period that the neutrophil count was below 1,000/mm3. Twelve patients (26 courses) were given granisetron, with significant reduction of the incidence of GI symptoms. These findings suggest that M-VAC therapy is effective and safe when used in combination with these drugs.


Road environment, crash type and hospitalisation of bicyclists and motorcyclists presented to emergency departments in Western Australia.
Emergency presentations by bicyclists and motorcyclists are often the result from a fall or non-motor vehicle collision that occurred in off-road locations. Consequently, they are unlikely to be captured by police records. If the injury is not severe enough to warrant hospitalisation, they will also not be captured by the hospital admission system. To ascertain the nature and type of these crash events, a 6-month prospective study was undertaken of bicyclists and motorcyclists who presented to emergency departments in Perth, Western Australia, due to involvement in a crash or non-motor vehicle collision. Of the 330 eligible presentations, 151 bicyclists and 104 motorcyclists agreed to participate in a structured interview, representing a response rate of 77.3%. Among them, 120 (79%) bicyclists and 71 (68%) motorcyclists had a fall or non-motor vehicle collision, and many of the crashes (88 (58%) and 47 (45%), respectively) occurred off-road. Moreover, 26.5% of bicyclist and 62.5% of motorcyclist presentations led to hospital admissions. Multivariate logistic regression analysis further showed that the crash location and road type affected hospitalisation for both groups. Although, crashes occurring in rural areas contributed 14% of the events overall, their adjusted risk of hospitalisation increased five times when compared to metropolitan locations. Crash preventive measures targeting rural areas should be considered to further improve the safety of bicyclists and motorcyclists.


Clinical applications of arterial stiffness; definitions and reference values.
Arterial stiffening is the most important cause of increasing systolic and pulse pressure, and for decreasing diastolic pressure beyond 40 years of age. Stiffening affects predominantly the aorta and proximal elastic arteries, and to a lesser degree the peripheral muscular arteries. While conceptually a Windkessel model is the simplest way to visualize the cushioning function of arteries, this is not useful clinically under changing conditions when effects of wave reflection become prominent. Many measures have been applied to quantify stiffness, but all are approximations only, on account of the nonhomogeneous structure of the arterial wall, its variability in different locations, at different levels of distending pressure, and with changes in smooth muscle tone. This article summarizes the methods and indices used to estimate arterial stiffness, and provides values from a survey of the literature, followed by recommendations of an international group of workers in the field who attended the First Consensus Conference on Arterial Stiffness, which was held in Paris during 2000, under the chairmanship of M.E. Safar and E.D. Frohlich.


Protein kinase C-delta is involved in induction of NOX1 gene expression by aldosterone in rat vascular smooth muscle cells.
In this study, we focused on the relationship between aldosterone and NOX1 expression in vascular smooth muscle cells (VSMCs). For the first time, with the use of specific inhibitors of protein kinase C (PKC), we report that PKCdelta mediates upregulation of NOX1 induced by 10 nM aldosterone in cultured VSMCs. Participation of PKC in the mediation of NOX1 regulation was further confirmed by the effect of diacylglycerol, a PKC agonist, on the NOX1 RNA in A7r5 cells with Northern blot analysis. To establish cause and effect, we next silenced the PKCdelta gene partly by RNA interference and found knockdown of PKCdelta gene attenuated aldosterone-induced NOX1 expression, generation of superoxide, as well as protein synthesis in VSMCs. Taken together, these data indicated PKCdelta might mediate aldosterone-dependent NOX1 upregulation in VSMCs. In addition, we showed that the cascade from aldosterone to PKCdelta activation had the participation of the mineralocorticoid receptor.


[Buerger's disease. Clinical and therapeutic review. Apropos of a case].
In reference to a typical case of Buerger's disease, the clinical, arteriographic, histological signs as well as diagnostic criteria are restated. The pathogenesis is still unknown; a genetic predisposition and an auto-immune process could be involved. The treatment, presently symptomatic, cannot arrest the evolution which is often mutilating, when the patient continues to smoke. An early diagnosis is possible in the presence of superficial phlebitis or Raynaud's phenomenon.


Looking for the Ideal Probiotic Healing Regime.
Wound healing is a multi-factorial response to tissue injury, aiming to restore tissue continuity. Numerous recent experimental and clinical studies clearly indicate that probiotics are applied topically to promote the wound-healing process. However, the precise mechanism by which they contribute to healing is not yet clear. Each strain appears to exert a distinctive, even multi-factorial action on different phases of the healing process. Given that a multi-probiotic formula exerts better results than a single strain, the pharmaceutical industry has embarked on a race for the production of a formulation containing a combination of probiotics capable of playing a role in all the phases of the healing process. Hence, the object of this review is to describe what is known to date of the distinctive mechanisms of each of the most studied probiotic strains in order to further facilitate research toward the development of combinations of strains and doses, covering the whole spectrum of healing. Eleven probiotic species have been analyzed, the only criterion of inclusion being a minimum of two published research articles.


Insoluble aggregates and protease-resistant conformers of prion protein in uninfected human brains.
Aggregated prion protein (PrPSc), which is detergent-insoluble and partially proteinase K (PK)-resistant, constitutes the major component of infectious prions that cause a group of transmissible spongiform encephalopathies in animals and humans. PrPSc derives from a detergent-soluble and PK-sensitive cellular prion protein (PrPC) through an alpha-helix to beta-sheet transition. This transition confers on the PrPSc molecule unique physicochemical and biological properties, including insolubility in nondenaturing detergents, an enhanced tendency to form aggregates, resistance to PK digestion, and infectivity, which together are regarded as the basis for distinguishing PrPSc from PrPC. Here we demonstrate, using sedimentation and size exclusion chromatography, that small amounts of detergent-insoluble PrP aggregates are present in uninfected human brains. Moreover, PK-resistant PrP core fragments are detectable following PK treatment. This is the first study that provides experimental evidence supporting the hypothesis that there might be silent prions lying dormant in normal human brains.


A systematic Analysis of Evaluation Methods for Inpatient Children and Adolescents Rehabilitation Programs.
Increasing chronic diseases in children and adolescents such as obesity, asthma bronchiale, or mental disorders, result in a growing importance of rehabilitation programs and subsequent program evaluations. We conducted a systematic literature search in 5 databases and provided an overview of methods for evaluating inpatient child and adolescent rehabilitation programs. This included an identification of generic and indication-specific outcomes and standardized measuring instruments. Eighteen studies covering 8 indication groups were considered for the qualitative analysis according to the pre-defined inclusion and exclusion criteria. Overall, the 18 studies included eight generic outcomes: (1) quality of life, (2) psychological well-being, (3) perceived changes, (4) functionality, (5) rehabilitation satisfaction, (6) self-management/-efficacy, (7) days of absence and (8) self-esteem. Eighteen standardized measuring instruments and 6 self-developed questionnaires were used to assess the 8 generic outcomes. Besides, indication-specific outcomes for 5 indications namely traumatic brain injury, obesity, diabetes, eating disorders and neurodermatitis, were reported. Overall, the present systematic review summarizes essential content (8 generic outcomes) and methods for evaluations of children and adolescent rehabilitation programs; however, before choosing a generic measuring tool during an evaluation, the psychometric validity of the instruments should be evaluated. In addition to the generic perspective, the evaluation of disease-specific outcomes is recommended.


Influence of HLA DRB1 alleles in the susceptibility of rheumatoid arthritis and the regulation of antibodies against citrullinated proteins and rheumatoid factor.
The purpose of this study was to investigate the association between HLA-DRB1 alleles with susceptibility to rheumatoid arthritis (RA) and production of antibodies against citrullinated proteins (ACPA) and rheumatoid factor (RF). We studied 408 patients (235 with RA, 173 non-RA) and 269 controls. ACPA, RF and HLA-DR typing were determined. We found an increased frequency of HLA DRB1 alleles with the shared epitope (SE) in ACPA-positive RA. Inversely, HLA DRB1 alleles encoding DERAA sequences were more frequent in controls than in ACPA-positive RA, and a similar trend was found for HLA DR3. However, these results could not be confirmed after stratification for the presence of the SE, probably due to the relatively low number of patients. These data may suggest that the presence of these alleles may confer a protective role for ACPA-positive RA. In RA patients we observed association between SE alleles and ACPA titers in a dose-dependent effect. The presence of HLA DR3 or DERAA-encoding alleles was associated with markedly reduced ACPA levels. No association between RF titers and HLA DR3 or DERAA-encoding alleles was found. HLA DRB1 alleles with the SE are associated with production of ACPA. DERAA-encoding HLA-DR alleles and HLA DR3 may be protective for ACPA-positive RA.


Multiple e-Pharmacophore Modeling Combined with High-Throughput Virtual Screening and Docking to Identify Potential Inhibitors of β-Secretase(BACE1).
β-Secretase (BACE1) is an aspartate protease involved in the production of amyloid-β a major peptide responsible for the pathogenesis of Alzheimer's disease. Given its role in the formation of amyloids leading to Alzheimer's disease, it has been a major therapeutic target for intervention and has been a challenge in the past and the progress has been very slow. More than hundred crystal structures with inhibitors are available in the protein data bank. Many strategies for drug design have been employed in the design of numerous diverse ligands for this target and many have failed due to undesirable drug properties primarily the inability to cross the blood-brain barrier. In the present work we attempted to consider multiple crystal structures with bound inhibitors showing affinity in the range of 2-210 nM efficacy and optimize the pharmacophoric requirement based on the energy involved in binding termed as e-pharmacophore mapping. A high throughput screening combined with molecular docking, ADMET predictions, logP values and in vitro assay led to the identification of 7 potential compounds showing inhibition at 10µM which could be further developed as novel inhibitors for β-secretase.


New approach toward nanosized ferrous ferric oxide and Fe(3)O(4)-doped titanium dioxide photocatalysts.
In this article, ferrous ferric oxide and Fe(3)O(4)-doped titanium dioxide have been synthesized by a low-temperature sol-gel process. In particular, we studied the structural characteristics of newly synthesized photosensitive catalysts and their photocatalytic abilities under UV and visible light irradiation. The elemental composition, surface area, crystallinity, and morphology of synthesized catalysts were characterized by Fourier transformation infrared spectrophotometry, X-ray photoelectron spectroscopy, X-ray diffraction, and transmission electron microscopy with selected area electron diffraction and energy-dispersive X-ray spectroscopy. The UV-vis transmission, zeta-potential, and cytotoxicity of the synthesized catalysts are also discussed. It is anticipated that the synthesized ferrous ferric oxide arrays a prospective photocatalyst substitute for titanium dioxide.


Urine creatinine in treatment-naïve HIV subjects in eastern Nigeria.
Human immunodeficiency virus (HIV) infection is a global healthcare problem. Some diseases and physiological states may be altered in HIV-infected individuals. Our objective was to evaluate urine creatinine and factors that influence urine creatinine in treatment-naïve HIV subjects in Nigeria. This was a cross-sectional study involving treatment-naïve HIV subjects in a tertiary hospital in Nigeria. Creatinine in spot and 24-hour urine samples and other relevant investigations were performed. Low urine creatinine or dilute urine was defined as 24-hour urine creatinine (24HUCr) <300mg, normal urine creatinine as 24HUCr 300-3000mg and high urine creatinine or concentrated urine as 24HUCr>3000mg.Theassociation of low urine creatinine and high urine creatinine with potential risk factors was determined. The mean spot urine creatinine (SUCr) of the treatment-naïve HIV subjects was 137.21± 98.47(mg/dl), minimum value 13.3mg/dl, maximum value 533.3mg/dl and range of values 520.0mg/dl. The mean 24HUCr was 1507±781mg, minimum value 206mg, maximum value 4849mg and range of values 4643mg. Twenty four-hour urine creatinine<300mg was observed in 2(0.5%) subjects, normal 24HUCr 300-3000mgin 349(93.1%) subjects and 24HUCr>3000mg in 24(6.4%) subjects. There was significant association between 24HUCr and serum low density lipoprotein cholesterol (LDL),serum high density lipoprotein cholesterol (HDL). There was high correlation between 24HUCr>3000mg and 24-hour urine osmolality (24HUOsm) (r=0.95), body mass index (BMI) (r=0.74), CD4 cells count (r=-0.71), serum HDL (r=-0.73). The prevalence of dilute urine and concentrated urine was low. Twenty-four hour urine osmolality. BMI, CD4 cells count and HDL were strong correlates of high urine creatinine. Lipid abnormalities were common in treatment-naïve HIV subjects with high urine creatinine. There is need for clinicians to routinely conduct urine creatinine and further search for abnormalities of serum lipids, weight changes, depressed immunity and anemia in HIV subjects with dilute or concentrated urine in the early stages of the infection.


[Comparative evaluation of acid-fast staining for the detection of Mycobacterium fortuitum--clinical performance of fluorescent and Ziehl-Neelsen staining].
Fluorescent staining is of paramount importance, not only for confirming the presence of mycobacteria in a given specimen but also for providing an estimated growth quantification. In this study, for rapidly growing Mycobacterium fortuitum, we evaluated the effectiveness of a rapid fluorescent staining method employing auramine-rhodamine (AR) fluorescent stain and acridine-orange (AO) fluorescent stain compared to that of the standard Ziehl-Neelsen (ZN) stain currently in use in our laboratory. We evaluated the acid-fast nature of M. fortuitum strain ATCC6841 and 42 clinical isolates from each patient diagnosed at NHO Kinki-chuo Chest Medical Center. These isolates were preliminarily identified as M. fortuitum using DNA-DNA hybridization (DDH Mycobacteria; Kyokuto Pharmaceutical, Tokyo, Japan). These isolates were further identified by comparative sequence analysis of the ITS regions and the partial 16S rRNA gene. A total of 26 M. fortuitum strains (61.9%) demonstrated the lack of an acid-fast nature by AR staining, and slightly fewer demonstrated the same by AO staining. Sequence analysis of these 42 clinical isolates led to the identification of 35 M. fortuitum subsp. acetamidolyticum isolates (83.3%) and 7 closely M. fortuitum isolates. This work reported the loss of the acid-fast nature of specific M. fortuitum strains. It is likely that both the specific cell envelope of M. fortuitum and the staining mechanics could have been responsible for the loss of the acid-fast nature since the 2 different fluorescent stains yielded the same results. M. fortuitum is a mycobacterium species that does not stain with the commonly used fluorescence microscopy technique. Therefore, we suggested the use of an identification scheme for these organisms that employs ZN staining and the study of cultural characteristics (growth rate, temperature, and pigment production).


A neurochemical hypothesis for the origin of hominids.
It has always been difficult to account for the evolution of certain human characters such as language, empathy, and altruism via individual reproductive success. However, the striatum, a subcortical region originally thought to be exclusively motor, is now known to contribute to social behaviors and "personality styles" that may link such complexities with natural selection. We here report that the human striatum exhibits a unique neurochemical profile that differs dramatically from those of other primates. The human signature of elevated striatal dopamine, serotonin, and neuropeptide Y, coupled with lowered acetylcholine, systematically favors externally driven behavior and greatly amplifies sensitivity to social cues that promote social conformity, empathy, and altruism. We propose that selection induced an initial form of this profile in early hominids, which increased their affiliative behavior, and that this shift either preceded or accompanied the adoption of bipedality and elimination of the sectorial canine. We further hypothesize that these changes were critical for increased individual fitness and promoted the adoption of social monogamy, which progressively increased cooperation as well as a dependence on tradition-based cultural transmission. These eventually facilitated the acquisition of language by elevating the reproductive advantage afforded those most sensitive to social cues.


Two-component system CbrA/CbrB controls alginate production in Azotobacter vinelandii.
Azotobacter vinelandii, belonging to the Pseudomonadaceae family, is a free-living bacterium that has been considered to be a good source for the production of bacterial polymers such as alginate. In A. vinelandii the synthesis of this polymer is regulated by the Gac/Rsm post-transcriptional regulatory system, in which the RsmA protein binds to the mRNA of the biosynthetic algD gene, inhibiting translation. In several Pseudomonas spp. the two-component system CbrA/CbrB has been described to control a variety of metabolic and behavioural traits needed for adaptation to changing environmental conditions. In this work, we show that the A. vinelandii CbrA/CbrB two-component system negatively affects alginate synthesis, a function that has not been described in Pseudomonas aeruginosa or any other Pseudomonas species. CbrA/CbrB was found to control the expression of some alginate biosynthetic genes, mainly algD translation. In agreement with this result, the CbrA/CbrB system was necessary for optimal rsmA expression levels. CbrA/CbrB was also required for maximum accumulation of the sigma factor RpoS. This last effect could explain the positive effect of CbrA/CbrB on rsmA expression, as we also showed that one of the promoters driving rsmA transcription was RpoS-dependent. However, although inactivation of rpoS increased alginate production by almost 100 %, a cbrA mutation increased the synthesis of this polymer by up to 500 %, implying the existence of additional CbrA/CbrB regulatory pathways for the control of alginate production. The control exerted by CbrA/CbrB on the expression of the RsmA protein indicates the central role of this system in regulating carbon metabolism in A. vinelandii.


Pupil-linked arousal is driven by decision uncertainty and alters serial choice bias.
While judging their sensory environments, decision-makers seem to use the uncertainty about their choices to guide adjustments of their subsequent behaviour. One possible source of these behavioural adjustments is arousal: decision uncertainty might drive the brain's arousal systems, which control global brain state and might thereby shape subsequent decision-making. Here, we measure pupil diameter, a proxy for central arousal state, in human observers performing a perceptual choice task of varying difficulty. Pupil dilation, after choice but before external feedback, reflects three hallmark signatures of decision uncertainty derived from a computational model. This increase in pupil-linked arousal boosts observers' tendency to alternate their choice on the subsequent trial. We conclude that decision uncertainty drives rapid changes in pupil-linked arousal state, which shape the serial correlation structure of ongoing choice behaviour.


[The sinus VK experimental apparatus for assisted circulation].
Presented in the paper is the unit for cardiosynchronized extra- and intracorporal control in the course of the intensive therapy of patients with acute cardiac failure. It provides several methods of the assisted circulation and can be used in clinical cardiology, surgery and reanimation departments. Main specifications, the structure and the principle of functioning are given. The prototype testing has shown that "CHHyc B K" can be used to advantage in clinical practice and should be introduced for the full - scale production.


Treatment of vitiligo with the 308-nm excimer laser: a pilot study.
Present vitiligo therapies require many months of treatment and often result in disappointing outcomes. Common therapeutic options include phototherapy with psoralens plus ultraviolet A (UVA) radiation and broadband or narrowband UVB radiation phototherapy. Some of these modalities require regular phototherapy sessions several times a week for up to a year to achieve a therapeutic response. Targeted phototherapy with single-wavelength laser light is a treatment alternative that may prove to be a time-efficient and effective therapeutic option for the management of vitiligo. This intervention study was designed as a before and after trial with a single arm. Twenty-nine patches of vitiligo from 18 patients (6 males and 12 females) were treated at the start of the study. Vitiligo patches were treated by using a 308-nm xenon-chloride excimer laser. Lesions were treated 3 times a week for a maximum of 12 treatments. Treatment was withheld if sunburn was observed and held until resolution. All patients had untreated vitiligo patches that served as control sites. Twenty-three vitiligo patches from 12 patients received at least 6 treatments and resulted in some repigmentation in 57% of the treated patches. Eleven vitiligo patches from 6 patients received all 12 treatments and resulted in some repigmentation in 82% of the treated patches. Untreated control patches remained unchanged. This degree of repigmentation in a period of 2 to 4 weeks is much higher than that achieved with any other present vitiligo therapy. The xenon-chloride excimer laser may represent a new treatment modality for the management of stable vitiligo.


The monoblock appliance.
While the use of this appliance would appear to require relatively litlle operative time, it must be clearly understood that the function of this appliance is not as predictable as that of other types of removable devices. Under such circumstances, it is of great importance to ensure that a correct diagnosis and evaluation of the original condition has been made relative to the listed criteria; all factors must be considered and no diagnosis should be made purely and simply on the basis of an obvious increase in overjet and a class II malocclusion. In addition, the personality and nature of the patient is of paramount importance, since it is his cooperation and interest that in the end will lead to successful therapy.


Regioselective Synthesis and Characterization of Tris- and Tetra-Prato Adducts of M3N@C80 (M = Y, Gd).
The tris- and tetra-adducts of M3N@Ih-C80 metallofullerenes were synthesized and characterized for the first time. The 1,3-dipolar cycloaddition (Prato reaction) of Y3N@Ih-C80 and Gd3N@Ih-C80 with an excess of N-ethylglycine and formaldehyde provided tris- and tetra-fulleropyrrolidine adducts in a regioselective manner. Purification by HPLC and analyses of the isolated peaks by NMR, MS, and vis-NIR spectra revealed that the major products were four tris- and one tetra-isomers for both Y3N@Ih-C80 and Gd3N@Ih-C80. Considering the large number of possible isomers (e.g., at least 1140 isomers for the tris-adduct), the limited number of isomers obtained indicated that the reactions proceeded with high regioselectivity. NMR analyses of the Y3N@Ih-C80 adducts found that the tris-adducts were all-[6,6]- or [6,6][6,6][5,6]-isomers and that some showed mutual isomerization or remained intact at room temperature. The tetra-adduct obtained as a major product was all-[6,6] and stable. For the structural elucidation of Gd3N@Ih-C80 tris- and tetra-adducts, density functional theory (DFT) calculations were performed to estimate the relative stabilities of tris- and tetra-adducts formed upon Prato functionalization of the most pyramidalized regions of the fullerene structure. The most stable structures corresponded to additions on the most pyramidalized (i.e., strained) bonds. Taking together the experimental vis-NIR spectra, NMR assignments, and the computed relative DFT stabilities of the potential tris- and tetra-adducts, the structures of the isolated adducts were elucidated. Electron resonance (ESR) measurements measurements of pristine, bis-, and tris-adducts of Gd3N@C80 suggested that the rotation of the endohedral metal cluster slowed upon increase of the addition numbers to C80 cage, which is favored for accommodating the Gd atoms of the relatively large Gd3N cluster inner space at the sp3 addition sites. This is presumably related to the high regioselectivity in the Prato addition reaction driven by the strain release of the Gd3N@C80 fullerene structure.


Time-resolved systems analysis of the induction of high photosynthetic capacity in Arabidopsis during acclimation to high light.
Induction of high photosynthetic capacity is a key acclimation response to high light (HL) for many herbaceous dicot plants; however, the signaling pathways that control this response remain largely unknown. Here, a systems biology approach was utilized to characterize the induction of high photosynthetic capacity in strongly and weakly acclimating Arabidopsis thaliana accessions. Plants were grown for 5 wk in a low light (LL) regime, and time-resolved photosynthetic physiological, metabolomic, and transcriptomic responses were measured during subsequent exposure to HL. The induction of high nitrogen (N) assimilation rates early in the HL shift was strongly predictive of the induction of photosynthetic capacity later in the HL shift. Accelerated N assimilation rates depended on the mobilization of existing organic acid (OA) reserves and increased de novo OA synthesis during the induction of high photosynthetic capacity. Enhanced sucrose biosynthesis capacity increased in tandem with the induction of high photosynthetic capacity, and increased starch biosynthetic capacity was balanced by increased starch catabolism. This systems analysis supports a model in which the efficient induction of N assimilation early in the HL shift begins the cascade of events necessary for the induction of high photosynthetic capacity acclimation in HL.


[Discrimination of Red Tide algae by fluorescence spectra and principle component analysis].
Fluorescence discrimination technology for 11 species of the Red Tide algae at genus level was constructed by principle component analysis and non-negative least squares. Rayleigh and Raman scattering peaks of 3D fluorescence spectra were eliminated by Delaunay triangulation method. According to the results of Fisher linear discrimination, the first principle component score and the second component score of 3D fluorescence spectra were chosen as discriminant feature and the feature base was established. The 11 algae species were tested, and more than 85% samples were accurately determinated, especially for Prorocentrum donghaiense, Skeletonema costatum, Gymnodinium sp., which have frequently brought Red tide in the East China Sea. More than 95% samples were right discriminated. The results showed that the genus discriminant feature of 3D fluorescence spectra of Red Tide algae given by principle component analysis could work well.


Pioneer neurog1 expressing cells ingress into the otic epithelium and instruct neuronal specification.
Neural patterning involves regionalised cell specification. Recent studies indicate that cell dynamics play instrumental roles in neural pattern refinement and progression, but the impact of cell behaviour and morphogenesis on neural specification is not understood. Here we combine 4D analysis of cell behaviours with dynamic quantification of proneural expression to uncover the construction of the zebrafish otic neurogenic domain. We identify pioneer cells expressing neurog1 outside the otic epithelium that migrate and ingress into the epithelialising placode to become the first otic neuronal progenitors. Subsequently, neighbouring cells express neurog1 inside the placode, and apical symmetric divisions amplify the specified pool. Interestingly, pioneer cells delaminate shortly after ingression. Ablation experiments reveal that pioneer cells promote neurog1 expression in other otic cells. Finally, ingression relies on the epithelialisation timing controlled by FGF activity. We propose a novel view for otic neurogenesis integrating cell dynamics whereby ingression of pioneer cells instructs neuronal specification.


Meta-analysis of drainage versus no drainage after laparoscopic cholecystectomy.
Routine drainage after laparoscopic cholecystectomy is still controversial. This meta-analysis was performed to assess the role of drains in reducing complications in laparoscopic cholecystectomy. An electronic search of Medline, Science Citation Index Expanded, Scopus, and the Cochrane Library database from January 1990 to June 2013 was performed to identify randomized clinical trials that compare prophylactic drainage with no drainage in laparoscopic cholecystectomy. The odds ratio for qualitative variables and standardized mean difference for continuous variables were calculated. Twelve randomized controlled trials were included in the meta-analysis, involving 1939 patients randomized to a drain (960) versus no drain (979). The morbidity rate was lower in the no drain group (odds ratio, 1.97; 95% confidence interval, 1.26 to 3.10; P = .003). The wound infection rate was lower in the no drain group (odds ratio, 2.35; 95% confidence interval, 1.22 to 4.51; P = .01). Abdominal pain 24 hours after surgery was less severe in the no drain group (standardized mean difference, 2.30; 95% confidence interval, 1.27 to 3.34; P < .0001). No significant difference was present with respect to the presence and quantity of subhepatic fluid collection, shoulder tip pain, parenteral ketorolac consumption, nausea, vomiting, and hospital stay. This study was unable to prove that drains were useful in reducing complications in laparoscopic cholecystectomy.


Pulmonary vein isolation by means of a novel cryoballoon technology in paroxysmal atrial fibrillation patients: 1-year outcome from a large Italian multicenter clinical registry.
Recently, a new cryoballoon (CB) technology (POLARx; Boston Scientific) has come onto the market. Preliminary data have shown that its acute safety and efficacy are similar to those of the first-generation CB. The aim of this study was to assess the medium-term outcome of pulmonary vein isolation (PVI) with the POLARxTM CB in a large multicenter registry. We prospectively collected data on 125 consecutive patients with paroxysmal atrial fibrillation (AF) who underwent PVI by means of a novel CB system. Two cases of transient phrenic nerve palsy occurred, with full recovery in the 48h post procedure; no major procedure-related adverse events were reported. During the 90-day blanking period, 4 (3.2%) patients experienced an early recurrence. After the blanking period, over a mean follow-up of 411 ± 62 days, 19 patients (15.2%) suffered an AF/atrial tachycardia (AT) recurrence. The 1-year freedom from AF/AT recurrence was 86.4% (n = 17): 10 (8%) patients had an AF recurrence, 6 (4.8%) had an AT occurrence and 1 (0.8%) suffered both events. Patients with AF/AT recurrences had both a shorter deflation time and total deflation time. Moreover, CB ablations with measured TTI < 90 s and TTI < 60 s were more frequent in patients without AF/AT recurrence (88.5% and 77.4%, respectively) than in those who experienced at least one AF/AT recurrence (67.5% and 55.0%, p = .001 and p = .005, respectively). The novel POLARx cryo-balloon system is safe and effective for PV isolation, displaying a 1-year freedom from atrial arrhythmia recurrence of 86.4%, which is in line to that reported with AFA-Pro CB or RF ablation. Catheter Ablation of Arrhythmias with a High-Density Mapping System in Real-World Practice (CHARISMA). URL: http://clinicaltrials.gov/ Identifier: NCT03793998. Registration date: January 4, 2019.


Vortex fluidic entrapment of functional microalgal cells in a magnetic polymer matrix.
Composite materials based on superparamagnetic magnetite nanoparticles embedded in polyvinylpyrrolidone (PVP) are generated in a continuous flow vortex fluidic device (VFD). The same device is effective in entrapping microalgal cells within this material, such that the functional cells can be retrieved from aqueous dispersions using an external magnet.


Analysis of pH-dependent elements in proteins: geometry and properties of pairs of hydrogen-bonded carboxylic acid side-chains.
A rather frequent but so far little discussed observation is that pairs of carboxylic acid side-chains in proteins can share a proton in a hydrogen bond. In the present article, quantum chemical calculations of simple model systems for carboxyl-carboxylate interactions are compared with structural observations from proteins. A detailed structural analysis of the proteins deposited in the PDB revealed that, in a subset of proteins sharing less than 90% sequence identity, 19% (314) contain at least one pair of carboxylic acids with their side-chain oxygen atoms within hydrogen-bonding distance. As the distance between those interacting oxygen atoms is frequently very short ( approximately 2.55 A), many of these carboxylic acids are suggested to share a proton in a strong hydrogen bond. When situated in an appropriate structural environment (low dielectric constant), some might even form a low barrier hydrogen bond. The quantum chemical studies show that the most frequent geometric features of carboxyl-carboxylate pairs found in proteins, and no or symmetric ligation, are also the most stable arrangements at low dielectric constants, and they also suggest at medium and low pH a higher stability than for isosteric amide-carboxylate pairs. The presence of these pairs in 119 different enzymes found in the BRENDA database is set in relation to their properties and functions. This analysis shows that pH optima of enzymes with carboxyl-carboxylate pairs are shifted to lower than average values, whereas temperature optima seem to be increased. The described structural principles can be used as guidelines for rational protein design (e.g., in order to improve pH or temperature stability).


Case-control study of genotypes in multiple chemical sensitivity: CYP2D6, NAT1, NAT2, PON1, PON2 and MTHFR.
Impaired metabolism of toxic chemicals is a postulated mechanism underlying multiple chemical sensitivity (MCS). Because genetic variation alters the rate of chemical metabolism, this study was designed to determine if MCS cases differed from controls for genetic polymorphisms in drug-metabolizing enzymes. Female Caucasian participants (203 cases and 162 controls) were drawn from a larger case-control study based on a reproducible and validated case definition. Common polymorphisms for CYP2D6, NAT1, NAT2, PON1, and PON2 were genotyped. Comparing cases and controls, significant differences were found in genotype distributions for CYP2D6 (P = 0.02) and NAT2 (P = 0.03). Compared with the referent homozygous inactive (CYP2D6) or slow (NAT2) metabolizers, the odds for being CYP2D6 homozygous active (OR = 3.36, P = 0.01) and NAT2 rapid (OR = 4.14, P = 0.01) were significantly higher in cases than controls. The odds for being heterozygous for PON1-55 (OR = 2.05, P = 0.04) and PON1-192 (OR = 1.57, P = 0.04) were also significantly higher in cases. A genetic predisposition for MCS may involve altered biotransformation of environmental chemicals. The CYP2D6 enzyme activates and inactivates toxins; the NAT2 enzyme bioactivates arylamines to protein-binding metabolites. A gene-gene interaction between CYP2D6 and NAT2 suggested that rapid metabolism for both enzymes may confer substantially elevated risk (OR = 18.7, P = 0.002). Our finding parallels others' observation of a link between PON1 heterozygosity and neurological symptoms in Gulf War syndrome. This first demonstration of genetic variation in drug-metabolizing enzymes in association with MCS requires replication. However, it suggests new research directions on genetically variable toxin pathways that might be important in MCS.


Narcissism and Attachment: The Importance of Early Parenting.
Research often shows that narcissism and attachment style are related. The present research examines the question of whether early parenting style contributes to adult attachment style, beyond the relation between narcissism and attachment. Parents reported on their parenting style at age 3, and offspring were assessed for attachment style and narcissism at age 23. The results indicated that early parenting style was not directly related to narcissism, but along with narcissism added to the prediction of attachment style. In combination with vulnerable narcissism, authoritarian parenting was positively related to secure attachment, and negatively related to preoccupied attachment, whereas permissive and responsive parenting were negatively related to secure attachment and positively related to preoccupied attachment.


An improved algorithm to reduce noise in high-order thermal ghost imaging.
A modified Nth-order correlation function is derived that can effectively remove the noise background encountered in high-order thermal light ghost imaging (GI). Based on this, the quality of the reconstructed images in an Nth-order lensless GI setup has been greatly enhanced compared to former high-order schemes for the same sampling number. In addition, the dependence of the visibility and signal-to-noise ratio for different high-order images on the sampling number has been measured and compared.


SARS-CoV-2-specific Cell-mediated Immunity in Kidney Transplant Recipients Recovered From COVID-19.
The magnitude and kinetics of severe acute respiratory syndrome coronavirus 2-specific cell-mediated immunity (SARS-CoV-2-CMI) in kidney transplant (KT) recipients remain largely unknown. We enumerated SARS-CoV-2-specific interferon-γ-producing CD69+ CD4+ and CD8+ T cells at months 4 and 6 from the diagnosis of coronavirus disease 2019 (COVID-19) in 21 KT recipients by intracellular cytokine staining. Overlapping peptides encompassing the SARS-CoV-2 spike (S) glycoprotein N-terminal 1- to 643-amino acid sequence and the membrane protein were used as stimulus. SARS-CoV-2 IgG antibodies targeting the S1 protein were assessed by ELISA at month 6. Detectable (≥0.1%) SARS-CoV-2-specific CD4+ T-cell response was found in 57.1% and 47.4% of patients at months 4 and 6. Corresponding rates for CD8+ T cells were 19.0% and 42.1%, respectively. Absolute SARS-CoV-2-specific T-cell counts increased from month 4 to month 6 in CD8+ (P = 0.086) but not CD4+ subsets (P = 0.349). Four of 10 patients with any detectable response at month 4 had lost SARS-CoV-2-CMI by month 6, whereas 5 of 9 patients mounted SARS-CoV-2-CMI within this period. All but 2 patients (89.5%) tested positive for SARS-CoV-2 IgG. Patients lacking detectable SARS-CoV-2-specific CD4+ response by month 6 were more likely to be under tacrolimus (100.0% versus 66.7%; P = 0.087) and to have received tocilizumab for the previous COVID-19 episode (40.0% versus 0.0%; P = 0.087). Although still exploratory and limited by small sample size, the present study suggests that a substantial proportion of KT recipients exhibited detectable SARS-CoV-2-CMI after 6 months from COVID-19 diagnosis.


The Organization and Function of the Phagophore-ER Membrane Contact Sites.
Macroautophagy is characterized by the de novo formation of double-membrane vesicles termed autophagosomes. The precursor structure of autophagosomes is a membrane cistern called phagophore, which elongates through a massive acquisition of lipids until closure. The phagophore establishes membrane-contact sites (MCSs) with the endoplasmic reticulum (ER), where conserved ATG proteins belonging to the ATG9 lipid scramblase, ATG2 lipid transfer and Atg18/WIPI4 β-propeller families concentrate. Several recent in vivo and in vitro studies have uncovered the relevance of these proteins and MCSs in the lipid supply required for autophagosome formation. Although important conceptual advances have been reached, the functional interrelationship between ATG9, ATG2 and Atg18/WIPI4 proteins at the phagophore-ER MCSs and their role in the phagophore expansion are not completely understood. In this review, we describe the current knowledge about the structure, interactions, localizations, and molecular functions of these proteins, with a particular emphasis on the yeast Saccharomyces cerevisiae and mammalian systems.


Development of interactive patient-based multimedia computer programs in veterinary orthopedic radiology.
Three computerized multimedia programs on large and small animal veterinary orthopedic radiology were developed and implemented for the radiology curriculum as an alternative to traditional film-based laboratory learning. Programs utilized "hot words" (colored text words that displayed an overlaid image label that highlighted lesions) and interactive quizzes which responded appropriately to selected answers. "Hot words" helped students develop confidence in accurate lesion detection and the interactive quizzes transformed learning from a passive to an active process. Multiple examples were provided for reinforcement and concepts were incorporated from other clinical disciplines for curriculum integration. Programs were written using a presentation software program, Toolbook for DOS based platform, and contained radiographic images made by laser-scanning digitization. Multiple students could simultaneously access the programs through a network server. These pilot programs were implemented successfully and computerized multimedia presentation proved to be well suited to teaching radiology. Development of the programs required attention to a number of hardware, software, time and cost factors.


Cardiac Transplant Postoperative Management and Care.
Heart failure impacts a multitude of individuals each year. Treatment is based on the progression of the disease and severity of symptoms. Cardiac transplant is the gold standard treatment of advanced heart failure, although the availability of organs limits the number of transplants received each year. Postoperative care and monitoring for cardiac transplant is complex and requires specialized nurses and providers at transplant centers for successful outcomes. This article outlines cardiac transplant from preoperative care through transplant, as well as posttransplant monitoring and care including discharge. Special attention is focused on management in the intensive care unit setting and potential complications that can occur in the immediate postoperative period. Interventions for potential complications are also highlighted.


Recent advances in the purification of the alpha 1-adrenoceptor.
In order to purify the alpha-adrenergic receptor labeled with [3H]POB, antibodies against the irreversible alpha-adrenergic antagonist, phenoxybenzamine (POB), have been raised in rabbits immunized with POB conjugated to immunoglobulin. The antigen was prepared by first reacting POB with the sulfhydryl group of cysteine, after which the POB complex was coupled to thyroglobulin in the presence of water soluble carbodimide. POB-specific antibodies were first detected 10 days after the first booster injection. These antibodies display a high degree of specificity. Slight cross-reactivity occurred only with compounds structurally related to POB, such as dibenamine and nitrogen mustards, while catecholamines and alpha-adrenergic compounds not structurally related to POB did not cross-react, even when present in 100,000-fold excess. Moreover, the antibodies are still capable of recognizing POB once bound to alpha-adrenergic receptor. When membranes, prelabeled with [3H]POB, are solubilized with digitonin and incubated with anti-POB antiserum, the radioactivity can be specifically precipitated upon addition of goat anti-rabbit gamma-globulin serum. Thus these antibodies may be a useful tool in the purification of the alpha-adrenergic receptor of rat liver covalently linked to POB.


Seasonality in the sudden infant death syndrome.
Distinctive epidemiological features of the sudden infant death syndrome (SIDS) are its age at death distribution and pronounced winter excess. Whether or not these effects are independent of the month of birth of the infant is uncertain. We used a loglinear model to separate the effects of age at death, month of death and month of birth amongst 6229 infants who died from SIDS in England and Wales during the period 1979-1983. The results suggest that month of birth and month of death independently influence the risk of the infant dying from SIDS, the risk related to month of death being much greater.


Microbial composition in bioaerosols of a high-throughput chicken-slaughtering facility.
The microbial composition of the air in various areas of a high-throughput chicken-slaughtering facility was investigated. Over a 4-mo period, 6 processing areas were sampled, and the influence of environmental factors was monitored. The highest counts of microorganisms were recorded in the initial stages of processing, comprising the receiving-killing and defeathering areas, whereas counts decreased toward the evisceration, air-chilling, packaging, and dispatch areas. Maximum microbial counts were as follows: coliforms, 4.9 x 10(3) cfu/m(3); Escherichia coli 3.4 x 10(3) cfu/m(3); Bacillus cereus, 5.0 x 10(4) cfu/m(3); Staphylococcus aureus, 1.6 x 10(4) cfu/m(3); Pseudomonas aeruginosa, 7.0 x 10(4) cfu/m(3); presumptive Salmonella spp., 1.5 x 10(4) cfu/m(3); Listeria monocytogenes, 1.6 x 10(4) cfu/m(3); and fungi, 1.4 x 10(4) cfu/m(3). Higher counts of airborne microorganisms found in the receiving-killing and defeathering areas indicate the importance of controlling microbial levels before processing to prevent the spread of organisms downstream. This should limit the risk of carrying over contaminants from areas known to generate high counts to areas where the final food product is exposed to air and surface contamination.


The motivational basis of concessions and compromise: archival and laboratory studies.
A content analysis system for measuring positive concessions (offering concessions) and negative concessions (rejecting offered concessions) was introduced and validated through an archival study of government-to-government documents from 4 crises, 2 of which escalated to war and 2 of which were peacefully resolved. In the archival documents, concession making was positively associated with affiliation motivation and negatively associated with power motivation. A 2nd, laboratory experimental study confirmed these relationships and demonstrated priming effects of motive imagery and concession making, in a received diplomatic letter, on participants' responses. Finally, the motive imagery and concessions scores in participants' responses were related in predicted ways to their policy choices.


Effect of insulin on basal and cholecystokinin-stimulated gallbladder motility in humans.
Acute hyperglycemia inhibits gallbladder contraction. In non-diabetic subjects this inhibitory effect may result from endogenous hyperinsulinemia. Therefore we investigated the effects of acute hyperglycemia and euglycemic hyperinsulinemia on basal and cholecystokinin-stimulated gallbladder motility. Gallbladder volume (ultrasonography) and duodenal bilirubin output were studied simultaneously in nine healthy volunteers (age 20-52 years) on 3 separate occasions in random order during: (a) saline infusion (control), (b) hyperglycemic hyperinsulinemic clamping (HG; plasma glucose at 15 mmol/l), and (c) euglycemic hyperinsulinemic clamping (HI; plasma insulin at 150 mU/l, glucose at 4-5 mmol/l). After a 2-h basal clamp period, cholecystokinin was infused intravenously for 60 min at 0.25 IDU x kg(-1) x h(-1), followed by another 60 min at 0.5 IDU x kg(-1) x h(-1). HI and HG significantly (p<0.05) reduced basal duodenal bilirubin output compared to control, while basal gallbladder volume did not change. At the low dose cholecystokinin, gallbladder emptying during HG (25+/-3%) and HI (39+/-4%) was significantly (p<0.01) reduced compared to control (61+/-4%). The inhibitory effect of HG was significantly (p<0.05) stronger compared to HI. Duodenal bilirubin output during the low dose cholecystokinin was significantly (p<0.05) reduced by HG, but not by HI. No inhibitory effect of HG and HI on gallbladder emptying and duodenal bilirubin output was observed with the high dose of cholecystokinin. In healthy subjects acute hyperglycemia and euglycemic hyperinsulinemia reduce basal duodenal bilirubin output and inhibit gallbladder emptying stimulated by low dose cholecystokinin. These results suggest that insulin is involved in the inhibitory effect of hyperglycemia on basal and cholecystokinin-stimulated gallbladder motility.


Breast and cervical cancer screening in Hispanic women: a literature review using the health belief model.
The aim of this study was to review published studies that examined factors influencing breast and cervical cancer screening behavior in Hispanic women, using the Health Belief Model (HBM). MEDLINE and PsycINFO databases and manual search were used to identify articles. Cancer screening barriers common among Hispanic women include fear of cancer, fatalistic views on cancer, linguistic barriers, and culturally based embarrassment. In addition, Hispanic women commonly feel less susceptible to cancer, which is an important reason for their lack of screening. Positive cues to undergo screening include physician recommendation, community outreach programs with the use of Hispanic lay health leaders, Spanish print material, and use of culturally specific media. Critical review of the literature using the theoretical framework of the Health Belief Model identified several culturally specific factors influencing cancer screening uptake and compliance among Hispanic women. Future interventions need to be culturally sensitive and competent.


Cannabinoid-1 (CB1) receptors regulate colonic propulsion by acting at motor neurons within the ascending motor pathways in mouse colon.
Cannabinoid-1 (CB(1)) receptors on myenteric neurons are involved in the regulation of intestinal motility. Our aim was to investigate CB(1) receptor involvement in ascending neurotransmission in mouse colon and to characterize the involved structures by functional and morphological means. Presence of the CB(1) receptor was investigated by RT-PCR, and immunohistochemistry was used for colabeling studies. Myenteric reflex responses were initiated by electrical stimulation (ES) at different distances, and junction potentials (JP) were recorded from circular smooth muscle cells by intracellular recording in an unpartitioned and a partitioned recording chamber. In vivo colonic propulsion was tested in wild-type and CB(1)(-/-) mice. Immunostaining with the cytoskeletal marker peripherin showed CB(1) immunoreactivity both on Dogiel type I and type II neurons. Further neurochemical characterization revealed CB(1) on choline acetyltransferase-, calretinin-, and 5-HT-immunopositive myenteric neurons, but nitrergic neurons appeared immunonegative for CB(1) immunostaining. Solitary spindle-shaped CB(1)-immunoreactive cells in between smooth muscle cells lacked specific markers for interstitial cells of Cajal or glial cells. ES elicited neuronally mediated excitatory JP (EJP) and inhibitory JP. Gradual increases in distance resulted in a wave-like EJP with EJP amplitudes being maximal at the location of stimulating electrode 6 and a maximal EJP projection distance of approximately 18 mm. The CB(1) receptor agonist WIN 55,212-2 reduced the amplitude of EJP and was responsible for shortening the oral spreading of the excitatory impulse. In a partitioned chamber, WIN 55,212-2 reduced EJP at the separated oral sites, proving that CB(1) activation inhibits interneuron-mediated neurotransmission. These effects were absent in the presence of the CB(1) antagonist SR141716A, which, when given alone, had no effect. WIN 55,212-2 inhibited colonic propulsion in wild-type mice but not in SR141716A-pretreated wild-type or CB(1)(-/-) mice. Activation of the CB(1) receptor modulates excitatory cholinergic neurotransmission in mouse colon by reducing amplitude and spatial spreading of the ascending electrophysiological impulses. This effect on electrophysiological spreading involves CB(1)-mediated effects on motor neurons and ascending interneurons and is likely to underlie the here reported in vivo reduction in colonic propulsion.


Coordinate regulation of DNA methylation and H3K27me3 in mouse embryonic stem cells.
Chromatin is separated into functional domains distinguished by combinatorial patterns of post-translational histone modifications and DNA methylation. Recent studies examining multiple histone modifications have found numerous chromatin states with distinct profiles of chromatin marks and functional enrichments. There are data showing coordinate regulation between DNAme and H3K27me3, which are both involved in the establishment and maintenance of epigenetic gene silencing, but the data are conflicting. Multiple studies have presented evidence to support the theory that PRC2 and DNAme cooperate to achieve silencing, or alternatively that H3K27me3 and DNAme act antagonistically. Here we examine the effect loss of either PRC2 or DNA methyltransferase activity has on the placement of the reciprocal mark in mouse ES cells. We find that DNAme is acting globally to antagonize the placement of H3K27me3, in accordance with recently published results. At least 471,011 domains in the mouse genome acquire H3K27me3 when DNAme is diminished. Of these 466,563 have been shown to be fully methylated in wildtype ES cells, indicating the effects of DNAme on H3K27me3 are direct. In a reciprocal experiment, we examine the effect loss of PRC2 has on the placement of DNAme. In contrast to the global antagonism DNAme has on the placement of H3K27me3, loss of H3K27me3 has a modest effect on DNAme, with only 4% of genes undergoing changes in DNAme, including 861 showing increases and 552 showing losses of overall DNAme. We anticipate that integrating genomic datasets where the effect of loss of a particular epigenetic mark has on the placement of other marks will help elucidate the rules governing epigenetic regulation and what role coordinate regulation of epigenetic marks plays in development and disease.