Sub-Inhibitory Concentration of Piperacillin-Tazobactam May be Related to Virulence Properties of Filamentous Escherichia coli.
Sub-inhibitory concentrations of antibiotics are always generated as a consequence of antimicrobial therapy and the effects of such residual products in bacterial morphology are well documented, especially the filamentation generated by beta-lactams. The aim of this study was to investigate some morphological and pathological aspects (virulence factors) of Escherichia coli cultivated under half-minimum inhibitory concentration (1.0 µg/mL) of piperacillin-tazobactam (PTZ sub-MIC). PTZ sub-MIC promoted noticeable changes in the bacterial cells which reach the peak of morphological alterations (filamentation) and complexity at 16 h of antimicrobial exposure. Thereafter the filamentous cells and a control one, not treated with PTZ, were comparatively tested for growth curve; biochemical profile; oxidative stress tolerance; biofilm production and cell hydrophobicity; motility and pathogenicity in vivo. PTZ sub-MIC attenuated the E. coli growth rate, but without changes in carbohydrate fermentation or in traditional biochemical tests. Overall, the treatment of E. coli with sub-MIC of PTZ generated filamentous forms which were accompanied by the inhibition of virulence factors such as the oxidative stress response, biofilm formation, cell surface hydrophobicity, and motility. These results are consistent with the reduced pathogenicity observed for the filamentous E. coli in the murine model of intra-abdominal infection. In other words, the treatment of E. coli with sub-MIC of PTZ suggests a decrease in their virulence.


Mice deficient in Mac-1 (CD11b/CD18) are less susceptible to cerebral ischemia/reperfusion injury.
Macrophage-1 antigen (Mac-1) (CD11b/CD18), a leukocyte beta2 integrin, facilitates neutrophil adhesion, transendothelial migration, phagocytosis, and respiratory burst, all of which may mediate reperfusion-induced injury to ischemic brain tissue in conditions such as stroke. To determine the role of Mac-1 during ischemia and reperfusion in the brain, we analyzed the effect of transient focal cerebral ischemia in mice genetically engineered with a specific deficiency in Mac-1. Transient focal ischemia/reperfusion was induced by occluding the left middle cerebral artery for 3 hours followed by a 21-hour reperfusion period in Mac-1-deficient (n=12) and wild-type (n=11) mice. Regional cerebral blood flow was determined with a laser-Doppler flowmeter. Brain sections were stained with 2% 2,3,5-triphenyltetrazolium chloride to determine the infarct volume. Neutrophil accumulation was determined by staining the brain sections with dichloroacetate esterase to identify neutrophils. Compared with the wild-type cohort, Mac-1-deficient mice had a 26% reduction in infarction volume (P<0.05). This was associated with a 50%, but statistically insignificant, reduction in the number of extravasated neutrophils in the infarcted areas of the brains in the mutant mice. There were no differences in regional cerebral blood flow between the 2 groups. Mac-1 deficiency reduces neutrophil infiltration and cerebral cell death after transient focal cerebral ischemia. This finding may be related to a reduction in neutrophil extravasation in Mac-1-deficient mice.


Cytokine response in children undergoing surgery for congenital heart disease.
Pediatric cardiac surgery with cardiopulmonary bypass (CPB) induces a complex inflammatory response that may cause multiorgan dysfunction. The objective of this study was to measure postoperative cytokine production and correlate the magnitude of this response with intraoperative variables and postoperative outcomes. Serum samples from 20 children (median age, 15 months) undergoing cardiac surgery with CPB were obtained preoperatively and on postoperative days (POD) 1-3. Serum levels of interleukin (IL)-6, IL-8, and IL-10 increased significantly on POD 1 (p < 0.01) vs pre-op values to 271 +/- 68, 44 +/- 9, 7.5 +/- 0.8 pg/ml, respectively, whereas serum IL-1beta, IL-12, and tumor neurosis factor -alpha were not significantly changed. The serum IL-6 and IL-8 levels correlated positively (p < 0.01) with the degree of postoperative medical intervention as measured by the Therapeutic Interventional Scoring System and indicated a greater need for inotropic support (p = 0.057). A negative correlation (p < 0.01) between IL-6, IL-8, and mixed venous oxygen saturation suggested compromised cardiopulmonary function. Patients with single ventricle anatomy had the highest levels of IL-6 and IL-8 (629 +/- 131 and 70 +/- 17 pg/ml, respectively), with a mean CPB time of 106 +/- 23 minutes. Thus, the proinflammatory response after surgery with CPB was associated with postoperative morbidity with increased need for medical intervention.


Special Section Guest Editorial: Advances in High-Dimensional Medical Image Processing.
Guest Editors Ivana Išgum, Bennett A. Landman, and Tomaž Vrtovec introduce the JMI Special Section on Advances in High-Dimensional Medical Image Processing.


Next generation sequencing methodologies--an overview.
Gene discovery has been one of the most important advances in our understanding of human disorders. Early linkage and positional cloning strategies have now given way to next generation sequencing (NGS) with age-old help from biostatistical and bioinformatical input. In this chapter, we present the importance of getting the basics right, namely, how the best phenotyping in the clinical domain will provide a higher chance of a successful NGS experiment. In addition, we show getting the correct submission of DNA samples to NGS providers is dependent on the type of inheritance pattern that may or may not be apparent. We discuss one of the most crucial decisions for investigators when designing a study, namely choosing a trio, quad or cohort for analysis. Following on from this, we compare and contrast the underlying technology adopted by provider companies as they vie for customers and submissions. Each platform has advantages and disadvantages based on false calls, coverage, and read depth; however, some of these issues may be solved with the third wave of sequencing technology development in early commercial roll-out. Lastly, we provide a bioinformatic filtering overview of a "quad"-based submission and show how 3 million SNPs and indels can be reduced to a biologically plausible and experimentally manageable n≤50 gene variants.


A case of multiple, bilateral Becker's Nevi of the trunk and lower extremities in a young girl: A case report.
A Becker's nevus typically appears as a single unilateral, well-demarcated, hyperpigmented patch over the upper trunk during adolescence. It uncommonly presents as multiple and bilateral patches and rarely involves a lower extremity. We describe the unusual case of a child with multiple, bilateral Becker's nevi of the trunk and lower extremities present since birth.


Unique features of early diastolic mitral annulus velocity in constrictive pericarditis.
In most primary myocardial diseases, early diastolic mitral annulus velocity (E') decreases with disease progression. To our knowledge, constrictive pericarditis (CP) is the only condition without this phenomenon. This study was performed to evaluate the diagnostic and pathophysiologic role of mitral annulus velocity in patients with CP. In all, 17 patients with CP (9 men; mean age 46.5 +/- 14.3 years), 8 patients with cardiac tamponade (Tamp) (2 men; mean age 44.5 +/- 15.0 years), and age- and sex-matched control subjects for CP and Tamp were recruited for the study. Early mitral inflow velocity and E' were obtained while simultaneously recording respiration. In 8 patients with CP and in all patients with Tamp, these measurements were repeated after the relief of constrictive physiology or after pericardiocentesis. In patients with CP, E' was significantly higher than it was for control subjects (12.9 +/- 3.0 cm/s vs 9.8 +/- 2.4 cm/s, P <.01). An E' of 2 cm/s higher than the predicted normal E' could differentiate patients with CP from control subjects with a sensitivity of 76% and specificity of 82%. In 12 of 17 patients (71%), inspiratory E' was higher than expiratory E'-the opposite of mitral inflow variation. In 8 patients, E' decreased significantly after the relief of constrictive physiology (13.8 +/- 2.5 cm/s vs 9.3 +/- 3.1 cm/s, P <.05). In contrast, E' in the Tamp group was significantly lower than in the control group (6.8 +/- 1.6 cm/s vs 10.2 +/- 2.5 cm/s, P <.01), did not show significant respiratory variation, and increased significantly after pericardiocentesis (6.8 +/- 1.6 cm/s vs 9.5 +/- 3.0 cm/s, P <.05). E' is exaggerated in CP, which is helpful for diagnosis. The opposite phenomenon was noted in Tamp, a dissimilarity that might contribute to different hemodynamics.


Re-caching by Western scrub-jays (Aphelocoma californica) cannot be attributed to stress.
Western scrub-jays (Aphelocoma californica) live double lives, storing food for the future while raiding the stores of other birds. One tactic scrub-jays employ to protect stores is "re-caching"-relocating caches out of sight of would-be thieves. Recent computational modelling work suggests that re-caching might be mediated not by complex cognition, but by a combination of memory failure and stress. The "Stress Model" asserts that re-caching is a manifestation of a general drive to cache, rather than a desire to protect existing stores. Here, we present evidence strongly contradicting the central assumption of these models: that stress drives caching, irrespective of social context. In Experiment (i), we replicate the finding that scrub-jays preferentially relocate food they were watched hiding. In Experiment (ii) we find no evidence that stress increases caching. In light of our results, we argue that the Stress Model cannot account for scrub-jay re-caching.


Spirituality/religiosity, substance use, and HIV testing among young black men who have sex with men.
Spirituality and religiosity may serve as both a resource and a barrier to HIV prevention with young black men who have sex with men (YBMSM). We examined indices of spirituality/religiosity as correlates of binge drinking, stimulant use, and recent HIV testing in a sample of YBMSM. From 2011-2013, annual venue-based surveys of sexually active YBMSM ages 18-29 were conducted in Dallas and Houston, Texas. Binge drinking and stimulant use were assessed in the past two months. Participants recently tested for HIV (i.e., within the past six months) were compared to those without recent HIV testing (i.e., never tested or tested more than six months ago). Among the 1565 HIV-negative or HIV-unknown YBMSM enrolled, more engagement in spiritual and religious activities was associated with greater odds of reporting stimulant use (Adjusted Odds Ratio [AOR]=1.20; 95% CI=1.04-1.40) while higher spiritual coping was associated with lower odds of reporting stimulant use (AOR=0.66; 95% CI=0.56-0.78). Binge drinking was independently associated with 29% lower odds of recent HIV testing (AOR=0.71; 95% CI=0.55-0.92), but lower odds of binge drinking did not mediate the association of engagement in spiritual and religious activities with 27% greater odds of recent HIV testing (AOR=1.27; 95% CI=1.11-1.46). Among YBMSM, culturally tailored approaches addressing spirituality/religiosity could support prevention of stimulant use and increase HIV testing. In particular, expanded efforts are needed to promote HIV testing in binge drinkers.


Coronary sinus septal defect. Surgical considerations.
The coronary sinus septal defect is an uncommon anomaly. If it is not suspected, found, and repaired at operation, a residual intracardiac shunt will result. The defect usually is associated with a left superior vena cava (LSVC), which necessitates special consideration in the conduct of cardiopulmonary bypass. Our recent treatment of three cases of coronary sinus septal defect stimulated us to review the embryology, anatomy, diagnosis, and surgical treatment of this malformation.


Platelet-activating factor and P-selectin activities in thrombotic and nonthrombotic Behçet's patients.
The aim of this study was to compare plasma Platelet-activating factor (PAF) and P-selectin (CD62P) activities in Behçet's disease patients with and without thrombosis. In this cross-sectional and descriptive study, 30 consecutive Behçet's patients were included, 15 of them with venous thrombosis. All patients were also divided into two subgroups according to the presence or absence of clinical activity. Plasma PAF levels, basal and Ca++ ionophore (A23187)-induced leukocyte (cellular) PAF activities, and platelet-rich plasma DeltaCD62P activity (the mean fluorescent density difference between CD62P phycoerythrin-positive and -negative stains) were evaluated. In the thrombotic group, plasma PAF (P=0.001), basal leukocyte PAF (P=0.017), induced leukocyte PAF (P=0.024), and DeltaCD62P (P=0.023) levels were significantly higher than in the nonthrombotic group. In the whole group of Behçet's patients, there was a positive correlation between plasma PAF and DeltaCD62P levels (r=0.533, P=0.002). When we compared clinically active and inactive patients with respect to the above parameters, there was no significant difference, irrespective of thrombosis. Plasma PAF (P=0.001), basal leukocyte PAF (P=0.004), and DeltaCD62P (P=0.038) levels were significantly higher in the presence of both clinical activity and thrombosis than of clinical activity alone. Platelet-activating factor and CD62P may contribute to endothelial injury and thrombosis development in Behçet's disease. These two parameters seem related to the presence of thrombosis rather than clinical activity.


Management of acute gallbladder disease in England.
Recent literature suggests that early laparoscopic cholecystectomy for acute gallbladder disease is safe and efficacious, but few data are available on the management of acute gallbladder disease in England. Hospital Episode Statistics data for the years 2003-2005 were obtained from the Department of Health. All patients admitted as an emergency with acute gallbladder disease during the period from April 2003 to March 2004 were included as a cohort. Repeat emergency admissions for acute gallbladder disease, and cholecystectomies performed during the first admission, an emergency readmission or an elective admission were followed up until March 2005. Some 25,743 patients were admitted as an emergency with acute gallbladder disease, of whom 3791 had an emergency cholecystectomy during the first admission (open cholecystectomy (OC) 29.8 per cent, laparoscopic conversion rate (LCR) 10.7 per cent) and 9806 patients had an elective cholecystectomy (OC 11.3 per cent, LCR 8.3 per cent) during the study period. Early cholecystectomy for acute gallbladder disease is not widely practised by surgeons in England. Open cholecystectomy is more commonly used in the emergency than in the elective setting. Early laparoscopic cholecystectomy following an emergency admission carries a higher conversion rate than elective cholecystectomy.


Enhancement of skin permeation of miconazole by phospholipid and dodecyl 2-(N,N-dimethyl amino)propionate (DDAIP).
Miconazole (MCZ) has very low solubility in both water and oil. Permeation rates through shed snakeskin from an aqueous suspension and a mineral oil suspension were 0.5 microg/cm(2)/h and almost none, respectively. When hydrogenated phosphatidylcholine (HPC) was added to mineral oil and heated to 95 degrees C, the solubility of MCZ increased in proportion to the HPC concentration. DSC measurements also indicated an interaction between them. Thus, a gel formed by hydrogenated phospholipid and mineral oil, as vehicle was prepared. The solubility of MCZ in the gel was around 1% and the permeation rate was 1.3 microg/cm(2)/h, which was about 2.5 times that from an aqueous suspension. As an alternative approach, a skin permeation enhancer, dodecyl 2-(N,N-dimethyl amino)propionate (DDAIP) was applied 2 h before a skin permeation study. The permeation from an aqueous suspension became 11 times that of the suspension without DDAIP pretreatment. The concentration of MCZ in the skin increased 8-fold, indicating that the enhancement effect involved high partition of MCZ into the skin. On the other hand, when a gel formulation was used, pretreatment with DDAIP was not as effective as incorporation of DDAIP in the gel formulation. Following pretreatment, permeation was only two times that of the gel without DDAIP pretreatment, and half that of the water suspension with DDAIP pretreatment. This suggested that release from the gel was the rate-limiting step with the gel formulation. When DDAIP was added to the gel, the permeation rate of MCZ was 3.3 microg/cm(2)/h. It was also a release limited type permeation. The gel with DDAIP is potentially a useful formulation, because of relatively high permeation while possibly avoiding overdosing.


Factors associated with delayed recognition of pulmonary tuberculosis in emergency departments in Taiwan.
To identify and evaluate factors associated with delayed recognition of pulmonary tuberculosis (TB) in the emergency department (ED). Delayed recognition of pulmonary TB in ED may precipitate mortality and morbidity. Medical records of newly diagnosed TB patients admitted to four hospitals in Taiwan were retrospectively reviewed. Patients were divided into two groups based on ED physicians' recognition or not of TB and statistically compared to identify differences in their characteristics. 310 newly diagnosed TB patients were identified; 150 were unrecognized in the ED. Cough, chest tightness, general malaise, and body weight loss were more common for those with recognized TB. Older age (≥65 yrs, P = 0.035) and chronic renal insufficiency (P = 0.005) were associated with delayed TB recognition. Older age and chronic renal insufficiency are risk factors for delayed TB while in the ED. Typical symptoms should heighten alertness for recognizing TB.


Expression of heat-shock protein-90 in non-Hodgkin's lymphomas.
Heat-shock protein-90 (HSP90) inhibitors are currently being used in phase I clinical trials for treating patients with a variety of neoplasms including lymphomas. Using immunohistochemical methods, we assessed for HSP90 expression in 412 cases of non-Hodgkin's lymphoma. In B-cell lymphomas, HSP90 was moderately to strongly expressed in all cases of Burkitt's lymphoma (5/5, 100%), and in subsets of follicular lymphoma (17/28, 61%), diffuse large B-cell lymphoma (27/46, 59%), nodal marginal zone B-cell lymphoma (6/16, 38%), plasma cell neoplasms (14/39, 36%), small lymphocytic lymphoma/chronic lymphocytic leukemia (3/9, 33%), mantle cell lymphoma (12/38, 32%) and lymphoplasmacytic lymphoma/Waldenstrom macroglobulinemia (3/10, 30%). HSP90 was weakly expressed in six of 14 (43%) cases of extranodal marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue. In T-cell lymphomas, HSP90 was moderately to strongly expressed in subsets of anaplastic large-cell lymphoma (14/24, 58%; 9/12 ALK+ and 5/12 ALK-), precursor-T-cell lymphoblastic leukemia/lymphoma (20/65, 31%), unspecified peripheral T-cell lymphoma (8/43, 23%) and angioimmunoblastic T-cell lymphoma (2/17, 12%). HSP90 was weakly expressed in seven of 58 (12%) cases of mycosis fungoides. We conclude that HSP90 is commonly expressed in a subset of many types of B- and T-cell lymphoma. These data suggest that many lymphoma types are suitable targets for modulation of HSP90 activity, and that HSP90 inhibitors are a potential investigational therapy for lymphoma patients.


Oxytocin induced labor causes region and sex-specific transient oligodendrocyte cell death in neonatal mouse brain.
Previous reports showed associations between oxytocin induced labor and mental disorders in offspring. However, those reports are restricted in epidemiological analyses and its mechanism remains unclear. In this study, we hypothesized that induced labor directly causes brain damage in newborns and results in the development of mental disorders. Therefore we aimed to investigate this hypothesis with animal model. The animal model of induced labor was established by subcutaneous oxytocin administration to term-pregnant C57BL/6J mice. We investigated the neonatal brain damage with evaluating immediate early gene expression (c-Fos, c-Jun and JunB) by quantitative polymerase reaction and TdT-mediated dUTP nick end labeling staining. To investigate the injured brain cell types, we performed double-immunostaining with TdT-mediated dUTP nick end labeling staining and each brain component specific protein, such as Oligo2, NeuN, GFAP and Iba1. Brain damage during induced labor led to cell death in specific brain regions, which are implicated in mental disorders, in only male offspring at P0. Furthermore, oligodendrocyte precursors were selectively vulnerable compared to the other cell types. This oligodendrocyte-specific impairment during the perinatal period led to an increased numbers of Olig2-positive cells at P5. Expression levels of oxytocin and Oxtr in the fetal brain were not affected by the oxytocin administered to mothers during induced labor. Oligodendrocyte cell death in specific brain regions, which was unrelated to the oxytocin itself, was caused by induced labor in only male offspring. This may be an underlying mechanism explaining the human epidemiological data suggesting an association between induced labor and mental disorders.


Critical Care of the Burn Patient.
Care of the critically ill burned patient must integrate a multidisciplinary care team composed of burn care specialists. As resuscitative mortality decreases more patients are surviving to experience multisystem organ failure relating to complications of their injuries. Clinicians must be aware of physiologic changes following burn injury and the implicated impacts on management strategy. Promoting wound closure and rehabilitation should be the backdrop for which management decisions are made.


Incremental utility of expanded mutation panel when used in combination with microRNA classification in indeterminate thyroid nodules.
Focused and expanded mutation panels were assessed for the incremental utility of using an expanded panel in combination with microRNA risk classification. Molecular results were reviewed for patients who underwent either a focused mutation panel (ThyGenX®) or an expanded mutation panel (ThyGeNEXT®) for strong and weak oncogenic driver mutations and fusions. microRNA results (ThyraMIR®) predictive of malignancy, including strong positive results highly specific for malignancy, were examined. Results of 12 993 consecutive patients were reviewed (focused panel = 8619, expanded panel = 4374). The expanded panel increased detection of strong drivers by 8% (P < .001), with BRAFV600E and TERT promoters being the most common. Strong drivers were highly correlated with positive microRNA results of which 90% were strongly positive. The expanded panel increased detection of coexisting drivers by 4% (P < .001), with TERT being the most common partner often paired with RAS. It increased the detection of weak drivers, with RAS and GNAS being the most common. 49% of nodules with weak drivers had positive microRNA results of which 33% were strongly positive. The expanded panel also decreased the number of nodules lacking mutations and fusions by 15% (P < .001), with 8% of nodules having positive microRNA results of which 22% were strongly positive. Using expanded mutation panels that include less common mutations and fusions can offer increased utility when used in combination with microRNA classification, which helps to identify high risk of malignancy in the cases where risk is otherwise uncertain due to the presence of only weak drivers or the absence of all drivers.


6-Substituted 2-Aminopurine-2'-deoxyribonucleoside 5'-Triphosphates that Trace Cytosine Methylation.
Gene expression is extensively regulated by the occurrence and distribution of the epigenetic marker 2'-deoxy 5-methylcytosine (5mC) in genomic DNA. Because of its effects on tumorigenesis there is an important link to human health. In addition, detection of 5mC can serve as an outstanding biomarker for diagnostics as well as for disease therapy. Our previous studies have already shown that, by processing O(6) -alkylated 2'-deoxyguanosine triphosphate (dGTP) analogues, DNA polymerases are able to sense the presence of a single 5mC unit in a template. Here we present the synthesis and evaluation of an extended toolbox of 6-substituted 2-aminopurine-2'-deoxyribonucleoside 5'-triphosphates modified at position 6 with various functionalities. We found that sensing of 5-methylation by this class of nucleotides is more general, not being restricted to O(6) -alkyl modification of dGTP but also applying to other functionalities.


Barriers to the utilization of thrombolysis for acute ischaemic stroke.
Thrombolysis is currently the only evidence-based pharmacological treatment available for acute ischaemic stroke (AIS); however, its current utilization is suboptimal (administered to <3% of AIS patients). The aim of this article was to identify the potential barriers to the use of thrombolysis via a review of the available literature. Medline, Embase, International Pharmaceutical Abstracts and Google Scholar were searched to identify relevant original articles, review papers and other literature published in the period 1995-2011. Several barriers to the utilization of thrombolysis in stroke have been identified in the literature and can be broadly classified as 'preadmission' barriers and 'post-admission' barriers. Preadmission barriers include patient and paramedic-related factors leading to late patient presentation for treatment (i.e. outside the therapeutic time window for the administration of thrombolysis). Post-admission barriers include in-hospital factors, such as suboptimal triage of stroke patients and inefficient in-hospital acute stroke care systems, a lack of appropriate infrastructure and expertise to administer thrombolysis, physician uncertainty in prescribing thrombolysis and difficulty in obtaining informed consent for thrombolysis. Suggested strategies to overcome these barriers include public awareness campaigns, prehospital triage by paramedics, hospital bypass protocols and prenotification systems, urgent stroke-unit admission, on-call multidisciplinary acute stroke teams, urgent neuroimaging protocols, telestroke interventions and risk-assessment tools to aid physicians when considering thrombolysis. Additionally, greater pharmacists' engagement is warranted to help identify the people at risk of stroke and support preventative strategies, and provide the public with information regarding the recognition of stroke, as well as facilitate the access and use of thrombolysis. The most effective interventions appear to be those comprising several strategies and those that target more than one barrier simultaneously. Therefore, optimal utilization of thrombolysis requires a systematic, integrated multidisciplinary approach across the continuum of acute care.


Indirect recognition and antibody production against a single mismatched HLA-A2-transgenic molecule precede the development of obliterative airway disease in murine heterotopic tracheal allografts.
Previous studies have implicated the allogeneic immune response in the development of obliterative bronchiolitis after lung transplantation. However, the progression of specific pathogenic events leading to this form of chronic allograft dysfunction have not been well characterized. We used a murine tracheal transplantation model in which a single mismatched HLA-A2-transgenic molecule is indirectly recognized by the recipient CD4(+) T cells to show that obliterative airway disease (OAD) that developed in these allografts was preceded by indirect recognition of the HLA-A2 molecule and subsequent development of anti-HLA-A2 antibodies. Tracheas from HLA-A2(+) C57BL/6 mice were heterotopically transplanted into C57BL/6 mice. Allograft histopathology as well as anti-HLA-A2 T-cell proliferative responses and anti-HLA-A2 antibody development were determined at days 5, 10, 20, and 28 after transplantation. All of the HLA-A2(+) tracheal allografts transplanted into C57BL/6 recipients demonstrated complete development of OAD by day 20. Spleen cells from the mice that underwent transplantation demonstrated significant proliferation against HLA-A2(+) cells by day 5. Indirect recognition of HLA-A2-derived peptides by spleen cells from allograft recipients was also higher on days 5 and 10 as compared with irrelevant peptides derived from HLA-A1, HLA-A3, and HLA-B44. Allograft recipients showed detectable levels of anti-HLA-A2 antibodies by day 5 and full development of anti-HLA-A2 antibodies by day 20. These results show that sensitization of CD4+ T cells against the mismatched HLA-A2 alloantigen precedes the development of anti-HLA antibodies as well as OAD, suggesting an important role for alloreactive CD4(+) T-cell activation and alloantibody development in the immunopathogenesis of OAD.


System-Wide Profiling of Protein Amino Termini from Formalin-Fixed, Paraffin-Embedded Tissue Specimens for the Identification of Novel Substrates.
Clinical tissues are used for histopathological diagnosis of many diseases including immunostaining and morphology subtyping as well as in molecular research such as for the analyses of DNA, RNA, and proteins. Formalin fixation and paraffin embedment (FFPE) of tissue specimens is routinely used for preserving clinical tissues for long-term storage, allowing histopathological diagnosis of many diseases. As such, FFPE tissues currently represent the most comprehensive collection of all clinical specimens, allowing great source of material for research opportunity, possibly due to the concern of protein integrity from antigen retrieval from fixation process. Hence, to date, very few studies have used FFPE specimens to look at the profiling of protein termini. Nevertheless in the field of protease research, the protein amino termini are particularly useful for the system-wide identification of substrates and for the characterization of protease-mediated cleavage sites . In this chapter, we outline a robust methodology for the extraction of proteins from FFPE specimens for the enrichment of protein amino termini. This approach enriches endogenous protein N-termini by removal of internal peptides using synthetic polymers of hyperbranched polyglycerol aldehyde. As a result, protein amino termini are analyzed using mass spectrometers to elucidate the biological regulation of protease-substrate interactions in healthy and diseased tissues.


Ileorectal anastomosis for ulcerative and Crohn's colitis.
Except in the presence of severe perineal suppuration or sphincter damage by previous surgery for fistulas, the rectum was preserved in all patients considered candidates for surgery for inflammatory disease of the bowel. A primary anastomosis with a single-layer 5-0 monofilament stainless steel wire was carried out when a relatively healthy rectum with erythema and granularity presented. For patients with more severe disease of the rectum, a two-stage operation with intensive interval treatment of the rectum stump with topical corticosteroids was carried out. Of a total of eighty-six patients with involvement of the colon and rectum with either Crohn's disease or chronic ulcerative colitis, fifty-six patients were treated by local abdominal colectomy and ileorectal anastomosis. Twenty-four had primary anastomosis and thiry-two had a two-stage operation. One anastomotic dehiscence developed. A mean follow-up of 8.4 years (6 months to 20 years) has been satisfactory. Only three anastomoses have been taken down for unsatisfactory results. With the proper selection of patients and with appropriate treatment of the diseased rectal segment, a large majority of patients with inflammatory disease of the bowel can have long-term salutory results after colectomy and ileorectal anastomosis.


Outline of a theory of manic-depressive illness.
Manic-depressive illness is characterized by locking into extreme states and alternation between those extremes. Locking into extreme states is also characteristic of a bistable, a basic electronic device of which the essential feature is a positive feedback loop with greater than unity gain. An argument is presented that under natural selection there has evolved a well-known and advantageous tendency for perceptions of successes to increase confidence, and that it is in the nature of confidence to increase perceptions of successes. Thus arises a positive feedback loop, in which perceptions of successes increase confidence, which further increases perceptions of successes, and so on, and the converse with failure and inconfidence. When the gain of the loop exceeds unity then locking into mania or depression will result. The feedback gain depends on both internal 'biological' factors and external situational factors. As a result of natural fluctuations the gain will occasionally fall temporarily below unity, releasing from the extreme state. Escape from mania will bias towards depression, whereas escape from depression will be elating. The physical embodiment of confidence may be serotonergic (or closely related) activity.


History of polar whaling: insights into the physiology of the great whales.
The sheer size and pelagic nature of the great whales has effectively precluded detailed studies of most of their physiological processes. The vast majority of all data for these species have come from anatomical studies conducted on specimens that were caught in commercial and native whaling operations. In both the polar regions, an incredible number of whales were hunted, but anatomical studies were not usually conducted until relatively recent times. However, the anatomical data that do exist provide a valuable insight into some of the physiological demands placed on the animals by their marine habitat. These include information on blubber and nutrition; baleen and feeding ecology; contaminant chemistry and tissue samples; diving chemistry and acoustics. Taken together, these anatomical data provide the only substantial information on how these animals dive and hunt. Recent breakthroughs in chemical techniques however, are providing even greater details on function (for example, fatty acid signature methods). Coupled with advanced methods for tracking these whales at sea (acoustic and satellite), future studies should provide significant new information on the general physiology of these difficult to study species.


Essential role of glucose transporter GLUT3 for post-implantation embryonic development.
Deletion of glucose transporter gene Slc2a3 (GLUT3) has previously been reported to result in embryonic lethality. Here, we define the exact time point of growth arrest and subsequent death of the embryo. Slc2a3(-/-) morulae and blastocysts developed normally, implanted in vivo, and formed egg-cylinder-stage embryos that appeared normal until day 6.0. At day 6.5, apoptosis was detected in the ectodermal cells of Slc2a3(-/-) embryos resulting in severe disorganization and growth retardation at day 7.5 and complete loss of embryos at day 12.5. GLUT3 was detected in placental cone, in the visceral ectoderm and in the mesoderm of 7.5-day-old wild-type embryos. Our data indicate that GLUT3 is essential for the development of early post-implanted embryos.


Acute cholecystitis in the elderly: use of computed tomography and correlation with ultrasonography.
Elderly patients diagnosed with acute cholecystitis (AC) may undergo both ultrasonography (US) and computed tomography (CT). A total of 475 patients (age, >64 y) with AC were included. Groups included US alone (n = 240), CT alone (n = 60), and CT + US (n = 168). Sixty patients (35.7%) in the US + CT group had inflammation in both studies, 34 (20.2%) had inflammation only on US, and 32 (19.0%) had inflammation only on CT. In the US + CT group, detection of cholelithiasis was not different, but mean common bile duct size did not correlate. There was no difference among the groups in age, sex, medical service admission, nonambulatory status, dementia, diabetes, or coronary artery disease. Peritonitis, leukocytosis, and acidosis were more frequent in the 2 groups undergoing CT. The cholecystectomy rate was lowest (and the complication rate was highest) in the CT + US group. CT often is used in the diagnosis of AC in the elderly, especially those with more acute presentations. CT and US findings may be complementary in AC.


Hirschsprung's disease studied with monoclonal antineurofilament antibodies on tissue sections.
In a retrospective study of Hirschsprung's disease two monoclonal antineurofilament antibodies were tested immunohistochemically. Biopsy samples of rectum and colon were fixed in formalin and embedded in paraffin. Tissue sections from normal controls and from unaffected colon of patients with Hirschsprung's disease showed partial staining of only some of the axon bundles in the myenteric and in the submucous plexus. In contrast, in all tissue sections from colon affected by Hirschsprung's disease there were heavily stained hyperplastic axon bundles. Sections of colon from patients with various other neurogenic disorders of the colon were studied by the same method; the characteristic Hirschsprung picture was never seen.


A self-crosslinking, double-functional group modified bacterial cellulose gel used for antibacterial and healing of infected wound.
Cellulose/chitosan composite, as a mature commercial antibacterial dressing, is an important type of wound repair material. However, how to achieve the perfect compound of two components and improve antibacterial activity is a major, lingering issue. In this study, a bifunctional group modified bacterial cellulose (DCBC) was prepared by carboxymethylation and selective oxidation. Further, the chitosan (CS) was compounded in the network of DCBC by self-crosslinking to form dialdehyde carboxymethyl bacterial cellulose/chitosan composites (S-DCBC/CS). The aldehyde group can react with amino of CS by Schiff base reaction. The carboxyl group of DCBC and the amorphous distribution of CS molecular chains increase the antimicrobial properties of composites. The bacteriostatic rate of composites could be higher than 95%. Bacteria can be attracted onto the surface of composites, what we call it "directional adhesion antibacterial effects". In particular, a kind of large animal wound model, deep Ⅱ degree infected scald of Bama miniature pig, was used to research the antimicrobial and healing properties of materials. The S-DCBC/CS can effectively inhibit bacterial proliferation of wound and kill the bacteria. The wound healing rate of S-DCBC/CS was up to 80% after three weeks. The composites show better antibacterial and promoting concrescence effects than traditional chitosan dressings.


Identification of bacterial infections and clinical manifestation associated with cytomegalovirus in liver transplantation patients.
Liver transplantation has become the most effective therapy for the treatment of patients with end-stage liver disease. With new immunosuppressive agents the incidence of acute rejection has been significantly reduced, but infection has become a serious problem. Our objective was to correlate cytomegalovirus (CMV) positivity of antigenemia and polymerase chain reaction (PCR) with clinical manifestations and bacterial infections among patients undergoing liver transplantation. This prospective study included patients monitored for 6 months for early detection of CMV infection. Sample collections were performed at the time of surgery and weekly until the second month followed by fortnightly in the third month, and monthly in the fourth to sixth month. CMV infection was defined by positive antigenemia (>3 cells) or 2 positive PCR tests associated or not with clinical symptoms. The methodology for the diagnosis of bacterial infection was through biochemical tests and the automated VITEK/bioMérieux (identification and antibiogram) using samples of urine and blood cultures. Chi-square test was used for dicotomic variables with significant differences when P < .05. Sixteen patients (32%) had CMV infections, including 13 (81%) with concomitant infections. Thirty-four patients (68%) did not have CMV infections and 8 of these (24%) had bacterial infection. There was a high correlation with bacterial infections among CMV-positive patients. Bacterial infections after liver transplantation were associated with CMV infection.


An ultrasensitive monoclonal antibody-based lateral flow immunoassay for the rapid detection of xylazine in milk.
In this study, a novel hapten was designed and successfully synthesized to prepared the artificial immunogen against xylazine (XYL), then the monoclonal antibody 1H2-2C6 was obtained and applied to a lateral flow immunoassay (LFI) strip for rapid detection of XYL in milk. The visual detection limit (cutoff value) of the LFI strip was 5 ng/mL, the linear regression formula was Y = -0.521lg(X) + 0.382 (R2 = 0.995, n = 6), the linear detection range was 0.16-2.23 ng/mL and the detection limit (LOD) was 0.1 ng/mL. The recovery rates were in the range from 88.95% to 91.45%, the highest coefficient of variation was 8.89% (n = 3). The LFI strip was also validated by LC-MS/MS, as revealed, there were no significance differences between the two methods (the confidence interval was 95%). In brief, the LFI strip provides a reliable, sensitive and portable testing tool for screening of XYL on-site.


Prediction of liquid chromatographic retention behavior based on quantum chemical parameters using supervised self organizing maps.
Self organizing maps (SOMs) in a supervised mode were applied for prediction of liquid chromatographic retention behavior of chemical compounds based on their quantum chemical information. The proposed algorithm was simple and required only a small alteration of the standard SOM algorithm. The application was illustrated by the prediction of the retention indices of bifunctionally substituted N-benzylideneanilines (NBA) and the prediction of the retention factors of some pesticides. Although the predictive ability of the supervised SOM could not be significantly greater than that of some previously established neural network methods, such as a radial basis function (RBF) neural network and a back-propagation artificial neural network (ANN), the main advantage of the proposed method was the ability to reveal non-linear structure of the model. The complex relationships between samples could be visualized using U-matrix and the influence of each variable on the predictive model could be investigated using component planes-which can provide chemical insight.


Late-onset hypocalcemia appearing years after thyroid surgery.
We revised four cases of hypocalcemia diagnosed yr after thyroid surgery, including 1 man and 3 women, operated 5-23 yr before the onset of symptoms, which ranged from mild paresthesia to convulsive seizures. Total serum calcium levels ranged from 1.1 to 2.05 mmol/l, and PTH levels were low in the 3 cases in which they were measured. Hypoparathyroidism appearing as late-onset hypocalcemia after extense thyroid surgery is a poorly understood condition, perhaps not as unfrequent as it is considered.


Does the frequency content of the surface mechanomyographic signal reflect motor unit firing rates? A brief review.
The purpose of this review is to examine the literature that has investigated the potential relationship between mechanomyographic (MMG) frequency and motor unit firing rates. Several different experimental designs/methodologies have been used to address this issue, including: repetitive electrical stimulation, voluntary muscle actions in muscles with different fiber type compositions, fatiguing and non-fatiguing isometric or dynamic muscle actions, and voluntary muscle actions in young versus elderly subjects and healthy individuals versus subjects with a neuromuscular disease(s). Generally speaking, the results from these investigations have suggested that MMG frequency is related to the rate of motor unit activation and the contractile properties (contraction and relaxation times) of the muscle fibers. Other studies, however, have reported that MMG mean power frequency (MPF) does not always follow the expected pattern of firing rate modulation (e.g. motor unit firing rates generally increase with torque during isometric muscle actions, but MMG MPF may remain stable or even decrease). In addition, there are several factors that may affect the frequency content of the MMG signal during a voluntary muscle action (i.e. muscle stiffness, intramuscular fluid pressure, etc.), independent of changes in motor unit firing rates. Despite the potential influences of these factors, most of the evidence has suggested that the frequency domain of the MMG signal contains some information regarding motor unit firing rates. It is likely, however, that this information is qualitative, rather than quantitative in nature, and reflects the global motor unit firing rate, rather than the firing rates of a particular group of motor units.


Ultrasensitive assay of RNA: application to 100-500 cells.
A new method has been developed to measure RNA over the range 0.5 to 25 ng. This procedure involves quantitative hydrolysis of RNA to 5'-monophosphates, conversion of the 5'-AMP to 5'-ATP, and measurement of the ATP by the firefly luciferase assay. This method can be applied to acid precipitated macromolecules without interference from DNA. The measurement of the RNA content of CHO cell colonies of less than 100 cells is described.


[Use of autoradiography. Studies of cell kinetics in normal and psoriatic epidermis as an aid in decision making in therapeutic management].
We report on cell kinetic data about in vivo and in vitro DNA-labelling of germinative keratinocytes in normal and psoriatic epidermis. In order to study the cell cycle and its component parts in psoriasis, information on duration of the DNA-synthesis phase, epidermal germinative growth fraction, as well as cell cycle time are of special interest. Comparison of various anti-psoriatics reveals that they have different effects on the cell cycle in psoriatic epidermis. This observation may be important for the development and analysis of new treatment schedules and their influence on psoriatic cell proliferation kinetics.


miRNA-223 upregulated by MYOD inhibits myoblast proliferation by repressing IGF2 and facilitates myoblast differentiation by inhibiting ZEB1.
Skeletal muscle differentiation can be regulated by various transcription factors and non-coding RNAs. In our previous work, miR-223 is differentially expressed in the skeletal muscle of chicken with different growth rates, but its role, expression and action mechanism in muscle development still remains unknown. Here, we found that MYOD transcription factor can upregulate miR-223 expression by binding to an E-box region of the gga-miR-223 gene promoter during avian myoblast differentiation. IGF2 and ZEB1 are two target genes of miR-223. The target inhibition of miR-223 on IGF2 and ZEB1 are dynamic from proliferation to differentiation of myoblast. miR-223 inhibits IGF2 expression only in the proliferating myoblast, whereas it inhibits ZEB1 mainly in the differentiating myoblast. The inhibition of IGF2 by miR-223 resulted in the repression of myoblast proliferation. During myoblast differentiation, miR-223 would be upregulated owing to the promoting effect of MYOD, and the upregulation of miR-223 would inhibit ZEB1 to promote myoblast differentiation. These results not only demonstrated that the well-known muscle determination factor MYOD can promote myoblast differentiation by upregulate miR-223 transcription, but also identified that miR-223 can influence myoblast proliferation and differentiation by a dynamic manner regulates the expression of its target genes.


Switching from onabotulinum toxin A to abobotulinum toxin A for treating detrusor overactivity in spinal cord injured patient, does it really work?
To assess the efficacy of switching to Abobotulinumtoxin A (ATA) intradetrusor injections (IDI) after failure of Onabotulinumtoxin A (OTA) IDI for the treatment of neurogenic detrusor overactivity in patients with spinal cord injury (SCI). A single-centre retrospective chart review study. All SCI patients who started OTA IDI after 2011 and had an ATA IDI switch were included. The primary outcome was the clinical and urodynamic efficacy of the switch to ATA IIDs at the last follow-up. Secondary outcomes were initial efficacy, duration of ATA treatment, and patient outcome including the occurrence of augmentation enterocystoplasty at last follow-up. Sixty-two patients were included. Eighteen patients (28.9%) were initially responders to ATA IDI. Nine patients (14.5%) remained responders at last follow-up after a median of 17 months (AE 8.8-29). Thirty-two patients (51.6%) had had or were awaiting augmentation enterocystoplasty with a follow-up time of 18.5 months (IQR 8-27). Eleven patients (17.7%) were on ATA IDI with low efficacy. Seven patients (11.3%) were switched back to OTA and 3 patients (4.8%) changed their voiding pattern. Switching from OTA to ATA toxin for IDI in the treatment of detrusor overactivity after spinal cord injury have long-term efficacy for a limited number of patients but may delay the need for surgery.


The structure of phenobarbital-inducible rat liver cytochrome P-450 isoenzyme PB-4. Production and characterization of site-specific antibodies.
Fifteen peptides corresponding in sequence to segments of the major phenobarbital-inducible forms of rat hepatic cytochrome P-450 (termed P-450 PB-4 and P-450 PB-5) were chemically synthesized, conjugated to carrier proteins, and used to prepare site-specific rabbit and/or mouse antipeptide antibodies. Four of the synthetic peptides were recognized by rabbit heterosera raised against purified P-450 PB-4. The titer of these heterosera measured against P-450 PB-4 was only partially reduced upon complete adsorption of antipeptide activity suggesting that these peptides represent minor antigenic determinants. Each of the antipeptide antibodies recognized purified P-450 PB-4 and the highly homologous P-450 PB-5 as demonstrated by a solid-phase enzyme-linked immunosorbent assay. Although each antipeptide immunoprecipitated both purified 125I-labeled P-450 PB-4 and also in vitro-synthesized apo-P-450 PB-4, the yields of immunoprecipitation were low relative to that obtained using anti-P-450 heterosera. Only one of the antipeptide antibodies gave a good signal in an immunoblot analysis of either microsomal or purified P-450s PB-4 and PB-5. Three antipeptide antibodies raised against hydrophilic segments located in the amino-terminal one-third of P-450 PB-4 markedly inhibited the P-450 PB-4-catalyzed O-deethylation of the model substrate 7-ethoxycoumarin. Four of the antipeptide antibodies were found to cross-react with P-450 beta NF-B, the major aromatic hydrocarbon-inducible rat hepatic P-450, suggesting that certain amino acid sequences or regions of secondary structure are conserved between the major phenobarbital-induced and polycyclic-induced rat liver P-450 isoenzymes. These studies demonstrate the utility of antipeptide antibodies for evaluation of antigenic sites exposed in native P-450 PB-4, for identification of specific amino acid sequences important for the interaction of P-450 PB-4 with its substrate and/or with cytochrome P-450 reductase in a reconstituted system and for elucidation of structural and immunochemical homologies between P-450 PB-4 and other P-450 isoenzymes present in rat liver endoplasmic reticulum.


Changes of immunological markers after autologous peripheral blood stem cell transplantation.
Recently high-dose chemotherapy with peripheral blood stem cell transplantation (PBSCT) has become an important treatment for hematological and solid tumors. Immunological parameters were examined before and after PBSCT in 9 patients with lung cancer and 13 patients with malignant lymphoma. Findings were compared with those for bone marrow transplantation (BMT). Peripheral blood cells were analyzed for phenotype and the levels of cytokines and soluble factors were measured. After PBSCT, activated T cells (CD3+HLA-DR+ cells, CD8+HLA-DR+ cells) and suppressor/cytotoxic T cells (CD8+CD11b- cells) were significantly higher in the patients with lung cancer than in those with malignant lymphoma. Serum levels of interleukin-4 and soluble interleukin-2 receptor were also significantly higher in the patients with lung cancer than in those with lymphoma. On the other hand, the serum levels of interferon gamma, tumor necrosis factor alpha, interleukin-6, soluble human leukocyte antigen class 1, and soluble thrombomodulin were significantly increased after bone marrow transplantation. The transfused peripheral stem cells of lung cancer and lymphoma patients had a similar number of granulocyte/macrophage-colony-forming units, but lung cancer patients had significantly more CD34-positive cells. By reinfusing large numbers of autologous immune cells, PBSCT may accelerate immune reconstitution, with T cells being likely to have a marked therapeutic potential. The changes after PBSCT were greater in patients with lung cancer than in lymphoma patients. These blood cells are potent mediators of anticancer activity and could play an important role in the elimination of autologous malignant cells.


Acute disinhibiting effects of alcohol as a factor in risky driving behavior.
Automobile crash reports show that up to 40% of fatal crashes in the United States involve alcohol and that younger drivers are over-represented. Alcohol use among young drivers is associated with impulsive and risky driving behaviors, such as speeding, which could contribute to their over-representation in alcohol-related crash statistics. Recent laboratory studies show that alcohol increases impulsive behaviors by impairing the drinker's ability to inhibit inappropriate actions and that this effect can be exacerbated in conflict situations where the expression and inhibition of behavior are equally motivating. The present study tested the hypothesis that this response conflict might also intensify the disruptive effects of alcohol on driving performance. Fourteen subjects performed a simulated driving and a cued go/no-go task that measured their inhibitory control. Conflict was motivated in these tasks by providing equal monetary incentives for slow, careful behavior (e.g., slow driving, inhibiting impulses) and for quick, abrupt behavior (fast driving, disinhibition). Subjects were tested under two alcohol doses (0.65 g/kg and a placebo) that were administered twice: when conflict was present and when conflict was absent. Alcohol interacted with conflict to impair inhibitory control and to increase risky and impaired driving behavior on the drive task. Also, individuals whose inhibitory control was most impaired by alcohol displayed the poorest driving performance under the drug. The study demonstrates potentially serious disruptions to driving performance as a function of alcohol intoxication and response conflict, and points to inhibitory control as an important underlying mechanism.


An unusual case of lower motor neuron disease.
An 11-year-old boy developed a rapidly progressive asymmetrical flaccid tetraparesis that ended fatally within 1 year. The classification of this unusual case is discussed.


A Rare Case of Duodenal Adenocarcinoma Presenting as a Subepithelial Lesion in a Patient Undergoing Investigation for Iron Deficiency Anaemia.
Adenocarcinomas account for approximately 40% of small bowel cancers. They are typically mucosal lesions with distinctive features on endoscopy. We describe a rare case of duodenal adenocarcinoma presenting as a subepithelial lesion which posed a diagnostic challenge. An 85-year-old male patient presented for investigation of iron deficiency anaemia. Initial upper endoscopy found a subepithelial duodenal lesion with central depression but otherwise normal appearing mucosa. Superficial biopsies of the duodenal lesion were unremarkable. Subsequent antegrade single balloon enteroscopy revealed active bleeding from the lesion which was refractory to endoscopic treatment. A complete local excision of the lesion via laparotomy was necessary to achieve haemostasis. Histopathology from the lesion showed a moderately differentiated duodenal adenocarcinoma with invasion into the submucosa but no evidence of lymphovascular spread. Duodenal adenocarcinomas are rare gastrointestinal tumours associated with a poor prognosis. This case report outlines a rare presentation of duodenal adenocarcinoma and highlights the importance of judicious assessment of lesions found on endoscopy. Advances in endoscopic diagnostic modalities could facilitate early diagnosis and improve therapeutic outcomes.


[A study through factor analysis of the connection between 7 variables concerning blood lipides, with a particular reference to the interpretation of the lipidograms (author's transl)].
The AA. examined 7 variables concerning blood lipides and studied their internal relationship by means of a method, the Factor Analysis, which aims at determining the least amount of Common Factors which can sum up the information contained in the same variables. In this case three Common Factors (F) could be obtained, two of which were connected to the interpretation of the data of the lipidogram. The AA. discuss the signification of the determined Factors; they have even calculated the scores of the same Factors for each single individual and derived from the frequence distributions of the scores of F1 and F2 the normal values of the same Factors.


The diameter of chromatin fibres depends on linker length.
We have studied the diameter of chromatin fibres embedded in epoxy resins for three different materials: mouse thymus, chicken erythrocytes and sea cucumber spermatozoa. We confirm that the diameter of chromatin fibres increases with linker length, both values being influenced by the protein composition of chromatin.


Further results of serological examination of domestic animals for leptospirosis in Afghanistan.
The authors examined serologically 329 specimens of domestic animals from 8 provinces of Afghanistan for the incidence of leptospirosis. They detected in 15.8% of the animals examined antibodies against Leptospira in titres of 1 : 800 and higher: in 6.4% with serotypes of the serogroup Hebdomadis, 5.5%--Tarassovi, 2.7%--Grippotyphosa, 2.4%--Pomona, 2.1% Javanica, 1.5%--Icterohaemorrhagiae, 0.6% each--Canicola, Ballum, Bataviae, 0.3%--Pyrogenes. Positivity was highest in the buffalo--55.0%, camel--10 of the 18 animals examined, and cattle--25.5%. It was considerably lower in sheep--2.3% and goat--3.2%; also positive was one of the 6 zebus examined.


Genomic rearrangements and the evolution of clusters of locally adaptive loci.
Numerous studies of ecological genetics have found that alleles contributing to local adaptation sometimes cluster together, forming "genomic islands of divergence." Divergence hitchhiking theory posits that these clusters evolve by the preferential establishment of tightly linked locally adapted mutations, because such linkage reduces the rate that recombination breaks up locally favorable combinations of alleles. Here, I use calculations based on previously developed analytical models of divergence hitchhiking to show that very few clustered mutations should be expected in a single bout of adaptation, relative to the number of unlinked mutations, suggesting that divergence hitchhiking theory alone may often be insufficient to explain empirical observations. Using individual-based simulations that allow for the transposition of a single genetic locus from one position on a chromosome to another, I then show that tight clustering of the loci involved in local adaptation tends to evolve on biologically realistic time scales. These results suggest that genomic rearrangements may often be an important component of local adaptation and the evolution of genomic islands of divergence. More generally, these results suggest that genomic architecture and functional neighborhoods of genes may be actively shaped by natural selection in heterogeneous environments. Because small-scale changes in gene order are relatively common in some taxa, comparative genomic studies could be coupled with studies of adaptation to explore how commonly such rearrangements are involved in local adaptation.


Myofibroblastoma of the male breast: a rare entity with radiologic-pathologic correlation.
A 73-year old man with a history of multiple genitourinary malignancies was found to have a left retroareolar soft tissue mass on CT assessment of disease, and dedicated breast imaging was recommended. Diagnostic mammography and ultrasonography confirmed a solid mass, for which biopsy was recommended. Pathologic analysis demonstrated a spindle cell neoplasm with an immunoreactivity pattern consistent with myofibroblastoma. While this entity is benign, nonspecific imaging features necessitate tissue sampling for pathologic diagnosis, and, given pathologic rarity, open communication between the radiologist and pathologist is important to establish the correct diagnosis and to recommend appropriate management.


Variation in the lactase dehydrogenase activity of the esophagus.
Quantitative assay and electrophoretic study of lactate dehydrogenase (LDH) from various tissues of the opossum esophagus were performed. On the basis of expression of the LDH isozymes, we concluded that the smooth muscle of the body of the esophagus carry on more anaerobic glycolysis than the striated muscle. The smooth muscle of the gastroesophageal junction carry on both anaerobic as well aerobic glycolysis.


Fluorometric documentation of increased cutaneous blood flow after topical application of a PGE2 analog in man.
The present study employed fiberoptic fluorometry, a noninvasive means of documenting delivery and removal of fluorescein dye, to evaluate the local circulatory changes elicited by topical application of DHV-PGE2 ME, an investigational PGE2 analog. On Day 1, inactive vehicle was applied to a 5 X 4 cm study site on each thigh of healthy volunteer subjects (n = 12). Symmetrical perfusion was confirmed by similar determinations of dye delivery and removal at each site. On Day 2, DHV-PGE2 ME, 30 or 120 micrograms, was applied to one site while inactive vehicle again was applied to the other. After administration of 120 micrograms in a petrolatum vehicle, fluorometry detected a pronounced increase in nutritive perfusion. There was significant acceleration of dye delivery and removal (p less than 0.05 by ANOVA). Less pronounced changes were noted after the lower dose of DHV-PGE2 ME and when the drug was applied in a triethyl citrate vehicle. The local circulatory changes were not accompanied by systemic effects; there were no changes in vital signs or in fluorometric indices at remote sites.


X-ray absorption studies of yeast copper metallothionein.
The local structures of the metal sites in copper metallothionein from Saccharomyces cerevisiae have been investigated by x-ray absorption spectroscopy at the copper and sulfur K edges. Analysis of the EXAFS (extended x-ray absorption fine structure) data indicates that each copper is trigonally coordinated to sulfur at a distance of 2.23 A. Cu-Cu interactions at 2.7 and 3.9 A have also been tentatively identified. Sulfur K edge data are compatible with cysteinyl thiolates bridging each of the eight Cu(I) ions. The data support a model for the copper cluster in yeast metallothionein consisting of a Cu8S12 core. EXAFS data on two specifically engineered carboxyl-terminal truncated mutants reveal that the copper coordination in the mutants is similar to that observed in the wild-type protein.


Intestinal obstruction in patients with previous laparotomy for non-malignancy.
Intestinal obstruction is one of the most common surgical emergencies. The aim of this study was to identify important management information from the evaluation of patients with intestinal obstruction who had undergone previous laparotomy for non-malignancy. Data from 176 patients with previous laparotomy for non-malignancy, and who were operated on for intestinal obstruction, were collected and analyzed retrospectively. Gastroduodenal operations, appendectomy, and obstetric/gynecologic procedures were the 3 most common previous abdominal surgeries. More than half of all bowel obstructions developed within 10 years after previous laparotomy, and particularly within the first 5 years. Most obstructions were related to adhesion, although their etiologies were diverse. The rate of bowel strangulation was much higher in patients with internal herniation, volvulus, intussusception, closed loop, and diaphragmatic hernia than in patients with simple adhesion, bezoar, tumor, and inflammation (48.3% vs 12.2%). The surgical mortality rate correlated significantly with bowel strangulation: the overall rate was 6.8%, that in patients with strangulation was 18.8%, and that in patients without strangulation was 4.2%. The etiologies of intestinal obstruction were not only significantly related to bowel strangulation, but were also an important determinant of therapeutic strategy.


Multiple risks analysis for aplastic anemia in Zhejiang, China: A case-control study.
To understand the risks associated with aplastic anemia (AA) in 4 cities of Zhejiang Province, China, with special focus on the joint contributions of multiple risks.Based on an Electronic Data Capture (EDC), a case control study was carried out. Data regarding socio-demographic, diseases history, living habits, and exposures to toxic substances, etc., were collected through survey questionnaires. t Test, chi-square test, or non-parametric rank sum test, and univariate and multivariate Logistic regression analysis were conducted to analyze data.The univariate logistic regression analysis results indicated that among all study participants (n = 1802), AA was associated with over 30 risks, in terms of their individual behaviors, daily and environmental exposures, diseases history, and family history. Multivariate logistic regression analysis further confirmed that the independent risks related to AA included presence of chemical factory within 3 km of living residence (odds ratio [OR] = 8.73, 95% CI: 1.42-53.74, P = .019), living in a newly decorated house/apartment (OR = 25.37, 95% CI: 4.44-144.81, P < .001), vegetarian diet (OR = 131.60, 95% CI: 3.45-5020.16, P = .009), preference of sugar (OR = 89.38, 95% CI: 7.22-1106.44, P < .001), preference of oily food (OR = 55.68, 95% CI: 5.12-605.26, P = .001), drinking lake water or pond water (OR = 58.05, 95% CI: 3.21-1049.81, P < .001), habit of staying up late (OR = 11.87, 95% CI: 3.43-41.02, P < .001), infection history (OR = 10.08, 95% CI: 2.75-36.93, P < .001). Result of receiver operating characteristic curve (ROC) analysis on the joint contribution of multiple risks indicated that AA was 13.835 times likely to occur when exposed to ≥1 risks than those exposed to 0 risks (95% CI: 9.995-19.149).Our study results demonstrated a comprehensive epidemiological pattern, in which the joint contributions of individual inherited health status, environment exposure, and individual behaviors lead to the occurrence of AA.


[Ligands possessing affinity to specific DNA base pair sequences. IX. Synthesis of netropsin and distamycin A analogs having sarcolysin residues or a platinum(II) atom].
In search for compounds capable of forming covalent bonds with DNA AT-pair clusters, distamycin A and netropsin analogues containing DL-sarcolysin or platinum (II) atom at the N-terminus of the molecule were synthesized, as well as bis-netropsin and bis-distamycin in which two netropsin- or distamycin-like fragments are bound via a cis-diammineplatinum (II) residue. It is shown that these substances can be used for the DNA selective cleavage.


Recognition of band-pass filtered hand-written numerals in foveal and peripheral vision.
The purpose of the present study was to find out what differences between foveal and peripheral pattern recognition remain unexplained by the inhomogeneities of retinal sampling and the optics of the eye. We measured contrast thresholds for pattern recognition at different eccentricities. The effects of retinal sampling were homogenised by using M-scaling of the stimuli, and the effects of the optics of the eye were by-passed either by using strong external noise (signal-to-noise ratio is not affected by optical attenuation) or by computing retinal image contrast by means of the optical modulation transfer function. The stimuli were hand-written numerals filtered to two-octave bands of various centre object spatial frequencies (c/object). The results were described as contrast thresholds and recognition efficiency. At all eccentricities, lowest contrast thresholds and highest recognition efficiencies were found at medium object spatial frequencies. At high object spatial frequencies the peripheral retinal contrast thresholds and recognition efficiencies were nearly as good as at the fovea, but at low object spatial frequencies most of the data showed superiority of the fovea to the periphery. Therefore, at high object spatial frequencies peripheral recognition performance could be explained relatively well by the retinal sampling gradient, or equivalently by the cortical magnification factor, together with the effects of the optics of the eye. Some eccentricity dependent deterioration of recognition at low object spatial frequencies remained unexplained.


Concentration- and rate-dependent electrophysiological effects of restacorin on isolated canine Purkinje fibres.
The cellular electrophysiological effects of restacorin, a new antiarrhythmic agent were studied using conventional microelectrode techniques in isolated dog cardiac Purkinje fibres. Restacorin (1-30 mumol/l) decreased the maximum rate of rise of the action potential upstroke and action potential amplitude while action potential duration measured at 90% of repolarization was shortened in a concentration-dependent manner during pacing at a constant basic cycle length of 500 ms. The effect of 10 mumol/l restacorin on maximal rate of rise of the action potential upstroke and on action potential duration measured at 90% of repolarization were also studied while varying the constant pacing cycle length between 300 and 5000 ms. The results of these studies indicated a rate-dependent effect of restacorin on the action potential characteristics examined. After abrupt changes in cycle length, 10 mumol/l restacorin slowed the fast component of the relation for restitution of action potential duration from 155.3 +/- 5.2 ms (control, n = 6) to 217.1 +/- 17.8 ms (n = 6, P less than 0.05). In the presence of restacorin (10 mumol/l), a second slow component for recovery of maximal action potential upstroke rising velocity was expressed having a time constants of 8.5 +/- 1.2 s. The range of premature action potential durations was significantly decreased (by 57.1%, P less than 0.01) by 10 mumol/l restacorin. These results indicate that the cellular electrophysiological effects produced by restacorin in dog cardiac Purkinje fibres best resemble those produced by recognized class Ic antiarrhythmic drugs.


Standard-b-Value Versus Low-b-Value Diffusion-Weighted Imaging in Hepatic Lesion Discrimination: A Meta-analysis.
We sought to determine the comparative diagnostic performance of standard-b-value (500-1000s/mm) versus low-b-value (≤500 s/mm) diffusion-weighted imaging (DWI) in the discrimination of hepatic lesions. A total of 1775 hepatic malignant lesions and 1120 benign hepatic lesions from 21 studies were included. (1) The global sensitivity was 0.86 (95% confidence interval [CI], 0.847-0.879), the specificity was 0.82 (95% CI, 0.797-0.842), the positive likelihood ratio (PLR) was 6.234 (95% CI, 4.260-9.123), the negative likelihood ratio (NLR) was 0.175 (95% CI, 0.135-0.227), and diagnostic odds ratio (DOR) was 42.836 (95% CI, 24.134-76.031). The area under the curve (AUC) and Q* index were 0.93 and 0.87. Publication bias was not present (P > 0.05). (2)The sensitivity of a subgroup meta-analysis of standard-b-value DWI was 0.858 (95% CI, 0.835-0.880), the specificity was 0.836 (95% CI, 0.807-0.863), the PLR was 6.527 (95% CI, 3.857-11.046), the NLR was 0.168 (95% CI, 0.123-0.239), and the DOR was 49.716 (95% CI, 22.897-107.98). The AUC and Q* index were 0.941 and 0.88. (3)The sensitivity of a subgroup meta-analysis of low-b-value DWI was 0.87 (95% CI, 0.84-0.89), the specificity was 0.80 (95% CI, 0.76-0.83), the PLR was 6.22 (95% CI, 3.29-11.76), the NLR was 0.19 (95% CI, 0.12-0.29), and the DOR was 37.14 (95% CI, 14.80-93.18). The AUC and Q* index were 0.922 and 0.86. Hepatic DWI is useful in differentiating between malignant and benign hepatic lesions. Standard-b-value DWI displayed an overall superior diagnostic accuracy over low-b-value DWI. Further trials needed to determine whether increasing b values beyond 1000 s/mm affects the diagnostic accuracy of hepatic lesion discrimination.


The role of policies in reducing the cost of capital for offshore wind.
Offshore wind will play a critical role in decarbonizing Europe's energy infrastructure. Nevertheless, according to recent financing cost surveys, its investment risk expressed as the cost of capital (CoC) is higher than for onshore wind and solar photovoltaics. This perspective elaborates on the possible reasons behind the offshore wind CoC premium and potential remedies. Our analysis discusses that the massive capital expenditures and construction complexity have concentrated European offshore wind ownership among utilities and oil & gas companies that owing to their legacy investments in fossil fuel infrastructure, have higher return expectations for offshore wind assets. Furthermore, these large-scale investors are bidding zero and negative in highly competitive auctions for offshore wind sites, increasing the project's merchant risks and CoC. We discuss possible policy solutions to alleviate these risks, including revenue stabilization, enabling a more liquid refinancing market, and creating more robust corporate Power Purchase Agreements via government guarantees.


Erythropoietin stimulates growth and STAT5 phosphorylation in human prostate epithelial and prostate cancer cells.
Erythropoietin (Epo), the principal regulator of erythroid progenitor survival, growth, and differentiation, initiates its action by binding to its cognate cell surface receptor (EpoR). EpoR have been identified on a variety of non-hematopoietic cells, both normal and malignant, however, little is known about the function of EpoR on malignant cells. RT-PCR, Western blotting, and immunohistochemistry were used to demonstrate that prostate cancer cells express EpoR at both the gene and protein level. Cell proliferation assays and STAT5 phosphorylation were used to demonstrate Epo's mitogenic action and intracellular signaling, respectively. We have demonstrated that transformed prostate epithelial and prostate cancer cell lines, as well as primary prostate tissue, express the EpoR. Importantly, the EpoR on prostate cells are functional, as demonstrated by the observation that each of the cell lines exhibited a dose-dependent proliferative response to Epo, and that Epo triggered STAT5b phosphorylation in the cells. Human prostatic epithelial cells and prostate cancer cells express functional EpoR, and Epo serves as a growth factor for these cells. These results have implications for our understanding of normal prostatic growth and development and of the pathobiology of human prostate cancer.


European allergen extract units and potency: review of available information.
There is considerable variability in how allergen extract potency is measured and reported worldwide. In Europe, where many sublingual immunotherapy studies have been conducted, manufacturers report allergen extract potency as units based on an in-house reference, making it difficult to understand the exact doses used and to compare studies. To describe the various methods of expressing extract potency that European allergen extract manufacturers use and to gather reports on the micrograms of major allergen of the in-house units of European allergen extract manufacturers. Information was derived from 3 sources: data on extract potency in micrograms of major allergen in articles on sublingual immunotherapy found by PubMed (references through October 2005) and in reference articles, brochures on allergen extracts from the manufacturers, and information provided by structured questionnaires e-mailed to the manufacturers. All but 1 of the European allergen extract manufacturers use in-house reference standards that are based on titrated skin prick testing of allergic patients. Subsequently, in vitro tests compare the potency of commercial batches with the in-house reference and potency is assigned as arbitrary units. Most manufacturers measure major allergen content of their standardized products but do not release this information with the package insert. Diversity in major allergen content was found. Micrograms of major allergens given in articles on sublingual immunotherapy to express the dose administered cannot be used to translate the dose to US extracts. Extract potency can only be compared if uniform test methods and reference extracts are used.


Long-term Effectiveness of Integrated Caries Preventive Programs in Thailand Among Preschool Children Using a System Dynamics Model.
To compare the effect of three caries preventive interventions provided by Thailand's Ministry of Public Health (MOPH): supervised toothbrushing (STB), fluoride varnish (FV), and combined STB+FV from 0 to 5 years old to the base case using the System Dynamics Model (SDM). The SDM was developed to compare the intervention scenarios: STB, FV, and combined STB+FV with the base case scenario. Meta-analyses determined the effective rates of the interventions included in the model. The model indicated that the population with no caries in deciduous teeth increased by 7.5%, 5.7%, and 4.2% at three years of age, and 12.8%, 9.9%, and 5.9% at five years of age, under STB+FV, STB, and FV respectively when compared to the base case. At three years old, the population with untreated caries in deciduous teeth decreased by 12.9% in STB+FV, 9.8% in STB, and 7.3% in FV, and by 10.9% in STB+FV, 8.5% in STB, and 4.9% in FV, at five years old. The increase in the population without caries is considered clinically significant compared to baseline. The model determined that an effective combination of STB and FV should be administered between 0 and 5 years of age to reduce caries in the primary dentition. The SDM could be applied as a "what-if" analysis in order to determine the most beneficial intervention or policy by comparing them.


Radiofrequency catheter ablation of idiopathic right ventricular outflow tract arrhythmias.
Idiopathic ventricular arrhythmias (VA) consist of various subtypes of VA that occur in the absence of clinically apparent structural heart disease. Affected patients account for approximately 10% of all patients referred for evaluation of ventricular tachycardia (VT). Arrhythmias arising from the outflow tract (OT) are the most common subtype of idiopathic VA and more than 70-80% of idiopathic VTs or premature ventricular contractions (PVCs) originate from the right ventricular (RV) OT. Idiopathic OT arrhythmias are thought to be caused by adenosine-sensitive, cyclic adenosine monophosphate (cAMP) mediated triggered activity and, in general, manifest at a relatively early age. Usually they present as salvos of paroxysmal ventricular ectopic beats and are rarely life-threatening. When highly symptomatic and refractory to antiarrhythmic therapy or causative for ventricular dysfunction, ablation is a recommended treatment with a high success rate and a low risk of complications.


Hypothalamic GABA system and plasma corticosterone in ether stressed rats.
Exposure of rats to the ether stress (2 X 2 min within 15 min) activated the hypothalamo-hypophyseal-adrenal (HHA) axis, as evidenced by the increased plasma corticosterone concentration, and affected the hypothalamic GABA system. The aminooxyacetic acid (AOAA) or L-cycloserine-induced accumulation of GABA was decreased, and the activity of glutamate decarboxylase (GAD), the enzyme responsible for GABA synthesis, was increased following ether stress. The concentration of GABA and the activity of GABA: 2-oxoglutarate aminotransferase (GABA-T), the enzyme responsible for GABA catabolism, remained unchanged under given conditions. Diazepam, a drug known to potentiate GABA-ergic transmission, elevated the concentration of plasma corticosterone, but prevented its further increase by ether stress. In spite of the diminished accumulation of GABA, the results might suggest that ether stress activates the hypothalamic GABA system, which is then able to prevent a further response of the HHA axis to stress.


What is harmful for male fertility: cell phone or the wireless Internet?
In this study, we aimed to assess the potential harmful effects of radiofrequency-electromagnetic radiation on sperm parameters. We requested semen for analyses from the male patients coming to our infertility division and also asked them to fill out an anonymous questionnaire. We queried their mobile phone and wireless Internet usage frequencies in order to determine their radiofrequency-electromagnetic radiation exposure. A total of 1082 patients filled the questionnaire but 51 of them were excluded from the study because of azoospermia. There was no significant difference between sperm counts and sperm morphology excluding sperm motility, due to mobile phone usage period, (p = 0.074, p = 0.909, and p = 0.05, respectively). The total motile sperm count and the progressive motile sperm count decreased due to the increase of internet usage (p = 0.032 and p = 0.033, respectively). In line with the total motile sperm count, progressive motile sperm count also decreased with wireless Internet usage compared with the wired Internet connection usage (p = 0.009 and p = 0.018, respectively). There was a negative correlation between wireless Internet usage duration and the total sperm count (r = -0.089, p = 0.039). We have also explored the negative effect of wireless Internet use on sperm motility according to our preliminary results.


Homogalacturonan enriched pectin based hydrogel enhances 6-gingerol's colitis alleviation effect via NF-κB/NLRP3 axis.
A nanolipidcarrier (NLC) loaded homogalacturonan enriched pectin (citrus modified pectin, MCP4) hydrogel was designed as a novel colon inflammation site-specific oral delivery system for 6-gingerol (6G) (6G-NLC/MCP4 hydrogel) administration, and its colitis alleviation effect were investigated. 6G-NLC/MCP4 exhibited typical "cage-like" ultrastructure with 6G-NLC embedded in the hydrogel matrix as observed by cryoscanning electron microscope. And due to the homogalacturonan (HG) domain in MCP4 specifically combined with Galectin-3, which is overexpressed in the inflammatory region, the 6G-NLC/MCP4 hydrogel targeted to severe inflammatory region. Meanwhile, the prolonged-release characteristics of 6G-NLC provided sustained release of 6G in severe inflammatory regions. The matrix of hydrogel MCP4 and 6G achieved synergistic alleviation effects for colitis through NF-κB/NLRP3 axis. Specifically, 6G mainly regulated the NF-κB inflammatory pathway and inhibited the activity of NLRP3 protein, while MCP4 regulated the expression of Galectin-3 and peripheral clock gene Rev-Erbα/β to prevent the activation of inflammasome NLRP3.


Di-butyl phthalate disrupts muscle, motor and sensory neuron development in embryonic zebrafish.
Phthalates are added to plastics to enhance its flexibility, durability and transparency. Phthalates are not covalently bound to plastics and leach into the environment. Phthalates are now pervasive and ubiquitously present in the atmosphere, soil and sediment, surface and wastewater. Phthalates are known endocrine disruptors and their effects on male and female reproduction are well noted. However, studies on the developmental effects of di-butyl phthalate (DBP) are limited. Here we investigated the developmental toxicity of DBP on motor and sensory neuron populations and the muscle structures motor neurons innervate using the zebrafish vertebrate model system. We investigated these effects during the time window of development spanning the period where embryonic patterning determines adult structures. We found that treatment with 2.5 μM DBP from 6 h post-fertilization (hpf) until 72hpf induces loss and disorganization of primary motor neuron innervation of the somatic tissue with concomitant disruptions to muscle fiber organization. Furthermore, we found disruptions to sensory motor neuron development including defects in dorsal root ganglion and their peripherally extending axons. Rohon-Beard sensory neurons were also disrupted showing loss of the bilateral soma positioning along the length of the spinal cord and their afferent axonal projections to the epithelium. Thus, we concluded that DBP is toxic to developing motor and sensory neurons during embryonic development.


Partial deficiency of galactose-1-phosphate uridyltransferase.
In screening programmes testing newborns for galactose-1-phosphate uridyltransferase and/or galactose, partial enzyme deficiency is frequently discovered. This is shown for one laboratory in Switzerland where 104 newborns were singled out from a total of 476,000. Of these, 72 had partial transferase deficiency below 9 mumol/h per g Hb and were assumed to be compound heterozygotes for "classical" galactosemia and the Duarte variant. Present day management of compound heterozygotes consisting of lactose-free diet for 4 months is described and discussed in the light of published opinion. The appropiateness of this pragmatic approach can at present not be judged according to objective criteria.


Beneficial effect of copper supplementation on deposition of fluoride in bone in fluoride- and molybdenum-fed rabbits.
Fluorosis is a major public health problem in the world, including India. The present study was undertaken to investigate the role of molybdenum (Mo) in the deposition of fluoride (F) in bone and whether copper (Cu) supplementation has any alleviating role when F and Mo are ingested together. For this purpose, four groups of rabbits were used [control (C), fluoride (F), fluoride + molybdenum (F + Mo), and fluoride + molybdenum + copper (F + Mo + Cu)] to find out the effect of these treatments on various bone-related parameters like intact parathyroid hormone (iPTH), alkaline phosphatase and Cu in serum, hydroxyproline and calcium (Ca) in urine, and minerals (F, Cu, manganese, and zinc) in femur bone ash. Bone mineral content (BMC), bone mineral density (BMD) [by dual energy X-ray absorptiometry (DXA)], and strength of femur bones were also assessed. F content in the femur was significantly higher (P < 0.01) in all experimental groups compared to control group. Mo supplementation increased F deposition in femur bone in the F + Mo group, whereas supplementation of Cu reduced F deposition in the F + Mo + Cu group compared to the F + Mo and F groups. Levels of Cu in femurs of the F + Mo and F + Mo + Cu groups were significantly higher (P < 0.05, P < 0.01, respectively) than in the C group, although serum Cu was significantly lower in the F and F + Mo than the C and F + Mo + Cu groups. Magnesium levels in the F + Mo group were significantly higher (P < 0.05) than in the F and F + Mo + Cu groups. Cu supplementation in the F + Mo + Cu group increased deposition of zinc significantly (P < 0.05) compared to the F and F + Mo groups. Serum iPTH, alkaline phosphatase, and urinary hydroxyproline and Ca were significantly higher (P < 0.01) in the F and F + Mo than in the C and F + Mo + Cu groups. However, serum iPTH and urinary hydroxyproline were higher in the F + Mo group than the F group. Alkaline phosphatase was significantly higher in the F + Mo group than the F and F + Mo + Cu groups. Levels of serum Cu in the F and F + Mo groups were lower than in the C group, though serum Cu was significantly higher in the F + Mo + Cu than in all other groups. DXA analysis of femur bone indicated that BMD in the F + Mo group was significantly higher than in the F (P < 0.05), C (P < 0.01), and F + Mo + Cu (P < 0.05) groups. However, there was no significant difference in BMC among the groups. Bone strength was significantly higher (P < 0.05) in the F + Mo group than in the C group. Results of the present study show that ingestion of Mo with F does not create secondary Cu deficiency (due to increased excretion of Cu through urine). However, Cu concentration was decreased in serum in this group (F + Mo) compared to the C and F + Mo + Cu groups. Deposition of F in femur bone was more (22%) when it was given along with Mo compared to F alone, while F deposition in femur bone was less in the F + Mo + Cu group by 80% compared to the F group. Also, deposition of F in the F + Mo + Cu group was 120% less compared to the F + Mo group. The increase in F level due to Mo addition appears to be offset by supplementation with Cu. Supplementation with Cu showed a beneficial effect on bone resorption as well as bone formation.


Evidence suggesting that the two forms of heme oxygenase are products of different genes.
Recently, we have reported on the presence of two forms of heme oxygenase in rat liver and testis microsomes, referred to as HO-1 and HO-2 (M. D. Maines, G. M. Trakshel, and R. K. Kutty (1986) J. Biol. Chem. 261, 411-419; G. M. Trakshel, R. K. Kutty, and M. D. Maines (1986) J. Biol. Chem. 261, 11131-11137). Although the two forms differed in several biochemical properties, we could not ascertain whether they represented two isozymes or whether they were isoforms of heme oxygenase. In the present study, we provide evidence suggesting that the two forms are isozymes and represent different gene products. We also provide data suggesting that HO-1 is the commonly known heme oxygenase form. The molecular weight and immunochemical properties of HO-1 and HO-2 did not vary depending on the tissue source examined, i.e. liver and testis. Major differences, however, were noted in the amino acid composition of the two forms including the presence of 3 cysteine/cystine residues in HO-2 only. Using antibody to HO-2, four testis clones and two liver clones were isolated, and one liver and one testis clone were sequenced. Both clones revealed a 274-base-pair insert, and the sequence of both inserts was the same. The validity of assignment was confirmed by matching a 14-amino-acid peptide obtained from purified HO-2 with the sequence. Approximately 43% amino acid homology was detected between the HO-2 insert and the published amino acid sequence of heme oxygenase. However, amino acid homology search revealed the presence of two regions of homology: one 22-mer sequence with only one unmatched amino acid, and one 10-mer sequence with one unmatched amino acid. Heme oxygenase appeared to be the HO-1 form, an assignment based on its amino acid sequence matching the sequence of 2 peptides obtained from purified HO-1 and the immunochemical properties of the cobalt-, hematin-, and bromobenzene-induced rat liver enzyme. The secondary structure prediction analysis revealed an area of 100% structural homology with only 72% sequence homology. We predict this region may represent the catalytic site of the enzyme.


The impact generated by publicly and charity-funded research in the United Kingdom: a systematic literature review.
To identify, synthesise and critically assess the empirical evidence of the impact generated by publicly and charity-funded health research in the United Kingdom. We conducted a systematic literature review of the empirical evidence published in English in peer-reviewed journals between 2006 and 2017. Studies meeting the inclusion criteria were selected and their findings were analysed using the Payback Framework and categorised into five main dimensions, namely knowledge, benefits to future research and research use, benefits from informing policy and product development, health and health sector benefits, and broader economic benefits. We assessed the studies for risk of selection, reporting and funding bias. Thirteen studies met the inclusion criteria. The majority of the studies (10 out of 13) assessed impact at multiple domains including the main five key themes of the Payback Framework. All of them showed a positive impact of funded research on outcomes. Of those studies, one (8%), six (46%) and six (46%) presented a low, moderate and high risk of bias, respectively. Empirical evidence on the impact of publicly and charity-funded research is still limited and subject to funding and selection bias. More work is needed to establish the causal effects of funded research on academic outcomes, policy, practice and the broader economy.


Autologous Cardiac Stem Cell Injection in Patients with Hypoplastic Left Heart Syndrome (CHILD Study).
Mortality in infants with hypoplastic left heart syndrome (HLHS) is strongly correlated with right ventricle (RV) dysfunction. Cell therapy has demonstrated potential improvements of RV dysfunction in animal models related to HLHS, and neonatal human derived c-kit+ cardiac-derived progenitor cells (CPCs) show superior efficacy when compared to adult human cardiac-derived CPCs (aCPCs). Neonatal CPCs (nCPCs) have yet to be investigated in humans. The CHILD trial (Autologous Cardiac Stem Cell Injection in Patients with Hypoplastic Left Heart Syndrome) is a Phase I/II trial aimed at investigating intramyocardial administration of autologous nCPCs in HLHS infants by assessing the feasibility, safety, and potential efficacy of CPC therapy. Using an open-label, multicenter design, CHILD investigates nCPC safety and feasibility in the first enrollment group (Group A/Phase I). In the second enrollment group, CHILD uses a randomized, double-blinded, multicenter design (Group B/Phase II), to assess nCPC efficacy based on RV functional and structural characteristics. The study plans to enroll 32 patients across 4 institutions: Group A will enroll 10 patients, and Group B will enroll 22 patients. CHILD will provide important insights into the therapeutic potential of nCPCs in patients with HLHS.Clinical Trial Registration https://clinicaltrials.gov/ct2/home NCT03406884, First posted January 23, 2018.


Solenopsis invicta transferrin: cDNA cloning, gene architecture, and up-regulation in response to Beauveria bassiana infection.
Transferrin genes from several insects have been shown to be induced in response to bacterial or fungal infection. We were interested to know whether transferrin genes in the red imported fire ant, Solenopsis invicta, are similarly induced by microbial challenge. Hence, the cDNA and structure of a gene exhibiting significant homology to insect transferrins were elucidated for S. invicta. The cDNA was comprised of 2417 nucleotides, excluding the poly(A) tail, with a large open reading frame of 2106 nucleotides. The predicted translation product of the S. invicta tranferrin (SiTf) gene was a 702 amino acid polypeptide with an estimated molecular mass of 77.3 kDa and a pI value of 5.66, characteristics consistent with transferrin proteins. Comparative analysis of genomic and cDNA sequences revealed that the SiTf gene was comprised of 8 exons. Quantitative real-time PCR was used to examine the expression of SiTf. Expression of SiTf was induced in worker ants exposed to Beauveria bassiana conidia. Autoclave-killed conidia did not elicit a SiTf induction response from worker ants. Genes, like SiTf, responding to microbe attack or infection may provide a unique approach to assist in the discovery of microbial control organisms for the target insect pest.


Thin liquid films from aqueous solutions of non-ionic polymeric surfactants.
The conditions of formation and stability of foam, emulsion, and wetting films from aqueous solutions of non-ionic polymeric surfactants have been established. Two types of polymeric surfactants - PEO-PPO-PEO three-block copolymers (A-B-A type) and hydrophobically modified inulin graft polymer (AB(n) type) - have been explored. Information about surface forces and nanoscale phenomena in aqueous films containing polymeric surfactants was obtained using the micro-interferometric technique and the Thin Liquid Film-Pressure Balance Technique. Two types of surface forces, which determine the stability of the foam and emulsion films, have been distinguished, namely: DLVO-forces at low electrolyte concentrations and non-DLVO-forces at high electrolyte concentrations. Non-DLVO-forces are steric surface forces of the brush-to-brush and loop-to-loop interaction type according to De Gennes. A substantial difference in the behavior of these two film types has been established and in the case of O/W emulsion films transitions to Newton black film (NBF) have been observed. These films are very stable and so are the respective emulsions. In contrast the wetting films are relatively thicker compared to emulsion films, and their thickness depends on the concentration of the AB(n) polymeric surfactant. The steric repulsion of the loops and tails of the polymeric surfactant determine the film thickness of wetting films on a hydrophilic solid surface. For solid surfaces with different degrees of hydrophobicity the wetting films are stable only at high polymer concentrations and low degree of hydrophobicity. Otherwise the films are unstable and rupture. Two types of bilayer emulsion films have been distinguished for the first time. One type is related to the brush-to-brush or loop-to-loop interactions according to De Gennes. The other type is a NBF where the forces are also steric between strongly hydrated brush and loops but they are short-range forces acting in a two-dimensional ordered system. They determine the high NBF stability.


Abdominal and miscellaneous applications of intraoperative ultrasound.
Intraoperative sonography is rapidly becoming a key diagnostic tool within the operative suite. Important applications are defined within the abdomen, vascular system, and other regions of the body. Intraoperative scanning may better define the normal anatomy and target pathology than preoperative studies. As with neurosurgical applications of intraoperative ultrasound, with the use of high-resolution, high-frequency real-time transducers, multiple transducer frequencies within one probe, and probe shapes specifically designed for certain types of operations, it is anticipated that the surgical use of intraoperative ultrasound will greatly increase in the future.