Chimerism, point mutation, and truncation dramatically transformed mast cell delta-tryptases during primate evolution.
Tryptases are serine peptidases stored in mast cell granules. Rodents express 2 soluble tryptases, mast cell proteases (MCPs) 6 and 7. Human alpha- and beta-tryptases are orthologs of MCP-6. However, much of the ancestral MCP-7 ortholog was replaced by parts of other tryptases, creating chimeric delta-tryptase. Human delta-tryptase's limited activity is hypothesized to be due to truncation and processing mutations. We sought to probe the origins and consequences of mutations in primate delta-tryptases. Prosimian (lemur), monkey (macaque), great ape (orangutan, gorilla, and chimpanzee), and human delta-tryptase genes were identified by means of data mining and genomic sequencing. Resulting genes were analyzed phylogenetically and structurally. The seminal conversion event generating the delta-tryptase chimera occurred early because all primates studied contain delta-tryptase genes. Truncation, resulting from a nonsense mutation of Trp206, occurred much later, after orangutans and other great apes last shared an ancestor. The Arg-3Gln propeptide mutation occurred most recently, being present in humans and chimpanzees but not in other primates. Surprisingly, the major active tryptase in monkeys is full-length delta-tryptase, not beta-tryptase, which is the main active tryptase in human subjects. Models of macaque delta-tryptase reveal that the segment truncated in human subjects contains antiparallel beta-strands coursing through the substrate-binding cleft, accounting for truncation's drastic effect on activity. Transformations in the ancestral MCP-7-like gene during primate evolution caused dramatic variations in function. Although delta-tryptases are nearly inactive in humans, they are active and dominant in monkeys.


Phosphorus as a limiting factor on sustainable greywater irrigation.
Water reuse through greywater irrigation has been adopted worldwide and has been proposed as a potential sustainable solution to increased water demands. Despite widespread adoption, there is limited domestic knowledge of greywater reuse. There is no pressure to produce low-level phosphorus products and current guidelines and legislation, such as those in Australia, may be inadequate due to the lack of long-term data to provide a sound scientific basis. Research has clearly identified phosphorus as a potential environmental risk to waterways from many forms of irrigation. To assess the sustainability of greywater irrigation, this study compared four residential lots that had been irrigated with greywater for four years and adjacent non-irrigated lots that acted as controls. Each lot was monitored for the volume of greywater applied and selected physic-chemical water quality parameters and soil chemistry profiles were analysed. The non-irrigated soil profiles showed low levels of phosphorus and were used as controls. The Mechlich3 Phosphorus ratio (M3PSR) and Phosphate Environmental Risk Index (PERI) were used to determine the environmental risk of phosphorus leaching from the irrigated soils. Soil phosphorus concentrations were compared to theoretical greywater irrigation loadings. The measured phosphorus soil concentrations and the estimated greywater loadings were of similar magnitude. Sustainable greywater reuse is possible; however incorrect use and/or lack of understanding of how household products affect greywater can result in phosphorus posing a significant risk to the environment.


Effect of sublingual misoprostol on severe postpartum haemorrhage in a primary health centre in Guinea-Bissau: randomised double blind clinical trial.
To evaluate whether routine administration of sublingual misoprostol 600 microg after delivery reduces postpartum haemorrhage. Randomised double blind placebo controlled trial. Primary health centre in Bissau, Guinea-Bissau, West Africa. 661 women undergoing vaginal delivery. Misoprostol 600 mug or placebo administered sublingually immediately after delivery. Postpartum haemorrhage, defined as a loss of > or = 500 ml and decrease in haemoglobin concentration after delivery. The incidence of postpartum haemorrhage was not significantly different between the two groups, the relative risk being 0.89 (95% confidence interval 0.76 to 1.04) in the misoprostol group compared with the placebo group. Mean blood loss was 10.5% (-0.5% to 20.4%) lower in the misoprostol group than in the control group. Severe postpartum haemorrhage of > or = 1000 ml or > or = 1500 ml occurred in 17% (56) and 8% (25) in the placebo group and 11% (37) and 2% (7) in the misoprostol group. Significantly fewer women in the misoprostol group experienced a loss of > or = 1000 ml (0.66, 0.45 to 0.98) or > or = 1500 ml (0.28, 0.12 to 0.64). The decrease in haemoglobin concentration tended to be less in the misoprostol group, the mean difference between the two groups being 0.16 mmol/l (-0.01 mmol/l to 0.32 mmol/l). Sublingual misoprostol reduces the frequency of severe postpartum haemorrhage.


Involvement of CD44, a molecule with a thousand faces, in cancer dissemination.
Tumor progression is substantially dependent on network of multiple factors, including adhesion and homing molecules, which support the malignant metastatic spread. CD44, one of the adhesion/homing molecules, has attracted much attention not only because it is expressed on many types of tumors, but also owing to its numerous functions, such as supporting cell migration and transmitting survival signals, thereby being pro-oncogenic by nature. We have used the mouse malignant LB lymphoma cell line as a model for comprehensive in vitro and in vivo analyses of the interaction between CD44 and hyaluronic acid (HA), and its relevance to tumor dissemination. The in vitro studies revealed that LB cells could not bind HA, either under static or dynamic (i.e., shear flow) conditions, unless their CD44 is activated by phorbol ester, deglycosylated (to increase the CD44 positive net charge) or transfected with CD44 variants. In parallel, in vivo experiments showed that LB cell dissemination could be controlled by injection of anti-CD44 monoclonal antibodies or hyaluronidase. Furthermore, LB cells transfected with CD44v4-v10 variant, rather than standard CD44, displayed enhanced invasion of the peripheral lymph nodes. This effect was completely lost if the HA binding site of CD44 were mutated. LB cell accumulation in the lymph nodes is caused by enhanced migration via the afferent lymphatics rather than by accelerated proliferation within the lymph node. This information can be exploited to tailor a "therapeutic suit" that should be maximally effective in inducing tumor resistance, while minimizing destructive side effects.


Clinical outcomes with a new microincisional diffractive multifocal IOL.
To evaluate the refractive outcomes and the optical performance as well as the quality of life in patients implanted with a new diffractive multifocal intraocular lens (IOL). Prospective, clinical study including 41 cases of patients who underwent cataract surgery and were divided in two groups: group 1, including 20 eyes implanted with the multifocal IOL SeeLens MF (Hanita Lenses, Israel); group 2, 21 eyes implanted with the Acrysof SA60AT IOL. Visual acuity, defocus curve, intraocular aberrations, contrast sensitivity function and quality of life were assessed during a follow up period of 6 months. Significant improvement was observed in the uncorrected distance visual acuity (UDVA) and corrected distance visual acuity (CDVA) in both groups (p < 0.02). The multifocal group showed better results in terms of uncorrected near visual acuity (UNVA) and distance-corrected near visual acuity (DCNVA) (p < 0.01). Comparison of both groups showed better visual acuities for the multifocal IOL group in defocus levels from -3.0 D to -1.50 D (p ≤ 0.01). At 6 months, there was a significant reduction of the internal higher order aberrations (p ≤ 0.04). A significant increase in scotopic contrast sensitivity was detected for 6 cycles/° spatial frequency during follow up (p = 0.04), but no significant changes were observed for the rest of spatial frequencies (p ≥ 0.06). Visual Functioning Index (VF-14) questionnaire showed that patients reported high levels of satisfaction when performing daily tasks. The SeeLens MF IOL is able to successfully restore distance, near and intermediate visions after cataract surgery. It also provides functional intermediate vision with optimal intraocular optical quality.


Thyroid hormone potentiates estrogen activation of vitellogenin genes and autoinduction of estrogen receptor in adult Xenopus hepatocytes.
Although the important role of thyroid hormones in regulating metamorphosis of amphibian larvae is well known, it has not been clearly established if thyroid hormones have any function in the activities of adult amphibian tissues. We now describe a strong effect of 3,3',5-triiodothyronine (T3) on adult Xenopus liver cells. Low doses of T3 rapidly (within 6-12 h) potentiate the activation of vitellogenin (Vit) genes by estradiol-17 beta (E2) in primary cultures of adult male and female Xenopus hepatocytes. This effect is developmentally regulated and is first manifested during metamorphic climax. In an attempt to explain this potentiation, we find that T3 also upregulates thyroid hormone receptor beta, but not alpha, transcripts and rapidly enhances the autoinduction of estrogen receptor (ER) mRNA in adult Xenopus hepatocytes. In transient transfection of the Xenopus cell line XTC-2 with an estrogen response element--chloramphenicol transacetylase (ERE-CAT) construct T3 was found to potentiate the transcription by E2 from the transfected ERE, thus suggesting that it enhances the accumulation of functional ER. We conclude that T3 can function in adult amphibian tissues, and discuss the significance of thyroid hormone potentiation of responses to estrogen in reproductive processes.


Prevalence of Hepatitis B Virus infection in the Gulf Cooperation Council: a systematic review and meta-analysis.
Hepatitis B virus (HBV) infection is a global public-health problem. Since the introduction of an effective vaccine, the epidemiology of HBV infection is changing. We aimed to estimate the prevalence of HBV infection in the Gulf Cooperation Council (GCC) region and delineate any variation in member-countries, special sub-groups, and over time. This is a systematic review and meta-analysis to review studies of HBV prevalence in the GCC region. Databases were searched and all studies from inception to July 31st, 2021, were considered for inclusion. The pooled HBV prevalence was analyzed using the random-effect model after assessment for heterogeneity. True prevalence was adjusted using the Rogan-Gladen estimator. Pre-defined subgroup analysis was performed, and publication bias was assessed. Overall, 99 studies (n = 1,944,200 participants) met the inclusion criteria. The overall HBV apparent prevalence was 3.05% (95% CI 2.60, 3.52) and the true prevalence was 1.67% (95% CI 1.66, 1.68). The apparent prevalence varied between subgroups. Over time, the apparent prevalence of HBV infection has declined from 9.38% (95% CI 7.26, 11.74) before 1990 to 1.56% (95% CI 1.07, 2.12) during the period 2010 to 2020. Over the last four decades the overall prevalence of HBV infection in the GCC region has decreased from high- to low-endemicity level. However, due to poor methodology of the included studies, further high-quality community-based studies are needed to obtain more precise estimate of HBV infection in this region.


[Critical limb ischemia in systemic sclerosis].
Vascular complications are common in systemic sclerosis (SSc). Critical limb ischemia leading to gangrene or amputation occurs in more than 10% of these patients and hence is a common emergency. This report highlights the different pathogenetic mechanisms leading to critical ischemic events and provides guidance for the diagnosis and therapy. Apart from SSc-associated vasculopathy and peripheral arterial disease, thromboembolic events and rarely also vasculitis may cause critical limb ischemia. An interdisciplinary approach to the diagnosis and therapy of these lesions is mandatory. Therapy goals are the prevention of further ischemia and, if possible, revascularization as well as optimal pain management.


Immunogenicity of meningococcal ACYW135 polysaccharide vaccine in Saudi children 5 to 9 years of age.
Meningococcal tetravalent polysaccharide vaccines were observed to be immunogenic in Saudi children 5 to 9 years of age, with >90% having serum bactericidal antibody titers of > or = 8 for serogroups A, Y, and W135; for serogroup C, 77% were putatively protected after vaccination.


Challenges and Opportunities of Genomic Approaches in Therapeutics Development.
The magnitude of all therapeutic responses is significantly determined by genome structure, variation, and functional interactions. This determination occurs at many levels which are discussed in the current review. Well-established examples of structural variation between individuals are known to dictate an individual's response to numerous drugs, as clearly illustrated by warfarin. The exponential rate of genomic-based interrogation is coupled with an expanding repertoire of genomic technologies and applications. This is leading to an ever more sophisticated appreciation of how structural variation, regulation of transcription and genomic structure, both individually and collectively, define cell therapeutic responses.


Iron Deficiency in Attention-Deficit Hyperactivity Disorder, Autism Spectrum Disorder, Internalizing and Externalizing Disorders, and Movement Disorders.
This article reviews the role of iron in brain development and function, with a focus on the association between iron deficiency (ID) and neuropsychiatric conditions. First, we describe how ID is defined and diagnosed. Second, the role of iron in brain development and function is summarized. Third, we review current findings implicating ID in a number of neuropsychiatric conditions in children and adolescents, including attention deficit hyperactivity disorder and other disruptive behavior disorders, depressive and anxiety disorders, autism spectrum disorder, movement disorders, and other situations relevant to mental health providers. Last, we discuss the impact of psychotropic medication on iron homeostasis.


A micro liquid column chromatography procedure for twelve anticonvulsants and some of their metabolites.
Solvent extracts of 50 microliter volumes of serum were sufficient for the determination of twelve anticonvulsant drugs. The liquid chromatography procedure utilized a C-18 reversed-phase column and isocratic elution with 15% acetonitrile in water. No derivatization was required. Eluted anticonvulsants were detected by UV absorption at 195 nm and quantitated by drug--internal standard peak area ratios. The procedure provided linear working curves over the concentration range from 1 to 100 mg/l of drug in the serum. The procedure for serum provided recoveries of the drugs from 92 to 101%. Within-day precision was about 4% and day-to-day precision was about 6.5%. The procedures has been applied to urine samples to facilitate bioavailability studies. Data are also given for several metabolites. There is a discussion of many practical aspects of the procedure to improve the reliability of the results.


Spatial interactions reveal inhibitory cortical networks in human amblyopia.
Humans with amblyopia have a well-documented loss of sensitivity for first-order, or luminance defined, visual information. Recent studies show that they also display a specific loss of sensitivity for second-order, or contrast defined, visual information; a type of image structure encoded by neurons found predominantly in visual area A18/V2. In the present study, we investigate whether amblyopia disrupts the normal architecture of spatial interactions in V2 by determining the contrast detection threshold of a second-order target in the presence of second-order flanking stimuli. Adjacent flanks facilitated second-order detectability in normal observers. However, in marked contrast, they suppressed detection in each eye of the majority of amblyopic observers. Furthermore, strabismic observers with no loss of visual acuity show a similar pattern of detection suppression. We speculate that amblyopia results in predominantly inhibitory cortical interactions between second-order neurons.


Usefulness of prior hysterectomy as an independent predictor of Framingham risk score (The Women's Health Initiative).
The association of hysterectomy with increased coronary risk is controversial, and previous studies have reached differing conclusions as to whether the excess risk is confined to women who have also undergone bilateral oophorectomy. This analysis uses the Framingham algorithm to evaluate the hypothesis that hysterectomy with or without ovarian preservation is associated with increased coronary risk, using a cross-sectional analysis of baseline data from 1,501 participants of the Women's Health Initiative. Framingham risk scores, derived from the algorithm in the National Cholesterol Education Program Adult Treatment Panel III guidelines, which include age, smoking, systolic blood pressure, total and high-density lipoprotein cholesterol, were determined in a subgroup of Women's Health Initiative participants with measured plasma lipids and known ovariectomy status. Women with hysterectomy had fewer years of education than those without hysterectomy (30% with college degree vs 41%, p <0.0001) and higher body mass index (29 vs 28 kg/m(2), p <0.0001), consumed less alcohol, exercised less, and had a higher Framingham risk of myocardial infarction or coronary death (46% vs 41% with 10-year risk >/=4%, p = 0.04). In multivariate analysis, hysterectomy with bilateral oophorectomy was an independent predictor of Framingham risk (p = 0.04), whereas hysterectomy with ovarian preservation was not.


Design, synthesis and computational evaluation of a novel intermediate salt of N-cyclohexyl-N-(cyclohexylcarbamoyl)-4-(trifluoromethyl) benzamide as potential potassium channel blocker in epileptic paroxysmal seizures.
The narrow therapeutic range and limited pharmacokinetics of available Antiepileptic drugs (AEDs) have raised serious concerns in the proper management of epilepsy. To overcome this, the present study attempts to identify a candidate molecule targeting voltage gated potassium channels anticipated to have superior pharmacological than existing potassium channel blockers. The compound was synthesized by reacting (S)-(+)-2,3-dihydro-1H-pyrrolo[2,1-c][1,4] benzodiazepine5,11(10H,11aH)-dione with 4-(Trifluoromethyl) benzoic acid (C8H5F3O2) in DMF and N,N'-dicyclohexylcarbodiimide (DCC) which lead to the formation of an intermediate salt of N-cyclohexyl-N-(cyclohexylcarbamoyl)-4-(trifluoromethyl)benzamide with a perfect crystalline structure. The structure of the compound was characterized by FTIR, 1H NMR and 13C NMR analysis. The crystal structure is confirmed by single crystal X-ray diffraction analysis. The Structure-Activity Relationship (SAR) studies revealed that substituent of fluoro or trifluoromethyl moiety into the compound had a great effect on the biological activity in comparison to clinically used drugs. Employing computational approaches the compound was further tested for its affinity against potassium protein structure by molecular docking in addition, bioactivity and ADMET properties were predicted through computer aided programs.


Treatment of renal carcinoma.
This paper examines diagnostic procedures for renal carcinoma and outlines surgical interventions, pre- and post-operative care and possible complications.


Present state bias in transition ratings was accurately estimated in simulated and real data.
Patient-reported transition ratings are supposed to reflect the change between a previous baseline health state and a present follow-up state, but may reflect the present state to a greater extent. This so-called "present state bias" (PSB) potentially threatens the validity of transition ratings. Several criteria have been proposed to assess PSB. We examined how well these criteria perform and to which extent confirmatory factor analysis (CFA) for categorical data provides an accurate assessment of the degree of PSB. We simulated multiple samples with baseline and follow-up item responses to a hypothetical questionnaire, and transition ratings. The samples varied with respect to various distributional characteristics and the degree of PSB. The performance of criteria proposed in the literature, and a new CFA-based criterion, were evaluated by the proportion of explained variance in PSB. In addition, four real datasets were analyzed. The known criteria explained 36-74% of the variance in PSB. A new CFA-based criterion, namely the ratio of the factor loadings of the transition ratings plus one, explained 81-98% of the variance in PSB across the samples. Present state bias in transition ratings can be estimated accurately using CFA.


Measurements of Exocytosis by Capacitance Recordings and Calcium Uncaging in Mouse Adrenal Chromaffin Cells.
Fusion of vesicles with the plasma membrane and liberation of their contents is a multistep process involving several proteins. Correctly assigning the role of specific proteins and reactions in this cascade requires a measurement method with high temporal resolution. Patch-clamp recordings of cell membrane capacitance in combination with calcium measurements, calcium uncaging, and carbon-fiber amperometry allow for the accurate determination of vesicle pool sizes, their fusion kinetics, and their secreted oxidizable content. Here, we will describe this method in a model system for neurosecretion, the adrenal chromaffin cells, which secrete adrenaline.


Inherited pulmonary surfactant metabolism disorders in Argentina: Differences between patients with SFTPC and ABCA3 variants.
Patients with inherited pulmonary surfactant metabolism disorders have a wide range of clinical outcomes and imaging findings. Response to current anti-inflammatory therapies has been variable and efficacy is unclear. To describe and compare genetic, clinical, histological, and computed tomography (CT) outcomes in a cohort of patients with variants in the genes encoding surfactant protein C (SP-C) or adenosine triphosphate-binding cassette transporter A3 (ABCA3) in Argentina. Observational cohort retrospective study. Patients carrying variants in genes encoding SP-C and ABCA3 proteins were included. Fourteen patients met the inclusion criteria: SFTPC n = 6, ABCA3 n = 8 (seven were heterozygous and one compound heterozygous). Neonatal respiratory distress was more frequent and severe in neonates with variants in the ABCA3 gene. The onset of the disease occurred in infancy before the age of 20 months in all cases. Patients with ABCA3 pathogenic variants had a severe clinical course, while long-term outcomes were more favorable in individuals with SFTPC variants. Initial CT findings were ground glass opacities and intraparenchymal cysts in both groups. Over time, signs of lung fibrosis were present in 57% of patients with ABCA3 variants and in 33% of the SFTPC group. The efficacy of anti-inflammatory interventions appears to be poor, especially for patients with ABCA3 pathogenic variants. Clinical, histological, and radiological features are similar in patients with SFTPC and ABCA3 variants; however, the latter have more severe clinical course. Current anti-inflammatory regimens do not appear to stop the progression of the disease.


Substrate specificity screening of oat (Avena sativa) seeds aminopeptidase demonstrate unusually broad tolerance in S1 pocket.
Aminopeptidases are proteolytic enzymes that remove one amino acid at a time from N-terminus of peptidic substrates. In plants, inhibitors of aminopeptidases can find potential applications in agriculture as herbicides. In this report we have used a library of fluorogenic derivatives of natural and unnatural amino acids for substrate specificity profiling of oat (Avena sativa) aminopeptidase. Interestingly, we have found that this enzyme recognizes effectively among the natural amino acids basic residues like Arg and Lys, hydrophobic Phe, Leu and Met, but also to some extent acidic residues Asp and Glu. In the case of unnatural amino acids hydrophobic residues (hPhe and hCha) and basic hArg were preferentially recognized.


ACTH 4-10 in the amelioration of neuropsychological symptomatology associated with senile organic brain syndrome.
Eighteen male and female volunteers over the age of sixty who exhibited mild senile organic brain syndrome were administered ACTH 4-10 (Org OI 63) (30 mg, s.c.) or saline in a 2 X 2 Latin square design. Subjects experienced a reduction in depression and confusion and an increase in vigor. This evidence of an increase in vigor was supported behaviorally by a delay in the onset of increased latency in reaction time. Data also indicated that retrieval from memory may be enhanced by this compound. The electroencephalogram evinced a shift to lower frequencies under ACTH 4-10, but this effect was primarily noted in the females who received drug followed by placebo. These effects of ACTH 4-10 are intriguing and suggest that further work in this area should be encouraged.


A method to correct for the influence of bovine serum albumin-associated vitamin D metabolites in protein extracts from neonatal dried blood spots.
We developed an assay to measure the concentration of 25 hydroxyvitamin D2 and D3 in protein extracts derived from stored neonatal dried blood spots. During this study, we postulated that these samples had been contaminated with exogenous vitamin D metabolites because of the addition of bovine serum albumin (BSA) as part of an extraction step undertaken 7 years earlier. The aim of the current study was to develop methods in order to adjust for this contamination. We identified between-plate variations in 25 hydroxyvitamin D2 and D3 concentrations which suggested the presence of three different BSA batches. Based on repeat extraction (without the addition of BSA) and testing of 395 samples, we developed models to correct for the exogenous 25 hydroxyvitamin D2 and D3. The regression models were Diff25OHD3 = - 8.2 + 1.8* Diff25OHD2 for low contamination, Diff25OHD3 = 23.8 + 1.7* Diff25OHD2 for middle contamination, and Diff25OHD3 = 14.3 + 3.0* Diff25OHD2 for high contamination. After these corrections, the three subsamples had comparable distributions within the expected range for both 25 hydroxyvitamin D2 and D3.


Vardenafil: update on clinical experience.
Vardenafil, a potent and selective phosphodiesterase 5 inhibitor, has entered phase 3 clinical trials. Pharmacodynamic studies showed that the maximum plasma concentration after oral administration of 20-40 mg of vardenafil occurred in 0.7-0.9 h, the half-life was 4-5 h, and negligible amounts remained in the circulation after 24 h. The efficacy of vardenafil compared with placebo was shown in RigiScan studies, a phase 2 study involving 601 men with mild-to-severe erectile dysfunction for at least 6 months, and a phase 3 study involving 452 diabetic men. Adverse effects were not severe and tended to decrease with time.


Laparoscopic versus open appendicectomy in pregnancy: a systematic review.
Acute appendicitis is the most common non-obstetric indication for surgical intervention in pregnant women. The benefits of a laparoscopic over an open approach to appendicectomy are well established in the non-pregnant population. Data on the optimal surgical approach to acute appendicitis in pregnant women are conflicting. A systematic review of reported cases of laparoscopic appendicectomy (LA) in pregnancy over the period 1990 to 2007. Twenty-eight articles documenting 637 cases of LA in pregnancy were included. Data on pregnancy outcome, patient characteristics, operative technique and peri-operative complications were analysed. The rate of fetal loss following LA in pregnancy approaches 6% and is significantly higher than that following open appendicectomy. Fetal loss was highest in cases of complicated appendicitis. Incidence of preterm delivery appears lower in the LA group although this complication is likely to be under-reported in a significant proportion of cases. Trimester at the time of LA does not appear to influence complication rates. The negative appendicectomy rate in this series was 27%, which is higher than in the non-pregnant population. Complication rates following LA with negative appendicitis are as high as with simple appendicitis. Rates of entry-related complications were 2.8% in the Veress needle group and 0% in the Hasson open entry group. The overall rate of conversion to laparotomy was 1%. No difference was found in the preterm delivery rate between women who received prophylactic tocolysis and those who were not tocolysed. Laparoscopic appendicectomy in pregnancy is associated with a low rate of intra-operative complications in all trimesters. However, LA in pregnancy is associated with a significantly higher rate of fetal loss compared to open appendicectomy. Rates of preterm delivery appear similar or slightly better following a laparoscopic approach. Open appendicectomy would appear to be the safer option for pregnant women for whom surgical intervention is indicated.


HhAntag, a Hedgehog Signaling Antagonist, Suppresses Chondrogenesis and Modulates Canonical and Non-Canonical BMP Signaling.
Chondrogenesis subtends the development of most skeletal elements and involves mesenchymal cell condensations differentiating into growth plate chondrocytes that proliferate, undergo hypertrophy, and are replaced by bone. In the pediatric disorder Hereditary Multiple Exostoses, however, chondrogenesis occurs also at ectopic sites and causes formation of benign cartilaginous tumors--exostoses--near the growth plates. No treatment is currently available to prevent or reverse exostosis formation. Here, we asked whether chondrogenesis could be stopped by targeting the hedgehog pathway, one of its major regulators. Micromass cultures of limb mesenchymal cells were treated with increasing amounts of the hedgehog inhibitor HhAntag or vehicle. The drug effectively blocked chondrogenesis and did so in a dose-dependent manner as monitored by: alcian blue-positive cartilage nodule formation; gene expression of cartilage marker genes; and reporter activity in Gli1-LacZ cell cultures. HhAntag blocked chondrogenesis even when the cultures were co-treated with bone morphogenetic protein 2 (rhBMP-2), a strong pro-chondrogenic factor. Immunoblots showed that HhAntag action included modulation of canonical (pSmad1/5/8) and non-canonical (pp38) BMP signaling. In cultures co-treated with HhAntag plus rhBMP-2, there was a surprising strong up-regulation of pp38 levels. Implantation of rhBMP-2-coated beads near metacarpal elements in cultured forelimb explants induced formation of ectopic cartilage that however, was counteracted by HhAntag co-treatment. Collectively, our data indicate that HhAntag inhibits not only hedgehog signaling, but also modulates canonical and non-canonical BMP signaling and blocks basal and rhBMP2-stimulated chondrogenesis, thus representing a potentially powerful drug-based strategy to counter ectopic cartilage growth or induce its involution.


Selection criteria of residents for residency programs in Kuwait.
In Kuwait, 21 residency training programs were offered in the year 2011; however, no data is available regarding the criteria of selecting residents for these programs. This study aims to provide information about the importance of these criteria. A self-administered questionnaire was used to collect data from members (e.g. chairmen, directors, assistants …etc.) of residency programs in Kuwait. A total of 108 members were invited to participate. They were asked to rate the importance level (scale from 1 to 5) of criteria that may affect the acceptance of an applicant to their residency programs. Average scores were calculated for each criterion. Of the 108 members invited to participate, only 12 (11.1%) declined to participate. Interview performance was ranked as the most important criteria for selecting residents (average score: 4.63/5.00), followed by grade point average (average score: 3.78/5.00) and honors during medical school (average score: 3.67/5.00). On the other hand, receiving disciplinary action during medical school and failure in a required clerkship were considered as the most concerning among other criteria used to reject applicants (average scores: 3.83/5.00 and 3.54/5.00 respectively). Minor differences regarding the importance level of each criterion were noted across different programs. This study provided general information about the criteria that are used to accept/reject applicants to residency programs in Kuwait. Future studies should be conducted to investigate each criterion individually, and to assess if these criteria are related to residents' success during their training.


Effects of dietary saturated, N-6 and N-3 polyunsaturated fats on blood pressure and prostaglandins outflow from perfused mesenteric vascular bed in rats.
Effects of the dietary administration of saturated fat and of n-6 and n-3 polyunsaturates on blood pressure, prostaglandin metabolism in small vessels, tissue fatty acid distribution and urinary PGE2 excretion were compared. Rats were divided into three groups. Diets contained 10% hydrogenated coconut oil (HCO), 10% safflower oil (SFO) or 10% cod liver oil (CLO) added to a basic fat free diet for 10 weeks. Systolic blood pressure was increased in the CLO group animals. Urinary PGE2 excretion was decreased in the HCO and CLO groups as compared to that in the SFO group animals. PGE2, 6-keto-PGF1 alpha and thromboxane (Tx) B2 outflow from isolated perfused mesenteric arterial beds were extremely decreased in the CLO group animals, and to a lesser extent in the HCO group as compared to the SFO animals. In the tissue phospholipid, 20:3n-9/20:4n-6 ratios were increased in the HCO group indicating essential fatty acid deficiency, and n-6 and n-3 polyunsaturates were elevated in the SFO and the CLO group animals respectively. Arachidonic acid concentration was highest in the SFO group, while there was no significant differences between the HCO and the CLO group. These results suggest that dietary fatty acid manipulation affects urinary PGE2 excretion and PGI2, PGE2 and TxA2 synthesis in mesenteric arterial beds and also changes the tissue fatty acid distribution. Furthermore, n-3 polyunsaturates caused an extreme reduction of 2-series PGs synthesis in small resistance vessels.


Degree of expression of inflammatory proteins in the amniotic cavity, but not prior obstetric history, is associated with the risk severity for spontaneous preterm birth after rescue cerclage for cervical insufficiency.
To examine whether the severity of spontaneous preterm birth (SPTB) risk after rescue cerclage for acute cervical insufficiency (CI) is linked to the degree of inflammatory response in the amniotic fluid (AF) based on the concentrations of various inflammatory proteins and prior obstetric history. We conducted a retrospective cohort study of 65 singleton pregnant women (17-25 weeks) who underwent rescue cerclage following the diagnosis of acute CI and were subjected to amniocentesis. EN-RAGE, IL-6, IL-8, and IP-10 as inflammatory mediators and kallistatin, MMP-2/8, and uPA as extracellular matrix remodeling-related molecules were assayed in the AF using ELISA. The level of each inflammatory mediator was divided into quartiles. Intra-amniotic inflammation (IAI; AF IL-6 level ≥2.6 ng/mL) was independently associated with SPTB after cerclage placement. The odds of SPTB at < 32 weeks, even after adjusting for confounders, increased significantly with each increasing quartile of baseline AF levels for each inflammatory mediator (p for trend < .05). Kaplan-Meier survival curves showed that the cerclage-to-delivery intervals were significantly shorter as the quartiles of AF EN-RAGE and MMP-8 increased (log-rank test, p < .01 each). Neither previous term birth nor prior PTB was associated with SPTB risk or cerclage-to-delivery interval after rescue cerclage. Multiparous women who experience CI after term birth showed significantly elevated levels of MMP-8 and reduced kallistatin levels in the AF. In patients with CI, SPTB risk (especially risk severity) after rescue cerclage is associated with the degree of the inflammatory response in AF as well as the presence of IAI but not with prior obstetric history.


Perinatal risk factors for pulmonary hemorrhage in extremely low-birth-weight infants.
Pulmonary hemorrhage (PH) is a life-threatening respiratory complication of extremely low-birth-weight infants (ELBWIs). However, the risk factors for PH are controversial. Therefore, the purpose of this study was to analyze the perinatal risk factors and short-term outcomes of PH in ELBWIs. This was a retrospective cohort study of live born infants who had birth weights that were less than 1000 g, lived for at least 12 hours, and did not have major congenital anomalies. A logistic regression model was established to analyze the risk factors associated with PH. There were 168 ELBWIs born during this period. A total of 160 infants were included, and 30 infants were diagnosed with PH. Risk factors including gestational age, small for gestational age, intubation in the delivery room, surfactant in the delivery room, repeated use of surfactant, higher FiO2 during the first day, invasive ventilation during the first day and early onset sepsis (EOS) were associated with the occurrence of PH by univariate analysis. In the logistic regression model, EOS was found to be an independent risk factor for PH. The mortality and intraventricular hemorrhage rate of the group of ELBWIs with PH were significantly higher than those of the group of ELBWIs without PH. The rates of periventricular leukomalacia, moderate-to-severe bronchopulmonary dysplasia and severe retinopathy of prematurity, and the duration of the hospital stay were not significantly different between the PH and no-PH groups. Although PH did not extend hospital stay or increase the risk of bronchopulmonary dysplasia, it increased the mortality and intraventricular hemorrhage rate in ELBWIs. EOS was the independent risk factor for PH in ELBWIs.


Effects of isomalt consumption on gastrointestinal and metabolic parameters in healthy volunteers.
The polyol isomalt (Palatinit) is a well established sugar replacer. The impact of regular isomalt consumption on metabolism and parameters of gut function in nineteen healthy volunteers was examined in a randomised, double-blind, cross-over trial with two 4-week test periods. Volunteers received 30 g isomalt or 30 g sucrose daily as part of a controlled diet. In addition to clinical standard diagnostics, biomarkers and parameters currently discussed as risk factors for CHD, diabetes or obesity were analysed. Urine and stool Ca and phosphate excretions were measured. In addition, mean transit time, defecation frequency, stool consistency and weight were determined. Consumption of test products was affirmed by the urinary excretion of mannitol. Blood lipids were comparable in both phases, especially in volunteers with hyperlipidaemia, apart from lower apo A-1 (P=0.03) for all subjects. Remnant-like particles, oxidised LDL, NEFA, fructosamine and leptin were comparable and not influenced by isomalt. Ca and phosphate homeostasis was not affected. Stool frequency was moderately increased in the isomalt phase (P=0.006) without changes in stool consistency and stool water. This suggests that isomalt is well tolerated and that consumption of isomalt does not impair metabolic function or induce hypercalciuria. In addition, the study data indicate that isomalt could be useful in improving bowel function.


[Construction and expression of the recombinant plasmid pcDNA3.1/Ts87 of Trichinella spiralis].
To construct and express DNA plasmid of Trichinella spiralis. The BamHI and HindIII enzyme sites and KOZAK sequences were introduced at both ends of Ts87 gene by PCR. Ts87 gene was ligated into pcDNA3.1(+) vector with T4 ligase. The recombinant plasmid pcDNA3.1/Ts87 and plasmid pcDNA3.1 were transfected into an eukaryotic cell line COS7 through Lipofectamine, respectively. The BALB/c mice were immunized with the purified plasmid DNA pcDNA3.1/Ts87 through two routes: intramuscular injection and gene-gun injection. The pcDNA3.1/Ts87 was expressed both in COS7 and in the BALB/c mice.


Combined use of supported liquid membrane and solid-phase extraction to enhance selectivity and sensitivity in capillary electrophoresis for the determination of ochratoxin A in wine.
This paper proposes a novel strategy to enhance selectivity and sensitivity in CE, using supported liquid membrane (SLM) and off-line SPE simultaneously. The determination of ochratoxin A (OA) in wine has been used to demonstrate the potential of this methodology. In the SLM step, the donor phase (either a 20 mL volume of a standard solution at pH 1 or a wine sample at pH 8) was placed in a vial, where a micromembrane extraction unit accommodating the acceptor phase (1 mL water, pH 11) in its lumen was immersed. The SLM was constructed by impregnating a porous Fluoropore Teflon (PTFE) membrane with a water-immiscible organic solvent (octanol). In the off-line SPE step, the nonpolar sorbent (C-18, 4 mg) selectively retained the target ochratoxin, enabling small volumes of acceptor phase (1 mL) to be introduced. The captured analytes were eluted in a small volume of methanol (0.1 mL). This procedure resulted in sample cleanup and concentration enhancement. The method was evaluated for accuracy and precision, and its RSD found to be 5%. The LODs for OA in the standard solutions and wine samples were 0.5 and 30 microg/L, respectively. The results obtained demonstrate that SLM combined with off-line is a good alternative to the use of immunoaffinity columns prior to CE analysis.


Male demographic constraints to extreme sex ratio in the twospotted spider mite.
Twospotted spider mites, Tetranychus urticae Koch are arrhenotokous. As a result of this genetic structure, primary and secondary sex ratios diverge from the 1:1 female:male ratio commonly found in diploid systems. Ratios vary, but 3:1 is the most common. The influence of life history parameters on spider mite sex ratio is unclear, although maternal genetic effects, resource quality and maternal age are known to play a role. An area that has not been studied is the relationship between male reproductive capabilities and spider mite sex ratio. A prior study on male reproduction in spider mites suggests that males have far too high a reproductive capacity to explain sex ratio patterns, but that study was not conducted under realistic mating conditions. Thus, this study was conducted to determine if there is a link between male reproduction and spider mite sex ratio. This was done by exposing males to various exposure regimes of females and recording the number of copulations, inseminations and daughters fathered by each male. Results include the following: i) males are most virile when one day old; ii) virgin males become nearly completely devoid of sperm (or other copulatory products) after about 15 matings and then take about four days to recover; and iii) the optimal number of matings per day (defined as that which produces the greatest number of daughters in the least number of inseminations) is four females per day. The principle conclusion is that although males have a high potential reproductive output, insemination quality is only high for the first four or five matings each day. Therefore, to ensure full inseminations of all daughters by sons, females are constrained to producing primary sex ratios of about 5:1 or less.


Young people's parental discussion about sexual and reproductive health issues and its associated factors in Awabel woreda, Northwest Ethiopia.
In Ethiopia besides the very low health seeking behavior of young people, they do not have access to sexual and reproductive health information and even the existing health services are adult-centered. Furthermore, health providers are not well equipped in addressing young people sexual and reproductive health needs. Therefore, parent-young people discussion about sexual and reproductive health issues are crucial in increasing their awareness and reduces their risky sexual behaviors. This study was aimed to assess young people's parental discussion about sexual and reproductive health issues and its associated factors in Awabel woreda, Northwest Ethiopia. A community based cross-sectional study was conducted among 781 young people aged 10-24 years in Awabel Woreda, Northwest Ethiopia. A pre-tested structured interview administered questionnaire was used for the data collection. The collected data were entered using Epi Data 3.1 and analyzed using SPSS for windows version 21. In the past 6 months, about one quarter, 25.3% of young people had a parental discussion about sexual and reproductive health issues. Young people who reside in urban areas were more likely to discuss on sexual and reproductive health issues with their parents [AOR = 2.44, 95% CI: 1.54-3.89]. Similarly, being male was more likely to have a parental discussion about sexual and reproductive health issues than females [AOR = 1.63, 95% CI: 1.11-2.38]. Furthermore, the odds of parent-young people discussion about SRH matters was more likely among young people aged 20-24 years [AOR = 4.57, 95% CI: 2.13-9.82], living with fathers [AOR = 2.46, 95% CI: 1.20-5.04] and had attained a primary level of education [AOR = 2.89, 95% CI: 1.22-6.87]. Parents lack of interest to discuss, feeling ashamed and culturally not acceptable to talk about sexual matters were found to deter young people's in discussing sexual and reproductive health matters. Parent-young people discussion about sexual and reproductive health is very low and there are different hindering factors. And therefore, young people's sexual and reproductive health programs or policies should be designed in addressing the cultural and societal factors besides the individual or behavioral factors.


Children living in areas with more street trees have lower prevalence of asthma.
The prevalence of childhood asthma in the USA increased by 50% from 1980 to 2000, with especially high prevalence in poor urban communities. Data on the prevalence of asthma among children aged 4-5 years and on hospitalisations for asthma among children less than 15 years old were available for 42 health service catchment areas within New York City. Street tree counts were provided by the New York City Department of Parks and Recreation. The proximity to pollution sources, sociodemographic characteristics and population density for each area were also measured. Controlling for potential confounders, an increase in tree density of 1 standard deviation (SD, 343 trees/km(2)) was associated with a lower prevalence of asthma (RR, 0.71 per SD of tree density; 95% CI, 0.64 to 0.79), but not with hospitalisations for asthma (RR, 0.89 per SD of tree density; 95% CI, 0.75 to 1.06). Street trees were associated with a lower prevalence of early childhood asthma. This study does not permit inference that trees are causally related to asthma at the individual level. The PlaNYC sustainability initiative, which includes a commitment to plant one million trees by the year 2017, offers an opportunity for a large prospective evaluation.


Effect of massaging babies on mothers: pilot study on the changes in mood states and salivary cortisol level.
The purpose of this pilot study was to evaluate the effects of baby massage for 3 months after delivery on mothers' mood status and salivary cortisol level. Study participants were a convenient sample of mothers who delivered their babies at a hospital in Japan, and were recruited at the time of the routine 5-6 weeks postnatal visit to the pediatric office. Thirty-nine mothers were randomly assigned to experimental and control groups. Nineteen mothers in the experimental group were examined before the first day of the baby massage, and 3 months after delivery. The psychological measurements used were profile of mood states (POMS). In the physiological measurements, the salivary cortisol level was analyzed. The result revealed that significant differences in the POMS score were seen in depression and vigor between the two groups at 3 months. There were no significant differences in the salivary cortisol levels. Baby massage was found to positively affect the mood status of the mothers. We propose that midwives and other health-care professionals should recommend mothers to do baby massage to improve their own mood status.


Multiwalled carbon nanotubes enter broccoli cells enhancing growth and water uptake of plants exposed to salinity.
Carbon nanotubes have been shown to improve the germination and growth of some plant species, extending the applicability of the emerging nano-biotechnology field to crop science. In this work, exploitation of commercial multiwalled carbon nanotubes (MWCNTs) in control and 100 mM NaCl-treated broccoli was performed. Transmission electron microscopy demonstrated that MWCNTs can enter the cells in adult plants with higher accumulation under salt stress. Positive effect of MWCNTs on growth in NaCl-treated plants was consequence of increased water uptake, promoted by more-favourable energetic forces driving this process, and enhanced net assimilation of CO2. MWCNTs induced changes in the lipid composition, rigidity and permeability of the root plasma membranes relative to salt-stressed plants. Also, enhanced aquaporin transduction occurred, which improved water uptake and transport, alleviating the negative effects of salt stress. Our work provides new evidences about the effect of MWCNTs on plasma membrane properties of the plant cell. The positive response to MWCNTs in broccoli plants opens novel perspectives for their technological uses in new agricultural practices, especially when 1plants are exposed to saline environments.


[Molecular aspects of chronic granulomatous disease. "the NADPH oxidase complex"].
Chronic granulomatous disease (CGD) is a hereditary illness generally occurring in childhood, in the form of recurrent severe infections. The main pathogens are staphylococci and aspergilli. It results from a failure of professional phagocytes, and particularly neutrophils, to produce superoxide ions O2- and their derivatives, which protect cells from bacterial, invasion through an oxidative and toxic defence mechanism. At an infection site. contact between the neutrophils and microorganisms or an inflammatory mediator triggers a respiratory burst, which results in the activation of the NADPH oxidase enzyme complex. NADPH depletes surrounding oxygen to yield O2-. In its active form. NADPH oxidase is an assembly of two components, namely the membrane cytochrome b558 (consisting o two subunits, gp91-phox and p22-phox) and soluble protein factors present in the resting neutrophil cytoplasm. Transfer of these cytosolic factors and their anchorage to cytochrome b558 determines the activity of NADPH oxidase. The respiratory burst lasts no more than a few minutes, but the precise mechanisms underlying its termination are not well known. In chronic granulomatous disease, neutrophils have lost their bactericidal capacity The most frequent form is hereditary and X-linked; in this case, the affected gene is CYBB, which encodes gp91-phox, the catalytic subunit of cytochrome b558. In autosomal and recessive forms of CGD the mutations affect the genes encoding p22-phox, p67-phox or p47-phox. We have unraveled the assembly mechanisms of the NADPH oxidase complex and have demonstrated that the cytosolic factor p67-phox is the determining element: it triggers both the assembly and the activation of NI4DPH oxidase. Binding of p67-phox to cytochrome b558 induces a gradual conformational change of cytochrome b558, which then becomes capable of transferring electrons produced in the cytoplasm from NADPH to oxygen, reducing the latter to O2-. The isolation of NADPH oxidase in its active and assembled form has allowed us to identify the activation partners of the oxidase complex. We also demonstrated that calcium-binding myeloid-related proteins (MRP). that are abundant in neutrophil cytoplasm, play a fundamental role in this activation. CGD patient management is essentially based on long-term high-dose prophylactic antibiotic administration. Gene therapy is promising but some distance away from practical application. We are currently investigating a new therapeutic concept that consists of transferring cytochrome b558 protein directly into deficient cells (initially the PLB 985 X cell line), encapsulated in proteoliposomes, which are hydrophobic.


A randomized controlled double blind trial comparing the effects of the prophylactic antibiotic, Cefazolin, administered at caesarean delivery at two different timings (before skin incision and after cord clamping) on both the mother and newborn.
Caesarean delivery (CD) increases the risk of postpartum infection by 5 to 20 fold. Prevention of surgical site infection (SSI) is the goal of antibiotic prophylaxis. This study was carried out to assess the optimum timing for prophylactic antibiotic administration and to assess the amount of the antibiotic crossing the placental barrier. Eligible mothers were recruited, after informed consent, once the decision for CD was made. Each mother received two injections, one prior to skin incision and one after cord clamping, (one being the study drug Cefazolin, and the other, a placebo) based on the randomization code. Demographic, maternal and neonatal monitoring data until discharge from hospital, and at the 6 weeks postpartum visit were collected. Levels of the prophylactic antibiotic were measured from the cord blood in every 8th neonate. The objective of the study was to compare the effects of the prophylactic antibiotic, intravenous Cefazolin 1 g, administered at Caesarean delivery (CD) at two different timings (before skin incision and after cord clamping) on both the mother and newborn. The secondary outcomes that were followed up were the number of maternal and neonatal readmissions. An appropriate test for significance, Fisher's exact test was used to find the association between risk variables and outcome. The total numbers of mothers enrolled were 1106, of whom 553 mothers received antibiotic prior to skin incision (pre-incision) and 543 mothers received antibiotic after cord clamping (post-incision). The pre-incision group had significantly less febrile illness (RR = 0.48, 95% CI: 0.29 - 0.80) and SSI (RR = 0.14, 95% CI: 0.04 - 0.53) when compared with the post- incision group. The post-incision group significantly had >7 days hospital stay when compared to the 4-7 days stay of the pre-incision group (p = 0.005).There were no differences in any of the neonatal outcomes. The quantity of the antibiotic in the cord blood was only 2-3%. Pre incision prophylactic antibiotic protected the mother from SSI and febrile illness and decreased the hospital stay significantly. The Clinical Trials Registry India (CTRI) was [ CTRI/2016/03/006710 dated, 04/03/2016].


The role of chloride in taurine transport across the human placental brush-border membrane.
Taurine, a sulfated beta-amino acid, is conditionally essential during development. A maternal supply of taurine is necessary for normal fetal growth and neurologic development, suggesting the importance of efficient placental transfer. Uptake by the brush-border membrane (BBM) in several other tissues has been shown to be via a selective Na(+)-dependent carrier mechanism which also has a specific anion requirement. Using BBM vesicles purified from the human placenta, we have confirmed the presence of Na(+)-dependent, carrier-mediated taurine transport with an apparent Km of 4.00 +/- 0.22 microM and a Vmax of 11.72-0.36 pmol mg-1 protein 20 s-1. Anion dependence was examined under voltage-clamped conditions, in order to minimize the contribution of membrane potential to transport. Uptake was significantly reduced when anions such as thiocyanate, gluconate, or nitrate were substituted for Cl-. In addition, a Cl(-)-gradient alone (under Na(+)-equilibrated conditions) could energize uphill transport as evidenced by accelerated uptake (3.13 +/- 0.8 pmol mg-1 protein 20 s-1) and an overshoot compared to Na+, Cl- equilibrated conditions (0.60 +/- 0.06 pmol mg-1 protein 20 s-1). A Cl(-)-gradient (Na(+)-equilibrated) also stimulated uptake of [3H]taurine against its concentration gradient. Analysis of uptake in the presence of varying concentrations of external Cl- suggested that 1 Cl- ion is involved in Na+/taurine cotransport. We conclude that Na(+)-dependent taurine uptake in the placental BBM has a selective anion requirement for optimum transport. This process is electrogenic and involves a stoichiometry of 2:1:1 for Na+/Cl-/taurine symport.


Identification of the First PPARα/γ Dual Agonist Able To Bind to Canonical and Alternative Sites of PPARγ and To Inhibit Its Cdk5-Mediated Phosphorylation.
A new series of derivatives of the PPARα/γ dual agonist 1 allowed us to identify the ligand ( S)-6 as a potent partial agonist of both PPARα and γ subtypes. X-ray studies in PPARγ revealed two different binding modes of ( S)-6 to the canonical site. However, ( S)-6 was also able to bind an alternative site as demonstrated by transactivation assay in the presence of a canonical PPARγ antagonist and supported from docking experiments. This compound did not activate the PPARγ-dependent program of adipocyte differentiation inducing a very less severe lipid accumulation compared to rosiglitazone but increased the insulin-stimulated glucose uptake in 3T3-L1 adipocytes. Finally, ( S)-6 inhibited the Cdk5-mediated phosphorylation of PPARγ at serine 273 that is currently considered the mechanism by which some PPARγ partial agonists exert antidiabetic effects similar to thiazolidinediones, without showing their typical side effects. This is the first PPARα/γ dual agonist reported to show this inhibitory effect representing the potential lead of a new class of drugs for treatment of dyslipidemic type 2 diabetes.


Detection and significance of the potentially pathogenic amoeboflagellate Naegleria italica in Australia.
Thermophilic amoeboflagellates in the genus Naegleria include both virulent and benign species. One of the less studied species, N. italica, has not been detected in the environment since the first reports from Italy in the 1980s; its virulence is known only from infection of laboratory mice. Two recent strains from recreational water in Western Australia (AWQC NG960, NG961) were tentatively identified as N. italica from the characteristic mobilities of seven isozymes. Sequences of the 5.8S rRNA gene and its flanking ITS aligned with a 380+bp length of the published sequence for N. italica with 98% identity. Differences from the type strain were confined to ITS2. Shorter alignments (<320 bp) were observed with other Naegleria species, corresponding to conserved regions of the 5.8S gene and ITS. Unlike the European type strain of N. italica, the Australian isolates failed to infect laboratory mice intranasally, confirming that infectivity of this species is variable and often lower than in N. fowleri.


Characterization of sulfur mustard resistant keratinocyte cell line HaCaT/SM.
The cell line HaCaT/SM was derived from the human keratinocyte cell line HaCaT. HaCaT/SM cells display a high resistance against sulfur mustard (SM). Intention of the presented study was to determine the cellular and molecular differences between HaCaT/SM and HaCaT so as to evaluate which changes might be responsible for being resistant against SM. Both cell lines HaCaT and HaCaT/SM were analyzed with respect to their cell growth, nuclei perimeter, clonogenicity and secretion profile. Moreover DNA alkylation pattern under presence of SM was investigated. In comparison to HaCaT, the HaCaT/SM showed a significant smaller nuclei perimeter. For DNA alkylation a significant difference was observed over time. The clonogenicity of HaCaT/SM was increased to 150%. The secretion profile of these cells demonstrated a strong increase of ANG, PDGF-AA, TIMP1, TIMP2, and a decrease of AREG, CCL5, CXC1, CXC2/3, CXCL6, CXCL7, CXCL8, CXCL10, MIF, Trappin-1. The sulfur mustard (SM) resistant cell line HaCaT/SM demonstrates a wide range of significant differences to their origin cell line HaCaT. These differences might be responsible to provide resistance against SM and might also be useful to establish treatment concepts for humans after SM exposure.


HIV-related knowledge, perceptions, attitudes, and utilisation of HIV counselling and testing: a venue-based intercept commuter population survey in the inner city of Johannesburg, South Africa.
HIV counselling and testing (HCT) and knowledge about HIV have been key strategies utilised in the prevention and control of HIV/AIDS worldwide. HIV knowledge and uptake of HCT services in sub-Saharan Africa are still low. This study was conducted to determine factors associated with HCT and HIV/AIDS knowledge levels among a commuter population in Johannesburg, South Africa. To identify the factors associated with HCT uptake among the commuter population. A simple random sampling method was used to select participants in a venue-based intercept survey at a taxi rank in the Johannesburg Central Business District. Data were collected using an electronic questionnaire. Logistic regression analysis assessed factors associated with HIV testing stratified by gender. 1,146 respondents were interviewed, the maority (n=579, 50.5%) were females and (n=780, 68.1%) were over 25 years of age. Overall HCT knowledge was high (n=951, 83%) with more females utilising HCT facilities. There was a significant difference in HIV testing for respondents living closer to and further away from health facilities. Slightly more than half of the respondents indicated stigma as one of the barriers for testing (n=594, 52%, p-value=0.001). For males, living with a partner (aOR: 1.68, 95% CI: 1.02-2.78, p-value: 0.041) and possessing a post-primary education were positively associated with testing (aOR: 2.00, 95% CI: 1.15-3.47, p-value: 0.014), whereas stigma and discrimination reduced the likelihood of testing (aOR: 0.40, 95% CI: 0.31-0.62, p-value: <0.001). For females, having one sexual partner (aOR: 2.65, 95% CI: 1.19-5.90, p-value: 0.017) and a low perceived benefit for HIV testing (aOR: 0.54, 95% CI: 0.30-0.96, p-value: 0.035) were associated with HIV testing. The overall HIV/AIDS knowledge was generally high. Gender-specific health education and HIV intervention programmes are needed for improved access to HCT services. One favourable intervention would be the use of home-based HCT programmes.


The diagnostic accuracy of spirometry as screening tool for adult patients with a benign subglottic stenosis.
There is a considerable diagnostic delay in the diagnosis 'benign acquired subglottic stenosis in adults' (SGS, diagnosed by the reference standard, i.e. laryngo- or bronchoscopy). Patients are frequently misdiagnosed since symptoms of this rare disease may mimic symptoms of 'asthma.' The 'Expiratory Disproportion Index' (EDI) obtained by spirometry, may be a simple instrument to detect an SGS-patient. The aim of this study was to evaluate the diagnostic accuracy of the EDI in differentiating SGS patients from asthma patients. We calculated the EDI from spirometry results of all SGS-patients in the Leiden University Medical Center (LUMC), who had not received treatment 2 years before their spirometry examination. We compared these EDI results with the EDI results of all true asthma patients between 2011 and 2019, who underwent a bronchoscopy (exclusion of SGS by laryngo- or bronchoscopy). Fifty patients with SGS and 32 true asthma patients were included. Median and IQR ranges of the EDI for SGS and asthma patients were 67.10 (54.33-79.18) and 37.94 (32.41-44.63), respectively. Area under the curve (ROC) of the accuracy of the EDI at discriminating SGS and asthma patients was 0.92 (95% CI = 0.86-0.98). The best cut-off point for the EDI was > 48 (i.e. possible upper airway obstruction), with a sensitivity of 88.0%% (95%CI = 77.2-95.0%%) and specificity of 84.4% (95%CI = 69.4-94.1%). The EDI has a good diagnostic accuracy discriminating subglottic stenosis patients from asthma patients, when compared to the reference standard. This measurement from spirometry may potentially shorten the diagnostic delay of SGS patients. Further studies are needed to evaluate clinical reproducibility.


Optimum threshold of the 4Kscore for biopsy in men with negative or indeterminate multiparametric magnetic resonance imaging.
The study aims to identify the optimal 4Kscore thresholds to determine the need for a prostate biopsy when multiparametric magnetic resonance imaging (MRI) (mpMRI) is negative or indeterminate. We analysed retrospective data from men in eight different institutions who underwent an mpMRI, 4Kscore and prostate biopsy for evaluation of prostate cancer. We selected men with a negative (PIRADS ≤2) or indeterminate (PIRADS 3) mpMRI. 4Kscore values were categorized into ranges of 1-7, 8-19, 20-32 and greater than 32. We evaluated the proportion of men with grade group 2 or higher (GG2+) cancer in groups defined by PIRADS and 4Kscore. We also evaluated the number of biopsies avoided and GG2+ cancer missed in each group reported depend on 4Kscore cutoff points. Among 1111 men who had an mpMRI, 4Kscore and biopsy, 625 of them had PIRADS ≤3 on mpMRI: 374 negative (PIRADS ≤2) and 251 indeterminate (PIRADS 3). In men with a negative mpMRI, we found a 4Kscore cut-point of 33 resulted in an increased risk of GG2+ cancer on biopsy. In patients with an equivocal lesion on mpMRI, men with a 4Kscore cutoff ≥8 had a greater risk of GG2+ cancer on biopsy. Decision curve analysis supported the proposed cut-points in each mpMRI group. In men with negative and indeterminate mpMRI, we found the best 4Kscore threshold to determine the need for biopsy to be 33 and 8 respectively. Future prospective studies in independent populations are needed to confirm these findings.


[Standardising sigmoidectomy by single umbilical incision].
The increase in single-incision endoscopic surgery has led to more and more procedures and surgical groups who perform them. Segmental resection is the procedure most likely to benefit from this approach since it achieves a significant minimisation in the parietal access; with umbilical mini-laparotomy the entrance ports are grouped together and extraction of the piece and preparation of the anastomosis can be performed. Among the colorectal procedures, sigmoidectomy seems to be the most appropriate area to start using these techniques by groups with laparoscopic colorectal experience. Our aim is the standardisation of single-incision sigmoidectomy, analysing the process with the support of video sequences, and placing emphasis on the factors where it differs from conventional laparoscopy and on the key points in order to avoid problems during the execution of the process.


Potent anti-calmodulin activity of cyclotetradepsipeptides isolated from Isaria fumosorosea using a newly designed biosensor.
Seven cyclotetradepsipeptides, namely beauverolides C (1), F (2), I (3), Ja (4), L (5), M (6), and N (7), were isolated from the entomopathogenic fungus Isaria fumosorosea. The beauverolides were evaluated as potential calmodulin (CaM) inhibitors using the newly designed CaM biosensor hCaM M124C-AF350; these peptides displayed high affinity to the protein with dissociation constants (Kd) ranging from 0.078 μM to 3.44 μM. Beauverolide Ja, the only one containing a tryptophan residue in its structure, showed the highest affinity. The docking study predicted that beauverolides could bind to CaM in the same site of interaction as chlorpromazine, a well-known calmodulin ligand.


Height at onset of insulin-dependent diabetes mellitus in high- and low-risk countries. Japan and Pittsburgh Childhood Diabetes Research Groups.
The relative height at the onset of insulin-dependent diabetes mellitus was recorded in 340 cases from Japan and 191 cases from Pittsburgh, U.S.A. In Pittsburgh, the children diagnosed after puberty were shorter than normal whereas those diagnosed prior to puberty were much taller than normal. However, in Japan, there was no association between age at onset and relative height; at every age the newly diabetic children had similar relative heights to the non-diabetic population. This discrepancy may be related to the 18-fold incidence difference between the two countries.


Biological factors influencing tissue compartmentalization of trace metals in the deep-sea hydrothermal vent bivalve Bathymodiolus azoricus at geochemically distinct vent sites of the Mid-Atlantic Ridge.
In this study, we investigated on concentrations of trace metals (Al, Cd, Mn, Co, and Hg) in the hydrothermal bivalve Bathymodiolus azoricus, a dominant species at most vent sites along the Mid-Atlantic Ridge (MAR), and in its endosymbiont bacteria and commensal parasite Branchipolynoe seepensis. Comparison of our results with data from the literature on non-hydrothermal bivalves suggests lack of "extreme" uptake of trace metals by B. azoricus, except for Hg concentration which exceeded manyfold previously reported values. Mussels collected from three geochemically distinct vent sites, Menez Gwen, Lucky Strike, and Rainbow, along the MAR showed significant differences in tissue concentration of metals. Proportionality of metals in soft tissues of mussels reflected variation of water chemistry at different vents, which in turn conserved the order of trace metal prevalence in undiluted fluids. There were significant tissue-specific differences in trace metal compartmentalization for all metals investigated. Byssus thread contained the highest metal concentration among examined tissues, and thus it is suggested to be an important detoxification route. Size-dependent differences in metal concentrations were detected only for Hg, revealing a general trend of small mussels accumulating more metal than big mussels. Endosymbiont bacteria are shown to exclusively sequester Al from the host gill and contribute to removal of other toxic metals in mussels from Menez Gwen. The commensal parasite present in all mussels from Lucky Strike had higher tissue concentrations of Mn, Al, and Co than the host gill, unlike Cd and Hg which were considerably lower in the former, and thus its role in detoxification remains unclear. Bioaccumulation potential of vent bivalves and associated organisms are quantified as concentration factors and compared to make inferences on the putative role of the endosymbiont bacteria and the commensal parasite in detoxification of trace metals.


Ovarian pure leiomyosarcoma: case report.
We report a case of a pure ovarian leiomyosarcoma, a rare tumor that comprises less than 0.1% out of all the ovarian malignant tumors.


Smoking as a risk factor for erectile dysfunction: data from the Andrology Prevention Weeks 2001-2002 a study of the Italian Society of Andrology (s.I.a.).
To analyse the relationship between smoking and erectile dysfunction (ED). To provide further data on this issue, we analysed information gathered from men attending a free andrologic consultation in 234 Italian medical centres, in the setting of a project focused on andrologic prevention. Men were asked about "their ability to achieve and maintain an erection sufficient for satisfactory sexual performance". If they were dissatisfied, they were defined as having ED. Out of 16724 subjects, ED was diagnosed in 4081 men (24.4%). After adjustment for age, marital status, education, alcohol consumption, physical activity and concomitant pathologies, in comparison with never smokers, men who currently smoked more than 10 cigarettes/day and former smokers showed significantly higher odds ratio (ORs 1.4 and 1.3, respectively) for ED. These results were confirmed performing analysis in strata of diabetes, hypertension, cardiovascular disease and hypercolesterolemia. This transversal observational study shows that the risk of ED is influenced by smoking. A dose- and duration-response effect is present; changes in smoking habits do not seem to significantly affect the risk to develop ED.


The repetition frequency of DNA in Balbiani ring 2 of Chironomus thummi.
The RNA of Balbiani ring BR2 of polytene chromosomes from Chironomus thummi salivary glands was microisolated and reassociated in the presence of an excess of total larval DNA. BR2 RNA reacts as a single component with a C0t 1/2 of 8.6. Ribosomal precursor RNA from microisolated nucleoli reassociates under identical conditions with a C0t 1/2 of 12.3. These C0t 1/2-values suggest repetition frequencies in the range of 35 and 50 for ribosomal DNA and Balbiani ring 2 DNA, respectively. The data presented here favour the view that the gene for BR2 RNA of C. thummi is internally repeated and contains only one type of DNA sequence.


Surgical Treatment of Small Bowel Neuroendocrine Tumors.
Neuroendocrine tumors of the small bowel are rare, slow-growing malignancies that commonly metastasize to nodes at the root of the mesentery and the liver. Liver metastases are associated with carcinoid syndrome. Mesenteric nodal masses can cause bowel obstruction, intestinal angina, or variceal hemorrhage. Patients die of liver failure or bowel obstruction. Primary resection is associated with improved survival rates. Selected patients may benefit from liver debulking operations. Liver resection has excellent survival rates even in the event of an incomplete resection, as well as improvement in hormonal symptoms. Radiofrequency ablation can help to preserve hepatic parenchyma during resection.


Use of a mouse model to elucidate the phenotypic effects of the von Willebrand factor cleavage mutants, Y1605A/M1606A and R1597W.
von Willebrand Factor (VWF) is tightly regulated by the metalloproteinase ADAMTS13, which cleaves VWF to reduce VWF multimer size and binding affinity for collagen and platelets. This study examines two VWF mutations, R1597W (enhanced cleavage) and Y1605A-M1606A (decreased cleavage), to determine their impact on VWF, in addition to ADAMTS13-mediated cleavage. In vitro mouse ADAMTS13 digestions were performed on recombinant proteins. VWF knockout mice received hydrodynamic injections of mouse Vwf cDNA, following which VWF antigen, multimer profile and VWF propeptide levels were determined. A ferric chloride injury model of thrombosis was also evaluated. In vitro ADAMTS13 digestion of full-length mouse VWF required > 97-fold higher ADAMTS13 levels for Y1605A/M1606A, and 68% lower ADAMTS13 levels for R1597W compared with wild type. In vivo, R1597W had reduced VWF:Ag and both mutations exhibited increased VWF propeptide/VWF:Ag ratios. R1597W multimers show a lower molecular weight profile compared with wild type and Y1605A/M1606A mice. When co-injected with Adamts13 cDNA, Y1605A/M1606A multimers were larger compared with wild type, and R1597W showed only a single multimer band and decreased clearance via VWFpp/VWF:Ag ratio. R1597W was associated with reduced thrombus formation but normal platelet accumulation in a ferric chloride injury model while Y1605A/M1606A had a loss of occlusive thrombi but increased platelet accumulation compared with wild type. This study demonstrates that mutations that alter ADAMTS13 cleavage also can affect VWF clearance, VWF antigen level, multimer structure and thrombotic potential in the VWF knockout hydrodynamic injection model.


Elaborative processing in the Korsakoff syndrome: context versus habit.
Using a procedure of Hay and Jacoby [Hay, J. F., & Jacoby, L. L. (1999). Separating habit and recollection in young and older adults: Effects of elaborative processing and distinctiveness. Psychology and Aging, 14, 122-134], Korsakoff patients' capacity to encode and retrieve elaborative, semantic information was investigated. Habits were created during initial training, whereupon cued-recall memory performance was examined, with habit opposing as well as facilitating recollection of earlier studied words. A first group of patients was instructed and tested in the same way as healthy controls and showed poor test performance. Nevertheless, when given more processing and response time, additional explanation, and explicit encouragement, a second group of patients performed similarly to healthy controls. The results suggest that, when given adequate support, Korsakoff patients are able to encode and make use of semantic, contextual, and sequential information. Word distinctiveness, however, only influenced performance of controls.


Development of extrahepatic arterial blood supply to the liver during hepatic arterial infusion chemotherapy.
The aim of this study was to evaluate the correlation of development of the collateral circulation to the liver during hepatic arterial infusion chemotherapy (HAIC) with the presence of hepatic tumours adjacent to the hepatic surface, and with pretreatment occlusion of aberrant hepatic arteries. In 102 patients with unresectable malignant hepatic tumours treated with HAIC using an implantable port system, development of collaterals to the liver was assessed with CT arteriography using the implantable port and pre- and postoperative angiography. Aberrant hepatic arteries, if present, were occluded prior to treatment for hepatic arterial redistribution. Collaterals to the liver were seen in 29 patients, who had 35 areas with collateral perfusion: 22 areas were in the right posterosuperior area, 6 in the left peripheral area and 7 in the right or left lobar area. Collaterals were revealed more frequently in patients with hepatic tumours adjacent to the hepatic surface than in those without hepatic tumours in peripheral areas in the liver (p < 0.0001). In addition, collaterals developed more frequently in patients with an aberrant hepatic arterial anatomy compared with those with conventional anatomy (p = 0.0007). Our results indicated that patients with hepatic tumours adjacent to the hepatic surface and with pretreatment occlusion of aberrant hepatic arteries had the potential to develop collaterals to the liver during HAIC.


A protein transistor made of an antibody molecule and two gold nanoparticles.
A major challenge in molecular electronics is to attach electrodes to single molecules in a reproducible manner to make molecular junctions that can be operated as transistors. Several attempts have been made to attach electrodes to proteins, but these devices have been unstable. Here, we show that self-assembly can be used to fabricate, in a highly reproducible manner, molecular junctions in which an antibody molecule (immunoglobulin G) binds to two gold nanoparticles, which in turn are connected to source and drain electrodes. We also demonstrate effective gating of the devices with an applied voltage, and show that the charge transport characteristics of these protein transistors are caused by conformational changes in the antibody. Moreover, by attaching CdSe quantum dots to the antibody, we show that the protein transistor can also be gated by an applied optical field. This approach offers a versatile platform for investigations of single-molecule-based biological functions and might also lead to the large-scale manufacture of integrated bioelectronic circuits.


Improvement in low back pain following spinal decompression: observational study of 119 patients.
Prospective clinical observational study of low back pain (LBP) in patients undergoing laminectomy or laminotomy surgery for lumbar spinal stenosis (LSS). To quantify any change in LBP following laminectomy or laminotomy spinal decompression surgery. 119 patients with LSS completed Oswestry Disability Index questionnaire (ODI) and Visual Analogue Scale for back and leg pain, preoperatively, 6 weeks and 1 year postoperatively. There was significant (p < 0.0001) reduction in mean LBP from a baseline of 5.14/10 to 3.03/10 at 6 weeks. Similar results were seen at 1 year where mean LBP score was 3.07/10. There was a significant (p < 0.0001) reduction in the mean ODI at 6 weeks and 1 year postoperatively. Mean ODI fell from 44.82 to 25.13 at 6 weeks and 28.39 at 1 year. The aim of surgery in patients with LSS is to improve the resulting symptoms that include radicular leg pain and claudication. This observational study reports statistically significant improvement of LBP after LSS surgery. This provides frequency distribution data, which can be used to inform prospective patients of the expected outcomes of such surgery.


A multiplex RT-PCR assay for detection and differentiation of avian H3, H5, and H9 subtype influenza viruses and Newcastle disease viruses.
Avian influenza viruses (AIVs) and Newcastle disease viruses (NDVs) co-circulate in the poultry population in China. These viruses cause repeated disease outbreaks that exhibit similar clinical symptoms and epidemiological patterns. H5 and H9 influenza viruses are the major pathogens infecting poultry stocks. Recently, H3 AIV (one of the main subtypes in waterfowl) has become endemic in chickens. A multiplex reverse-transcriptase polymerase chain reaction (mRT-PCR) assay was designed for simultaneous detection and differentiation of avian H3, H5, H9 subtype AIVs and NDVs. Four primer sets were evaluated, three of which specifically targeted the hemagglutinin genes of H3, H5 and H9 AIVs, while the other targeted the NDV fusion gene. The sensitivity and specificity of the mRT-PCR assay was determined. The assay detected the major clades or genotypes of all of the reference AIVs and NDVs currently circulating in China. In addition, the mRT-PCR results obtained from screening 380 clinical swabs and 12 experimental tracheal samples were consistent with those obtained using conventional virus isolation methods. The mRT-PCR assay was established successfully for the detection and differentiation of avian H3, H5, and H9 subtype AIVs and NDVs. The method should, therefore, provide a valuable diagnostic tool for these infections.


Treatment of massive posterior epistaxis by detachable coil embolization of a cavernous internal carotid artery aneurysm.
We present a case of massive posterior epistaxis caused by a cavernous internal carotid artery aneurysm. This lesion was treated with endovascular placement of electrolytically detachable platinum embolization coils. The treatment resulted in cessation of epistaxis until the patient's death 3 months following embolization. We discuss aspects of using these above coils for this condition.


Hydrocortisone differentially alters lesion-induced axon sprouting in male and female rats.
Hydrocortisone was administered to young adult male or female rats after removal of the entorhinal cortex. Lesion-induced outgrowth of the commissural-associational afferent fibers in the hippocampus was quantitated. The glucocorticoids caused a significant decline in axon sprouting in the male subjects and a significant increase in outgrowth in female subjects. Depending on the sex, the hormonal effects on lesion-induced axonal growth are markedly different in the rat.


Using artificial intelligence languages for the calculation of inbreeding coefficients--new tools for an old problem.
A computer program was written for the calculation of inbreeding coefficients in pedigrees. Trees and recursivity allow a very efficient codification into LISP or PROLOG.


Monoclonal antibody G10 against microtubule-associated protein 1x distinguishes between growing and regenerating axons.
Monoclonal antibody G10 binds to an epitope on a microtubule-associated protein and specifically labels elongating axons in the developing rat central nervous system. In this paper the expression of this microtubule-associated protein epitope has been studied in developing and regenerating rat peripheral neurons and sprouting and transplanted neurons in the central nervous system. During development bright immunofluorescent fibres were seen from embryonic day 12 (the earliest age studied) to birth, but immunoreactivity declined postnatally and was only barely detectable in adult sciatic nerves. In contrast to the situation in developing axons, immunostaining could not be demonstrated in axons regenerating after crushing of the sciatic nerve nor in axons growing from grafts of septal tissue into the adult hippocampus, despite good evidence for extensive reinnervation in both cases. Immunostaining was only seen at early stages within the septal grafts, and not as axons grew out to reinnervate the host. No staining was seen in areas of deafferented hippocampus where collateral sprouting takes place. These results suggest a molecular difference between the cytoskeletons of developing and regenerating or sprouting nerves.


Ophthalmic Aspects of Stevens-Johnson Syndrome and Toxic Epidermal Necrolysis: A Narrative Review.
The aim of our review article was to summarize the current literature on Stevens-Johnson syndrome (SJS) and its severe form, toxic epidermal necrolysis (TEN). SJS/TEN is a serious, rare multi-system, immune-mediated, mucocutaneous disease with a significant mortality rate that can lead to severe ocular surface sequelae and even to bilateral blindness. Restoration of the ocular surface in acute and chronic SJS/TEN is challenging. There are only limited local or systemic treatment options for SJS/TEN. Early diagnosis, timely amniotic membrane transplantation and aggressive topical management in acute SJS/TEN are necessary to prevent long-term, chronic ocular complications. Although the primary aim of acute care is to save the life of the patient, ophthalmologists should regularly examine patients already in the acute phase, which should also be followed by systematic ophthalmic examination in the chronic phase. Herein, we summarize actual knowledge on the epidemiology, aetiology, pathology, clinical appearance and treatment of SJS/TEN.


Highly efficient redox-driven reversible color switching of dye molecules via hydrogenation/oxygenation.
We report a novel reversible color switching system based on one-pot hydrogenation/oxygenation reactions over Pd/CeO2-x catalysts and fast interconversion of thionine (TH+) and leuco thionine (LTH). Oxygen vacancies produced by Pd-catalyzed instant hydrogenation of CeO2 and strong metal-support interaction (SMSI) could lead to fast color switching.


Skeletal muscle regulatory proteins enhance F-actin in vitro motility.
Using an in vitro motility assay, we have investigated the effects of rabbit skeletal muscle regulatory proteins, troponin and tropomyosin, on the gliding of F-actin filaments or F-actin filaments containing these regulatory proteins. We demonstrate that Ca2+ does not affect the motility of F-actin gliding on HMM, but does in the presence of skeletal muscle tropomyosin and troponin. We conclude that Ca2+ affects motility through troponin because, like F-actin, F-actin-Tm filaments show no Ca(2+)-dependence to their gliding speeds. Furthermore, there is a large enhancement of the gliding speed (about 75%) in the presence of skeletal muscle tropomyosin, troponin + saturating Ca2+ over that seen with F-actin filaments. This enhancement is not due to the action of tropomyosin alone as skeletal muscle tropomyosin without troponin enhances the speed little (about 5%) over that of F-actin. Thus troponin confers Ca2+ sensitivity to the motility and, additionally, potentiates motility greatly along with tropomyosin in the presence of saturating Ca2+. When [HMM] is varied, the decline in speed of F-actin seen at low HMM density is changed little by tropomyosin in the F-actin-Tm filaments. These data show that the skeletal regulatory proteins interact with F-actin to enhance the interaction with HMM particularly in the presence of troponin and saturating Ca2+ and enhance the gliding speed in the in vitro motility assay as they potentiate the ATPase activity in the isolated proteins. This enhancement of speed in the motility assay cannot be ascribed to tropomyosin alone.


Contributions of early-life cognitive reserve and late-life leisure activity to successful and pathological cognitive aging.
The identification of factors that specifically influence pathological and successful cognitive aging is a prerequisite for implementing disease prevention and promoting successful aging. However, multi-domain behavioral factors that characterize the difference between successful and pathological cognitive aging are not clear yet. A group of community-dwelling older adults (N = 1347, aged 70-88 years) in Beijing was recruited in this cross-sectional study, and a sub-cohort was further divided into successful cognitive aging (SCA, N = 154), mild cognitive impairment (MCI, N = 256), and cognitively normal control (CNC, N = 173) groups. Analyses of variance, regression models with the Shapley value algorithm, and structural equation model (SEM) analyses were conducted to determine specific influencing factors and to evaluate their relative importance and interacting relationships in altering cognitive performance. We found that abundant early-life cognitive reserve (ECR, including the level of education and occupational attainment) and reduced late-life leisure activity (LLA, including mental, physical, and social activities) were distinct characteristics of SCA and MCI, respectively. The level of education, age, mental activity, and occupational attainment were the top four important factors that explained 31.6% of cognitive variability. By SEM analyses, we firstly found that LLA partially mediated the relationship between ECR and cognition; and further multi-group SEM analyses showed ECR played a more direct role in the SCA group than in the MCI group: in the SCA group, only the direct effect of ECR on cognition was significant, and in the MCI group, direct effects between ECR, LLA and cognition were all significant. Results of this large-sample community-based study suggest it is important for older adults to have an abundant ECR for SCA, and to keep a high level of LLA to prevent cognitive impairment. This study clarifies the important rankings of behavioral characteristics of cognitive aging, and the relationship that ECR has a long-lasting effect on LLA and finally on cognition, providing efficient guidance for older adults to improve their cognitive function and new evidence to explain the heterogeneity of cognitive aging.


Typical, dysplastic, congenital, and Spitz nevi: a comparative immunohistochemical study.
Nevus cell components have been observed in up to 40% of melanomas, but little is known of the pathobiology of these components in relation to their malignant potential. We studied 44 nevi of the typical, dysplastic, congenital, and Spitz types with a battery of monoclonal and polyclonal antibodies that react on formalin-fixed, paraffin-embedded tissues (HMB.45, S-100 protein, RAP-5, epithelial membrane antigen [EMA], and neuron-specific enolase [NSE]) by avidin-biotin immunohistochemical methods. EMA and RAP-5 (which detects the ras oncogene-associated P21 protein) were negative in all cases. Melanoma-specific HMB.45 was strongly reactive with the epidermal component and had a weak to negative reaction with the dermal component in the typical nevi. However, the reaction seen with HMB.45 in the junctional component of dysplastic nevi, congenital nevi, and some Spitz nevi was heterogeneous. One Spitz nevi showed HMB.45 staining in a pattern near to that of melanoma. In contrast to HMB.45, S-100 protein labeled nevomelanocytes, regardless of whether they were within the epidermis or dermis, although, in half of the dysplastic nevi, the reaction was heterogeneous, with some atypical cells failing to stain. But, with cytologically atypical junctional component (dysplastic-appearing), congenital nevi also stained heterogeneously for S-100 protein compared with the dermal component. NSE stained the central component of some Spitz nevi more intensely than the lateral component. Junctional nevomelanocytic subsets of some congenital nevi revealed HMB.45 and S-100 reactivity similar to dysplastic nevi.


Computer vision quantitation of erythrocyte shape abnormalities provides diagnostic, prognostic, and mechanistic insight.
Examination of red blood cell (RBC) morphology in peripheral blood smears can help diagnose hematologic diseases, even in resource-limited settings, but this analysis remains subjective and semiquantitative with low throughput. Prior attempts to develop automated tools have been hampered by their poor reproducibility and limited clinical validation. Here, we present a novel, open-source machine-learning approach (denoted as RBC-diff) to quantify abnormal RBCs in peripheral smear images and generate an RBC morphology differential. RBC-diff cell counts showed high accuracy for single-cell classification (mean AUC, 0.93) and quantitation across smears (mean R2, 0.76 compared with experts, interexperts R2, 0.75). RBC-diff counts were concordant with the clinical morphology grading for 300 000+ images and recovered the expected pathophysiologic signals in diverse clinical cohorts. Criteria using RBC-diff counts distinguished thrombotic thrombocytopenic purpura and hemolytic uremic syndrome from other thrombotic microangiopathies, providing greater specificity than clinical morphology grading (72% vs 41%; P < .001) while maintaining high sensitivity (94% to 100%). Elevated RBC-diff schistocyte counts were associated with increased 6-month all-cause mortality in a cohort of 58 950 inpatients (9.5% mortality for schist. >1%, vs 4.7% for schist; <0.5%; P < .001) after controlling for comorbidities, demographics, clinical morphology grading, and blood count indices. RBC-diff also enabled the estimation of single-cell volume-morphology distributions, providing insight into the influence of morphology on routine blood count measures. Our codebase and expert-annotated images are included here to spur further advancement. These results illustrate that computer vision can enable rapid and accurate quantitation of RBC morphology, which may provide value in both clinical and research contexts.


Torsion of the penis: a gross anatomic study.
A prospective gross anatomic study of the elements that permit correction of torsion of the penis is presented. Torsion most frequently is corrected by reflection of the skin and dartos tunic only. In some cases resection of Buck's fascia will provide correction. Careful alignment of the skin during closure gives an excellent cosmetic result even when the torsion is not corrected by reflection of the penile investments.


Kinetics of the oxidation of p-aminobenzoic acid catalyzed by horseradish peroxidase compounds I and II.
The kinetics of p-aminobenzoic acid oxidation catalyzed by horseradish peroxidase Compounds I and II was investigated intensively as a function of pH at 25 degrees in aqueous solutions of ionic strength 0.11. All of the rate data were collected from single turnover experiments involving reactions of a single enzyme compound. In reactions of both compounds, deviations from first order behavior with respect to the enzyme were observed at high pH values which were explained in terms of a free radical interaction of product with the enzyme. The effect could be eliminated with sufficient excess of substrate. Kinetic behavior which deviated from first order in substrate, observed at low pH, was explained by a mechanism involving an enzyme-substrate complex which reacted with an additional molecule of substrate but at a slower rate. The pH dependence of the second order rate constants for the reaction of p-aminobenzoic acid with free Compounds I and II is similar to results obtained for the comparable reactions of ferrocyanide, suggesting similar proton-transfer mechanisms for both reducing substrates. The reduction of Compound II by p-aminobenzoic acid appeared to be influenced by two ionizable groups on the enzyme which affect the electronic environment of the heme. The lack of influence of substrate ionizable groups on the rate of the Compound II reaction indicated that potential differences in reactivities of NH2C6H4COO- and NH2C6H4COOH were levelled by the diffusion-controlled limit in the acid region of pH. The reduction of Compound I by p-aminobenzoic acid was not diffusion-controlled and the rate-pH profile could be explained in terms of three acid ionizations, two on the substrate and one on Compound I.


Prevention of contrast media-associated nephropathy: randomized comparison of 2 hydration regimens in 1620 patients undergoing coronary angioplasty.
The administration of radiographic contrast agents remains an important cause of acute renal failure. The optimal infusion for hydration has not been evaluated. To compare the incidence of contrast media-associated nephrotoxicity with isotonic or half-isotonic hydration. Prospective, randomized, controlled, open-label study. Patients scheduled for elective or emergency coronary angioplasty were randomly assigned to receive isotonic (0.9% saline) or half-isotonic (0.45% sodium chloride plus 5% glucose) hydration beginning the morning of the procedure for elective interventions and immediately before emergency interventions. An increase in serum creatinine of at least 0.5 mg/dL (44 micromol/L) within 48 hours was defined as contrast media-associated nephrotoxicity. Secondary end points were cardiac and peripheral vascular complications. A total of 1620 patients were assigned to receive isotonic (n = 809) or half-isotonic (n = 811) hydration. Primary end point analysis was possible in 1383 patients. Baseline characteristics were well matched. Contrast media-associated nephropathy was significantly reduced with isotonic (0.7%, 95% confidence interval, 0.1%-1.4%) vs half-isotonic (2.0%, 95% confidence interval, 1.0%-3.1%) hydration (P =.04). Three predefined subgroups benefited in particular from isotonic hydration: women, persons with diabetes, and patients receiving 250 mL or more of contrast. The incidence of cardiac (isotonic, 5.3% vs half-isotonic, 6.4%; P =.59) and peripheral vascular (isotonic, 1.6% vs half-isotonic, 1.5%, P =.93) complications was similar between the 2 hydration groups. Isotonic hydration is superior to half-isotonic hydration in the prevention of contrast media-associated nephropathy.