Hepatic S-adenosylmethionine after maternal alcohol exposure on offspring rats.
S-adenosylmethionine (SAM) is an universal methyl donor for biological systems in transmethylation reactions. Another metabolic pathway involving S-adenosylmethionine is initiated with the release of -CH3 from the molecule and the formation of S-adenosylhomocysteine and then homocysteine and cysteine, a precursor of the main cellular antioxidant glutathione. Chronic ethanol consumption could affect the bioavailability of amino acids such as methionine. Our purpose was to determine the effect of chronic alcohol feeding during gestation or lactation on hepatic S-adenosyl-methionine, S-adenosylhomocysteine, DNA methylation and homocysteine serum concentration at the end of the lactation period (21-day-old offspring). Wistar dam rats were fed with alcohol during periconceptional, gestation and lactation periods (alcohol-fed rats). This study was conducted with three groups of offspring with different periods of alcohol exposure: control offspring (C), no treatment; and gestation (G) and lactation (L) offspring, exposed to alcohol only during gestation or lactation, respectively. To obtain these last two groups of offspring, on parturition day control newborn rats were cross-fostered to alcohol-fed dams (L) and alcohol new-born rats were cross-fostered to control dams (G). In conclusion, these results indicate that exposure of rats to ethanol during the lactation period affects SAM values more severely than ethanol exposure only during gestation.


A 3D Monte Carlo code for the modeling of plasma dynamics and beam formation mechanism in electron cyclotron resonance ion sources.
The code here presented is the first part of a Monte Carlo (MC) self-consistent 3D plasma simulator. It is yet able to solve the equation of motion for thousands of independent charged particles. The procedure allows to understand the consequences of each phenomenon introduced in the evolution steps of the code. MC random selection of starting parameters is used for each particles; the environmental conditions enclosed in the simulation are ECRIS magnetic field, resonant electromagnetic wave, initial plasma density distribution and MC calculation of Spitzer collision. The results of the first simulations explain some typical effects as the hollow beam formation and the main plasma deconfinement mechanism.


Linear versus non-linear measures of temporal variability in finger tapping and their relation to performance on open- versus closed-loop motor tasks: comparing standard deviations to Lyapunov exponents.
The nature of temporal variability during speeded finger tapping was examined using linear (standard deviation) and non-linear (Lyapunov exponent) measures. Experiment 1 found that right hand tapping was characterised by lower amounts of both linear and non-linear measures of variability than left hand tapping, and that linear and non-linear measures of variability were often negatively correlated with one another. Experiment 2 found that increased non-linear variability was associated with relatively enhanced performance on a closed-loop motor task (mirror tracing) and relatively impaired performance on an open-loop motor task (pointing in a dark room), especially for left hand performance. The potential uses and significance of measures of non-linear variability are discussed.


Localized giant pseudopolyposis of the colon in ulcerative and granulomatous colitis.
The occurrence of localized large clusters of colonic pseudopolyps in 4 patients is described. The underlying disease was chronic ulcerative colitis in 2 cases and granulomatous colitis in 2 others. Each lesion presented as a bulky polypoid mass causing partial or total occlusion of the transverse segment or splenic flexure of the colon. The radiographic and pathologic features of this rare entity are illustrated, and the previously reported 10 cases are briefly reviewed.


Diagnostic utility of PRAME, p53 and 5-hmC immunostaining for distinguishing melanomas from naevi, neurofibromas, scars and other histological mimics.
High expression of PRAME (PReferentially expressed Antigen in MElanoma) and p53 (a proposed marker of desmoplastic melanoma) and low expression of 5-hydroxymethylcytosine (5-hmC) have each been reported in melanoma. However, their combined diagnostic utility for distinguishing melanomas, including uncommon variants, from histological mimics is unknown. This study sought to determine the utility of PRAME, p53 and 5-hmC immunostains for diagnosing melanocytic tumours. A total of 333 cutaneous melanocytic tumours (melanoma n=280, naevi n=53), 20 cutaneous neurofibromas, and 15 scars were evaluated using multiplex immunofluorescence (n=313) or single-plex chromogenic immunohistochemical staining (n=55). Immunostaining for PRAME and 5-hmC were each scored using a previously described semiquantitative scale 0 (absent) to 4+ (diffuse). p53 was scored using a previously described immunoreactive score (range 0-300). PRAME expression was significantly higher in melanomas than in naevi (p<0.0001), with the lowest PRAME expression found in low-grade desmoplastic melanomas compared to the other melanoma subtypes. In non-desmoplastic melanomas, 38% showed 4+ staining (>75% positive tumour cells) and 70% showed 3+ or 4+(>50% positive tumour cells). Conversely, 96% of naevi showed 0, 1+ or 2+ expression. 5-hmC expression was significantly lower in melanomas than in naevi (p<0.0001). However, acral melanomas were not significantly associated with loss of 5-hmC expression (p=0.84). Compared with using PRAME in isolation, combining PRAME and 5-hmC scores increased sensitivity (64%-84%) for detecting melanoma. With respect to desmoplastic melanoma compared to scar or neurofibroma, strong PRAME or p53 staining were almost exclusively found in high-grade desmoplastic melanomas; low-grade desmoplastic melanomas, neurofibromas and scars were negative. 5-hmC was not useful in distinguishing desmoplastic melanomas from neurofibromas or scars. Our data support the use of PRAME as a highly specific ancillary investigation in the diagnosis of melanoma, however PRAME should be considered 'positive' if there is 3+ or 4+ staining (rather than the widely recommended 4+ threshold). 5-hmC, PRAME and p53 appear to have a limited role in the diagnosis of low-grade desmoplastic melanomas.


Automatic plaque characterization employing quantitative and multicontrast MRI.
Multicontrast magnetic resonance imaging (MRI) has shown promise in identifying and characterizing atherosclerotic plaques. One of the limitations of this technique is the lack of a practical automated plaque characterization scheme. In the current study, a prior-information-enhanced clustering (PIEC) technique that utilizes both multicontrast MR images and quantitative T(2) maps is proposed to characterize atherosclerotic plaque components automatically. The PIEC algorithm was assessed on computationally simulated images and multicontrast MRI data of coronary arteries. Multicontrast (T(1)-, T(2)-, partial T(2)-, and proton density-weighted) MR images were acquired from freshly excised human coronary arteries using a 4.7T small-animal scanner. The T(2) distribution for each plaque constituent was measured by exponentially fitting the signal from multiple MR images with different TEs and the same TR. The calculated T(2) distributions were used as the a priori information and combined with the Fuzzy C-Means (FCM)-based clustering algorithm to characterize plaque constituents. The proposed PIEC technique appears to be a promising algorithm for accurate automated plaque characterization.


Endoscopic Management of Tympanic Membrane Retraction Pockets: A Two Handed Technique with Endoscope Holder.
The tympanic membrane retraction pocket is a pathological invagination of tympanic membrane into the middle ear space. The most common sites for formation of retraction pocket are pars flaccida and postero-superior parts. Decision about the procedure and the timing of the treatment of retraction pockets is debatable and depends on the functional and anatomic condition of the ear. To evaluate the results of the technique of two handed endoscopic management of retraction pockets with sliced tragal cartilage. Prospective Non Randomized Clinical Study. The study included 41 ears operated with the technique of two handed endoscopic cartilage tympanoplasty for retraction pockets with endoscope holders from November 2013 to May 2016 with a follow up period ranging from 22 to 53 months. Cases of cholesteatoma and tympanic membrane perforation were excluded from the study. Pre and postoperative symptoms and air-bone gaps were recorded. The average preoperative air-bone gap in the study group was 24.53 ± 4.326 dB. 28 ears were of pars tensa retractions (stage II-4, stage III-15, and stage IV-9) and 13 were pars flaccida retractions (stage III-8 and stage IV-5). 24 ears had ossicular erosion. The follow up revealed that, the results of two handed endoscopic sliced cartilage tympanoplasty for retraction pockets were stable and there was no recurrence of the retraction and the post-operative air-bone-gap closure was achieved to 13.62 ± 4.78, 14.13 ± 5.91 dB, 14.32 ± 3.46 and 14.57 ± 3.88 dB at 6 months, 1 year, 2 years, 3 years respectively. Though, indications for surgery are based mostly on anatomic status and observation of behaviour of retraction pocket over time, we recommend early management of retraction pockets by the technique of endoscopic sliced tragal cartilage tympanoplasty with tragal cartilage of 0.5 mm thickness using endoscope holder. Level of Evidence: Level 4.


Structural and molecular genetic insight into a widespread sulfur oxidation pathway.
Many environmentally important photo- and chemolithoautotrophic bacteria accumulate globules of polymeric, water-insoluble sulfur as a transient product during oxidation of reduced sulfur compounds. Oxidation of this sulfur requires the concerted action of Dsr proteins. However, individual functions and interplay of these proteins are largely unclear. We proved with a DeltadsrE mutant experiment that the cytoplasmic alpha2beta2gamma2-structured protein DsrEFH is absolutely essential for the oxidation of sulfur stored in the intracellular sulfur globules of the purple sulfur bacterial model organism Allochromatium vinosum. The ability to degrade stored sulfur was fully regained upon complementation with dsrEFH in trans. The crystal structure of DsrEFH was determined at 2.5 A resolution to assist functional assignment in detail. In conjunction with phylogenetic analyses, two different types of putative active sites were identified in DsrE and DsrH and shown to be characteristic for sulfur-oxidizing bacteria. Conserved Cys78 of A. vinosum DsrE corresponds to the active cysteines of Escherichia coli YchN and TusD. TusBCD and the protein TusE are parts of sulfur relay system involved in thiouridine biosynthesis. DsrEFH interacts with DsrC, a TusE homologue encoded in the same operon. The conserved penultimate cysteine residue in the carboxy-terminus of DsrC is essential for the interaction. Here, we show that Cys78 of DsrE is strictly required for interaction with DsrC while Cys20 in the putative active site of DsrH is dispensable for that reaction. In summary, our findings point at the occurrence of sulfur transfer reactions during sulfur oxidation via the Dsr proteins.


Solar cheilosis: an ominous precursor part II. Therapeutic perspectives.
The differential diagnosis of SC includes malignant, premalignant, metastatic, inflammatory, and eczematoid disorders, along with photodermatoses and a few rare but important disorders of the lower lip. Current treatment options include topical, ablative, and surgical therapies. Several clinical challenges are also addressed, including the issue of obtaining a high-yield diagnostic biopsy specimen while minimizing patient morbidity, field-directed treatment for SC, and strategies for combination therapy.


Economics of alcohol-involved traffic crashes in the USA: an input-output analysis.
Preventing traffic crashes reduces crash costs paid by employers and employees. The related savings filter through the economy, impacting its performance. This study is the first to measure the impact of traffic crash reduction on a national economy. It focuses on impaired driving crashes. We analysed the impact of the almost 50% alcohol-involved driving crash rate reduction from 1984-1986 to 2010 and the impact if such crashes in 2010 had not occurred. The analysis entered published estimates of costs that employers, consumers and governments paid because of US impaired-driving crashes as production costs and demand changes in Rutgers University's input-output model of the US economy. For example, reducing medical costs paid by employers lowers the cost of labour inputs to production while reducing vehicle repairs raises demand for other goods. Running the model at current and alternative crash rates revealed the impacts of crash reductions on economic output, gross domestic product (GDP), national income and employment. Alcohol-involved crash reductions since 1984-1986 increased economic output in 2010 by an estimated $20 billion, raised GDP by $10 billion, increased US income by $6.5 billion, and created 215 000 jobs. GDP gains from alcohol-involved crash reduction contributed 5% of the $200 million compounded average annual growth in US GDP from 1985 to 2013. Eliminating remaining alcohol-involved crashes would result in similar economic gains. Alcohol-involved crashes drag down the US economy. On average, each of the 25.5 billion miles Americans drove impaired in 2010 reduced economic output by $0.80. Those losses are preventable.


A boy with Burkitt lymphoma associated with Noonan syndrome due to a mutation in RAF1.
This article reports on an association between Burkitt lymphoma and Noonan syndrome (NS) due to a RAF1 gene mutation. The patient was a 7-year-old boy with NS, who was included in the first series reporting the association between Noonan and RAF1, and who later presented with a 2-week history of asymptomatic unilateral tonsillar swelling and ipsilateral cervical lymphadenopathy. Histological and biological examinations of the tonsillar biopsy led to the diagnosis of Burkitt lymphoma. While there is a well-established association between NS and solid cell tumors, this is the first case described in the literature of Burkitt lymphoma in a patient with NS, and adds to the growing list of data supporting neoplasia's association with NS.


Germ cell tumors of the ovary arising after dermoid cyst resection: a long-term follow-up study.
To study the long-term ovarian neoplastic consequences of resection of a dermoid cyst. The study population comprised 99 patients who were operated on for an ovarian dermoid cyst. Follow-up information was obtained for 91 women for a mean period of 5.06 +/- 2.46 years. Of the 99 women, 18 had bilateral dermoid cysts. Multiple dermoid cysts in a single ovary were found in nine of the women with bilateral cysts and in one of the remaining patients. Two patients developed malignant germ cell tumors, and three developed a recurrent dermoid cyst in an ovary from which a dermoid cyst had previously been extracted. Bilateral or multiple ovarian dermoid cysts were present at the initial operation in four (80%) of these patients. Women with bilateral or multiple dermoid cysts may include a subgroup of patients with a greater tendency to develop future ovarian germ cell neoplasms.


srBERT: automatic article classification model for systematic review using BERT.
Systematic reviews (SRs) are recognized as reliable evidence, which enables evidence-based medicine to be applied to clinical practice. However, owing to the significant efforts required for an SR, its creation is time-consuming, which often leads to out-of-date results. To support SR tasks, tools for automating these SR tasks have been considered; however, applying a general natural language processing model to domain-specific articles and insufficient text data for training poses challenges. The research objective is to automate the classification of included articles using the Bidirectional Encoder Representations from Transformers (BERT) algorithm. In particular, srBERT models based on the BERT algorithm are pre-trained using abstracts of articles from two types of datasets, and the resulting model is then fine-tuned using the article titles. The performances of our proposed models are compared with those of existing general machine-learning models. Our results indicate that the proposed srBERTmy model, pre-trained with abstracts of articles and a generated vocabulary, achieved state-of-the-art performance in both classification and relation-extraction tasks; for the first task, it achieved an accuracy of 94.35% (89.38%), F1 score of 66.12 (78.64), and area under the receiver operating characteristic curve of 0.77 (0.9) on the original and (generated) datasets, respectively. In the second task, the model achieved an accuracy of 93.5% with a loss of 27%, thereby outperforming the other evaluated models, including the original BERT model. Our research shows the possibility of automatic article classification using machine-learning approaches to support SR tasks and its broad applicability. However, because the performance of our model depends on the size and class ratio of the training dataset, it is important to secure a dataset of sufficient quality, which may pose challenges.


Psychological factors predict symptoms of severe recurrent genital herpes infection.
In a group of patients with severe recurrent genital herpes infection, we measured symptoms (pain and itching), psychological factors (depression, anxiety, somatisation, interpersonal sensitivity, and life change) and objective indices of disease (number of recurrences in the previous year, total number of recurrences, duration of recurrences, number of lesions per recurrence). Psychological factors were more predictive of pain and itching than were somatic indices, even after correcting for sex.


Effects of tumour cell culture supernatants on some biochemical activities of macrophages.
The following enzyme activities were measured in cell lysates and supernatants from mouse peritoneal macrophages incubated with products of cultured tumour and other cells: acid phosphatase, beta-D-glucuronidase, N-acetyl-D-glucosaminidase, muramidase (lysozyme), lactic dehydrogenase (supernatant only) and plasminogen activator (supernatant only). There were no noteworthy changes in enzyme activities. Hydrogen peroxide production by appropriately stimulated mouse and/or guinea-pig peritoneal exudate macrophages was variably inhibited by supernatants from some tumour cells and some normal cells. Changes in these biochemical activities of macrophages do not appear to be closely related to the anti-inflammatory activity of tumour cell products.


Healthy knee KT1000 measurements of anterior tibial translation have significant variation.
The most commonly used arthrometer for measuring and reporting anterior tibial translation after anterior cruciate ligament reconstruction is the KT1000. Reports on its consistency and reproducibility vary in the literature, but it remains the "gold standard". The purpose of this study was to assess agreement of KT1000 measurements in a daily clinical setting. A retrospective analysis of anterior knee translation in the healthy knee of 770 patients over a 17-year time period was performed. In this cohort, a total of 24 investigators performed 1890 measurement sets at 89 Newtons (N), 134N and at maximum manual force (MMax) level. To assess the inter- and intra-observer agreement, the intraclass-correlation coefficient (ICC) was calculated. The "investigator effect" was a difference between two examiners in the same patient and the "device effect'' a difference within one examiner in the same patient. Minimally important difference (MID) was calculated as 0.5 of the standard deviation. Thirteen investigators were female, performing 1099 measurements and 11 were male, performing 791 measurements. ICC ranged between 0.558 and 0.644. At the MMax level, male investigators had a higher mm reading than female investigators (p < 0.001). Increased experience did not correlate with a higher ICC. MID ranged between 0.85 and 1.65 mm. This study investigated the KT1000 arthrometer in a clinical setting with a large number of investigators. This device delivers moderate agreement of results. Both the device and investigator effect are present. The MMax level has shown the lowest agreement and a dependency on the investigator gender. Level III diagnostic study.


[Effect of the time between symptom onset, swabbing and testing on the detection of influenza virus].
Influenza surveillance requires the collection of nasopharyngeal swabs in Primary Care for testing in reference laboratories. We evaluated the influence on the laboratory results of the time since the onset of symptoms to swabbing (TSS) and from then until laboratory processing (TSL). We analysed swabs collected in the Sentinel Network of Navarra during the 2009-2010 influenza season. The samples were kept refrigerated until analysed by RT-PCR and viral culture. We analysed the percentage of positive swabs to influenza virus in accordance with the TSS and TSL by logistic regression. From a total of 937 swabs, 373 (40%) were positive for influenza by RT-PCR. The TSS ranged from 0-15 days. In the adjusted analysis by period, laboratory and age, having a positive influenza culture decreased to less than half when the TSS was 4-5 days (OR=0.47; 95% CI, 0.24-0.94), and having a positive RT-PCR decreased when the TSS was 5 days or more (OR=0.24, 95% CI, 0.09-0.65). TSL does not significantly affect the result of the RT-PCR (OR by each day=0.96; 95% CI, 0.88-1.04), or the result of the viral culture (OR by each day=0.97, 95% CI, 0.89-1.06). A TSS over 3 days reduced the likelihood of confirmation of influenza, affecting the viral culture more than the RT-PCR. A TSL within a range of two weeks had no significant effect on the results of the RT-PCR or the viral culture.


Antidromic vs orthodromic sensory median nerve conduction studies.
Median sensory nerve conduction studies are arguably the most often performed electrodiagnostic tests worldwide. Routine tests in clinical practice are done using either antidromic or orthodromic techniques type of stimulation, with no universal agreement on the use of one or the other technique. We review the advantages and drawbacks of antidromic and orthodromic as well as their particularities for clinical application and research. The two techniques differ on how physical and physiological changes affect the action potential. Near-nerve recording is better suited for the orthodromic than for the antidromic technique, while studies of nerve excitability are better suited for the antidromic than for the orthodromic technique. Both techniques are equally suitable for routine tests but research studies may specifically demand one or the other.


Adsorption of nonylphenol on coastal saline soil: Will microplastics play a great role?
Microplastics widely exist in diverse matrices to become important hosts of pollutants. Little information regarding adsorption of emerging contaminants on coastal saline soils influenced by co-existing microplastics is available. Thus, the adsorption behaviors of nonylphenol (NP) on coastal saline soil influenced by microplastics were discussed. Polyvinyl chloride (PVC, 4.7 mm), polyethylene (4.85 mm), and polypropylene (4.51 mm) with addition dose of 10% were used to discuss the effect of microplastic type on adsorption of NP by coastal saline soil while PVC samples with size of 4.7 mm and 0.11 mm were used to explore the effect of microplastic size on NP adsorption. The NP adsorption capacity of the saline soil containing 10% of PVC (4.7 mm) was twice that of soil without PVC. Smaller-size PVC (0.11 mm) with addition amount of 10% enhanced the NP adsorption capacity of the coastal saline soil by 117% to reach 8.91 μg g-1. The desorption capacity of NP on saline soil decreased from 40% to 30% of total adsorption capacity with co-existing PVC. Adsorption/desorption kinetics of NP on coastal saline soil with PVC microplastics could be well explained by pseudo second order model while Freundlich model could better fit the isotherm data of NP adsorption/desorption to show possible occurrence of the multiple-layer adsorption. This study will provide new information regarding the environmental behaviors of typical emerging contaminants on coastal saline soil containing microplastics.


Chronic mastopathy and breast cancer. A follow-up study.
Over an average period of seven years 2,9000 cases of benign breast lesions diagnosed by biopsy between 1948 and 1973 in the Department of Pathology, Kaiser Foundation Hospital, Oakland, were followed for breast cancer development. When classified according to traditional diagnostic categories, the cancer incidence per 1,000 person-years varies between 2.7 and 7.9 and appears to be elevated in comparison to expectations obtained from the Third National Cancer Survey, San Francisco Bay Area. Two thousand four hundred biopsies were also scored by the Black-Chabou method. There is an upward trend in the breast cancer incidence as the atypia score rises, a finding which confirms conclusions from a retrospective case-control study by Black et al.


Brunner's gland adenoma: a rare cause of vomiting.
Upper gastrointestinal system adenoma is generally seen amongst elderly patients and quite rarely seen during the childhood. A 14-y-old female patient was referred to our hospital with complaints of vomiting and abdominal pain. She had been followed up for 6 y with the diagnosis of familial intermittent fever and chronic renal failure due to amyloidosis. Endoscopic examination of the upper gastrointestinal system revealed mild hyperaemia in the corpus and antrum, and a polyp of 0.5x0.5 cm with an ulcerated and pedunculated top in the bulbus. Brunner's gland adenoma was diagnosed by the histopathological examination of the lesion following polypectomy. Brunner's gland adenoma is usually asymptomatic; however, it may reveal clinical findings such as obstruction, bleeding or intussusception, especially in uraemic patients. Thus, we would like to emphasize that, in patients with chronic renal failure and acute onset vomiting and abdominal pain, Brunner's gland adenoma should be considered in the differential diagnosis.


Correlation between musculoskeletal mass and perfusion in patients with gastrointestinal malignancy: a preliminary study based on quantitative CT and CT perfusion.
To investigate the correlation between musculoskeletal mass and perfusion using quantitative computer tomography (QCT) and CT perfusion (CTP) in patients with gastrointestinal malignancy. In this prospective study, 96 patients (mean age 66 years, range 25-90; 63.5% male) with gastrointestinal malignancy underwent QCT and CTP between May 2019 and February 2021. Bone mineral density (BMD) and body composition [perivertebral muscular mass index (PMI), skeletal muscular mass index (SMI) and muscular fat fraction] were evaluated through QCT. Musculoskeletal perfusion parameters were measured by CTP. Differences in these parameters between (or among) two (or three) groups (grouped by BMD, SMI, and TNM staging) were analyzed. There were significant differences in PMI and muscular fat fraction among normal (n = 30), osteopenia (n = 43), and osteoporosis (n = 23) groups (both P < 0.001). Blood flow (r = 0.336, P = 0.001; adjusted for age and gender, r = 0.383, P < 0.001), blood volume (r = 0.238, P = 0.011; adjusted for age and gender, r = 0.329, P = 0.001), and flow extraction product (r = 0.217, P = 0.034; adjusted for age and gender, r = 0.320, P = 0.002) vaules of vertebral perfusion showed positive correlation with BMD. However, the relationships between PMI and perfusion parameters of perivertebral muscle were not significant. No significant differences were found in musculoskeletal mass and perfusion parameters between different TNM staging. The changes of bone mass and perivertebral muscular mass in patients with gastrointestinal malignancy are synchronous. Decreased vertebral bone mass is accompanied with reduced perivertebral muscular mass, increased muscular fat, and decreased bone perfusion. However, the changes of perfusion in vertebra and perivertebral muscles are asynchronous. Musculoskeletal mass and perfusion have no correlation with TNM staging of the patients with gastrointestinal malignancy. SHSY-IEC-4.1/20-242/01 (Registered 09-12-2020, Retrospectively registered).


Inhibition of PI3K/C/EBPβ axis in tolerogenic bone marrow-derived dendritic cells of NOD mice promotes Th17 differentiation and diabetes development.
Dendritic cells (DCs) are key regulators of the adaptive immune response. Tolerogenic dendritic cells play a crucial role in inducing and maintaining immune tolerance in autoimmune diseases such as type 1 diabetes in humans as well as in the NOD mouse model. We previously reported that bone marrow-derived DCs (BM.DCs) from NOD mice, generated with a low dose of GM-CSF (GM/DCs), induce Treg differentiation and are able to protect NOD mice from diabetes. We had also found that the p38 MAPK/C/EBPβ axis is involved in regulating the phenotype, as well as the production of IL-10 and IL-12p70, by tolerogenic GM/DCs. Here, we report that the inhibition of the PI3K signaling switched the cytokine profile of GM/DCs toward Th17-promoting cytokines without affecting their phenotype. PI3K inhibition abrogated the production of IL-10 by GM/DCs, whereas it enhanced their production of IL-23 and TGFβ. Inhibition of PI3K signaling in tolerogenic GM/DCs also induced naive CD4+ T cells differentiation toward Th17 cells. Mechanistically, PI3K inhibition increased the DNA-binding activity of C/EBPβ through a GSK3-dependent pathway, which is important to maintain the semimature phenotype of tolerogenic GM/DCs. Furthermore, analysis of C/EBPβ-/- GM/DCs demonstrated that C/EBPβ is required for IL-23 production. Of physiological relevance, the level of protection from diabetes following transfusion of GM/DCs into young NOD mice was significantly reduced when NOD mice were transfused with GM/DCs pretreated with a PI3K inhibitor. Our data suggest that PI3K/C/EBPβ signaling is important in controlling tolerogenic function of GM/DCs by limiting their Th17-promoting cytokines.


hsa_circ_0001955 Enhances In Vitro Proliferation, Migration, and Invasion of HCC Cells through miR-145-5p/NRAS Axis.
Increasing circular RNAs (circRNAs) have been reported to act as key players in human malignancies. However, the expression, role, and mechanism of circRNAs in HCC are not well elucidated. In this study, some differentially expressed circRNAs (DECs) between hepatocellular carcinoma (HCC) and normal tissues were identified using three circRNA microarrays (Gene Expression Omnibus [GEO]: GSE78520, GSE94508, and GSE97332). Twenty-one DECs were found to be commonly upregulated in all the three datasets. Among the 21 DECs, hsa_circ_0001955 ranked as the top three most upregulated DECs in GEO: GSE78520, GSE94508, and GSE97332. Moreover, hsa_circ_0001955 expression in HCC cells and tissues was significantly higher than that in corresponding normal controls. Functional experiments revealed that knockdown of hsa_circ_0001955 markedly inhibited proliferation, migration, and invasion of HCC, and its overexpression led to the opposite effects. hsa_circ_0001955 was mainly located in the cytoplasm, in which hsa_circ_0001955 could directly bind to miR-145-5p. miR-145-5p was downregulated in HCC, and its expression was negatively linked to hsa_circ_0001955 expression. Furthermore, we identified that NRAS was a downstream direct target of the hsa_circ_0001955/miR-145-5p axis in HCC. Collectively, our findings demonstrate the oncogenic roles of the hsa_circ_0001955/miR-145-5p/NRAS axis in HCC, which may represent a potential therapeutic target for HCC.


Intrinsically directed pruning as a mechanism regulating the elimination of transient collateral pathways.
In neonatal cats, neurons in frontoparietal areas of the cerebral cortex have axons which branch, some collaterals project transiently to the cerebellum, whereas others project by way of the pyramidal tract to the brainstem and spinal cord and persist into the adult. If cerebrocerebellar collaterals are eliminated simply because they are exuberant, then experimentally removing the collaterals in the pyramidal tract should cause the normally ephemeral projections to the cerebellum to persist. To test this hypothesis, the pyramidal tract was cut unilaterally at the pontomedullary junction in 5-9-postnatal-day-old (PND) cats, and 35-68 days later the frontoparietal cortex ipsilateral to the pyramidotomy was injected with tritiated amino acids. From the end of the lesioned pyramidal tract, labeled axons were traced into pathways that descended aberrantly into the caudal medulla and spinal cord, but there was never any transported label in the cerebellum. In a second series of experiments, the fluorescent dye Fast blue (FB) was injected into the spinal cord (2-5 PND) prior to cutting the contralateral pyramidal tract (9-12 PND) to determine if the pyramidotomy caused the axotomized cortical neurons to die. There were no neurons labeled with FB in the frontoparietal cortex on the side of the pyramidotomy, but many retrogradely labeled neurons were present contralaterally in the cortex, suggesting that the pyramidotomy caused the death of all axotomized cortical neurons. In a final set of experiments, FB was injected into the spinal cord and the cerebellar cortex was ablated (2-3 PND) prior to cutting the pyramidal tract (9-72 PND). Cerebellar decortication results in the persistence of cerebrocerebral projections to the partially deafferented deep nuclei, therefore injections of Nuclear yellow (NY) or Diamidino yellow (DY) were made later (32-86 PND) into the cerebellar nuclei on the side of the decortication to determine if these projections persist in pyramidotomized cats. After pyramidotomies at 9 PND, there were no neurons labeled with fluorescent dyes in the ipsilateral frontoparietal cortex, indicating that the cerebrocerebellar collaterals, even under experimental conditions which normally cause them to persist, could not sustain the axotomized cortical neurons. Pyramidotomies at 24 PND or later did not cause all axotomized neurons to die since neurons labeled with FB were present in the ipsilateral cortex. These findings suggest that during development of corticosubcortical pathways there is a hierarchical.(ABSTRACT TRUNCATED AT 400 WORDS)


Cerebral microvascular reactivity to carbon dioxide during isoflurane anesthesia as assessed by laser-Doppler flowmetry.
To determine the effects of carbon dioxide on the cerebral microcirculation during isoflurane anesthesia. We studied 10 rabbits using laser Doppler flowmetry on the parietal cortex. After surgical preparation, the end-tidal concentration of isoflurane was maintained at one MAC (2.05%). Mean arterial pressure, rectal temperature and hematocrit were kept constant. The arterial carbon dioxide tension (PaCO2) was randomly adjusted to three levels: 20-25 mmHg (hypocapnia); 35-40 mmHg (normocapnia); and 45-50 mmHg (hypercapnia). Cerebral blood flow (CBF) was measured after target PaCO2 had been achieved. CBF at normocapnia was 27.4 +/- 13.5 (mean +/- SD) ml/100 gm/min, which increased to 42.4 +/- 14.3 ml/100 g/min (p < 0.01) and decreased to 15.3 +/- 7.5 ml/100 g/min (p < 0.05) at hypercapnia and hypocapnia respectively. The slopes of CBF versus PaCO2 were 1.60 +/- 0.63 ml/100 g/min/mmHg from normocapnia to hypercapnia and -0.79 +/- 0.51 ml/100 g/min/mmHg from normocapnia to hypocapnia. The corresponding percentage of alteration in CBF on the respective change of PaCO2 was 7.2 +/- 5.1 and -3.0 +/- 1.2%/mmHg. These data indicate that the changes of the cerebral microvascular blood flow reflect the CO2 effects during isoflurane anesthesia.


Development of novel formulations for Chagas' disease: Optimization of benznidazole chitosan microparticles based on artificial neural networks.
Benznidazole (BZL) is one of the two therapeutic agents used for the treatment of Chagas' disease. However, the use of BZL in most pharmaceutical preparations and research experiments is still limited due to its low water solubility (0.4mg/mL). To overcome the dissolution rate-limiting step in oral absorption, chitosan microparticles prepared by the coacervation method were chosen, owing to non-toxicity of the polymer and mild conditions of the method. The influence of process parameters such as encapsulation efficiency, size, yield, and dissolution rate was optimized by using artificial neural networks (ANNs). The optimal conditions were found to be 1.5% (w/v) for the polymer concentration, 6.0% (w/v) for the coacervation agent concentration, 1400.0rpm for the stirring rate, and 5.0mL/min for the spraying rate. Confirmation experiments showed good agreement between predicted and experimental values of the optimized properties. These results indicate that ANNs is a valuable tool for the development of optimized BZL chitosan microparticles. To our knowledge it is the first report based on the development of optimized BZL microparticles.


Expression of cDNA clones by coupled in vitro transcription/translation and transfection into COS-1 cells: protein mapping in two-dimensional gels.
We present two procedures that can be used to map proteins in two-dimensional gels if the cDNA is at hand. The first procedure, which is illustrated with the expression of cDNAs encoding the fatty acid binding protein 5 (FABP 5), psoriasin and stratifin, makes use of the in vitro transcription/translation assay marketed by Promega. The procedure is simple and allows the mapping of the primary translation product in a very short time. The second method--which faithfully reproduces post-translational modifications--is based on the expression of cDNAs transfected into COS-1 cells using a eukaryotic expression plasmid. This procedure is illustrated with the expression of cDNAs encoding Ha-ras p21 and rab 11, two small GTP-binding proteins known to undergo complex modifications.


Evaluation of biomarkers for cardiotoxicity of anthracyclin-based chemotherapy.
The clinical assessment of the myocardial damage caused by anthracyclin (ANT)-therapy is difficult. Therefore a study was performed to evaluate non-invasive markers of anthracyclin-induced cardiac effects, with emphasis on course-to-course variation. Eligible for study participation were patients, without known cardiologic abnormalities who did not use cardiotoxic medication (except for ANT-therapy), who had previously completed at least three cycles of anthracyclin-containing chemotherapy (n = 14) and patients who were ANT-naïve and who were scheduled to receive doxorubicin-containing chemotherapy (n = 12). Seven patients in this last group also completed at least three cycles and were available for follow-up assessments; thus a total population of 21 patients (12F/9M) completed at least three courses ANT-chemotherapy. In these patients blood samples and ECG-recordings were taken within 6 months after completion of ANT-therapy. In 12 patients (10F/2M) assessments were also done before, immediately afterwards and at 24 h after each course of ANT. In the patients who completed chemotherapy, NT-proBNP was 277% (n = 21; 95% CI: 86-661%, P < 0.001) higher compared to healthy volunteers. During the first course NT-proBNP rose 269% (n = 12; 167-409%, P < 0.0001) at 24 h post-administration. The linear corrected QT (QTcL) directly after the first administration of ANT increased by 9.56 ms (n = 12; 3.85-15.27, P < 0.001) and this prolongation was still present at 24 h, 11.48 ms (n = 12; 5.61-17.34, P < 0.0001). Both NT-proBNP and QTcL returned to baseline before the start of the next course and a similar pattern was observed during each course. NT-proBNP and QTcL may be useful markers for course-to-course evaluation of anthracyclin-induced cardiotoxicity.


Pre-operative point-of-care assessment of left ventricular diastolic dysfunction, an observational study.
Left ventricular (LV) diastolic dysfunction is an acknowledged peri-operative risk factor that should be identified before surgery. This study aimed to evaluate a simplified echocardiographic method using e' and E/e' for identification and grading of diastolic dysfunction pre-operatively. Ninety six ambulatory surgical patients were consecutively included to this prospective observational study. Pre-operative transthoracic echocardiography was conducted prior to surgery, and diagnosis of LV diastolic dysfunction was established by comprehensive and simplified assessment, and the results were compared. The accuracy of e'-velocities in order to discriminate patients with diastolic dysfunction was established by calculating accuracy, efficiency, positive (PPV) and negative predictive (NPV) values, and area under the receiver operating characteristic curve (AUROC). Comprehensive assessment established diastolic dysfunction in 77% (74/96) of patients. Of these, 22/74 was categorized as mild dysfunction, 43/74 as moderate dysfunction and 9/74 as severe dysfunction. Using the simplified method with e' and E/e', diastolic dysfunction was established in 70.8% (68/96) of patients. Of these, 8/68 was categorized as mild dysfunction, 36/68 as moderate dysfunction and 24/68 as severe dysfunction. To discriminate diastolic dysfunction of any grade, e'-velocities (mean < 9 cm s- 1) had an AUROC of 0.901 (95%CI 0.840-0.962), with a PPV of 55.2%, a NPV of 90.9% and a test efficiency of 0.78. The results of this study indicate that a simplified approach with tissue Doppler e'-velocities may be used to rule out patients with diastolic dysfunction pre-operatively, but together with E/e' ratio the severity of diastolic dysfunction may be overestimated. Clinicaltrials.gov, Identifier: NCT03349593 . Date of registration 21/11/2017. https://clinicaltrials.gov .


Evolution of implantable cardioverter defibrillator indications: comparison of guidelines in the United States and Europe.
The rate of implantation of implantable cardioverter defibrillators (ICD) is growing much faster in the United States than in western Europe. Among several factors, education and awareness of the problem of sudden cardiac death might account for this discrepancy. Guidelines for ICD implantation developed in the United States and various European countries are compared, including the one recently published by the Working Groups on Arrhythmias and Cardiac Pacing of the European Society of Cardiology. Although all guidelines fully concur in their recommendations for patients after cardiac arrest due to ventricular fibrillation and ventricular tachycardia, small but noticeable differences in opinion can be traced for patients with sustained ventricular tachycardia or syncope, and for primary prevention. In general, European guidelines are more restrictive for the latter indications. Whereas U.S. guidelines were quickly updated over the years, this process took much longer in Europe, and several countries only recently adopted their first guidelines. Although different in details, the recent release of ICD guidelines in major western European countries will undoubtedly lead to a pronounced increase of patients protected by this form of therapy in Europe in the coming years.


Genome sequencing of Cladophialophora exuberans, a novel candidate for bioremediation of hydrocarbon and heavy metal polluted habitats.
Cladophialophora exuberans is a filamentous fungus related to black yeasts in the order Chaetothyriales. These melanized fungi are known for their 'dual ecology', often occurring in toxic environments and also being frequently involved in human infection. Particularly Cladophialophora exuberans, C. immunda, C. psammophila, and Exophiala mesophila have been described with a pronounced ability to degrade aromatic compounds and xenobiotic volatiles, such as benzene, toluene, ethyl-benzene, and xylene, and are candidates for bioremediation applications. The objective of the present study is the sequencing, assembly, and description of the whole genome of C. exuberans focusing on genes and pathways related to carbon and toxin management, assessing the tolerance and bioremediation of lead and copper, and verifying the presence of genes for metal homeostasis. Genomic evaluations were carried out through a comparison with sibling species including clinical and environmental strains. Tolerance of metals was evaluated via a microdilution method establishing minimum inhibitory (MIC) and fungicidal concentrations (MFC), and agar diffusion assays. Heavy metal bioremediation was evaluated via graphite furnace atomic absorption spectroscopy (GFAAS). The final assembly of C. exuberans comprised 661 contigs, with genome size of 38.10 Mb, coverage of 89.9X and a GC content of 50.8%. In addition, inhibition of growth was shown at concentrations of 1250 ppm for copper and at 625 ppm for lead, using the MIC method. In the agar tests, the strain grew at 2500 ppm of copper and lead. In GFAAS tests, uptake capacities were observed of 89.2% and 95.7% for copper and lead, respectively, after 21 experimental days. This study enabled the annotation of genes involved in heavy metal homeostasis and also contributed to a better understanding of the mechanisms used in tolerance of and adaptation to extreme conditions.


2,2'-(Carbono-thio-yldisulfanedi-yl)bis-(2-methyl-propanoic acid).
The mol-ecular structure of the title compound, C9H14O4S3, exhibits intra-molecular C-H⋯S hydrogen bonds. In the crystal, pairs of O-H⋯O hydrogen bonds lead to the formation of centrosymmetric dimers, which are in turn connected by weak C-H⋯O inter-actions. The combination of these inter-actions generates edge-fused R 2 (2)(8) and R 2 (2)(20) rings running along [211].


Experimentally induced postinhibitory rebound in rat nucleus ambiguus is dependent on hyperpolarization parameters and membrane potential.
Postinhibitory rebound (PIR), a transient depolarization subsequent to release from experimental hyperpolarization, was identified and characterized in 81% of the cells studied in the nucleus ambiguus in slices from medulla of rat. Hyperpolarizing current pulses were administered via the recording microelectrode in the bridge-balanced mode to test for PIR. The voltage trajectory was characterized by a depolarizing sag during the pulse, rebound depolarization (PIR) after the pulse and increased input resistance during rebound. The amplitude and time course of PIR were dependent on prepulse membrane potential, pulse amplitude and pulse duration. These results suggest a potential role of PIR in respiratory rhythmogenesis.


Identification of S-nitrosylation sites based on multiple features combination.
Protein S-nitrosylation (SNO) is a typical reversible, redox-dependent and post-translational modification that involves covalent modification of cysteine residues with nitric oxide (NO) for the thiol group. Numerous experiments have shown that SNO plays a major role in cell function and pathophysiology. In order to rapidly analysis the big sets of data, the computing methods for identifying the SNO sites are being considered as necessary auxiliary tools. In this study, multiple features including Parallel correlation pseudo amino acid composition (PC-PseAAC), Basic kmer1 (kmer1), Basic kmer2 (kmer2), General parallel correlation pseudo amino acid composition (PC-PseAAC_G), Adapted Normal distribution Bi-Profile Bayes (ANBPB), Double Bi-Profile Bayes (DBPB), Bi-Profile Bayes (BPB), Incorporating Amino Acid Pairwise (IAAPair) and Position-specific Tri-Amino Acid Propensity(PSTAAP) were employed to extract the sequence information. To remove information redundancy, information gain (IG) was applied to evaluate the importance of amino acids, which is the information entropy of class after subtracting the conditional entropy for the given amino acid. The prediction performance of the SNO sites was found to be best by using the cross-validation and independent tests. In addition, we also calculated four commonly used performance measurements, i.e. Sensitivity (Sn), Specificity (Sp), Accuracy (Acc), and the Matthew's Correlation Coefficient (MCC). For the training dataset, the overall Acc was 83.11%, the MCC was 0.6617. For an independent test dataset, Acc was 73.17%, and MCC was 0.3788. The results indicate that our method is likely to complement the existing prediction methods and is a useful tool for effective identification of the SNO sites.


Incidence of Simonsiella in the oral cavity of dogs.
The gliding bacterium Simonsiella (Cytophagales, Simonsiellaceae) was found in palate samples from 66 out of 67 dogs. It is considered a common resident in the oral cavities of dogs.


Parallel functional category deficits in clauses and nominal phrases: The case of English agrammatism.
Individuals with agrammatic aphasia exhibit restricted patterns of impairment of functional morphemes, however, syntactic characterization of the impairment is controversial. Previous studies have focused on functional morphology in clauses only. This study extends the empirical domain by testing functional morphemes in English nominal phrases in aphasia and comparing patients' impairment to their impairment of functional morphemes in English clauses. In the linguistics literature, it is assumed that clauses and nominal phrases are structurally parallel but exhibit inflectional differences. The results of the present study indicated that aphasic speakers evinced similar impairment patterns in clauses and nominal phrases. These findings are consistent with the Distributed Morphology Hypothesis (DMH), suggesting that the source of functional morphology deficits among agrammatics relates to difficulty implementing rules that convert inflectional features into morphemes. Our findings, however, are inconsistent with the Tree Pruning Hypothesis (TPH), which suggests that patients have difficulty building complex hierarchical structures.


Normal fetal growth in women with antiphospholipid syndrome treated with high-dose intravenous immunoglobulin (IVIG).
Antiphospholipid antibodies are associated with fetal distress and fetal death. Although different therapeutic regimens have been used, the incidence of fetal growth retardation varies between 30 and 60 per cent of reported cases. We report the evolution of fetal growth in patients with antiphospholipid antibody syndrome treated with high-dose intravenous immunoglobulins (IVIG). Fourteen patients with a history of recurrent spontaneous abortion and immunological diagnosis of antiphospholipid syndrome were followed longitudinally. Intravenous immunoglobulin at a dose of 0.5 g/kg body weight for two consecutive days was started from the fifth week of pregnancy and repeated every 4 weeks until the 33rd week of gestation. Fetal biometry was evaluated longitudinally from the appearance of the gestational sac at 4 weekly intervals. In the period between 26 and 34 weeks, the frequency of evaluation was increased to every 14 days. Data obtained were compared with a control group of 70 fetuses with uneventful pregnancies matched for gestational age. Neonatal weight is shown in relation to the centiles for the normal population. One patient out of 14 (7.1 per cent) developed gestational hypertension and abruptio placentae. No other pregnancy complications were seen. No proteinuria was found. The mean maternal age was 31.2 +/- 3.8 years. Median birth weight was 3433 g +/- 287. The median centile of the birth weight was 65.3 +/- 18.6. Mean gestational age at delivery was 1.3 weeks. No fetal or neonatal growth retardation was seen.(ABSTRACT TRUNCATED AT 250 WORDS)


Altered flecainide disposition in healthy volunteers taking quinine.
The pharmacokinetics of flecainide and its two sequential metabolites, both free and conjugated, its pharmacodynamics, and the influence of simultaneously administered quinine, have been studied in 10 healthy subjects. The study comprised two, 48-h open phases at an interval of 1 week. Flecainide acetate 150 mg was given as a 30-min i.v. infusion and quinine sulphate orally 500 mg x 3 over 24 h. Quinine administration did not change the apparent volume of distribution or the renal clearance of flecainide, but it significantly reduced its systemic clearance (9.1 vs 7.6 ml.kg-1.min-1), thus increasing the elimination half-life (9.6 vs 11.5 h). The amount of flecainide transformed to its first, meta-O-dealkylated metabolite (MODF) fell with no change in the renal excretion of the latter, either in its free or conjugated forms. This finding, in association with a fall in amount of the second, meta-O-dealkylated lactam metabolite (MODLF) recovered in its conjugated forms in the urine, suggests that quinine inhibits both the first and the second steps of the sequential metabolism of flecainide. When the subjects received quinine in addition to flecainide, the PR interval in the ECG was slightly more prolonged than with flecainide alone. Due to the study design, an effect of quinine per se and the consequence of increased serum flecainide levels could not be distinguished.


Development of Contact Lens-Shaped Crosslinked Amniotic Membranes for Sutureless Fixation in the Treatment of Ocular Surface Diseases.
To develop a new method of manufacturing contact lens-shaped crosslinked amniotic membranes (AMs) using glutaraldehyde (GA) and dialdehyde starch (DAS) as crosslinking agents. Amniotic membranes were placed on a curved plastic mold and crosslinked with either 4.5% DAS or 1% GA, after which their physical properties and biological safety were evaluated. The tensile strength of the GA- and DAS-crosslinked samples was much increased compared with that of normal AMs. Neither crosslinking process affected AM transparency. Although the GA-crosslinked AM showed better enzymatic resistance, its physiological structure was severely damaged after the crosslinking process. On the other hand, compared with the GA-crosslinked AM, the DAS-crosslinked AM showed higher growth factor concentrations and better biocompatibility, similar to normal AMs. In addition, the DAS-crosslinked AM was effective in the recovery of corneal epithelial wounds and was well maintained over 3 days without decentration or degradation on the ocular surface in human subjects. Contact lens-shaped AMs were successfully prepared with crosslinking agents. Crosslinking with DAS did not affect the structural properties or biological activity of the AMs, and the improved mechanical properties helped the AM to maintain its curved shape. This crosslinking method allowed us to transplant AMs into patients' eyes without sutures. Sutureless fixation of contact lens-shaped AMs would be very convenient and safe for the treatment of corneal surface disease.


Determination of hypoxia-inducible factor-1 by using a ratiometric colorimetric test based on click-mediated growth of gold nanoparticles.
The authors describe a significantly improved colorimetric nanoprobe for the determination of transcription factors (TFs). It is making use of click-mediated growth of gold nanoparticles (AuNPs) to amplify the signal-to-noise ratio. Hypoxia-inducible factor-1 (HIF-1) is an important TF that acts as a mediator of cell response to hypoxia. So, the detection of HIF-1 was chosen as the model analyte. Specifically, target HIF-1 is designed to bind to the hypoxia response element within DNA duplex. The click chemistry between the DNA duplex and alkynyl-functionalized AuNPs (AF-AuNPs) is then inhibited because of significant steric hindrance. As a result, the AF-AuNPs grow into larger-sized highly-aggregated irregular nanostructures, which in turn enable colorimetric determination. The ratio of absorbances at 620 and 560 nm increases in the 0.5 to 10 nM HIF-1 concentration range, and the detection limit is 0.27 nM. This is better by a factor of 100 than that of aggregation-based colorimetric assays. The nanoprobe is selective and can be used in complex samples. Conceivably, it may also be extended to the determination of other TFs by simply changing the used DNA duplex. Graphical abstract Schematic of a nanoprobe for detecting hypoxia-inducible factor-1 (HIF-1). Three concepts are involved: the binding of HIF-1 and hypoxia response element, the Cu+-catalyzed click chemistry between P1/P2 duplex and alkynyl-functionalized AuNPs (AF-AuNPs), and the AuNPs growth with hydroxylamine and HAuCl4.


Predictors of Neonatal mortality in Neonatal intensive care unit at referral Hospital in Southern Ethiopia: a retrospective cohort study.
The first one month of life; the neonatal period is the most risky time for child survival. In Ethiopia, neonatal mortality is unacceptably high, and the trend in reduction is slower as compared to infant and child mortality. The magnitude and associated factors of neonatal mortality in a tertiary care facility were not well documented. Therefore, the aim of this study was to determine neonatal mortality and predictors among neonates admitted to neonatal intensive care unit of Wolaita Sodo University Teaching and Referral Hospital, South Ethiopia. A retrospective cohort study design was done among neonates admitted to neonatal intensive care unit of a University Teaching and Referral Hospital from 2015 to 2017. Data were collected using data extraction checklist from the medical registry. The main outcome was the occurrence of death within the first four weeks. The survival time was calculated in days between the date of admission and the date of death. Kaplan-Meier survival was used to depict the pattern of death in 28 days and Cox-Proportional model was used to identify the predictors of the neonatal mortality. A total of 964 neonates which contributed to 5889 neonates-days were included in the study. There were 159 neonatal deaths during the follow-up time. Overall, the neonatal mortality incidence was 27 per 1000 neonates-days. Predictors of neonatal mortality were: multiple birth, mothers who did not attend antenatal care visits, neonates born by cesarean section, not initiated breast feeding within 1 h of birth, neonates resuscitated, hyaline membrane disease and perinatal asphyxia. Neonatal mortality at neonatal intensive care unit was high. Managing neonatal complications, initiating breast feeding within 1 h of birth, promoting antenatal care visits, improving quality of services and ensuring continuum of care are recommended to increase survival of neonates.


Purification and characterization of two soluble Cl(-)-activated arginyl aminopeptidases from human brain and their endopeptidase action on neuropeptides.
Two closely related Cl(-)-activated arginyl aminopeptidases (I and II) were purified from a soluble extract of postmortem human cerebral cortex by anion-exchange chromatography and preparative gel electrophoresis. The electrophoretic mobility of II was approximately 80% that of I; the molecular mass of both enzymes was approximately 70 kilodaltons (kDa) (gel filtration). The aminopeptidase action of I and II on aminoacyl-7-amido-4-methylcoumarin (AMC) substrates was restricted to the Arg and Lys derivatives. Both enzymes had significant endopeptidase activity, hydrolysing several biologically active peptides including neurotensin, bradykinin, angiotensin-I, substance P, luliberin, and somatostatin at internal bonds. Other peptides [Leu-enkephalin, proctolin, thyroliberin, adrenocorticotropin18-39 (ACTH18-39), ACTH11-24, and dynorphin (1-13)] were not appreciably hydrolysed. The amino- and endopeptidase activities had pH optima at 6.5 and 7, respectively, and were both inhibited by metal ion chelators and sulphydryl group blocking agents. The aminopeptidase activity was stimulated 20-fold by Cl- ions, whereas the endopeptidase activity was unaffected by the latter. Km values for neurotensin degradation were 20 microM (I) and 37 microM (II) and for Arg-AMC hydrolysis they were 167 microM (I) and 125 microM (II). The endopeptidase activity was not inhibited by the aminopeptidase inhibitors arphamenine or bestatin (IC50 = 9 nM and 0.1 microM, respectively, with Arg-AMC substrate).


Intracellular ATP is required for mitochondrial apoptotic pathways in isolated hypoxic rat cardiac myocytes.
The present study examined the possibility that intracellular ATP levels dictate whether hypoxic cardiac myocytes die by apoptosis or necrosis. Although apoptosis and necrosis may appear to be distinct forms of cell death, recent studies suggest that the two may represent different outcomes of a common pathway. In ischemic myocardium, apoptosis appears early, while energy stores are presumably still available, followed only later by necrosis. Neonatal rat cardiac myocytes were exposed to continuous hypoxia, during which the intracellular ATP concentration was modulated by varying the glucose content in the medium. The form of cell death was determined at the end of the hypoxic exposure. Under total glucose deprivation, ATP dropped precipitously and cell death occurred exclusively by necrosis as determined by nuclear staining with ethidium homodimer-1 and smearing on DNA agarose gels. However, with increasing glucose concentrations (10, 20, 50, 100 mg/dl) cellular ATP increased correspondingly, and apoptosis progressively replaced necrosis until it became the sole form of cell death, as determined by nuclear morphology, DNA fragmentation on agarose gels, and caspase-3 activation. The data showed a significantly positive correlation between myocyte ATP content and the percentage of apoptotic cells. Hypoxia resulted in lactate production and cellular acidification which stimulates apoptosis. However, acidification-induced apoptosis was also increased in an ATP-dependent fashion. Loss of mitochondrial membrane potential and cytochrome c release from the mitochondria was observed in both the apoptotic and necrotic cells. Furthermore, translocation of Bax from cytosol into mitochondria preceded these events associated with mitochondrial permeability transition. Increased lactate production and a lack of effect by the mitochondrial inhibitor oligomycin indicated that ATP was generated exclusively through glycolysis. We demonstrate that ATP, generated through glycolysis, is a critical determinant of the form of cell death in hypoxic myocytes, independently of cellular acidification. Our data suggest that necrosis and apoptosis represent different outcomes of the same pathway. In the absence of ATP, necrosis prevails. However, the presence of ATP favors and promotes apoptosis.


Using sequence logos and information analysis of Lrp DNA binding sites to investigate discrepancies between natural selection and SELEX.
In vitro experiments that characterize DNA-protein interactions by artificial selection, such as SELEX,are often performed with the assumption that the experimental conditions are equivalent to natural ones. To test whether SELEX gives natural results, we compared sequence logos composed from naturally occurring leucine-responsive regulatory protein (Lrp) binding sites with those composed from SELEX-generated binding sites. The sequence logos were significantly different, indicating that the binding conditions are disparate. A likely explanation is that the SELEX experiment selected for a dimeric or trimeric Lrp complex bound to DNA. In contrast, natural sites appear to be bound by a monomer. This discrepancy suggests that in vitro selections do not necessarily give binding site sets comparable with the natural binding sites.


Genetic transformation of the figwort, Scrophularia buergeriana Miq., an Oriental medicinal plant.
Scrophularia buergeriana Miq. (figwort) contains a diverse group of bioactive natural products and is used to treat a variety of ailments, including fever, constipation, neuritis, and laryngitis. A transformation protocol was established for S. buergeriana using Agrobacterium tumefaciens. Kanamycin-resistant plants were regenerated from leaf explants co-cultivated with A. tumefaciens strain GV3101. The shoot regeneration medium was supplemented with 2 mg l(-1) 6-benzylaminopurine and 70 mg l(-1) putrescine to improve the efficiency of organogenesis. Detection of the neomycin phosphotransferase gene, the presence of high levels of beta-glucuronidase (GUS) transcripts and enzyme activity, and the histochemical localization of GUS confirmed the genetic transformation of S. buergeriana. This work demonstrates the potential of using A. tumefaciens to efficiently transfer foreign genes into a commercially and culturally important Oriental medicinal plant.


Cellular and cytokine responses of the human central nervous system to intracranial administration of tumor necrosis factor alpha for the treatment of malignant gliomas.
To elucidate the role of tumor necrosis factor alpha (TNF alpha) as a biological response modifier, we studied cellular and cytokine responses of the central nervous system to TNF alpha administered intracranially in a phase I clinical trial for patients with malignant gliomas. Six patients received injections of TNF alpha (1.25 x 10(3)-10 x 10(3) U/injection) into the tumor cavities, and regional fluids (RF) and lumbar cerebrospinal fluids (CF) were serially sampled before and after the injections. Recruitment of neutrophils occurred, mostly peaking 8 h after TNF alpha injection, and fewer numbers of CD4+ T cells and monocytes/macrophages migrated, subsequently peaking at 24 h. The CF leukocytosis persisted for 48 h and was associated with an increased level of neutrophil chemotactic activity in the CF. This neutrophil chemotactic activity was attributed to interleukin-8 (IL-8) by HPLC. The level of IL-6 activity in the CF and RF consistently increased; beginning 2 h after TNF alpha injection and reaching the maximum between 8 h and 12 h. It returned to the basal level within 48 h. IL-1 beta was detected in the CF of three patients, its level peaking at 8 h. Prostaglandin E2 also increased after injection of TNF alpha, peaking between 4 h and 12 h and then gradually decreasing. Transforming growth factor beta was found in all cases tested and one patient showed a significant change after TNF alpha injection. IL-2 activity, interferon alpha (INF alpha) activity, IFN beta, and granulocyte/macrophage-colony-stimulating factor were not detected in the CF or RF. In conclusion, TNF alpha is biologically effective in inducing migration of immune cells and generating multiple cytokine responses in the human central nervous system.


The LIN28B-let-7-PBK pathway is essential for group 3 medulloblastoma tumor growth and survival.
Children with Group 3 medulloblastoma (G3 MB) have a very poor prognosis, and many do not survive beyond 5 years after diagnosis. A factor that may contribute to this is the lack of available targeted therapy. Expression of protein lin-28 homolog B (LIN28B), a regulator of developmental timing, is upregulated in several cancers, including G3 MB, and is associated with worse survival in this disease. Here, we investigate the role of the LIN28B pathway in G3 MB and demonstrate that the LIN28B-lethal-7 (let-7; a microRNA that is a tumor suppressor)-lymphokine-activated killer T-cell-originated protein kinase (PBK; also known as PDZ-binding kinase) axis promotes G3 MB proliferation. LIN28B knockdown in G3-MB-patient-derived cell lines leads to a significant reduction in cell viability and proliferation in vitro and in prolonged survival of mice with orthotopic tumors. The LIN28 inhibitor N-methyl-N-[3-(3-methyl-1,2,4-triazolo[4,3-b]pyridazin-6-yl)phenyl]acetamide (1632) significantly reduces G3 MB cell growth and demonstrates efficacy in reducing tumor growth in mouse xenograft models. Inhibiting PBK using HI-TOPK-032 also results in a significant reduction in G3 MB cell viability and proliferation. Together, these results highlight a critical role for the LIN28B-let-7-PBK pathway in G3 MB and provide preliminary preclinical results for drugs targeting this pathway.


Outcome of patients requiring tracheostomy in a pediatric intensive care unit.
Although tracheostomy is a commonly performed procedure, there is a lack of studies in the pediatric intensive care unit (PICU) setting that describe its association with patient outcome and especially hospital mortality. Our goal was to evaluate the outcome of patients receiving a tracheostomy, while on mechanical ventilation (MV), in a PICU. Records of 260 children were reviewed retrospectively regarding PICU mortality, PICU length of stay (PICU LOS), duration of MV and a cost indicator (weighted hospital days; WHD). Nineteen patients received tracheostomy (7.3%). The mortality of patients submitted to tracheostomy in the longer term was significantly higher compared to patients who were not (52.6%vs. 27.6%; P = 0.04) despite having a significantly lower severity of illness at admission (Pediatric Risk of Mortality score--PRISM) (10.9 vs. 13.7; P < 0.001). The mortality of patients without tracheostomy, however, was significantly higher within 30 days (24.8%vs. 5.2%, P < 0.001). Tracheostomized patients had significantly higher mean PICU LOS (68 days vs. 8 days; P < 0.001), duration of MV (62 days vs. 4 days; P < 0.001) and higher WHD (171.5 vs. 21.5; P < 0.001). Contrary to findings in critically ill adult patients, ventilated children receiving a tracheostomy had less favorable outcomes compared with non-tracheostomized patients. In view of the greater use of resources, further studies are needed to confirm and to identify the subgroups of mechanically ventilated patients who will benefit most from this procedure.


Focal adhesions and cell-matrix interactions.
Focal adhesions are areas of cell surfaces where specializations of cytoskeletal, membrane and extracellular components combine to produce stable cell-matrix interactions. The morphology of these adhesions and the components identified in them are discussed together with possible mechanisms of their formation.


Mitogenic properties of insulin-like growth factors I and II, insulin-like growth factor binding protein-3 and epidermal growth factor on human breast stromal cells in primary culture.
Insulin-like growth factors I and II (IGF-I and IGF-II) are growth factors implicated in both normal mammary gland development and breast cancer. We have previously reported on the effects of components of the IGF system on breast epithelial cells. Since data suggests that stromal-epithelial interactions play a crucial role in breast cancer, we have now investigated the mitogenic properties of IGF-I, IGF-II, insulin-like growth factor binding protein-3 (IGFBP-3) and epidermal growth factor (EGF) on human breast stromal cells in primary culture. We show that, under serum-free conditions, stromal cells are stimulated to grow in response to IGF-I and IGF-II in a dose-dependent manner. IGF-I and EGF, a potent stimulator of human breast epithelial cell growth in primary culture and also associated with breast cancer, appear to stimulate stromal cell growth in a synergistic manner. IGFBP-3 does not inhibit the stimulation of growth by IGF-I, or IGF-I plus EGF. However, IGFBP-3 does inhibit the stimulation of growth by IGF-II. In contrast to our previous results with human breast epithelial cells, IGFBP-3 does not have an IGF-independent inhibitory effect on stromal cell growth. This study is the first to address the effects of IGF-I, IGF-II and IGFBP-3 alone and in combination with EGF on human breast stromal cell growth in primary culture. Characterizing the role of the IGF system in both normal breast epithelial cells and stromal cells will aid in our understanding of the mechanisms behind the role of the IGF system in breast cancer.


Sum uncertainty relations for arbitrary N incompatible observables.
We formulate uncertainty relations for arbitrary N observables. Two uncertainty inequalities are presented in terms of the sum of variances and standard deviations, respectively. The lower bounds of the corresponding sum uncertainty relations are explicitly derived. These bounds are shown to be tighter than the ones such as derived from the uncertainty inequality for two observables [Phys. Rev. Lett. 113, 260401 (2014)]. Detailed examples are presented to compare among our results with some existing ones.


[Carbon monoxide poisoning of the fetus. Apropos of a clinico-pathological case].
Anatomical and clinical observations are described in a case of carbon monoxide poisoning which caused the death of the baby rapidly. The anatomical lesions are comparable to those observed in 8 other cases reported in the literature. The physiopathological mechanism involved is discussed in relation to previously reported clinical, toxicological, and experimental data.


Contributing factors for intraocular pressure control in patients with mostly normal-tension glaucoma after initial Ex-PRESS drainage device implantation.
To investigate the postoperative intraocular pressure (IOP) control and identify the factors associated with failure of initial Ex-PRESS surgery in patients with open-angle glaucoma for 3 years. A total of 79 patients with medically uncontrolled open-angle glaucoma (55 normal-tension glaucoma and 24 primary open-angle glaucoma) were enrolled. All patients underwent Ex-PRESS implantation (including combined cataract surgery). The outcome measure was the survival rate using life table analysis, the failure was defined as IOP of > 18 mmHg (criterion A), > 15 mmHg (criterion B) or > 12 mmHg (criterion C) and/or IOP reduction of < 20% from baseline (each criterion) without any glaucoma medications. The Cox proportional hazards model was used to identify risk factors for IOP management defined as the above criterion. RESULTS: The mean preoperative IOP was 19.3 ± 5.8 mmHg. At 36 months, the mean IOP was 11.8 ± 3.6 mmHg with a mean IOP change of 7.5 mmHg (reduction rate 39.0%). The cumulative probability of success was 58% (95%CI: 42-64%) (criterion A), 48% (95%CI: 37-59%) (criterion B) and 30% (95%CI: 20-40%) (criterion C). In multivariate analyses, factors that predicted poor IOP control included the intervention of bleb needling after 6 months after the surgery (HR: 2.43; 95%CI: 1.35-4.37; P = 0.032). Transient hypotony was observed in 4 patients. The implementation of bleb needling after Ex-PRESS surgery in the late postoperative period was suggested to be the main risk factor for achieving lower IOP.


Bilateral tympanic myxoma: A CARE case report.
Ear myxoma is a rare benign tumor sometimes located on the pinna and the external auditory meatus, associated with Carney Complex (CNC). However, tympanic membrane myxoma has never been described. We present here a case of bilateral tympanic membrane myxoma, following CARE guidelines. A 35-year-old woman presented to our department with right otalgia. Otoscopy showed non-specific bilateral tissular masses in the posterior quadrant of the tympanic membranes, with normal hearing thresholds. CT-scan showed a tissular mass without osteolysis. Right-side resection confirmed the lesion as being a myxoma, ruling out differential diagnoses. The patient was then screened for extra-otologic lesions typically associated with ear myxoma in CNC. Only perilabial lesions similar to lentigos suggested CNC. Cardiac, endocrine and thyroid assessment were normal. Genetic testing for a PKRAR1A gene mutation was negative. This is to our knowledge the first reported case of tympanic membrane myxoma. It is of particular interest, being bilateral and showing spontaneous involution of the left lesion over the years. Genetic screening was negative; nevertheless, thorough evaluation is essential due to the life-threatening nature of cardiac myxoma and the frequently associated malignant tumors. Potential new mutations associated with CNC should be considered in the future.


Selective removal of pharmaceuticals and personal care products from water by titanium incorporated hierarchical diatoms in the presence of natural organic matter.
Natural organic matter (NOM), such as humic acids, fulvic acids, and tannic acids, is ubiquitous in water bodies and hinders the photodegradation of pharmaceuticals and personal care products (PPCPs). We prepared titanium incorporated hierarchical diatoms as a novel photocatalyst to selectively remove PPCPs (triclosan, bisphenol A or BPA, and N, N-Diethyl-meta-toluamide or DEET) in the presence of NOM (humic acid). Diatom (Stephanodiscus hantzschii) grown in a titanium(IV) bis(ammonium lactato) dihydroxide solution integrated 7.2% ± 1.4% (mass fraction) of titanium in their cell wall and formed silica-titania frustules. The photodegradation of triclosan, BPA, and DEET by both silica-titania frustules and titania nanopowder (a control photocatalyst) follows pseudo-first-order kinetics. Under ultraviolent light irradiation, the titanium-content-normalized pseudo-first-order removal rate constants of triclosan, BPA, and DEET by silica-titania frustules were 3, 4, and 4-times those by titania nanopowder, respectively, at a humic acid concentration of 10 mg•L-1. Incorporation of titanium did not alter the morphology and hierarchical nano/microstructures of the diatom. The silica-titania frustules were rich in nanopores with a diameter of 20 ± 4 nm (mean ± standard deviation), allowing PPCPs with a small molecular weight (typically < 600 g•mol-1) to pass through while efficiently rejecting NOM with high molecular weights. The silica-titania frustules with hierarchical nano/microstructures served as a prefiltration unit by selectively allowing PPCPs to pass through the nanopores and are therefore promising for photodegradation and environmental remediation applications.


A sensitive and label-free impedimetric biosensor based on an adjunct probe.
A highly sensitive and label-free impedimetric biosensor is achieved based on an adjunct probe attached nearby the capture probe. In this work, the adjunct probe was co-assembled on the surface of gold electrode with the capture probe hybridized with the reporter probe, and then 6-mercapto-1-hexanol was employed to block the nonspecific binding sites. When target DNA was added, the adjunct probe functioned as a fixer to immobilize the element of reporter probe displaced by the target DNA sequences and made the reporter probe approach the electrode surface, leading to effective inhibition of charge transfer. The increase in charge transfer resistance is related to the quantity of the target DNA in a wide range. The linear range for target DNA with specific sequences was from 0.1 nM to 0.5 μM with a good linearity (R=0.9988) and a low detection limit of 6.3 pM. This impedimetric biosensor has the advantages of simplicity, sensitivity, good selectivity, and large dynamic range.


Effect of maxillary expansion on orthodontics.
To explore the effect of maxillary expansion on orthodontics. Eight beagle dogs were randomly divided into two groups, with 4 dogs in each group. Dogs in group 1 were executed immediately and received the direct physical measurement. The magnetic expansion appliance was used in group 2 for the maxillary expansion. After the expansion, the model was taken again and they were executed after cone beam CT (CBCT) scanning. The model measurement method was adopted in group 1 to measure the dental measurement indicators and width of base bone arch. The CBCT measurement method was employed to measure the above dental indicators and bone indicators. The difference in the indicators measured by different methods was compared and analyzed. Before the expansion, there was no significant difference in the bone measurement indicators between the CBCT measurement method and direct physical measurement method. After the expansion, there was no significant difference in indicators between the CBCT measurement method and direct physical measurement. But there was significant difference among the model measurement method, CBCT measurement method and direct physical measurement method. There was the significant difference in the dental indicators between the CBCT measurement method and model measurement, as well as the bone indicators of posterior marginal spacing of greater palatine foramen, posterior marginal spacing of incisive foramen, width of base bone arch and spacing of implant anchorage. There is no significant difference between the effect of CBCT measurement method and direct physical measurement method, but CBCT is significantly better than the model measurement.


The cytotoxicity of [(N-alkyl-1H,3H-1-oxoisoindoline-5-yl-oxyl alkanoates and related benzamides in murine and human tissue cultured cell lines.
Substituted oxoisoindolines are effective cytotoxic agents, causing cell death in a number of tissue culture lines, e.g. L1210, Tmolt-3, and HeLa-S3. In general these agents were not active against the solid cell growth, i.e. KB, skin, HCT-8 ileum, colon, bronchogenic lung, osteosarcoma and glioma. The mode of action of the derivatives involves inhibition of de novo purine synthesis of Tmolt-3 cells, which reduces DNA and RNA syntheses. Purine synthesis was reduced by compound 16 at both regulatory enzymes, i.e. PRPP amido transferase, IMP dehydrogenase and dihydrofolate reductase. The agent lowered d(GTP) and d(CTP) pool levels, further reducing DNA synthesis. DNA strand scission was evident after incubation with Compound 16 for 24 hr at 100 microM and some undefined interaction between the drug and the nucleoside bases appeared to occur, lowering DNA synthesis and causing cell death.


Involvement of angiotensin II in development of spontaneous nephrosis in Dahl salt-sensitive rats.
We investigated the effect of angiotensin-converting enzyme inhibition on spontaneous nephrosis in Dahl salt-sensitive (Dahl/S) rats. Dahl/S rats fed on a normal sodium diet spontaneously developed nephrosis and mild hypertension from a young age. In young Dahl/S rats, an angiotensin-converting enzyme inhibitor, imidapril, attenuated the development of proteinuria accompanied by a decrease in blood pressure. Methylprednisolone, a potent therapeutic agent for proteinuria, did not affect the development of nephrosis. An angiotensin AT1 receptor antagonist, losartan, but not a Ca2+ channel blocker, verapamil, inhibited the development of nephrosis while both agents decreased blood pressure to a similar extent as imidapril. In mature Dahl/S rats, imidapril suppressed not only the development of proteinuria but also the glomerular lesions. It is concluded that the development of spontaneous nephrosis in Dahl/S rats is mediated by angiotensin II.


Precise clinicopathologic findings for application of genetic testing in pediatric kidney transplant recipients with focal segmental glomerulosclerosis/steroid-resistant nephrotic syndrome.
Establishing a molecular genetic diagnosis of focal segmental glomerulosclerosis (FSGS)/steroid-resistant nephrotic syndrome (SRNS) can be useful for predicting post-transplant recurrence. Monogenic causes are reportedly present in approximately 20-30% of patients with FSGS/SRNS. However, the characteristics of patients who are likely to have a monogenic cause remain to be determined. Pediatric recipients with SRNS and/or biopsy-proven FSGS who underwent their first kidney transplantation at our center between 1999 and 2019 were analyzed. Patients with secondary FSGS/SRNS were excluded. The recipients were divided into three groups: familial/syndromic, presumed primary, and undetermined FSGS/SRNS. Patients who met all of the following criteria were categorized as having presumed primary FSGS/SRNS: (i) nephrotic syndrome, (ii) complete or partial remission with initial steroid therapy and/or additional immunosuppressive therapies, and (iii) diffuse foot process effacement on electron microscopy in the native kidney biopsy. All patients underwent genetic testing using next-generation sequencing. Twenty-four patients from 23 families were analyzed in this study. Pathogenic or likely pathogenic variants in FSGS/SRNS-related genes were identified in four of four families, zero of eight families, and 10 of 11 families with familial/syndromic, presumed primary, and undetermined FSGS/SRNS, respectively. Post-transplant recurrence only occurred in patients with presumed primary FSGS/SRNS. Our systematic approach based on precise clinicopathological findings including nephrotic syndrome, treatment responses, and diffuse foot process effacement might be useful to differentiate pediatric kidney transplant recipients with FSGS/SRNS who are likely to have a monogenic cause from patients who are not, and to predict post-transplant recurrence. A higher resolution version of the Graphical abstract is available as Supplementary information.


Acid-base disorders: application of total body carbon dioxide titration in anesthesia.
Every anesthesiologist is aware of the importance of acid-base determinations, but the multiplicity of duties required in clinical anesthesia often leave time for blood-gas measurements at most. The author presents a relatively simple four-step system for more complete interpretation, with biomathematical back-grounding. In summary, he points out that the dominant parameter remains the patient's clinical history in relation to hydrogen ion, Paco2, and bicarbonate.


Recent work on Alzheimer's disease transgenics.
The most significant feature of the current transgenic models of Alzheimer's disease continues to be the amyloid phenotype. In the past year, mice have been more extensively characterized in terms of the effect of amyloid accumulation on downstream events, such as neurodegeneration and behavioral changes, but the results have been complex. Genetic crosses have shown that apolipoprotein E and TGF-beta1 influence the deposition event and that the presenilins act synergistically with the amyloid precursor protein in pathology development. The mice have great utility in amyloid modulation studies but are still not complete models of Alzheimer's disease.


Gender, migration and violence among third country nationals accessing the sexual health clinic in Malta.
In 2020, the number of displaced people worldwide reached 41.3 million (IOM, 2020). Among them, are many migrants and refugees at risk of sexual and gender-based violence (SGBV). Healthcare providers have a key role to play in identifying migrant victims/survivors of violence. This paper seeks to assess STIs prevalence, sexual health and sexual violence among third country nationals (TCNs) attending the GUC in Malta. This is a mixed methods study carried out at the Genitourinary Clinic (GUC), which is the only public sexual health clinic in Malta. Demographic data, sexual history and diagnoses of patients attending the GUC between January 2018 and December 2019 were collected and retrospectively analysed. A SGBV risk assessment was performed through a semi-structured questionnaire. In the 24-month study period, a total of 12 654 patients accessed the GUC in Malta. Demographic data were collected on age, gender, nationality, marital status and sexual orientation. 16.4% (n = 2064) of these were extra-European migrants, predominantly male. 80 different nationalities were recorded, with the 5 most common being Nigerian, Filipino, Libyan, Syrian and Brazilian. The average age was 32.6 years. Over 110 sex workers were visited at the GUC in the study period - 20 were foreign, primarily from China. The presence of a 'massage parlour owner' during consultation, lack of control over passports and other factors were identified as warning signs of trafficking. 5 cases of sexual violence and forced prostitution involving girls from Sub-Saharan Africa and, in 2 cases, boys recently arrived in Malta by boat, were encountered. 6 African women accessing the service exhibited a type of female genital mutilation (FGM). Migration, sexual health and SGBV overlap in important ways. Further research and training in SGBV and migration in the healthcare setting and awareness-raising about existing services among the migrant population are required.


Sensitive detection of cancer cells using light-mediated apta-PCR.
Apta-PCR is an ultrasensitive assay in which aptamers are exploited not only as biomolecular recognition elements, but also as reporter labels for amplification via real-time PCR. This methodology has been successfully applied to the detection of proteins, achieving limits of detection in the picomolar range. The introduction of caged aptamers that bear photo-labile groups, so called cages, at strategic positions so that their tertiary structure and thus their binding properties can be controlled by light, facilitates a more robust and attractive assay in terms of sample conservation and reusability. In this work, we report for the first time the use of caged aptamers for cell detection in an apta-PCR assay. Specifically, a sandwich format is used combining the capture of B-cells by an antibody with the specific detection of Burkitt's lymphoma cancer cells by a caged aptamer, acting as a reporter probe. Elution of the aptamer bound to the cancer cells is performed by light and the number of cells is then correlated with the amount of eluted caged aptamer using real-time PCR analysis. The reported technique shows an excellent sensitivity, achieving detection of as few as 77 cells, and due to the inherent robustness of the assay, this detection platform can be reused for further analyses, demonstrating potential applicability in proteomics and clinical diagnostics.


Ophthalmic features of Turner's syndrome.
Turner's syndrome is one of the most common of all chromosomal abnormalities and is associated with significant ophthalmic morbidity. Turner's 1938 account included two patients with strabismus, and hitherto the condition has generated more interest among orthoptists than ophthalmologists. This systematic review of the literature seeks to redress the balance. Based on the pooled data of 274 patients with Turner's syndrome, it is the most complete evaluation so far of the prevalence and severity of ophthalmic problems in this population. This includes both a systematic review of the ophthalmic literature (via Medline) and the much larger body of work available in the orthoptic literature. Finally, we consider recent progress that enables the ophthalmologist to progress from the simple recognition of a phenotype to the correlation of genotypic variations with embryogenesis and consequent features of that phenotype.


Digit ratio (2D:4D) and psychopathic traits moderate the effect of exogenous testosterone on socio-cognitive processes in men.
Recent evidence suggests that testosterone is negatively correlated with empathic processes in both men and women. Also, administration of testosterone to young women impairs socio-cognitive performance as assessed using the "Reading the Mind in the Eyes Task", especially among those exposed to elevated testosterone concentrations prenatally. However, the extent to which testosterone plays a similar causal role in socio-cognitive abilities in men is currently unknown. Here, using a crossover, double-blind, placebo-controlled, within-subject design, we investigated the extent to which a single administration of testosterone to healthy young men (N=30) would impair socio-cognitive abilities assessed using the "Reading the Mind in the Eyes Task" (RMET). Also, we investigated whether individual differences in 2D:4D ratio and psychopathic traits would moderate the effect of testosterone on task performance. Results indicated that testosterone administration on its own did not impair RMET performance. However, variability in both 2D:4D ratio and psychopathic traits moderated the effect of testosterone on task performance. Specifically, testosterone impaired RMET performance among individuals with relatively low (i.e., masculinized) 2D:4D ratio and among individuals scoring relatively low on the interpersonal/affective facet (i.e., Factor 1) of psychopathy. Our findings highlight the importance of considering theoretically- and empirically-based individual difference factors when attempting to characterize the neuroendocrine mechanisms underlying socio-cognitive processes.


HLA-D/DR restriction of Langerhans cell-dependent antigen activation of T lymphocytes. The same D/DR determinants are restriction elements on monocytes and Langerhans cells.
In previous studies we have shown that the response of T cells to antigen presented by epidermal Langerhans cells (LC) is restricted by products of the HLA-D region. An optimal antigen-specific response required that the LC used for antigen presentation shared both, or at least one, of the D/DR determinants of the T-cell donor. These studies were, however, disturbed by a strong allogeneic response induced by the foreign D/DR determinants of the LC. We report here that by separating the antigen-specific T cells from those that are alloreactive, a clearer picture of the D/DR restriction phenomenon may be obtained. Furthermore, the present studies demonstrate that the same D/DR determinants function as restriction elements on peripheral blood monocytes and LC.


Thirty-day recurrence after ischemic stroke or TIA.
Incidence of recurrent stroke is highest within 30 days after the initial ischemic stroke (IS) or TIA, but knowledge about early recurrence is lacking. We aimed to identify etiological groups with highest risk of early recurrence and assess how the TOAST classification identified index stroke etiology. Medical records of 1874 IS and TIA patients in the Bergen NORSTROKE registry were retrospectively reviewed for identification of recurrent IS or TIA within 30 days after index IS or TIA. Stroke etiology was determined by review of electronical medical journals. Logistic regression was used to calculate odds ratios (OR) for 30-day recurrence. Thirty-three patients (1.8%) were readmitted with recurrent IS or TIA within 30 days after index stroke. By using TOAST, 12 patients were initially classified with stroke of unknown etiology (SUE). Etiologies behind recurrent IS or TIA were after the recurrent episode identified as extracranial large artery atherosclerosis (LAA) in 14 patients (42.4%), intracranial arterial pathology in seven patients (21.2%), active malignancy in six patients (18.2%), and cardio embolism in four patients (12.1%). Small vessel occlusion and SUE were the causes in one patient each. Logistic regression showed that patients with stroke of other determined etiology (SOE) and LAA had increased risk of 30-day recurrence (OR = 9.72, 95% CI 1.84-51.3, p < 0.01 and OR = 4.36, 95% CI 2.01-9.47, p < 0.01, respectively). Patients with LAA and SOE had increased risk of recurrent IS or TIA within 30 days. TOAST was inadequate at identifying exact etiologies behind recurrent stroke at index event.


The ryh1 gene in the fission yeast Schizosaccharomyces pombe encoding a GTP-binding protein related to ras, rho and ypt: structure, expression and identification of its human homologue.
A gene, ryh1, of the fission yeast Schizosaccharomyces pombe encoding a GTP-binding protein of 201 amino acids and belonging to the ras superfamily was isolated using the protein-coding region of the cloned Saccharomyces cerevisiae YPT1 gene as hybridization probe. The ryh1 gene is interrupted by three introns. ryh1 null mutants are viable but unable to grow at temperatures greater than 35.5 degrees C. Invertase of ryh1- cells is properly secreted but has a faster electrophoretic mobility compared to that of wild-type cells. The temperature-sensitive phenotype of ryh1 null mutants is complemented by the human rab6 cDNA expressed either under transcriptional control of the S.pombe adh or the SV40 early promoter.


Influence of CYP3A4, CYP3A5, and ABCB1 genotype and expression on budesonide pharmacokinetics: a possible role of intestinal CYP3A4 expression.
Budesonide treatment of chronic inflammatory bowel disease commonly leads to non-response or adverse reactions, possibly because of alterations in efflux transport mediated by the ABCB1 gene product P-glycoprotein or metabolism by CYP3A isoenzymes. Two groups, each consisting of nine healthy volunteers, one with the CYP3A5(*)1/(*)3 genotype (expressors) and the other with the CYP3A5(*)3/(*)3 genotype (non-expressors), were given a single oral dose of 9 mg budesonide. Plasma and urine concentrations of budesonide and its major metabolites were determined using liquid chromatography-tandem mass spectrometry. Subsequently, rectosigmoidal biopsies were taken for analysis of messenger RNA (mRNA) expression. Budesonide pharmacokinetics did not differ between genotype groups. However, intestinal CYP3A4 expression was shown to correlate directly with partial metabolic clearances of 16-hydroxy-prednisolone (r(2) = 0.30; P = 0.010) and 6-hydroxy-budesonide (r(2) = 0.25; P = 0.016), but inversely with budesonide AUC(0-24 h) (r(2) = 0.18; P = 0.040). Interestingly, a strong correlation was found between CYP3A5 and ABCB1 expression in CYP3A5 expressors (r(2) = 0.79; P = 0.001). This study suggests that intestinal CYP3A4 expression has an impact on budesonide pharmacokinetics. Moreover, CYP3A5 and ABCB1 expression appears to be coregulated.