Novel mutations of HSD17B3 in three Chinese patients with 46,XY Disorders of Sex Development.
17β-Hydroxysteroid dehydrogenase type 3 (17β-HSD3) converts the inactive Δ4-androstenedione (A) to testosterone (T). Its deficiency is the most common testosterone biosynthesis defect that results in 46,XY Disorders Of Sex Development (DSD). However, the disease is difficult to distinguish from other 46,XY DSD for similar clinical phenotypes. Therefore, genetic testing provides good criteria for the diagnosis of the disease. In this study, HSD17B3 gene was examined in 3 unrelated Chinese patients with 46,XY DSD. Direct sequencing and quantitative PCR of HSD17B3 gene revealed the presence of a compound heterozygous mutation (p.I60T/exon1 deletion) in Patient 1, a homozygous (p.I60T) mutation in Patient 2 and a frameshift mutation (p.V25Efs∗54) and an exon1 deletion in Patient 3. All of the mutations have not been reported previously. These novel mutations may expand the mutation database of HSD17B3 gene and provide us new insights into the molecular mechanism of 17β-HSD3 deficiency. It is noteworthy that when direct sequence analysis showed a rare homozygous mutation in patients with non-consanguineous parents, "apparent homozygosity" should be taken into an account and the intragenic deletion should be screened. In addition, when single mutation was found in patients with disease in recessive heredity mode, the intragenic deletion should also be screened.


Geriatric benign paroxysmal positional vertigo: a single-center study.
To compare the clinical features, risk factors, distribution of Benign Paroxysmal Positional Vertigo (BPPV) subtypes, and effectiveness of canalith repositioning between geriatric and non-geriatric patients with BPPV. A total of 400 patients with BPPV were enrolled. Canalith repositioning was performed according to the semicircular canals involved. Patients were divided by age into a geriatric group (≥60 years) and a non-geriatric group (20-59 years). Clinical characteristics, potential age-related risk factors, distribution of subtypes, and effectiveness of canalith repositioning were compared between the groups. Female sex was significantly more common in all age groups, with a peak female-to-male ratio of 5.1:1 in the group aged 50-59 years. There was a higher proportion of men in the geriatric group. A history of disease associated with atherosclerosis was significantly more common in the geriatric group (p < 0.05). Migraine was significantly more common in the non-geriatric group (p = 0.018), as was posterior canal BPPV. The horizontal canal BPPV (especially horizontal canal BPPV-cupulolithiasis), and multicanal BPPV subtypes were more common in the geriatric group, whereas anterior canal BPPV was more common in the non-geriatric group. Two canalith repositioning sessions were effective in 58.0% of the geriatric cases and in 72.6% of the non-geriatric cases (p = 0.002). There was a tendency for the effectiveness of canalith repositioning to decrease with increasing age. BPPV was more common in women. However, the proportion of men with BPPV increased with age. Elderly patients often had a history of diseases associated with atherosclerosis (i.e., hypertension, diabetes, and hyperlipidemia). The horizontal canal BPPV (particularly horizontal canal BPPV-cupulolithiasis) and multicanal BPPV subtypes were more common and the anterior canal BPPV subtype was less common in elderly patients. The effectiveness of canalith repositioning may decrease with age. Therefore, older patients should receive more comprehensive medical treatment.


Regulation of the p300 HAT domain via a novel activation loop.
The transcriptional coactivator p300 is a histone acetyltransferase (HAT) whose function is critical for regulating gene expression in mammalian cells. However, the molecular events that regulate p300 HAT activity are poorly understood. We evaluated autoacetylation of the p300 HAT protein domain to determine its function. Using expressed protein ligation, the p300 HAT protein domain was generated in hypoacetylated form and found to have reduced catalytic activity. This basal catalytic rate was stimulated by autoacetylation of several key lysine sites within an apparent activation loop motif. This post-translational modification and catalytic regulation of p300 HAT activity is conceptually analogous to the activation of most protein kinases by autophosphorylation. We therefore propose that this autoregulatory loop could influence the impact of p300 on a wide variety of signaling and transcriptional events.


Involvement of secretory and cytosolic phospholipases A2 during infection of THP1 human monocytic cells with Toxoplasma gondii. Effect of interferon gamma.
Toxoplasma gondii is an intracellular, obligate protozoan parasite that actively invades host cells. Phospholipases A2 (PLA2) are widely distributed enzymes that exist in many isoforms [secretory (sPLA2), cytosolic (cPLA2)] in monocytic and other human cells (pancreatic, kidney, etc) and also in parasites. We examined the effects of inhibitors of sPLA2 type II and cPLA2 on the invasion of human monocytic cells (THP1) by T. gondii (RH strain). We also measured sPLA2 type II and cPLA2 enzyme activities and their modulation by interferon gamma (IFN gamma) in extracts of host cells and parasite. Inhibition of both parasite and THP1 sPLA2 type II, and of parasite cPLA2 reduced the number of infected cells. Enzyme assays and immunoblot analyses demonstrated T. gondii sPLA2 type II and cPLA2 activities and indicated that T. gondii increased the activity of THP1 sPLA2 type II. Incubation with IFN gamma (1,000 units/ml) for 20 h reduced the activities of sPLA2 and cPLA2 in infected and non-infected cells. Thus, IFN gamma blocks the activities of both THP1 and parasite sPLA2 and cPLA2 in membrane fractions, resulting in protection against active invasion by T. gondii.


Post-anesthetic cortical blindness in cats: twenty cases.
The medical records of 20 cats with post-anesthetic cortical blindness were reviewed. Information collected included signalment and health status, reason for anesthesia, anesthetic protocols and adverse events, post-anesthetic visual and neurological abnormalities, clinical outcome, and risk factors. The vascular anatomy of the cat brain was reviewed by cadaver dissections. Thirteen cats were anaesthetised for dentistry, four for endoscopy, two for neutering procedures and one for urethral obstruction. A mouth gag was used in 16/20 cats. Three cats had had cardiac arrest, whereas in the remaining 17 cases, no specific cause of blindness was identified. Seventeen cats (85%) had neurological deficits in addition to blindness. Fourteen of 20 cats (70%) had documented recovery of vision, whereas four (20%) remained blind. Two cats (10%) were lost to follow up while still blind. Ten of 17 cats (59%) with neurological deficits had full recovery from neurological disease, two (12%) had mild persistent deficits and one (6%) was euthanased as it failed to recover. Four cats (23%) without documented resolution of neurological signs were lost to follow up. Mouth gags were identified as a potential risk factor for cerebral ischemia and blindness in cats.


Texture analysis and multiple-instance learning for the classification of malignant lymphomas.
Malignant lymphomas are cancers of the immune system and are characterized by enlarged lymph nodes that typically spread across many different sites. Many different histological subtypes exist, whose diagnosis is typically based on sampling (biopsy) of a single tumor site, whereas total body examinations with computed tomography and positron emission tomography, though not diagnostic, are able to provide a comprehensive picture of the patient. In this work, we exploit a data-driven approach based on multiple-instance learning algorithms and texture analysis features extracted from positron emission tomography, to predict differential diagnosis of the main malignant lymphomas subtypes. We exploit a multiple-instance learning setting where support vector machines and random forests are used as classifiers both at the level of single VOIs (instances) and at the level of patients (bags). We present results on two datasets comprising patients that suffer from four different types of malignant lymphomas, namely diffuse large B cell lymphoma, follicular lymphoma, Hodgkin's lymphoma, and mantle cell lymphoma. Despite the complexity of the task, experimental results show that, with sufficient data samples, some cancer subtypes, such as the Hodgkin's lymphoma, can be identified from texture information: in particular, we achieve a 97.0% of sensitivity (recall) and a 94.1% of predictive positive value (precision) on a dataset that consists in 60 patients. The presented study indicates that texture analysis features extracted from positron emission tomography, combined with multiple-instance machine learning algorithms, can be discriminating for different malignant lymphomas subtypes.


Comparative effects of three 1,4-dihydropyridine derivatives [OSI-1210, OSI-1211 (etaftoron), and OSI-3802] on rat liver mitochondrial bioenergetics and on the physical properties of membrane lipid bilayers: relevance to the length of the alkoxyl chain in positions 3 and 5 of the DHP ring.
The 1,4-dihydropyridines OSI-1210, OSI-1211 (etaftoron), and OSI-3802 are compounds with similar chemical structure. They differ by the length of the alkoxyl chain in positions 3 and 5 of the dihydropyridine (DHP) ring and by their pharmacological action characteristics. However, as far as we know, a clear relationship between the effects of these compounds and the length of the alkoxyl chain in positions 3 and 5 of the DHP has not been established. The goal of this study was to compare the influence of OSI-1210, OSI-1211 (etaftoron), and OSI-3802 on rat liver mitochondrial bioenergetics and on the physical properties of membrane lipid bilayers, correlating their actions with the length of the alkoxyl chain in positions 3 and 5 of the DHP ring. Using either glutamate/malate or succinate as respiratory substrates, all the compounds, in concentrations of up to 500 microM, depressed state 3 and uncoupled respiration, respiratory control (RCR) and ADP/O ratios, and phosphorylation rate, whereas state 4 respiration was stimulated. However, the stimulatory effect on state 4 induced by OSI-3802, the compound with the longest chain in positions 3 and 5 of the DHP ring, as well as its inhibitory effects on RCR and ADP/O ratios and phosphorylation rate were more pronounced than that induced by OSI-1210 and OSI-1211 (etaftoron), the compounds with the shortest and intermediate chains, respectively. Moreover, OSI-3802 maximized state 4 stimulation and minimized RCR and ADP/O ratios, and phosphorylation rate at a concentration of 100 microM, whereas low graduate effects were detected with OSI-1210 and OSI-1211 (etaftoron) for concentrations of up to 500 microM. At low concentrations (< or =30 microM), OSI-3802, like its analogue OSI-1212 (cerebrocrast), reduced the phase transition temperature, the cooperative unit size, and the enthalpy associated with the phase transition temperature of dimyristoylphosphatidylcholine (DMPC) membrane bilayers. A good correlation was established between the effects of 200 microM OSI-1210, OSI-1211 (etaftoron), and OSI-3802 on glutamate/malate- and succinate-dependent RCR of rat liver mitochondria and on the enthalpy change (Delta H) for the thermotropic profile of DMPC membrane bilayers at a 0.2 drug/DMPC molar ratio, indicating that the changes induced by these compounds on both mitochondrial membrane integrity and physical properties of DMPC membrane bilayers are strongly related to the length of the alkoxyl chain in positions 3 and 5 of the DHP ring. A putative relationship between membrane physical perturbation, bioenergetics impairment and the molecular characteristics of the compounds will be established as an approach to better understand their differentiated toxicological and pharmacological actions.


Examining the symptom of fatigue in primary care: a comparative study using electronic medical records.
The symptom of fatigue is one of the top five most frequently presented health complaints in primary care, yet it remains underexplored in the Canadian primary care context. The objective of this study was to examine the prevalence and impact of patients presenting with fatigue in primary care, using the only known electronic database in Canada to capture patient-reported symptoms. Data were extracted from the Deliver Primary Healthcare Information (DELPHI) database, an electronic medical record database located in Ontario, Canada. Patients were identified using the International Classification of Primary Care, Revised Second Edition coding system. Two groups of patients (fatigue or non-fatigue symptom) were followed for one year and compared. Both descriptive and multivariable analyses were conducted. A total of 103 fatigue symptom patients, and 103 non-fatigue symptom patients, were identified in the DELPHI database. The period prevalence of fatigue presentation was 8.2%, with the majority of patients being female and over 60 years of age. These patients experienced numerous co-occurring morbidities, in addition to the fatigue itself. During the one year follow-up period, fatigue symptom patients had significantly higher rates of subsequent visits (IRR = 1.19, p = 0.038) and investigations (IRR = 1.68, p < 0.001), and markedly high levels of referrals following their index visit. This research used an electronic database to examine the symptom, fatigue. Using these data, fatigue symptom patients were found to have higher rates of health care utilisation, compared to non-fatigue symptom patients.


Establishment and characterization of a clonal line of parathyroid endothelial cells.
We have isolated endothelial cells derived from bovine parathyroid tissue. These cells have been cloned and maintained by serial passage for more than 40 months without showing signs of senescence. Prolonged culture was accomplished by using a medium favoring endothelial cell growth and methods for enriching endothelial cells in primary culture. The cloned parathyroid endothelial cells contained factor VIII-related antigen, took up acetylated low-density lipoproteins and parathyroid hormone, and showed morphological features comparable to other endothelial cells. Bovine parathyroid endothelial cells replicated with a mean doubling time of 65 h. Fibroblast growth factors, platelet-derived growth factor, and calcium acted as mitogens for parathyroid endothelial cells, whereas transforming growth factor beta inhibited proliferation.


The use of nanomaterials to treat bone infections.
A new era of osteomyelitis treatment has been taking strides towards efficient, local administration of antibiotics at the site of infection. By having them localized to the site of infection, this toxicity is no longer an issue and actually has shown to be a more productive treatment for osteomyelitis. Researchers have focused the production of non-biodegradable, antibiotic, infused bone cements specifically designed for proficient osteocyte binding, useful antibiotic release over a desirable period of time, and promotion of bone regeneration. These cements are then surgically placed on the infected site following debridement and irrigation. The problem, however, is that the use of ineffective cements and the overuse of antibiotics has led to the development of resistant bacteria. Due to this, further research is being done in the field of antibiotic discovery and delivery. Specifically, the development of biodegradable materials capable of efficiently delivering antibiotics and also eliminating the need for follow-up surgery to remove the delivery material is being done, thus reducing exposure risk. Nanoparticles have been developed in the forms of scaffolds and injections to deliver a higher degree and longer lasting duration of antibiotic release, while promoting bone regeneration.


[Discovery of a new species of the pentastomid genus Porocephalus (Humboldt, 1811) from Taiwan, China and its pathogenic features].
To describe the morphological characteristics of Porocephalus taiwana sp. nov., discuss its pathogenic features and the method of etiological diagnosis of the new disease. Fecal sedimentation concentration was used to collect nymphs from the patient's watery stool for species identification. Clinical information was collected for determining the pathogenic features of the new infection. A new pathogenic pentastomid Porocephalus taiwana sp. nov. is discovered and a new disease, porocephaliasis taiwana, is nominated. With the findings from this case it is proposed that the traditional visceral pentastomiasis should be divided into two subtypes, Encystic and Excystic. According to the pathological features, this case belongs to the excystic visceral pentastomiasis. Porocephalus taiwana sp. nov. is a new pathogenic pentastomid infecting humans. Porocephaliais taiwana belongs to a novel type (excystic) of visceral pentastomiasis.


Akt is required for Axl-Gas6 signaling to protect cells from E1A-mediated apoptosis.
Adenovirus type 5 E1A protein (E1A) associates with anti-tumor activities by reversing the transformed phenotype, inhibiting metastasis, and inducing apoptosis. We have previously identified that E1A suppresses the expression of Axl, a transforming tyrosine kinase and that Axl-Gas6 receptor-ligand interaction prevents E1A transfectants from apoptosis induced by serum deprivation. To determine how the Axl-Gas6 interaction prevents E1A-mediated apoptosis, we analysed the expression of anti-apoptotic molecules and found that the activated form of Akt was suppressed in the E1A transfectant ip 1-E1A and that Gas6 was able to activate Akt in ip 1-E1A cells reexpressing Axl (ip 1-E1A-Axl). To determine whether activated Akt is required to prevent E1A-mediated apoptosis, ip 1-E1A-Axl cells were treated with the phosphatidylinositol-3'-OH kinase inhibitor wortmannin or transfected with a dominant negative Akt mutant. In both cases, Gas6 no longer protected the cells from serum deprivation-induced apoptosis. Thus, we conclude that activated Akt is required for Axl-Gas6 signaling to prevent E1A-mediated apoptosis induced by serum deprivation. Downstream molecules of Akt, including NF-kappaB, Fas ligand, and BAD were examined, among which phosphorylation of BAD by Axl-Gas6 signaling is associated with the anti-apoptotic activity of Akt in our study.


Prediction of biodegradation rate constants of hydroxylated polychlorinated biphenyls by fungal laccases from Trametes versicolor and Pleurotus ostreatus.
Quantitative structure-activity relationship (QSAR) models for fungal laccase-catalyzed degradation of different hydroxylated polychlorinated biphenyls (OH-PCBs) were developed using some fundamental quantum chemical descriptors. The cross-validated Q(2)(cum )values for the two optimal QSAR models are as high as 0.958 and 0.961 for laccases from Trametes versicolor and Pleurotus ostreatus, respectively, indicating good predictive abilities for laccase-catalyzed degradation of OH-PCBs. Results from this study show that increasing heat of formation (DeltaH(f)) and frontier molecular orbital energy (i.e. E(LUMO) + E(HOMO)) values or decreasing frontier molecular orbital energy (i.e. E(HOMO-1)) and core-core repulsion energy (CCR) values leads to the increase of OH-PCB degradation rates by laccases.


Priming of measles virus-specific humoral- and cellular-immune responses in macaques by DNA vaccination.
Although the currently used live attenuated measles vaccines are safe and effective, they are dependent on cold chain maintenance and are often ineffective in young infants due to interference by maternal antibody. Therefore, besides vector-based vaccines, different new generation non-replicating candidate measles vaccines are being considered, including nucleic acid vaccines. We have vaccinated cynomolgus macaques transdermally with DNA plasmids encoding measles virus (MV) proteins. Following two vaccinations, low serum antibody responses were detected. Wild-type measles virus challenge 1 year after vaccination showed reduced viraemia in some animals. However, accelerated humoral- and cellular-immune responses were observed in all vaccinated macaques, demonstrating successful priming by the DNA vaccines.


Positional therapy for obstructive sleep apnea: an objective measurement of patients' usage and efficacy at home.
Positional therapy that prevents patients from sleeping supine has been used for many years to manage positional obstructive sleep apnea (OSA). However, patients' usage at home and the long term efficacy of this therapy have never been objectively assessed. Sixteen patients with positional OSA who refused or could not tolerate continuous positive airway pressure (CPAP) were enrolled after a test night study (T0) to test the efficacy of the positional therapy device. The patients who had a successful test night were instructed to use the device every night for three months. Nightly usage was monitored by an actigraphic recorder placed inside the positional device. A follow-up night study (T3) was performed after three months of positional therapy. Patients used the device on average 73.7 ± 29.3% (mean ± SD) of the nights for 8.0 ± 2.0 h/night. 10/16 patients used the device more than 80% of the nights. Compared to the baseline (diagnostic) night, mean apnea-hypopnea index (AHI) decreased from 26.7 ± 17.5 to 6.0 ± 3.4 with the positional device (p<0.0001) during T0 night. Oxygen desaturation (3%) index also fell from 18.4 ± 11.1 to 7.1 ± 5.7 (p = 0.001). Time spent supine fell from 42.8 ± 26.2% to 5.8 ± 7.2% (p < 0.0001). At three months (T3), the benefits persisted with no difference in AHI (p = 0.58) or in time spent supine (p = 0.98) compared to T0 night. The Epworth sleepiness scale showed a significant decrease from 9.4 ± 4.5 to 6.6 ± 4.7 (p = 0.02) after three months. Selected patients with positional OSA can be effectively treated by a positional therapy with an objective compliance of 73.7% of the nights and a persistent efficacy after three months.


Blockade of the nootropic action of piracetam-like nootropics by adrenalectomy: an effect of dosage?
The present experiments demonstrate that the absence of any memory-improving action of nootropics in adrenalectomized animals cannot be ascribed to an effect of dosage. Doses of 1, 10, 100, 1000 and 3000 mg/kg p.o. of piracetam, oxiracetam, aniracetam or pramiracetam are ineffective in adrenalectomized mice.


Calcium absorption from small soft-boned fish.
The prevalence of osteoporosis in developing countries is low compared to most industrialised countries despite an apparent low Ca intake. It is possible, however, that food surveys have overlooked important Ca sources in developing countries. Small fish eaten with the bones can be a rich source of Ca, even though Ca from bone may be considered unavailable for absorption. In the present study, absorption of Ca from indigenous Bengali small fish was compared with the Ca absorption from milk. Ca absorption from single meals was determined in 19 healthy men and women (21-28 y). Each subject received two meal types on two separate occasions. Both meals consisted of white wheat bread, butter and ultra pure water with the main Ca source being either small Bengali fish (397 mg Ca in total) or skimmed milk (377 mg Ca in total). The meals were extrinsically labelled with 47Ca, and whole-body retention was measured on day 8, 12, 15 and 19 after intake of each meal. The labelling procedure was evaluated by an in vitro method. The calculated absorption of Ca as measured with 47Ca whole-body retention was 23.8 +/- 5.6% from the fish meal and 21.8 +/- 6.1% from the milk meal (mean +/- SD), which was not significantly different (p = 0.52). Even after correction for an incomplete isotope exchange, as indicated by the in vitro study, Ca absorption was similar from the two meal types. It was concluded that Ca absorption from small Bengali fish was comparable that from skimmed milk, and that these fish may represent a good source of Ca.


Activation of the estrogen receptor from N-nitrosomethylurea-induced rat mammary tumors.
The activation of the estrogen receptor (ER) from N-nitrosomethylurea (NMU)-induced rat mammary tumors was studied in vitro. The activation of the receptor induced by heating of the cytosol containing occupied ER was measured by a 3-4-fold increase of receptor binding to nuclei in comparison with the nuclear binding of the nonactivated ER. The activation of the ER was further shown by alteration of the elution profile from DEAE-cellulose. A shift of the receptor peak from 234 mM (Peak II, nonactivated ER) to 70 mM (Peak I, activated ER) phosphate buffer could be obtained. The overall recoveries of activated ER following chromatography on DEAE-cellulose were significantly lower than the recoveries of the nonactivated ER, 71 and 85%, respectively. Binding of the activated ER to nuclei and chromatography of the supernatant which is not able to bind to nuclei on DEAE-cellulose resulted in a decrease of Peak I and in an increase of the overall recovery. These findings suggest that the nuclear bound ER consists of two parts. One is represented partially by Peak I of the elution profile and the other one by that part of the receptor which can not be eluted from the column under the conditions used. Furthermore, the dissociation of tritiated estradiol (E3H) from the nonactivated ER followed a two component exponential function whereas after activation a monophasic dissociation curve could be observed. The mean half times for the dissociation of E3H from the activated and nonactivated ER were 101 and 7.2 min, respectively. Finally, the nonactivated molybdate stabilized ER sedimented in 5-20% sucrose density gradients as two peaks, one at 9.5 S and the other at 4 S. After activation of the ER only the smaller 4 S peak was evident. Molybdate inhibited the activation of the ER measured by nuclear binding assays, sucrose density gradient analysis, dissociation kinetics or ion exchange chromatography but not completely in every case.


Amino acids: chemistry, functionality and selected non-enzymatic post-translational modifications.
The ultimate goal of proteomics is determination of the exact chemical composition of protein species, including their complete amino acid sequence and the identification of each modified side chain, in every protein in a biological sample and their quantification. We are still far from achieving this goal due to limitations in analytical methodology and data analysis but also due to the fact that we surely have not discovered all amino acid modifications that occur in nature. To detect modified side chains and to discover new, still unknown amino acid derivatives, an understanding of the chemistry of the reactive groups of amino acids is mandatory. This tutorial focuses on the chemistry of the amino acid side chains and addresses non-enzymatic modifications. By highlighting some exemplary reactions a glimpse of the huge diversity of modified amino acids provides the reader with sufficient insight into amino acid chemistry to raise the awareness for unexpected side chain modifications. We further introduce the reader to a terminology, which enables the comprehensive description of the exact chemical composition of a protein species, including its full amino acid sequence and all modifications of its amino acid side chains. This Tutorial is part of the International Proteomics Tutorial Programme (IPTP number 10).


Parents' experiences of having their children take part in participatory action research creating a serious game about radiotherapy.
Radiotherapy (RT) is one of several treatment modalities used for children diagnosed with cancer. Several studies have designed interventions aimed to alleviate the stress that can occur in conjunction with RT. To include children in the design of interventions is rare and dependent on the parents giving consent to participation in research on their children's behalf. The aim was to illuminate, from the parents' perspective, the experience of their children being part in the co-creation of a serious game and their previous experiences of RT. Ten parents of children taking part in a participatory action research study of the development of a serious game were invited to an interview and seven parents consented. An inductive, manifest content analysis was performed. The analysis resulted in an overarching theme: Parents' pre-understanding from their child's cancer treatment created a sensitivity to their child's wishes and a willingness to contribute to science. Four categories are presented: Intrinsic factors influenced the intent to participate, Extrinsic factors that had an effect on participation, Parents role in the game development, and Radiotherapy impinged the child and the parent. To be able to pay it forward to healthcare and other families with a child diagnosed with cancer was a contributing factor for parents' willingness to consent to participation after their children had undergone RT, especially since their children wanted to do so. Parents indicated interest in the developmental process and in following their children being part of the creation of a serious game about RT. To understand why parents allow their children to participate in studies creates opportunities for the research community to structure studies that facilitate participation.


Sex determination from the pubis by discriminant function analysis.
A total of 122 adult human pubes (66 males and 56 females) are used in this study of a discriminant function analysis for determining sex from four pubic measurements: the angle formed by the middle line of the superior ramus and inferior ramus of pubis (in degrees, X1), subpubic angle (in degrees, X2), the minimum distance from the symphyseal surface to the obturator (in millimeters, X3), and the minimum thickness of the ischiopubic ramus (in millimeters, X4). The method introduced by Fisher is used. Using the formula Y = 0.270 mean1-0.985mean2-0.761mean3 + 1.806mean4,F = 200 F = 200 (F > F0.01(4,117), P < 0.01), section point -32.47, an accuracy of 100% could be achieved for 230 pubes (130 males, 100 females).


Do migrainous headaches become more consistently lateralized?
Analysis of information collected from 468 patients with common or classical migraine does not suggest that those with a longer history are more likely to have headaches fixed to a single side of the head. This finding does not support the notion that successive unilateral headache attacks on one side cause damage that predisposes the patient to further attacks on the same side.


Diabetes mellitus presenting as "acute osteomyelitis" of the index finger.
A case of previously undiagnosed maturity onset diabetes in a fifty-eight-year-old man with a "septic index finger" is reported. Initial X-rays showed gross destruction of the distal phalanx, suggestive of acute osteomyelitis. Control of the diabetes combined with minimal debridement of the finger resulted in healing of the lesion. X-rays taken four weeks later showed complete reconstitution of the distal phalanx, confirming the need to be conservative in the management of lesions associated with diabetes. In spite of a careful and extensive search into the literature covering unusual presentations of diabetes mellitus, the author is unable to trace a similar recorded case involving the digit of upper limb.


Web- and Artificial Intelligence-Based Image Recognition For Sperm Motility Analysis: Verification Study.
Human sperm quality fluctuates over time. Therefore, it is crucial for couples preparing for natural pregnancy to monitor sperm motility. This study verified the performance of an artificial intelligence-based image recognition and cloud computing sperm motility testing system (Bemaner, Createcare) composed of microscope and microfluidic modules and designed to adapt to different types of smartphones. Sperm videos were captured and uploaded to the cloud with an app. Analysis of sperm motility was performed by an artificial intelligence-based image recognition algorithm then results were displayed. According to the number of motile sperm in the vision field, 47 (deidentified) videos of sperm were scored using 6 grades (0-5) by a male-fertility expert with 10 years of experience. Pearson product-moment correlation was calculated between the grades and the results (concentration of total sperm, concentration of motile sperm, and motility percentage) computed by the system. Good correlation was demonstrated between the grades and results computed by the system for concentration of total sperm (r=0.65, P<.001), concentration of motile sperm (r=0.84, P<.001), and motility percentage (r=0.90, P<.001). This smartphone-based sperm motility test (Bemaner) accurately measures motility-related parameters and could potentially be applied toward the following fields: male infertility detection, sperm quality test during preparation for pregnancy, and infertility treatment monitoring. With frequent at-home testing, more data can be collected to help make clinical decisions and to conduct epidemiological research.


Comparative spectroscopic studies on drug binding characteristics and protein surface hydrophobicity of native and modified forms of bovine serum albumin: possible relevance to change in protein structure/function upon non-enzymatic glycation.
The interaction between serum albumin (SA) and drugs has provided an interesting ground for understanding of drug effects, especially in drug distribution and drug-drug interaction on SA, in the case of multi-drug therapy. Determination of the impact of various factors on drug-protein interaction is especially important upon significant binding of drug to albumin. In the present study, the interaction of two drugs (furosemide and indomethacin) with native and modified albumins were investigated by using various spectroscopic methods. Fluorescence data indicated that 1:1 binding of drugs to bovine serum albumin (BSA) is associated with quenching of albumin intrinsic fluorescence. The Job's plot also confirmed that drug binds to BSA via mentioned stoichiometry. Analysis of the quenching and thermodynamic parameters indicated that intermolecular interactions between drug and albumin may change upon protein modification. The theoretical analyses also suggested some conformational changes of interacting side chains in subdomain IIA binding site (at the vicinity of W237), which were in good agreement with experimental data. Decrease of protein surface hydrophobicity (PSH) was also observed upon both albumin modification and drug binding.


Baclofen-induced, calcium-dependent stimulation of in vivo release of D-[3H]aspartate from rat hippocampus monitored by intracerebral microdialysis.
The release of D-[3H]aspartate (used as a tracer for endogenous glutamate and aspartate) was studied at high K+ (100 mM) and under ischemia in rats implanted with 0.3 mm diameter dialysis tubing through the hippocampus. The effect on the D-[3H]aspartate release of the two gamma-aminobutyric acid (GABA) agonists 4,5,6,7-tetrahydroisoxazolo[5,4-c]-pyridin-3-ol (THIP) and (+/-)-beta-(p-chlorophenyl)GABA (baclofen), which specifically activate GABAA and GABAB receptors, respectively, was studied. Initial experiments employing HPLC analysis showed a coincident increase in the amounts of glutamate, aspartate and the amount of radioactivity following introduction of K+ (100 mM) or a period of ischemia suggesting that the D-[3H]aspartate labels the transmitter pools of the two amino acids under the present experimental conditions. The presence of 10 mM baclofen or 10 mM THIP in the perfusion medium did not inhibit ischemia induced D-[3H]aspartate release. On the contrary, 10 mM baclofen alone (but not 0.1 or 1 mM) in the perfusion medium induced release of D-[3H]aspartate in a calcium dependent manner, whereas 10 mM THIP had no significant releasing effect.


FOXJ2 expression in rat spinal cord after injury and its role in inflammation.
Foxj2 (forkhead box J2), a novel member of the forkhead/HNF3 family, binds DNA with a dual sequence specificity. It may play a role in maintenance and survival of developing and adult neurons. However, its expression and function in the central nervous system lesion are still unclear. In this study, we performed a spinal cord injury (SCI) model in adult Sprague-Dawley rats and investigated the dynamic changes of Foxj2 expression in the spinal cord. Western blot analysis revealed that Foxj2 was present in normal spinal cord. It gradually increased, reached a peak at day 5 after SCI, and then declined during the following days. Double immunofluorescence staining revealed wide expression of Foxj2, which is detected in neurons and astrocytes. After injury, Foxj2 expression was increased predominantly in astrocytes, which highly expressed proliferating cell nuclear antigen, a marker for proliferating cells. And knockdown of Foxj2 in cultured primary astrocytes by siRNA showed that Foxj2 played an important role in lipopolysaccharide-induced inflammatory responses. These results suggested that Foxj2 may be involved in the pathophysiology of SCI, and further research is needed to have a good understanding of its function and mechanism.


Molecular characterization and functional analysis of tumor necrosis factor receptor-associated factor 2 in the Pacific oyster.
Tumor necrosis factor receptor (TNFR)-associated factors (TRAFs) are a family of crucial adaptors, playing vital roles in mediating signal transduction in immune signaling pathways, including RIG-I-like receptor (RLR) signaling pathway. In the present study, a new TRAF family member (CgTRAF2) was identified in the Pacific oyster, Crassostrea gigas. Comparison and phylogenetic analysis revealed that CgTRAF2 could be a new member of the invertebrate TRAF2 family. Quantitative real-time PCR revealed that CgTRAF2 mRNA was highly expressed in the digestive gland, gills, and hemocytes, and it was significantly up-regulated after Vibrio alginolyticus and ostreid herpesvirus 1 (OsHV-1) challenge. The CgTRAF2 mRNA expression profile in different developmental stages of oyster larvae suggested that CgTRAF2 could function in early larval development. CgTRAF2 mRNA expression pattern, after the silence of CgMAVS (Mitochondrial Antiviral Signaling) -like, indicated that CgTRAF2 might function downstream of CgMAVS-like. Moreover, the subcellular localization analysis revealed that CgTRAF2 was localized in cytoplasm, and it may play predominately important roles in signal transduction. Collectively, these results demonstrated that CgTRAF2 might play important roles in the innate immunity and larval development of the Pacific oyster.


Extraction of high inverted mesiodentes via the labial, palatal and subperiostal intranasal approach:A clinical prospective study.
The aim of this study is to provide criteria for the choice of the surgical approach for extraction of high inverted mesiodens. The operation statistics, life quality of postoperative patients, and the operative injury/recovery were compared and analysed. The laser Doppler blood flowmetry, laser speckle contrast imaging, and electric pulp testing were explored to detect the postoperative pulp and gingiva blood supply of adjacent teeth. For the clinician's primary concerns, the surgical time, the volume of osteotomy, and the amount of bleeding in the labial approach group (The p values are 0.0001, <0.0001, and 0.0131, respectively.) and intranasal approach group (All p values were <0.0001.) were significantly less than that in the palatal approach group. However, from the patient's perspective, the postoperative swelling in the labial approach was far more than that in the intranasal approach group (p =0.0044), with unsurprisingly lower satisfaction (p <0.0001). There were no significant differences in pulp and gingival blood supply of adjacent teeth and jaw development. Trauma was manageable in all patients. Within the limitations of the study it seems that extraction of mesiodens by the intranasal approach achieves a delicate balance between reducing surgical trauma and optimizing postoperative recovery.


Aggressive Crystalloid Resuscitation Outcomes in Low-Severity Pediatric Trauma.
Trauma is the leading cause of death among children. Studies have found that insufficient intravenous (IV) fluid resuscitation contributes significantly to morbidity and mortality in pediatric trauma. While large-volume resuscitation represents a potential solution, overly aggressive fluid management may complicate hospitalizations and recovery. Through this study, we aim to evaluate the impact of aggressive fluid resuscitation on outcomes in pediatric trauma. This is a retrospective review utilizing our level I trauma center registry for pediatric patients aged <16 y admitted from 2014 to 2017. Patients transferred from our center within 24 h and those who arrived from outside hospitals were excluded. Patients who received blood product transfusions were excluded. Included patients were divided into two crystalloid groups: <60 mL/kg/24 h and ≥60 mL/kg/24 h. Outcome measures included ICU length-of-stay, length-of-hospitalization, complications, and mortality rate. Study sample included 320 patients (<60 mL/kg/24 h = 219; ≥60 mL/kg/24 h = 101). The ≥60 mL/kg/24 h group was younger (9.95 versus 5.27, P = 0.0001). There were no significant differences in GCS on arrival, injury severity score, Abbreviated Injury Scale, Revised Trauma Scores, traumatic brain injury, and operative intervention between groups. Outcome measures showed there was no significant difference in 30-day readmission rate, complications, or mortality. Large-volume crystalloid resuscitation was associated with longer mean ICU length-of-stay (1.5 d versus 0.8 d, P = 0.004). In this single-institution retrospective database analysis, large-volume crystalloid resuscitation (≥60 mL/kg) was associated with a significant increase in ICU length-of-stay without survival benefit. More research in the form of randomized trials will help determine the optimal rate for fluid resuscitation in pediatric trauma patients while weighing potential critical care complications.


The crux of helix curvature: A potential surface landmark for the anterior border of the sigmoid sinus in minimally invasive presigmoid approaches.
Surgical landmarks are widely used across all surgical specialties to assist surgeons in accurately estimating the deep anatomical structures. We describe the crus of helix curvature as a potential indicator for the anterior border of the sigmoid sinus (SS) in the setting of minimally invasive presigmoid approaches. Anatomy investigations were performed on injected cadaveric heads to identify consistent surface surgical landmarks estimating the curse of the SS. Presigmoid mastoid was noted in the center of the skin incision in 100% of the specimens. The crus of helix curvature was superimposed on the anterior border of the SS on 6 sides (60%). On the other four sides, the curvature lays within 5 mm of the anterior SS border (40%). For the minimally invasive presigmoid approaches, the "crus of helix curvature" can be used as a landmark for the anterior border of the SS, which can aid in the speed and safety of the procedure. The ease of use of this landmark makes it particularly convenient for physicians early in training.


Mortality in hepatitis C virus-infected patients with a diagnosis of AIDS in the era of combination antiretroviral therapy.
Before the introduction of combination antiretroviral therapy (cART), patients infected with the human immunodeficiency virus (HIV) rarely died of liver disease. In resource-rich countries, cART dramatically increased longevity. As patients survived longer, hepatitis C virus (HCV) infection became a leading cause of death; however, because patients with AIDS continue to have 5-fold greater mortality than non-AIDS patients, it is unclear whether HCV infection increases mortality in them. In this investigation, which is part of the Longitudinal Studies of the Ocular Complications of AIDS, plasma banked at enrollment from 2025 patients with AIDS as defined by the Centers for Disease Control and Prevention were tested for HCV RNA and antibodies. Three hundred thirty-seven patients had HCV RNA (chronic infection), 91 had HCV antibodies and no HCV RNA (cleared infection), and 1597 had no HCV markers. Median CD4(+) T-cell counts/µL were 200 (chronic), 193 (cleared), and 175 (no markers). There were 558 deaths. At a median follow-up of 6.1 years, patients with chronic HCV had a 50% increased risk of mortality compared with patients with no HCV markers (relative risk [RR], 1.5; 95% confidence interval [CI], 1.2-1.9; P = .001) in an adjusted model that included known risk factors. Mortality was not increased in patients with cleared infection (RR, 0.9; 95% CI, .6-1.5; P = .82). In patients with chronic HCV, 20.4% of deaths were liver related compared with 3.8% in patients without HCV. Chronic HCV infection is independently associated with a 50% increase in mortality among patients with a diagnosis of AIDS, despite competing risks. Effective HCV treatment may benefit HIV/HCV-coinfected patients with AIDS.


Control of grip and stick in cell adhesion through lateral relationships of membrane glycoproteins.
Although cell adhesion to substrate must ultimately depend on 'sticking' by physical forces at the outer surface, we suggest that control is exercised through the 'grip' of the cytoskeleton. Our experiments indicate that grip can be relaxed by agents which release surface glycoproteins from some form of side-to-side organisation, and can be consolidated by suitable cross-linking to augment this organisation.


Aberrant nerve fibres within the central nervous system.
Three cases of aberrant nerve fibres in the spinal cord and medulla oblongata are described. The literature on these fibres is discussed and their possible role in regeneration. Different views on the possibility of regeneration or functional recovery of the central nervous system are mentioned in the light of recent publications, which are more optimistic than before.


Cultural intelligence and proactive service performance: mediating and moderating role of leader's collaborative nature, cultural training and emotional labor.
Drawing on cultural intelligence and social exchange theories, this study examines cultural intelligence and its effects on proactive service performance and the mediating role of leader's collaborative nature and the moderating role of cultural training and emotional labor, particularly deep acting and surface acting, in the relationship between cultural intelligence and proactive service performance. The study sample comprised 510 healthcare practitioners. Structural equation modeling was used to examine the hypotheses. The results show that cultural intelligence positively influences proactive service performance. Additionally, leadership's collaborative nature influences proactive service performance. The moderating effect of cultural training and deep acting positively influences the relationship between cultural intelligence and proactive service performance. In contrast, surface acting reveals a reverse effect, thus exhibiting a positive effect on this relationship. These findings suggest that public healthcare organizations should pay more attention in improving deep acting, cultural training and leadership's collaborative nature for optimal service performance. The novelty of this study lies in its presentation of an integrated framework based on cultural intelligence and social exchange theories that can solve the contemporary challenges facing healthcare firms operating in emerging markets in integrating cultural intelligence and service performance.


Circumcision Status is Not Associated with Condom Use and Prevalence of Sexually Transmitted Infections Among Young Black MSM.
This study investigated whether intact young Black MSM differed from their circumcised counterparts regarding condom use behaviors and perceptions and HIV/Chlamydia/gonorrhea. Young Black MSM completed a self-interview, including a pictorial item assessing circumcision status and measures of condom use. Twenty-seven percent of 388 participants reported not being circumcised. With one exception, no associations tested approached significance. The mean frequency of unprotected insertive anal sex for circumcised men was about twice as high compared to those intact (P = .04). Intact young Black MSM did not differ from circumcised men relative to prevalence of STIs (including HIV) or condom use behaviors as reported only by insertive partners.


Reduction in C-reactive protein indicates successful targeting of the IL-1/IL-6 axis resulting in improved survival in early stage multiple myeloma.
We report the long-term follow-up results of a phase II trial of IL-1 receptor antagonist and low-dose dexamethasone for early stage multiple myeloma (MM). Patients were eligible if they had smoldering multiple myeloma (SMM) or indolent multiple myeloma (IMM) without the need for immediate therapy. Forty seven patients were enrolled and subsequently treated with IL-1Ra; in 25/47 low-dose dexamethasone (20 mg weekly) was added. The primary endpoint was progression-free survival (PFS). In the clinical trial, three patients achieved a minor response (MR) to IL-1Ra alone; five patients a partial response (PR) and four patients an MR after addition of dexamethasone. Seven patients showed a decrease in the plasma cell labeling index (PCLI) which paralleled a decrease in the high sensitivity C-reactive protein (hs-CRP). The median PFS for the 47 patients was 1116 days (37.2 months). The median PFS for patients without (n = 22) and with (n = 25) a decrease in their baseline hs-CRP was 326 days (11 months) vs. 3139 days (104 months) respectively (P <0.0001). The median overall survival (OS) for the 47 patients was 3482 days (9.5 years). The median OS for patients without and with a decrease in their baseline hs-CRP was 2885 days (7.9 years) vs. median not reached, respectively (P = 0.001). In SMM/IMM patients at risk for progression to active myeloma, reduction in the hs-CRP indicates successful targeting of the IL-1/IL-6 axis resulting in improved PFS and OS. (Clinical Trials.gov Identifier: NCT00635154) Am. J. Hematol. 91:571-574, 2016. © 2016 Wiley Periodicals, Inc.


Phenylalanine ammonia-lyase family is closely associated with response to phosphate deficiency in rice.
Phenylalanine ammonia-lyase (PAL) catalyzes the first step in the biosynthetic phenylpropanoid pathway (PPP) via deamination of phenylalanine to trans-cinnamic acid, a precursor for the lignin and flavonoid biosynthetic pathways. Although its role is well-established in various plants, the functional significance of PAL genes in rice remains poorly understood. This study aims to find out the global feature of rice PAL genes associated with phosphate use efficiency. To identify the biological functions of individual rice PAL genes, we performed meta-expression profiling analysis based on phylogenomics of rice PAL genes and confirmed the expression patterns using Quantitative real-time PCR (qPCR). We identified nine genes that were remarkably up-regulated during long-term phosphate (Pi) starvation and recovery processes through RNA-Seq data analysis. Expression patterns of the nine rice PAL genes under Pi starvation were further confirmed by qPCR, indicating that the function of PAL genes is strongly associated with Pi starvation response in rice. Our study reports the functional significance of rice PAL genes involved in adaptation to low Pi growth conditions and provides useful information to improve Pi use efficiency in crop plant.


Quantitative detection of crystalline lysine supplementation in poultry feeds using a rapid bacterial bioluminescence assay.
Lysine is an essential amino acid for both humans and animals; and it is usually the first or second limiting amino acid in most formulated diets. In order to estimate the lysine content in feeds and feed sources, rapid amino acid bioassays have been developed. The objective of this work is to assess a rapid assay for lysine supplementation in chicken feeds, using a luminescent Escherichia coli lysine-auxotrophic strain, to avoid prior thermal sterilization. An E. coli lysine auxotroph carrying a plasmid with lux genes was used as the test organism. The lysine assay was conducted using depleted auxotrophic cells in lysine samples. Luminescence was measured with a Dynex MLX luminometer after addition of the aldehyde substrate. Growth response (monitored as optical density at 600 nm) and light emission response of the assay E. coli strain were monitored to generate standard curves. Bioluminescent analysis of feed samples indicated that the method works well in the presence of a complex feed matrix. Comparison of both optical density and luminescent-based methods indicated that, when the assay takes place under optimal conditions, both methodologies correlated well ( r(2)=0.99). Except for the 0.64% lysine-supplemented feed, estimates for lysine based on the bacterial assay were over 80% (82-97%) of the theoretical values. Animal data showed that the bacterial bioluminescent method correlated well with the chick bioassay when diets with different levels of lysine supplementation were assayed for lysine bioavailability ( r(2)=0.97). Luminescent methodology coupled with a bacterial growth assay is a promising technique to assess lysine availability in supplemented animal feeds.


Hepatitis C as a metabolic disease: Implication for the pathogenesis of NASH.
In addition to the link with development of hepatocellular carcinoma (HCC), hepatitis C virus (HCV) infection is associated with several extrahepatic manifestations such as essential mixed cryoglobulinemia, porphyria cutanea tarda or Sjögren's syndrome. A role of hepatic steatosis in the pathogenesis of chronic hepatitis C has also been known, implying hepatitis C as a metabolic disease. In addition, recent epidemiological studies have suggested a linkage between type 2 diabetes and chronic HCV infection. However, the presence of additional factors in patients, such as obesity, aging or cirrhosis, prevents the establishment of a definite relationship between HCV infection and these two conditions, lipid metabolism disturbance and diabetes. In addition to the data indicating the presence of dyslipidemia and diabetes or insulin resistance in our cohort of chronic hepatitis C patients, we found a series of evidence showing the association between the conditions and HCV infection in mouse models that are transgenic for the HCV genes. In patients with chronic hepatitis C, a significant decrease in the serum levels of total cholesterol and apolipoproteins C2 and C3 was observed compared to those with chronic hepatitis B that were comparable in liver function. In an animal model, C18:1 mono-unsaturated fatty acids were significantly increased in the liver from HCV core gene transgenic mice, which was similarly observed in the liver from human hepatitis C patients. Thus, a disturbance in lipid metabolism was observed in both humans and an HCV mouse model, supporting that it is a specific event in HCV infection. A significant increase in the value of an indicator for homeostasis model assessment of insulin resistance (HOMA-IR), was observed in patients with chronic hepatitis C, even at the very early stage of chronic hepatitis. In the animal model, a marked insulin resistance was exhibited from a very young age in HCV core gene transgenic mice. Insulin resistance observed in the core gene transgenic mice was chiefly due to the shortage of insulin action on the suppression of glucose production in the liver. Thus, the ability of insulin to lower the plasma glucose level in the HCV transgenic mice was impaired, as observed in chronic hepatitis C patients. These results provide a direct experimental evidence for the contribution of HCV in the development of insulin resistance in human HCV infection, which finally leads to the development of type 2 diabetes. Insulin resistance may be a critical factor in the pathogenesis of chronic hepatitis C as recently suggested in non-alcoholic steatohepatitis (NASH), along with impairment in lipid metabolism. Our results would provide a clue for further understanding of pathobiology of HCV infection, and may provide an implication for the pathogenesis of NASH.


On the beneficial role of silicon to organisms: a case study on the importance of silicon chemistry to metal accumulation in yeast.
Silicon is involved in numerous important structural and functional roles in a wide range of organisms, including diatoms, plants, and humans, but clear mechanisms have been discovered only in diatoms and sponges. Silicate availability influences metal concentrations within various cell- and tissue-types, but a mechanism has not been discovered so far. In an earlier study on Baker's yeast Saccharomyces cerevisiae it was proposed that a chemical mechanism, rather than a biological one, is important. In the present study, the interaction of silicon with Baker's yeast is further investigated by studying the influence of zinc and magnesium on Si accumulation both at a low and a high silicate concentration in the medium. Si accumulation fitted well with Freundlich adsorption and Si release followed depolymerization kinetics, indicating that silicate adsorbs to the surface of the cell rather than being transported over the cell membrane. Subsequently, adsorbed silicate interacts with metal ions and, therefore, alters the cell's affinity for these ions. Since several metals are nutritional, these Si interactions can significantly change the growth and viability of organisms. In conclusion, the results show that chemistry is important in Si and metal accumulation in Baker's yeast, and suggest that similar mechanisms should be studied in detail in other organisms to unravel essential roles of Si.


Sustainable yarn production using leftover fabric from apparel industries.
A huge amount of waste was generated from the apparel industries. This study aims to develop the process of producing recycled yarn from apparel waste. The apparel leftover fabric was converted to fiber, and the fiber was mixed with virgin cotton in different ratios to produce sustainable 6/1 Ne rotor yarn. The produced yarn qualities viz. count strength product (CSP), elongation percentage, total quality index (TQI) and tenacity were decreased linearly, and opposite scenario observed for thick and thin places, neps, imperfection index (IPI) and hairiness (H) attributes with increasing the amount of waste addition with virgin cotton. The leftover fabric (LOF) can be utilized to develop a sustainable yarn and to zero waste management.


Preoperative and early postoperative magnetic resonance imaging in two cases of childhood choroid plexus carcinoma.
We present and illustrate the MRI appearances of two children with choroid plexus carcinoma. The MRI characteristics of these rare tumours are reviewed. Since total surgical resection is a significant prognostic factor, early postoperative MRI was performed in both cases to ensure surgical clearance. In one case a complete resection was documented and this patient remains well at short-term follow-up. Residual tumour was noted in the second case, but despite "second look" surgery there was subsequent local relapse.


Synthetic protein-binding DNA sponge as a tool to tune gene expression and mitigate protein toxicity.
Versatile tools for gene expression regulation are vital for engineering gene networks of increasing scales and complexity with bespoke responses. Here, we investigate and repurpose a ubiquitous, indirect gene regulation mechanism from nature, which uses decoy protein-binding DNA sites, named DNA sponge, to modulate target gene expression in Escherichia coli. We show that synthetic DNA sponges can be designed to reshape the response profiles of gene circuits, lending multifaceted tuning capacities including reducing basal leakage by >20-fold, increasing system output amplitude by >130-fold and dynamic range by >70-fold, and mitigating host growth inhibition by >20%. Further, multi-layer DNA sponges for decoying multiple regulatory proteins provide an additive tuning effect on the responses of layered circuits compared to single-layer sponges. Our work shows synthetic DNA sponges offer a simple yet generalizable route to systematically engineer the performance of synthetic gene circuits, expanding the current toolkit for gene regulation with broad potential applications.


Transfection of the cloned human excision repair gene ERCC-1 to UV-sensitive CHO mutants only corrects the repair defect in complementation group-2 mutants.
The human DNA-excision repair gene ERCC-1 is cloned by its ability to correct the excision-repair defect of the ultraviolet light- and mitomycin-C-sensitive CHO mutant cell line 43-3B. This mutant is assigned to complementation group 2 of the excision-repair-deficient CHO mutants. In order to establish whether the correction by ERCC-1 is confined to CHO mutants of one complementation group, the cloned repair gene, present on cosmid 43-34, was transfected to representative cell lines of the 6 complementation groups that have been identified to date. Following transfection, mycophenolic acid was used to select for transferants expressing the dominant marker gene Ecogpt, also present on cosmid 43-34. Cotransfer of the ERCC-1 gene was shown by Southern blot analysis of DNA from pooled (500-2000 independent colonies) transformants of each mutant. UV survival and UV-induced UDS showed that only mutants belonging to complementation group 2 and no mutants of other groups were corrected by the ERCC-1 gene. This demonstrates that ERCC-1 does not provide an aspecific bypass of excision-repair defects in CHO mutants and supports the assumption that the complementation analysis is based on mutations in different repair genes.


Association of folate deficiency and selected tumor marker concentrations in long-term hexavalent chromium exposed population.
Both hexavalent chromium [Cr (VI)] exposure and folate deficiency have been associated with increased cancer risks. Our previous studies have found folate deficiency in Cr (VI) exposed population. Here the relationship between some tumor markers and folate status in long-term Cr (VI) exposure was investigated carefully to show the multiple aspects of Cr (VI) carcinogenesis. A group of 115 workers occupationally exposed to chromate and 60 matched, unexposed controls in Shandong province of China were recruited. Environmental and biological exposure assessments including personal exposure to airborne Cr and Cr contents in erythrocytes were performed. Serum folate, plasma total homocysteine (tHcy) and plasma carcinoembryonic antigen (CEA), neuron specific enolase (NSE), squamous cell carcinoma antigen (SCC), cytokeratin fragment antigen 21-1 (CYFRA 21-1), cancer antigen 72-4 (CA72-4), as well as α-fetoprotein (AFP) were measured. Smoking index (SI) was also calculated to discriminate possible confounding effects of smoking status. Serum folate level decreased significantly, while plasma tHcy, CEA, NSE, SCC, CYFRA21-1, CA72-4 and AFP concentrations increased significantly after Cr (VI) exposure. Meanwhile, plasma CEA, NSE and SCC were negatively correlated with serum folate. SI was negatively correlated with serum folate but positively correlated with plasma tHcy, CEA and NSE levels. Present study suggests that folate deficiency was associated with increased cancer risks and might be affected by smoking in Cr (VI) exposed population. Folate might play a key role in Cr (VI) carcinogenesis although further detailed investigations are needed to clarify the mechanism of this process.


Analgesia produced by microinjection of baclofen and morphine at brain stem sites.
Microinjection of either baclofen (1.5 microgram) or morphine (2.5 microgram), in equimolar doses (14 mM), at sites located in the caudal periaqueductal gray (PAG) resulted in a delay in tail flick latency (analgesia). The relative analgesic potency of baclofen among caudal PAG sites, however, did not correlate with that of morphine. Application of either drug into the caudal aspect of the cerebral aqueduct also produced analgesia, but neither agent caused analgesia when applied at PAG sites rostral to the interaural line. Baclofen also produced analgesia when microinjected in the lower brain stem at sites lateral to the midline in or near the nucleus gigantocellularis, but did not produce analgesia when applied on the midline at sites located within or near the raphe magnus. Conversely, morphine produced analgesia when applied locally at midline sites but not at sites located lateral to the midline. These data suggest that the analgesia produced by systemic administration of baclofen and morphine involves activation of different neuronal substrates.


Effects of upper respiratory tract illnesses and stress on alertness and reaction time.
It has been shown that stress increases susceptibility to the upper respiratory tract illnesses (URTIs) such as the common cold. Compared to healthy individuals, those with URTIs also report reduced alertness and have slower reaction times. The present study investigated whether those with an URTI who had been exposed to stressful events showed greater impairments than either individuals without a cold or those with an illness and low stress exposure. A prospective cohort study was conducted. The volunteers (360 young adults) were recruited when healthy and completed questionnaires measuring negative life events, personality and health-related behaviours. They also rated their alertness and performed a simple reaction task. If volunteers developed an upper respiratory illness they returned to the laboratory and completed a symptom check list and had nasal secretion and sub-lingual temperature recorded. They also completed a questionnaire measuring recent daily hassles. Alertness and simple reaction time were also recorded again. Those who did not develop a cold were recalled as controls 12 weeks after the start of the study. Analyses of covariance were carried out comparing colds/no colds and high/low stress groups. Baseline measures were included as covariates. 356 participants completed the study. 120 developed URTIs and 236 were re-tested as controls. The frequency and severity of daily hassles were associated with symptom severity. Alertness was reduced and simple reaction time was slower in the URTIs group and the high stress/ill group showed the biggest impairments. These effects remained significant when health related behaviours and personality were covaried. The difference between the high and low stress URTI groups did not reflect symptom severity. The behavioural impairments induced by the common cold are greater when the person has been exposed to stressful events.


Students' acceptance of online learning in developing nations: scale development and validation.
Most education systems were severely impacted by the COVID-19 pandemic, and as a result, learning shifted from face-to-face to online in higher education institutions. This unprecedented shift in the learning environment caused substantial challenges for students. The situation was more severe in developing nations such as Bangladesh, which lacked available resources and knowledge of online education to support their students. Recent studies suggest that students resisted online learning in various developing nations. To support online learning in developing nations, this study develops the Acceptance of Online Learning (AOL) scale comprised of both institutional and student-related factors. To validate the AOL scale, the study collected data from 441 students across 30 higher education institutions in Bangladesh to determine the factors explaining students' acceptance of online learning using AOL measurements. The results showed that institutional factors, such as technological sufficiency, instructor efficiency, and technical assistance play significant roles in students' acceptance of online learning in developing nations. These findings will help education policymakers and administrators in developing nations to assess the needs of students with respect to online learning, and the AOL scale will assist in the evaluation of students' acceptance of online learning in these nations.


Carrier detection in X-linked recessive (Duchenne) muscular dystrophy: pyruvate kinase isoenzymes and creatine phosphokinase in serum and blood cells.
Allosterism allows individual assay of both isoenzymes, one abundant in muscle, of pyruvate kinase (PK), recently reported superior to serum creatine phosphokinase (CPK) in detecting patients with and female carriers of X-linked recessive (Duchenne) muscular dystrophy (DMD). Extensive comparative studies did not support these findings and confirmed the marked superiority of CPK over rariants of PK or other enzymes in sensitivity, stability and convenience. Deducting the adenylate kinase increment (AKI) further refined the CPK assay, eliminating the effect of haemolysis in diagnosis and enabling studies of blood cell content. Both leucocytes and erythrocytes liberated PK and lactate dehydrogenase (LDH) after brief chilling or disruption. Only erythrocytes showed a CPK content, however, constantly adjusted to match that of serum as if by free cell membrane passage, but less accomodating to a sudden large influx of CPK than of LDH, where an apparent buffering effect could account for differences in clinical response.


A double-stranded RNA unwinding activity introduces structural alterations by means of adenosine to inosine conversions in mammalian cells and Xenopus eggs.
Amphibian eggs and embryos as well as mammalian cells have been reported to contain an activity that unwinds double-stranded RNA. We have now found that adenosine residues have been modified in the RNA products of this unwinding activity. Although the modified RNA remains double-stranded, the modification causes the RNA to be susceptible to single-strand-specific RNase and to migrate as a retarded smear on a native polyacrylamide electrophoresis gel. The modification is specific for double-stranded RNA. At least 40% of the adenosine residues can be modified in vitro in a given random sequence RNA molecule. By using standard two-dimensional TLC and HPLC analyses, the modified base has been identified as inosine. Mismatched base-pairing between inosine and uridine appears to be responsible for the observed characteristics of the unwound RNA. The biological significance of this modifying activity and also of the modified double-stranded RNA is discussed.


Hypericin phototoxicity induces different modes of cell death in melanoma and human skin cells.
Hypericin, the major component of St. John's Wort, absorbs light in the UV and visible ranges whereupon it becomes phototoxic through the production of reactive oxygen species. Although photodynamic mechanisms (i.e. through endogenous photosensitizers) play a role in UVA phototherapy for the treatment of skin disorders such as eczema and psoriasis, photodynamic therapy employing exogenous photosensitizers are currently being used only for the treatment of certain forms of non-melanoma skin cancers and actinic keratoses. There are few reports however on its use in treating melanomas. This in vitro study analyses the phototoxic effect of UVA (400-315 nm) - activated hypericin in human pigmented and unpigmented melanomas and immortalised keratinocytes and melanocytes. We show that neither hypericin exposure nor UV irradiation alone reduces cell viability. We show that an exposure to 1 microM UVA-activated hypericin does not bring about cell death, while 3 microM activated hypericin induces a necrotic mode of cell death in pigmented melanoma cells and melanocytes and an apoptotic mode of cell death in non-pigmented melanoma cells and keratinocytes. We hypothesis that the necrotic mode of cell death in the pigmented cells is possibly related to the presence of melanin-containing melanosomes in these cells and that the hypericin-induced increase in reactive oxygen species leads to an increase in permeability of melanosomes. This would result in toxic melanin precursors (of an indolic and phenolic nature) leaking into the cytoplasm which in turn leads to cell death. Hypericin localisation in the endoplasmic reticulum in these cells shown by fluorescent microscopy, further support a disruption in cellular processing and induction of cell death. In contrast, this study shows that cells that do not contain melanosomes (non-pigmented melanoma cells and keratinocytes) die by apoptosis. Further, using a mitochondrial-specific fluorescent dye, we show that intracellular accumulation of hypericin induces a mitochondrial-associated caspase-dependent apoptotic mode of cell death. This work suggests that UVA is effective in activating hypericin and that this phototoxicity may be considered as treatment option in some cases of lentigo maligna or lentigo maligna melanoma that are too large for surgical resection.


Revisional Surgery After One Anastomosis/Minigastric Bypass: an Italian Multi-institutional Survey.
Efficacy and safety of OAGB/MGB (one anastomosis/mini gastric bypass) have been well documented both as primary and as revisional procedures. However, even after OAGB/MGB, revisional surgery is unavoidable in patients with surgical complications or insufficient weight loss. A questionnaire asking for the total number and demographics of primary and revisional OAGB/MGBs performed between January 2006 and July 2020 was e-mailed to all S.I.C. OB centres of excellence (annual caseload > 100; 5-year follow-up > 50%). Each bariatric centre was asked to provide gender, age, preoperative body mass index (BMI) and obesity-related comorbidities, previous history of abdominal or bariatric surgery, indication for surgical revision of OAGB/MGB, type of revisional procedure, pre- and post-revisional BMI, peri- and post-operative complications, last follow-up (FU). Twenty-three bariatric centres (54.8%) responded to our survey reporting a total number of 8676 primary OAGB/MGBS and a follow-up of 62.42 ± 52.22 months. A total of 181 (2.08%) patients underwent revisional surgery: 82 (0.94%) were suffering from intractable DGER (duodeno-gastric-esophageal reflux), 42 (0.48%) were reoperated for weight regain, 16 (0.18%) had excessive weight loss and malnutrition, 12 (0.13%) had a marginal ulcer perforation, 10 (0.11%) had a gastro-gastric fistula, 20 (0.23%) had other causes of revision. Roux-en-Y gastric bypass (RYGB) was the most performed revisional procedure (109; 54%), followed by bilio-pancreatic limb elongation (19; 9.4%) and normal anatomy restoration (19; 9.4%). Our findings demonstrate that there is acceptable revisional rate after OAGB/MGB and conversion to RYGB represents the most frequent choice.


Increased metabolic rate associated with immune stimulation of heat-killed Vibrio anguillarum at different temperatures in zebrafish (Danio rerio).
The action of the immune response in zebrafish (Danio rerio) has been a target of many studies. However, the energetic demands involved in the immune response are poorly understood in ectothermic poikilotherms, such as fish. This research aims to characterize the energetic response of zebrafish to an immune challenge of heat-killed Vibrio anguillarum at 22 °C and 27.5 °C. Zebrafish were either not injected, injected intraperitoneally with 10 μl of saline and Freund's incomplete adjuvant (sham), or heat-killed Vibrio anguillarum & Freund's incomplete adjuvant (1.21 × 1010 cfu/ml). Respirometry was then performed on these zebrafish for a period of 27 h. Following this, spleen was collected for quantitative PCR analysis of the catalytic subunit of AMPK (ampka1 & ampka2), the nuclear factor kappa-light-chain-enhancer of activated B cells (nf-kb), and several cytokines (tnfa, il-1b, il-8, il-10). While there was no increase in oxygen consumption with any treatment at 22 °C, there was a marked 30% increase in oxygen consumption in zebrafish injected with heat-killed Vibrio at 27.5 °C. Furthermore, temperature had a strong effect on the timing of the immune response. At 22 °C, there was a 2-3-fold increase in the cytokines measured associated with heat-killed Vibrio injection, whereas there were no differences found at 27.5 °C. Furthermore, while there was an increase in ampka2 at 22 °C, there was a sharp decrease in ampka2 at 27.5 °C, although the changes in ampka2 transcript abundance could not be solely attributed to heat-killed Vibrio, as there were similar changes associated with the sham group. The results of this study demonstrate some of the first evidence that zebrafish increase routine metabolic rate associated with immune stimulation.


Keratosis follicularis and familial benign chronic pemphigus in the same patient.
A case is described with features both of Hailey-Hailey and Darier's disease in a male patient who presented skin lesions since he was 8 years old. The present case is discussed in relation to data obtained from the literature, and a differential diagnosis from Grover's disease is made.


A molecular approach for the rapid, selective and sensitive detection of Exophiala jeanselmei in environmental samples: development and performance assessment of a real-time PCR assay.
Exophiala jeanselmei is an opportunistic pathogenic black yeast growing in humid environments such as water reservoirs of air-conditioning systems. Because this fungal contaminant could be vaporized into the air and subsequently cause health problems, its monitoring is recommended. Currently, this monitoring is based on culture and microscopic identification which are complex, sometimes ambiguous and time-demanding, i.e., up to 21 days. Therefore, molecular, culture-independent methods could be more advantageous for the monitoring of E. jeanselmei. In this study, we developed a SYBR®green real-time PCR assay based on the internal transcribed spacer 2 from the 18S ribosomal DNA complex for the specific detection of E. jeanselmei. The selectivity (100 %), PCR efficiency (95.5 %), dynamic range and repeatability of this qPCR assay were subsequently evaluated. The limit of detection for this qPCR assay was determined to be 1 copy of genomic DNA of E. jeanselmei. Finally, water samples collected from cooling reservoirs were analyzed using this qPCR assay to deliver a proof of concept for the molecular detection of E. jeanselmei in environmental samples. The results obtained by molecular analysis were compared with those of classical methods (i.e., culture and microscopic identification) used in routine analysis and were 100 % matching. This comparison demonstrated that this SYBR®green qPCR assay can be used as a molecular alternative for monitoring and routine investigation of samples contaminated by E. jeanselmei, while eliminating the need for culturing and thereby considerably decreasing the required analysis time to 2 days.


Single-cell A/B testing for cell-cell communication.
A new method developed by Francisco Quintana's group, systematic perturbation of encapsulated associated cells followed by sequencing (SPEAC-seq), applies a CRISPR screen to co-cultured interacting cells to identify the ligands mediating cell-cell communication. Using this approach, the authors discover the molecular basis of a microglia-astrocyte feedback loop that suppresses neuroinflammatory disease.


Geochemical characterization of elements in Vitis vinifera cv. Negroamaro grape berries grown under different soil managements.
The present geochemical study concerns the impact of viticultural practices in the chemical composition of the grape cultivar "Negroamaro" in Apulia, a southern Italian region renowned for its quality wine. Three types of soil management (SM), two cover cropping with different mixtures, and a soil tillage were considered. For each SM, the vines were irrigated according to two irrigation levels. Chemical composition of soil and of berries of Vitis vinifera cultivar "Negroamaro" were analyzed by X-ray fluorescence, inductively coupled plasma-mass spectrometry and multivariate statistics (linear discrimination analysis). In detail, we investigated major and trace elements behavior in the soil according to irrigation levels, the related index of bioaccumulation (BA) and the relationship between trace element concentration and soil management in "Negroamaro" grapes. The results indicate that soil management affects the mobility of major and trace elements. A specific assimilation of these elements in grapes from vines grown under different soil management was confirmed by BA. Multivariate statistics allowed to associate the vines to the type of soil management. This geochemical characterization of elements could be useful to develop fingerprints of vines of the cultivar "Negroamaro" according to soil management and geographical origin.


On plotting amplitude-transition functions for voluntary eye saccades.
The first of two oppositely directed saccades may be small if the time interval between the two saccades is short. We have examined the merits of plotting the amplitude of the first saccade against saccadic interval. This plot provides a useful supplement to other approaches in the literature, as it is insensitive to variation due to extraneous factors or to task. Reasons for this are discussed.


Simultaneous estimation and variable selection in median regression using Lasso-type penalty.
We consider the median regression with a LASSO-type penalty term for variable selection. With the fixed number of variables in regression model, a two-stage method is proposed for simultaneous estimation and variable selection where the degree of penalty is adaptively chosen. A Bayesian information criterion type approach is proposed and used to obtain a data-driven procedure which is proved to automatically select asymptotically optimal tuning parameters. It is shown that the resultant estimator achieves the so-called oracle property. The combination of the median regression and LASSO penalty is computationally easy to implement via the standard linear programming. A random perturbation scheme can be made use of to get simple estimator of the standard error. Simulation studies are conducted to assess the finite-sample performance of the proposed method. We illustrate the methodology with a real example.


Sperm DNA Methylation Epimutation Biomarkers for Male Infertility and FSH Therapeutic Responsiveness.
Male factor infertility is increasing and recognized as playing a key role in reproductive health and disease. The current primary diagnostic approach is to assess sperm quality associated with reduced sperm number and motility, which has been historically of limited success in separating fertile from infertile males. The current study was designed to develop a molecular analysis to identify male idiopathic infertility using genome wide alterations in sperm DNA methylation. A signature of differential DNA methylation regions (DMRs) was identified to be associated with male idiopathic infertility patients. A promising therapeutic treatment of male infertility is the use of follicle stimulating hormone (FSH) analogs which improved sperm numbers and motility in a sub-population of infertility patients. The current study also identified genome-wide DMRs that were associated with the patients that were responsive to FSH therapy versus those that were non-responsive. This novel use of epigenetic biomarkers to identify responsive versus non-responsive patient populations is anticipated to dramatically improve clinical trials and facilitate therapeutic treatment of male infertility patients. The use of epigenetic biomarkers for disease and therapeutic responsiveness is anticipated to be applicable for other medical conditions.


Western and dot immunoblotting analysis of viral antigens and antibodies: application to murine hepatitis virus.
Viral proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred quantitatively to nitrocellulose by electroblotting in SDS-containing buffer. Monoclonal antibodies directed against previously defined epitopes on the viral proteins were used as probes to detect viral protein synthesis and processing, as well as expression in animal tissues. Circulating polyclonal antibodies were also probed and characterized for their polypeptide specificities. Under appropriate conditions, this Western immunoblotting technique was quantitative. Finally, a highly sensitive dot immunoblotting assay was used to analyze the sensitivity to denaturation of various epitopes on the viral proteins. This assay detected picogram quantities of viral antigens and antibodies.


Synthesis of N-substituted indole-3-carboxylic acid derivatives via Cu(I)-catalyzed intramolecular amination of aryl bromides.
A variety of N-alkylated and N-arylated derivatives of methyl 1 H-indole-3-carboxylate were synthesized efficiently via Ullmann-type intramolecular arylamination, using the CuI-K 3PO 4-DMF system. This catalytic amination procedure can be performed with good to high yields under mild conditions under an air atmosphere.


Inhibition of hyaluronan synthesis alters sulfated glycosaminoglycans deposition during chondrogenic differentiation in ATDC5 cells.
In chondrogenic differentiation, expression and collaboration of specific molecules, such as aggrecan and type II collagen, in extracellular matrix (ECM) are crucial. However, few studies have clarified the roles of hyaluronan (HA) in proteoglycan aggregation during chondrogenic differentiation. We assessed the roles of HA in sulfated glycosaminoglycans deposition during chondrogenic differentiation by means of 4-methylumbelliferone (4-MU), an HA synthase inhibitor, using ATDC5 cells. ATDC5 cells were treated with 0.5 mM 4-MU for 7 or 21 days after induction of chondrogenic differentiation with insulin. Depositions of sulfated glycosaminoglycans were evaluated with Alcian blue staining. mRNA expression of ECM molecules was determined using real-time RT-PCR. The deposition of aggrecan and versican was investigated with immunohistochemical staining using specific antibodies. Effects of 4-MU on HA concentrations were analyzed by HA binding assay. 4-MU suppressed the positivity of Alcian blue staining, although this delay was reversible. Interestingly, stronger positivity of Alcian blue staining was observed at day 21 in cultures with 4-MU discontinuation than in the control. 4-MU significantly increased the mRNA expression of aggrecan, versican, and type II collagen, which was consistent with increased deposition of aggrecan and versican. The HA concentration in ECM and cell-associated region was significantly suppressed with 4-MU treatment. We conclude that the inhibition of HA synthesis slows sulfated glycosaminoglycans deposition during chondrogenic differentiation despite the increased deposition of other ECM molecules. Transient starvation of HA with 4-MU accelerates chondrogenic ECM formation, suggesting its potential to stimulate chondrogenic differentiation with adequate use.


Dietary counseling versus dietary supplements for malnutrition in chronic pancreatitis: a randomized controlled trial.
Up to 50% of patients with chronic pancreatitis (CP) are malnourished. There are limited data on the role of dietary intervention in improving the nutritional status of such patients. The aim was to compare the efficacy of medium chain triglyceride (MCT)-enriched commercial dietary supplements with dietary counseling for homemade food in the management of malnutrition in patients with CP. In a randomized controlled trial, consecutive undernourished patients with CP (body mass index [BMI] <18.5 kg/m(2)) at a tertiary care hospital were randomized to receive either dietary counseling for regular homemade food or commercial MCT-enriched dietary supplements for a period of 3 months to compensate for the dietary calorie deficit. All patients received standard management for CP including pancreatic enzyme supplements. Primary outcome measure was improvement in BMI. Sixty malnourished patients with CP were randomized to counseling group (n = 29; mean age, 32 +/- 10 years; male, 83%) and supplementation group (n = 31; mean age, 28 +/- 10 years; male, 84%). BMI increased in both the counseling group and supplementation group (17.2 +/- 1.7 vs 18.1 +/- 1.8 kg/m(2), P = .001; 16.7 +/- 1.6 vs 18.2 +/- 1.6 kg/m(2), P = .001). There were similar improvements in triceps skinfold thickness, dietary intake, fecal fat, and pain score during a period of 3 months in both groups. There was, however, no significant difference between the counseling and supplementation groups with regard to any of the outcome measures. Dietary counseling for a balanced homemade diet is as good as commercial food supplements in improving malnutrition in patients with CP.


An unholy alliance: substance abuse and social exclusion among assertive outreach patients.
To investigate the relationship between social exclusion and outcomes of mentally ill patients with substance abuse problems receiving assertive outreach treatment in London. Analysis was conducted on data on 580 patients from the 'Pan-London Assertive Outreach Study (PLAO)'. Data were collected using clinician-rated scales of alcohol and drug abuse in the last 6 months before baseline. Outcomes - hospitalization and compulsory hospitalization- were assessed over a 9-month follow-up period. The analysis identified a group of patients with substance abuse who suffer from social exclusion and forensic problems (n = 77, 15.8%), and had poorer outcomes than the rest of the patients in terms of hospitalization (52% vs. 36% respectively) and compulsory (39% vs. 22% respectively) hospitalization. There is a distinct group of patients whose treatment requires social inclusion and forensic expertise. Future research into which model of service is suitable for these patients and in what setting is recommended.


Detailed insight in intraoperative eABR measurements to assist auditory brainstem implantation in a patient with neurofibromatosis type 2.
Auditory brainstem implant (ABI) is used to provide auditory sensations in patients with neurofibromatosis type 2 who lost their hearing due to a surgical removal of the tumor. ABI surgery, implant activation and follow-up sessions present unique challenges including the exact placement of the electrode pad in the lateral recess of the IVth ventricle, identification of electrodes that trigger non-auditory sensation and their deactivation which lowers the number of electrodes responsible for hearing, changes of T- and C-levels across follow-up sessions. We present a complete procedure using an example case starting from the surgical part with the detailed description of intraoperative eABR measurement as a guidance for pad placement to the ABI activation and first fitting sessions with auditory sensation assessment. Since the first ABI electrode pad position presented non-satisfactory intraoperative eABR results it was decided to move the pad slightly which resulted in better eABR (more electrodes with auditory responses). The discussed patient demonstrated great auditory and speech perception results after the first ABI fitting (which included three sessions over 2 consecutive days). Repositioning of the ABI electrode pad during the surgery was carried out taking into account the intraoperative eABR results and this led to an overall positive outcome for the patient. The placement of ABI electrode pad is crucial for later auditory results. This study provides detailed insight in this very specialized procedure that is not performed in every clinic and adds to the knowledge of intraoperative navigation using eABR measurements during ABI surgery.


Insights into the Biosynthesis of Dehydroalanines in Goadsporin.
Dehydroalanines in goadsporin are proposed to be formed by GodF and GodG, which show slight homology to the N-terminal glutamylation and C-terminal elimination domains, respectively, of LanB, a class I lanthipeptide dehydratase. Although similar, separated-type LanBs are conserved among thiopeptides and indispensable for their biosynthesis and biological activities, these enzymes had not yet been characterized. Here, we identified goadsporin B, which has unmodified Ser4 and Ser14, from both godF and godG disruptants. The godG disruptant also produced goadsporin C, a glutamylated-Ser4 variant of goadsporin B. These results suggested that dehydroalanines are formed by glutamylation and glutamate elimination. NMR analysis revealed for the first time that the glutamyl group was attached to a serine via an ester bond, by the catalysis of LanB-type enzymes. Our findings provide insights into the function of separated-type LanBs involved in the biosynthesis of goadsporin and thiopeptides.


Effect of a gonadotropin-releasing hormone agonist on luteinizing hormone and testosterone secretion and testicular histology in male rhesus monkeys.
Three adult male rhesus monkeys were treated for 20 weeks with a gonadotropin-releasing hormone (GnRH) agonist (Ag; Wy-40972; Wyeth Laboratories, Philadelphia, PA) using osmotic minipumps. Ag administration resulted in a transient increase and then a precipitous decrease in the concentration of luteinizing hormone (LH) and testosterone (T). After 5 weeks, serum levels of LH were undetectable (less than 0.2 microgram/ml), while serum T was always detectable, but continued to fall throughout the period of Ag administration. The serum LH and T response to 50 micrograms of GnRH was abolished by 4 weeks of Ag treatment, and this effect persisted through the treatment period. Testicular histology at 20 weeks of Ag treatment exhibited diffuse atrophy of seminiferous tubules, suppressed germinal cell division, and the absence of spermatids or spermatozoa. There was no evidence of testicular necrosis or calcification of the seminiferous tubules. Following the termination of Ag infusion, serum LH and T concentrations rebounded to levels that exceeded pretreatment values for a 5-week period before falling back to baseline levels. A complete restoration of spermatogenesis and testicular volume occurred by 12 weeks after treatment. These data suggest that continuous Ag administration is an effective method of reversibly disrupting spermatogenesis in the male rhesus monkey.


Endoscopic Management of Vascular Sinonasal Tumors, Including Angiofibroma.
The greatest challenge in the surgical treatment of angiofibromas is dealing with the hypervascularity of these tumors. Staging systems that take into account the vascularity of the tumor may be more prognostic. A variety of treatment strategies are used to deal with the vascularity of angiofibromas, including preoperative embolization, segmentation of the tumor into vascular territories, use of hemostatic tools, and staging of surgery. Even large angiofibromas with intracranial extension and residual vascularity can be successfully managed by a skull base team using endoscopic techniques.


Effectiveness and safety of endoscopic treatment for duodenal variceal bleeding: a systematic review.
Duodenal varix is a rare condition that involves massive bleeding, diagnostic difficulties, and a high rate of rebleeding and mortality. The purpose of this study was to systematically review endoscopic treatment for duodenal variceal bleeding to evaluate its effectiveness and safety. We searched PubMed, Embase, Web of Science, and the Cochrane Library up to 21 November 2019. Ninety-two studies containing 156 patients were finally included, and individual data from 101 patients (mean age: 52.67 ± 13.82 years, male: 64.4%) were collected and further analyzed. We used an analysis of variance and χ2 or Fisher's exact tests to analyze individual data from 101 patients. The cause of duodenal variceal bleeding was cirrhosis-related intrahepatic portal hypertension (IPH) in 76.2% of patients. The overall rates of initial hemostasis and treatment success of endoscopic treatment for duodenal variceal bleeding were 89.1 and 81.2%, respectively. The median duration of follow-up was 4.5 (1.0, 12.0) months. The overall rates of rebleeding and mortality were 8.9 and 13.9%, respectively. Among a variety of endoscopic treatments available, only the initial hemostasis rate was significantly different between the endoscopic injection sclerotherapy and endoscopic tissue adhesive (ETA) groups (72.7 vs. 94.7%, P = 0.023); differences in treatment success, rebleeding, mortality, and adverse events were not statistically significant among the four groups. Endoscopic intervention is a feasible, well tolerated, and effective modality for the treatment of duodenal variceal bleeding. Among the variety of endoscopic treatments available, ETA with cyanoacrylate may be preferable for duodenal variceal bleeding.


Combination of hand-assisted and laparoscopic proctocolectomy (HALP): Technical aspects, learning curve and early postoperative results.
Various techniques for laparoscopic proctocolectomy have been reported worldwide. We evaluated the technical aspects and early postoperative results of hand-assisted laparoscopic proctocolectomy (HALP) with construction of an ileal pouch-anal anastomosis through a Pfannenstiel incision. Between June 2004 and May 2006, 20 patients (median age 28 years) underwent combined HALP at our institution. Preoperative diagnosis included ulcerative colitis (n = 16), indeterminate colitis (n = 1), familial adenomatous polyposis (n = 2), and carcinoma of the rectum associated with ulcerative colitis (n = 1). All patients were under immunosuppressive therapy. Laparoscopic mobilisation of rectum, sigmoid and descending colon was performed first. Subsequently, hand-assisted laparoscopic mobilization of the transverse and ascending colon as well as creation of an ileal J-pouch were performed through a Pfannenstiel incision. Ileal pouch-anal anastomosis was completed by transrectal stapling device and protected by a loop ileostomy. The ileal pouch-anal anastomosis could be achieved in 19 cases (95%). There was one conversion (5%) to open surgery with construction of an end-ileostomy. No intraoperative blood transfusions were necessary. The median operating time was 210 minutes (range 180 min to 330 min). It was longer for the first five procedures but then remained constant. Two patients (10%) developed anastomotic leakage, which could be treated conservatively. Mean length of hospital stay was 11 days (range 7-32 days). Combined HALP with construction of an ileal J-pouch-anal anastomosis can be performed safely and effectively. The Pfannenstiel incision proved to be advantageous for hand-assisted mobilisation of the transverse colon. Additionally, it was useful for the specimen removal and the J-pouch construction. Our new technique not only proved to be safe, but also resulted in a shortened total operation-time after a learning curve of about five procedures.