Nitrous oxide levels during retinal cryosurgery.
Nitrous oxide levels were monitored before, during, and after retinal cryotherapy. These units produced an average elevation of the nitrous oxide concentration in the operating room to the level of 963 ppm. Nitrous oxide levels in a control operating room using general endotracheal anesthesia with nitrous oxide ranged from 75 to 200 ppm. Nitrous oxide has been reported to be associated with various teratogenic effects in experimental animals and may be associated with other human ailments. The use of a scavenger system specifically designed to handle the exhaust from ophthalmic nitrous oxide units can reduce the nitrous oxide concentration by 50% or more and is, therefore, recommended.


Navicular tenosuspension with anterior tibialis tendon (Young procedure) associated to calcaneo-stop for the treatment of paediatric flexible flatfoot: clinical and ultrasound study.
Flexible flatfoot is one of the most common deformities in pediatric orthopaedics. Arthroeresis procedures are designed to correct this deformity. Among them, calcaneo-stop is a procedure with both biomechanical and proprioceptive properties. There could be other surgical procedure combined, such as a percutaneous Achilles tendon lengthening and the Gould tibialis posterior retension or Young tibialis anterior navicular tenosuspension. This study analyzed the clinical and sonographic results of 36 patients following flexible flatfoot surgical treatment with a calcaneo-stop arthroeresis combined with Achilles lengthening and a Young procedure. From March 2001 to August 2014, 36 patients (54 feet) were treated with calcaneo-stop arthroeresis, percutaneous Achilles tendon lengthening and Young's tenosuspension. The clinical assessment and a sonography of the anterior tibialis tendon (ATT) were performed in all patients. The average follow-up was 7.4 years (range 8 months-14 years) with a satisfactory outcome in 51 feet (94.5%). No major and minor complications were observed. In four cases the calcaneo-stop was removed for pain and low tolerance of the patient. The AOFAS score and the talocalcaneal angle did not have statistically significant in case of ATT was or not still inserted in the navicular at the follow-up. The calcaneo-stop procedure is a simple, reliable and minimally invasive procedure for the treatment of pediatric flexible flatfoot. Although the indications for the Young tenosuspension as an isolated procedure is very narrow, it can still be an effective procedure when combined to calcaneo-stop. The key to appropriate utilization is a thorough understanding of the biomechanics of the foot function and a specific appreciation of the function of the ATT.


Ribosomal resistance to the 12,13-epoxytrichothecene antibiotics in the producing organism Myrothecium verrucaria.
An extract of Myrothecium verrucaria, a fungus which produces a range of 12,13-epoxytrichothecene toxins, was found to be resistant to T-2 toxin, one of its products. The epoxytrichothecenes are inhibitors of eukaryotic protein synthesis and normally bind to the 60S ribosomal subunit so as to inhibit peptidyltransferase activity. Ribosomes from M. verrucaria contain 60S subunits which are not subject to inhibition by T-2 toxin and are also resistant to certain other drugs such as anisomycin and homoharringtonine, but not sparsomycin or cycloheximide.


Studies of quality control of 99mTc-labelled macroaggregated albumin--Part 1. Aggregation of non-mercaptalbumin and its conformation.
The aggregative condition of albumin was investigated using bovine serum albumin (BSA) as a model for quality control of 99mTc-macroaggregated albumin (99mTc-MAA). Uniformalized aggregates were obtained from the oxidized non-mercapt-type of BSA by heating. The size of the aggregates was affected by the pH and the types of buffer solutions used as well as the concentrations of albumin and buffers. The beta form structure of the albumin was more stable on heating and this may contribute to its aggregation. Aggregation of oxidized non-mercaptalbumin afforded a portion of smaller sized particles in MAA, this being an inappropriate factor for scintiscanning of lungs. Our results suggest that it is necessary to remove oxidized type albumin from human serum albumin as the starting material, in order to prepare MAA with a uniform and larger particle size.


Late-onset epilepsy and the risk of dementia: a systematic review and meta-analysis.
Patients with dementia have higher risk of epilepsy. However, it remains not comprehensively evaluated if late-onset epilepsy (LOE) is associated with higher risk of dementia. We, therefore, performed a meta-analysis to systematically evaluate the association. Relevant cohort studies were identified by search of electronic databases including PubMed, Embase, and Web of Science. A randomized-effect model incorporating the possible between-study heterogeneity was used to pool the results. Overall, seven cohort studies including 873,438 adults were included, and 16,036 (1.8%) of them had LOE. With a mean follow-up duration of 8.7 years, 33,727 of them were diagnosed as dementia. Pooled results showed that LOE was associated with a higher risk of dementia (risk ratio [RR] 2.39, 95% confidence interval [CI] 2.04-2.81, p < 0.001, I2 = 67%). Results of subgroup analysis showed that the association between LOE and the risk of dementia was stronger in hospital-derived participants (RR 4.23, 95% CI 2.67-6.70, p < 0.001) than that in community-derived population (RR 2.25, 95% CI 1.93-2.63, p < 0.001; p for subgroup difference = 0.01). Pooled results of three studies showed that LOE was associated with a higher risk of Alzheimer's disease (RR 2.35, 95% CI 1.08-5.08, p = 0.03, I2 = 85%). One study suggested a significant association between LOE and risk of vascular dementia (RR 2.0, 95% CI 1.77-2.26, p < 0.001). Evidence from cohort studies suggests that LOE may be a risk factor of dementia.


[Development of fluorogenic probes for the detection of histone deacetylase activity].
Fluorescence imaging using synthetic probes is becoming increasingly popular as a chemistry-based technique for the analysis of biomolecules. Real-time visual information on various biological molecules can be obtained by designing probes with high flexibility. The focus of our research is to design fluorogenic probes which are non-fluorescent in their initial intact form, but exhibit enhanced fluorescence intensity upon reactions with target biomolecules. The fluorescence switch of these probes allows the detection of the function and the localization of biomolecules with a high signal-to-noise ratio. Thus far, we have succeeded in designing and synthesizing fluorogenic probes that visualize molecules involved in epigenetics. Histone deacetylases (HDACs) play an important role in the epigenetic regulation of gene expression. A significant amount of epigenetic information can be obtained by the detection of enzyme activity. However, the existing methods require complicated multistep procedures. To overcome this limitation, we developed fluorogenic probes for the detection of HDAC activity in a one-step procedure. In this review, the details of the strategy for probe design and the detection method have been described.


Failure to diagnose recent hepatitis C virus infections in London injecting drug users.
Hepatitis C virus (HCV) infection results in chronic liver disease in a substantial proportion of those infected. Most new cases of HCV infection in the UK are associated with intravenous drug use. It is important to identify these infections because of the implications for the future health of the individuals concerned and for the control of further spread of infection. However, as hepatitis C infection is characterised by a relatively long asymptomatic period of seronegative viraemia, a laboratory diagnostic protocol that does not test HCV seronegative samples for the presence of HCV RNA may wrongly designate HCV viraemic seronegative individuals as uninfected. Amongst 424 injecting drug users whose serum was sent to our diagnostic laboratory for "HCV screening" over a 14-month period, the prevalence of HCV seropositivity was 48.4%. We retrospectively identified seven individuals for whom there was evidence of recent acquisition of HCV infection. Three of these infections were identified using our routine diagnostic protocol: testing for the presence of HCV-specific antibody and performing HCV RNA testing only on seropositive and seroindeterminate specimens. However, four cases were only identified by HCV RNA testing of HCV seronegative serum. On the basis of these observations, we estimate the incidence of HCV infection amongst London injecting drug users as being 14.3 per 100 person-years. We advocate that all HCV seronegative blood samples obtained from injecting drug users should be tested for the presence of HCV RNA, and suggest that this could be done efficiently by nucleic acid testing the specimens in small pools. .


Gene therapy and cell reprogramming for the aging brain: achievements and promise.
In the central nervous system, cholinergic and dopaminergic (DA) neurons are among the cells most susceptible to the deleterious effects of age. Thus, the basal forebrain cholinergic system is known to undergo moderate neurodegenerative changes during normal aging as well as severe atrophy in Alzheimer's disease (AD). Parkinson's disease (PD), a degeneration of nigro-striatal DA neurons is the most conspicuous reflection of the vulnerability of DA neurons to age. Overall, there is growing evidence that a progressive decline in cognitive function and central DA activity represents basic features of normal aging both in humans and laboratory rodents. Spontaneous or environmental neurotoxin-mediated exacerbation of these processes contributes to the symptoms of AD and PD, respectively. In this context, neurotrophic factors that can prevent or delay the decline in cognitive function and central DA activity are of clinical interest. Among them, Insulin-like Growth Factor I and Glial cell line-Derived Neurotrophic Factor are emerging as powerful neuroprotective molecules. This article discusses the experimental evidence supporting the neuroprotective relevance of these and related factors in the aging brain. The availability of induced pluripotent stem cells offers a new promise for the treatment of pathologies associated with the loss of specific cell types as for instance, nigral DA neurons (in PD) or basal forebrain cholinergic neurons (BFCN) in the early stages of AD. Recent studies documenting the use of cell reprogramming for the generation of multipotent neuronal precursors as well as functional BFCN and DA neurons are reviewed.


Satisfaction with health care among dermatological inpatients.
Measuring patient satisfaction is regarded as one of the principal methods for obtaining patients' evaluation of services they receive. During the last decades more interest has been devoted to inpatients' preferences and needs. Surveys on patient satisfaction may provide information to hospitals about areas where improvement is needed. In this survey, 648 questionnaires were completed by dermatological inpatients and were analyzed for overall satisfaction and seven satisfaction determinants. Background factors and clinical parameters were considered. Emotional support was recognized as a critical area. "Coordination of care" and "information and education" were identified as the most relevant determinants for overall patient satisfaction. The Hospital Management, taking into account the results, planned training in order to improve personnel's communication skills. This study confirms that patient satisfaction analysis is a useful instrument also among dermatological inpatients, and satisfaction is a valid measure of quality of health care.


A Redesign Approach for Improving Animal Care Services for Researchers.
Because a research institution's animal care and use program oversees the provision of services specified in approved protocols designed by researchers, the effective provision of services within these programs is paramount to ensuring the humane care and treatment of research animals in accordance with federal, state, and local regulations and institutional policies. To improve the services provided to researchers through animal care and use programs, we investigated the relationship between the researchers who conduct these types of studies and the veterinary operations that provide care and treatment for research animals. Through a case study conducted at a leading public research university, we used an action-research approach to redesign aspects of the process through which researchers and the veterinary operations interact by using the Design for Six Sigma methodology. Using this structured approach for building quality into the design of a process to better serve customers, we identified and prioritized researchers' expectations regarding the role of veterinary operations in supporting their animal research activities. In addition, ideas for addressing researchers' top-rated needs were generated through focus groups. By updating online resources, creating checklists and newsletters, and hiring additional veterinary staff, the services provided were amended to provide researchers with increased access to valuable information, improved clarity regarding the process for obtaining access to research facilities, and enhanced support for animal care services.


The expression of Drosha, DGCR8, Dicer and Ago-2 genes are upregulated in human umbilical vein endothelial cells under hyperglycemic condition.
It has been shown that dysregulation of miRNAs expression contributes to the pathogenesis and progression of the diabetes and diabetes-related complications. Drosha, DGCR8, Dicer, and Ago-2 are involved in the miRNA maturation. The aim of the present study was to investigate the mRNA expression levels of these genes in the human umbilical vein endothelial cells (HUVECs) under hyperglycemic condition. HUVECs were cultured in normo-(5 mM) and hyperglycemic (25 mM) conditions for 24 h. As osmotic control, cells were treated with D-mannitol (25 mM, for 24 h). The mRNA expression levels of Drosha, DGCR8, Dicer and Ago-2 were evaluated using quantitative real-time PCR. The expression level of Drosha, DGCR8, Dicer, and Ago-2 were increased in hyperglycemic HUVECs compared to the control group. Our results show that under hyperglycemic condition, expression of genes involved in the miRNA maturation was significantly increased in HUVECs. Upregulation of these genes may have role in diabetic complications through the dysregulation of the miRNA expression.


Pitch for nonsimultaneous successive harmonics in quiet and noise.
Pitch perception was studied with respect to nonsimultaneous successive harmonics of fundamentals in quiet and noise. Stimuli consisted of three nonoverlapping sine waves each of 40-msec duration. Pitch perception, examined by discrimination and matching, was found to correspond to spectral frequency in quiet and fundamental frequency in noise. The results suggest a pitch processor that integrates over time (synthetic mode) in noise and analyzes spectral components (analytic mode) in quiet. The findings are discussed with respect to current theories of pitch perception and auditory integrative functions. The findings are thought to be particularly relevant to analytic and synthetic modes of pitch perception.


Antigen recognition is facilitated by invadosome-like protrusions formed by memory/effector T cells.
Adaptive immunity requires that T cells efficiently scan diverse cell surfaces to identify cognate Ag. However, the basic cellular mechanisms remain unclear. In this study, we investigated this process using vascular endothelial cells, APCs that possess a unique and extremely advantageous, planar morphology. High-resolution imaging revealed that CD4 memory/effector T cells dynamically probe the endothelium by extending submicron-scale, actin-rich "invadosome/podosome-like protrusions" (ILPs). The intimate intercellular contacts enforced by ILPs consistently preceded and supported T cell activation in response to endothelial MHC class II/Ag. The resulting calcium flux stabilized dense arrays of ILPs (each enriched in TCR, protein kinase C-θ, ZAP70, phosphotyrosine, and HS1), forming what we term a podo-synapse. Similar findings were made using CD8 CTLs on endothelium. Furthermore, careful re-examination of both traditional APC models and professional APCs suggests broad relevance for ILPs in facilitating Ag recognition. Together, our results indicate that ILPs function as sensory organelles that serve as actuators of immune surveillance.


A Promising New Anti-Cancer Strategy: Iron Chelators Targeting CSCs.
Iron is a trace but vital element in the human body and is necessary for a multitude of crucial processes in life. However, iron overload is known to induce carcinogenesis via oxidative stress. Cancer cells require large amounts of iron for their rapid division and cell growth. Iron was recently found to play a role in cancer stem cells (CSCs); it maintains stemness during development. Iron also plays an important role in stemness by moderating reactive oxygen species. Thus, iron metabolism in CSCs is a promising therapeutic target. In this review, we summarize the roles of iron in cancer cells and CSCs. We also summarize anti-cancer therapeutic studies with iron chelators and describe our expectation of a new therapeutic strategy for CSCs on the basis of our findings.


The spectrum of non alcoholic fatty liver disease in morbidly obese patients: prevalence and associate risk factors.
To determine the prevalence of non alcoholic fatty liver disease (NAFLD) and non alcoholic steatohepatitis (NASH) in morbidly obese patients undergoing bariatric surgery and to identify risk factors associated with the disease spectrum. Liver biopsy was performed in 60 patients who underwent bariatric surgery, after other causes of liver disease were excluded. Clinical, biochemical and histological features were evaluated. NAFLD was detected in fifty-seven patients (95%) of the sample and forty patients (66.7%) of the total sample met the criteria for NASH. Perisinusoidal fibrosis was only found in three (7.5%) patients with NASH. The γGT was an independent predictive factor associated with the degree of hepatic steatosis. The variables such as dyslipidemia and ALT were independently associated with the presence of Mallory's corpuscles with the following values, respectively, OR 0, 05, 95% CI 0.002 to 0.75, P = 0.031 and OR 10, 99, 95% CI 1.44 to 83.93, P = 0.021. Non alcoholic fatty liver disease seems to be an obese-related condition with approximately half of asymptomatic morbidly obese patients having histological NASH. The γGT was an independent predictor of the degree of steatosis.


The PML-RAR alpha fusion mRNA generated by the t(15;17) translocation in acute promyelocytic leukemia encodes a functionally altered RAR.
We have previously shown that the t(15;17) translocation specifically associated with acute promyelocytic leukemia (APL) fuses the retinoic acid receptor alpha (RAR alpha) locus to an as yet unknown gene, initially called myl and now renamed PML. We report here that this gene product contains a novel zinc finger motif common to several DNA-binding proteins. The PML-RAR alpha mRNA encodes a predicted 106 kd chimeric protein containing most of the PML sequences fused to a large part of RAR alpha, including its DNA- and hormone-binding domains. In transient expression assays, the hybrid protein exhibits altered transactivating properties if compared with the wild-type RAR alpha progenitor. Identical PML-RAR alpha fusion points are found in several patients. These observations suggest that in APL, the t(15;17) translocation generates an RAR mutant that could contribute to leukemogenesis through interference with promyelocytic differentiation.


ORAI1 establishes resistance to SARS-CoV-2 infection by regulating tonic type I interferon signaling.
ORAI1 and STIM1 are the critical mediators of store-operated Ca 2+ entry by acting as the pore subunit and an endoplasmic reticulum-resident signaling molecule, respectively. In addition to Ca 2+ signaling, STIM1 is also involved in regulation of a cytosolic nucleic acid sensing pathway. Using ORAI1 and STIM1 knockout cells, we examined their contribution to the host response to SARS-CoV-2 infection. STIM1 knockout cells showed strong resistance to SARS-CoV-2 infection due to enhanced type I interferon response. On the contrary, ORAI1 knockout cells showed high susceptibility to SARS-CoV-2 infection as judged by increased expression of viral proteins and a high viral load. Mechanistically, ORAI1 knockout cells showed reduced homeostatic cytoplasmic Ca 2+ concentration and severe impairment in tonic interferon signaling. Transcriptome analysis showed downregulation of multiple cellular defense mechanisms, including antiviral signaling pathways in ORAI1 knockout cells, which are likely due to reduced expression of the Ca 2+ -dependent transcription factors of the activator protein 1 (AP-1) family and MEF2C . Our results identify a novel role of ORAI1-mediated Ca 2+ signaling in regulating the baseline type I interferon level, which is a determinant of host resistance to SARS-CoV-2 infection.


Organized kidney retrieval on a regional basis. A new approach.
A network of small to medium-sized communities has been developed to increase the supply of cadaveric kidneys for renal allotransplantation. After a training period, community physicians and the community hospitals were designated as members of the transplant team to retrieve kidneys. A threefold increase in the number of cadaveric kidneys has resulted from this project. This mechanism for organized kidney retrieval on a regional basis could be used to augment the supply of cadaveric organs in other areas of the United States.


The effects of methamphetamine self-administration on cortical monoaminergic deficits induced by subsequent high-dose methamphetamine administrations.
Preclinical models suggest that repeated high-dose methamphetamine (METH) exposures, administered in a "binge-like" pattern, acutely decrease norepinephrine (NE), and acutely and persistently decrease serotonin (5-hydroxytryptamine; 5HT) content in the frontal cortex. However, the impact of METH self-administration on this region is unknown. Because of the importance of the monoaminergic neurons in the frontal cortex to a variety of cognitive and addictive processes, effects of METH self-administration on cortical NE and 5HT content were assessed. Results revealed several novel findings. First, METH self-administration decreased cortical NE content as assessed 24 h after last exposure. Consistent with previous preclinical reports after a binge METH regimen, this decrease was reversed 8 days after the final METH exposure. Second, and in contrast to our previous reports involving the hippocampus or striatum, METH self-administration caused persistent decreases in 5HT content as assessed 8 days after the final METH exposure. Of note, the magnitude of this decrease (≈ 20%) was less than that observed typically after a binge METH treatment. Third, prior METH self-administration attenuated METH-induced serotonergic deficits as assessed 7 days, but not 1 h, following a neurotoxic METH regimen. No protection was observed when the binge exposure occurred 15 days after the last self-administration session. Taken together, these data demonstrate important and selective alterations in cortical serotonergic neuronal function subsequent to METH self-administration. These data provide a foundation to investigate complex questions involving "resistance" to the persistent deficits caused by neurotoxic METH exposure and frontal cortical function.


Studies on the oestrogen sulphatase and arylsulphatase C activities of rat liver.
Detailed studies on the hydrolysis of p-acetylphenyl sulphate and oestrone sulphate by rat liver preparations strongly indicate that arylsulphatase C and oestrogen sulphatase are the same enzyme. Liver is the richest source of both enzymes, which have identical intracellular distributions, being localized mainly in the microsomal fraction. Low oestrogen sulphatase and arylsulphatase C activities were present in foetal liver and these increased at a similar rate after birth. The activities of the enzymes in an ethionine-induced hepatoma were similarly low. Results of heat inactivation, mixed-substrate and competitive-inhibition experiments employing liver microsomal fractions were also consistent with one enzyme being involved. Oestradiol-17beta 3-sulphate was also hydrolysed by microsomal preparations and activity towards both this substrate and oestrone sulphate was inhibited by oestrone and oestradiol-17beta. The physiological significance of this inhibition is discussed.


[The surgical treatment of urethral strictures].
The authors reviewed the results of surgical management of 233 patients aged 8-76 years who had suffered urethral strictures. A total of 266 operations were made in the period of 1970-1989. Urethral tunneling which had been widely used in clinical practice for strictures of the urethra until 1980 yielded 91.2% recurrences of the disease, which made surgeons modify the surgical policy. During the past decade 127 patients with urethral strictures and obliterations have undergone 138 surgical interventions: 91 (65.9%) urethral resections, 34 (24.6%) internal optical urethrotomies and transurethral resections of scarring tissue, 7 (5.1%) cutaneous urethral plastic surgeries and only 6 (4.4%) urethral tunnelings. The number of relapses was reduced to 13.4%. The authors consider that urethral resection should be the operation of choice in urethral strictures and obliterations. Endoscopic treatment techniques (urethrotomy and transurethral resection) are indicated in short strictures and scarring deformity of the posterior urethra after prostate operation. It is advisable to apply cutaneous urethral plastic surgery in case of extended strictures.


Reintroducing the Sodium-Iodide Symporter to Anaplastic Thyroid Carcinoma.
Anaplastic thyroid carcinoma (ATC), the most aggressive form of thyroid cancer, is unresponsive to radioiodine therapy. The current study aimed to extend the diagnostic and therapeutic application of radioiodine beyond the treatment of differentiated thyroid cancer by targeting the functional sodium-iodide symporter (NIS) to ATC. The study employed nanoparticle vectors (polyplexes) based on linear polyethylenimine (LPEI), shielded by polyethylene glycol (PEG) and coupled to the synthetic peptide GE11 as an epidermal growth factor receptor (EGFR)-specific ligand in order to target a NIS-expressing plasmid (LPEI-PEG-GE11/NIS) to EGFR overexpressing human thyroid carcinoma cell lines. Using ATC xenograft mouse models, transfection efficiency by 123I scintigraphy and potential for systemic radioiodine therapy after systemic polyplex application were evaluated. In vitro iodide uptake studies in SW1736 and Hth74 ATC cells, and, for comparison, in more differentiated follicular (FTC-133) and papillary (BCPAP) thyroid carcinoma cells demonstrated high transfection efficiency and EGFR-specificity of LPEI-PEG-GE11/NIS that correlated well with EGFR expression levels. After systemic polyplex injection, in vivo 123I gamma camera imaging revealed significant tumor-specific accumulation of radioiodine in an SW1736 and an Hth74 xenograft mouse model. Radioiodine accumulation was found to be higher in SW1736 tumors, reflecting in vitro results, EGFR expression levels, and results from ex vivo analysis of NIS staining. Administration of 131I in LPEI-PEG-GE11/NIS-treated SW1736 xenograft mice resulted in significantly reduced tumor growth associated with prolonged survival compared to control animals. The data open the exciting prospect of NIS-mediated radionuclide imaging and therapy of ATC after non-viral reintroduction of the NIS gene. The high tumor specificity after systemic application makes the strategy an attractive alternative for the treatment of highly metastatic ATC.


[Micavibrio admirandus gen. et sp. nov].
Micavibrio gen. nov. (M.L. fem. n. mica--a tiny thing and M.L. masc. n. Vibroia generic name, M.L. masc. n. Micavibrio a tiny vibrio). Cells are single, small curved rods, in motile stage with a single polar not sheathed flagellum. Reproduced by binary division, gram-negative. Cells attach to host-bacteria cells and grow as exoparasites destroying the host, may exhibit a host-range specificity. The G + C content of the DNA of the strain examined is 57,1% M. Isolated from sewage waters. The type species is M. admirandus sp. nov. M. admirandus sp. nov (M.L. adj. admirable). Curved gram-negative rods 0,25-0,35X0,6-1,0 micron with a single flagellum 13-15 nm in width. Motile cells attach to the cell wall of the bacterium. Pseudomonas maltophilia, lose the motility and grow as exoparasites destroying the host. Develops only on. P. maltophilia cells and is unable to multiply in the absence of a host organism. Resistant to a vibriostatic agent pteridine 0/129. The G + C content of the DNA of the strain examined is 57,1% M. The type strain ARL-14 is maintained in the culture collection of the Institute of Biochemistry and Physiology of Microorganisms, USSR Academy of Sciences. The type strain has been isolated from sewage waters of the town of Pushchino.


Effect of inhibiting N-glycosylation or oligosaccharide processing on vasoactive intestinal peptide receptor binding activity and structure.
We used inhibitors of four steps of the glycosylation pathway to examine the contribution of carbohydrate moieties to the ligand-binding activity, cell-surface expression and apparent molecular mass of the human vasoactive intestinal peptide (VIP) receptor. Human melanoma IGR 39 cells, incubated for 60 h with the inhibitors tunicamycin, castanospermine, swainsonine or deoxymannojirimycin, under conditions where cell viability and protein synthesis were not affected, expressed VIP receptor species with different VIP-binding properties. The most pronounced effects on VIP binding were obtained with tunicamycin and deoxymannojirimycin, which respectively caused 80% and 67% inhibition. Treatment with either swainsonine or castanospermine resulted in only a 25-32% decrease in VIP specific binding. Based on Scatchard analyses of data from competition experiments, the decrease in VIP-binding activity in either swainsonine- or deoxymannojirimycin-treated cells was due to a decrease in ligand affinity; the cell-surface number of VIP-binding sites remained unchanged. In contrast, tunicamycin and castanospermine caused decreases in the cell-surface number of functional VIP receptors without affecting affinity. Besides, the drug-treated cells produced VIP-binding proteins with different molecular masses and endoglycosidase H (Endo H) sensitivities. When compared with their counterpart synthesized in control cells, VIP-binding proteins produced by deoxymannojirimycin- or swainsonine-treated cells were smaller in size and exhibited the expected sensitivity to Endo H. No modification in the apparent molecular mass was observed in the presence of either castanospermine or tunicamycin. In addition, after Endo F digestion, all of the deglycosylated proteins migrated with the same electrophoretic mobility. Finally, processing in the presence of castanospermine led to an Endo H-resistant receptor species which showed an unexpected neuraminidase-sensitivity, indicating that, as in control cells, these receptors carry V-linked oligosaccharides with terminal sialic acid residues.


Highly enantioselective adsorption of small prochiral molecules on a chiral intermetallic compound.
Intrinsically chiral surfaces of intermetallic compounds are shown to be novel materials for enantioselective processes. Their advantage is the significantly higher thermal and chemical stability, and therefore their extended application range for catalyzed chiral reactions compared to surfaces templated with chiral molecular modifiers or auxiliaries. On the Pd1 -terminated PdGa(111) surface, room-temperature adsorption of a small prochiral molecule (9-ethynylphenanthrene) leads to exceptionally high enantiomeric excess ratios of up to 98 %. Our findings highlight the great potential of intrinsically chiral intermetallic compounds for the development of novel, enantioselective catalysts that can be operated at high temperatures and potentially also in harsh chemical environments.


Dual mechanisms of metabolite acquisition by the obligate intracytosolic pathogen Rickettsia prowazekii reveal novel aspects of triose phosphate transport.
Rickettsia prowazekii is an obligate intracytosolic pathogen and the causative agent of epidemic typhus fever in humans. As an evolutionary model of intracellular pathogenesis, rickettsiae are notorious for their use of transport systems that parasitize eukaryotic host cell biochemical pathways. Rickettsial transport systems for substrates found only in eukaryotic cell cytoplasm are uncommon among free-living microorganisms and often possess distinctive mechanisms. We previously reported that R. prowazekii acquires triose phosphates for phospholipid biosynthesis via the coordinated activities of a novel dihydroxyacetone phosphate transport system and an sn-glycerol-3-phosphate dehydrogenase (K. M. Frohlich et al., J. Bacteriol. 192:4281-4288, 2010). In the present study, we have determined that R. prowazekii utilizes a second, independent triose phosphate acquisition pathway whereby sn-glycerol-3-phosphate is directly transported and incorporated into phospholipids. Herein we describe the sn-glycerol-3-phosphate and dihydroxyacetone phosphate transport systems in isolated R. prowazekii with respect to kinetics, energy coupling, transport mechanisms, and substrate specificity. These data suggest the existence of multiple rickettsial triose phosphate transport systems. Furthermore, the R. prowazekii dihydroxyacetone phosphate transport systems displayed unexpected mechanistic properties compared to well-characterized triose phosphate transport systems from plant plastids. Questions regarding possible roles for dual-substrate acquisition pathways as metabolic virulence factors in the context of a pathogen undergoing reductive evolution are discussed.


Causal reasoning identifies mechanisms of sensitivity for a novel AKT kinase inhibitor, GSK690693.
Inappropriate activation of AKT signaling is a relatively common occurrence in human tumors, and can be caused by activation of components of, or by loss or decreased activity of inhibitors of, this signaling pathway. A novel, pan AKT kinase inhibitor, GSK690693, was developed in order to interfere with the inappropriate AKT signaling seen in these human malignancies. Causal network modeling is a systematic computational analysis that identifies upstream changes in gene regulation that can serve as explanations for observed changes in gene expression. In this study, causal network modeling is employed to elucidate mechanisms of action of GSK690693 that contribute to its observed biological effects. The mechanism of action of GSK690693 was evaluated in multiple human tumor cell lines from different tissues in 2-D cultures and xenografts using RNA expression and phosphoproteomics data. Understanding the molecular mechanism of action of novel targeted agents can enhance our understanding of various biological processes regulated by the intended target and facilitate their clinical development. Causal network modeling on transcriptomic and proteomic data identified molecular networks that are comprised of activated or inhibited mechanisms that could explain observed changes in the sensitive cell lines treated with GSK690693. Four networks common to all cell lines and xenografts tested were identified linking GSK690693 inhibition of AKT kinase activity to decreased proliferation. These networks included increased RB1 activity, decreased MYC activity, decreased TFRC activity, and increased FOXO1/FOXO3 activity. AKT is involved in regulating both cell proliferation and apoptotic pathways; however, the primary effect with GSK690693 appears to be anti-proliferative in the cell lines and xenografts evaluated. Furthermore, these results indicate that anti-proliferative responses to GSK690693 in either 2-D culture or xenograft models may share common mechanisms within and across sensitive cell lines.


[Sociodemographic research on family planning: the case of the Health Department].
The Head Office for Family Planning of the Mexican Ministry of Health implemented a program of sociodemographic research for the purpose of studying family planning. On the basis of the sociodemographic profile drawn up in 1983--the year in which said research program was initiated--research needs were gradually identified and then met by means of surveys and monographs for individual states in Mexico. This paper comments on the main features of the research projects that have been carried out. We order to show the data bank which the Head Office for Family Planning has at the present time for studying the phenomenon all of its activities are centered around.


Absolute two-point quantification of proteins using dimethylated proteotypic peptides.
Absolute targeted proteomics typically employs known amounts of synthetic stable isotopically labeled peptides which are mixed with the analyte and analysed by LC-MS to determine the concentration of proteins. In order to obtain more data, we evaluated the use of two different stable isotopes of the same peptide as spike-in for absolute quantification. For this purpose, peptide labeling by reductive amination was applied, which is a mild reaction for dimethylation of amine groups with very high yield. Three different forms can be generated with e.g., light and heavy labels for spike-in peptides, and medium label for endogenous peptides. The method was studied with peptides of apolipoprotein A-I, apolipoprotein B-100, and leucine-rich alpha-2-glycoprotein without and with serum. In serum, the endogenous protein concentrations were measured across four orders of magnitude by the two-point quantification method. Less than 20% of coefficient of variation (CV) values and strong correlation with R2 of 0.99 across three analytical replicates was observed. Most importantly, the two-point quantification method allows an internal quality control of the spike-in peptide as strong deviations in ratios calculated between the first and second reference indicate a methodical error. Because of the significant lower costs than synthetically stable isotopically labeled peptides, this approach might be particularly interesting for the absolute quantification of multiple proteins.


[Tarsal gland of ixodid ticks (Ixodidae)].
In the tarsi of all legs of Ixodes persulcatus, Dermacentor daghestanicus, Rhipicephalus turanicus and Hyalomma asiaticum the multicellular gland was observed. The gland is formed by a single layer of glandular cells that form a central cavity where two ciliary structures in a cuticular (scolopoid) sheath are situated. A possible function of the gland is discussed in the article.


Spheroid formation of hepatocytes using synthetic polymer.
It is known that hepatocytes show the different morphology depending on the surface condition of cultural substratum. The cells form a floating cell-aggregate called spheroid on proteoglycan-coated or positively charged dishes. The liver cell functions are generally high and maintained for the long-term in the spheroid culture. Since spheroid morphology is preferable for the construction of a bioartificial liver, it is important to develop an effective method of preparing spheroids. In this regard, we examined a preparation method of functional spheroid-like cell-aggregates, in which a synthetic polymer, Eudragit was added to culture medium for inducing liver cell-aggregation. The cell-cell attachment of the aggregate was loose at the beginning of the culture, but it became tight and spheroids were formed 2-3 days after inoculation. When 0.1% Eudragit was added to the medium, the liver functions such as albumin secretion, ammonia removal and urea synthesis were enhanced compared with monolayer and conventional spheroid cultures. The spheroid formation was also performed with suspension culture in a spinner flask. Approximately 80% of the cells inoculated formed spheroids by the addition of the polymer. Moreover, the polymer showed a protective effect from cell damage by agitation. Since this procedure does not require surface for cell attachment, a large amount of spheroids can be prepared in suspension culture.


No Impact of the Analytical Method Used for Determining Cystatin C on Estimating Glomerular Filtration Rate in Children.
Measurement of inulin clearance is considered to be the gold standard for determining kidney function in children, but this method is time consuming and expensive. The glomerular filtration rate (GFR) is on the other hand easier to calculate by using various creatinine- and/or cystatin C (Cys C)-based formulas. However, for the determination of serum creatinine (Scr) and Cys C, different and non-interchangeable analytical methods exist. Given the fact that different analytical methods for the determination of creatinine and Cys C were used in order to validate existing GFR formulas, clinicians should be aware of the type used in their local laboratory. In this study, we compared GFR results calculated on the basis of different GFR formulas and either used Scr and Cys C values as determined by the analytical method originally employed for validation or values obtained by an alternative analytical method to evaluate any possible effects on the performance. Cys C values determined by means of an immunoturbidimetric assay were used for calculating the GFR using equations in which this analytical method had originally been used for validation. Additionally, these same values were then used in other GFR formulas that had originally been validated using a nephelometric immunoassay for determining Cys C. The effect of using either the compatible or the possibly incompatible analytical method for determining Cys C in the calculation of GFR was assessed in comparison with the GFR measured by creatinine clearance (CrCl). Unexpectedly, using GFR equations that employed Cys C values derived from a possibly incompatible analytical method did not result in a significant difference concerning the classification of patients as having normal or reduced GFR compared to the classification obtained on the basis of CrCl. Sensitivity and specificity were adequate. On the other hand, formulas using Cys C values derived from a compatible analytical method partly showed insufficient performance when compared to CrCl. Although clinicians should be aware of applying a GFR formula that is compatible with the locally used analytical method for determining Cys C and creatinine, other factors might be more crucial for the calculation of correct GFR values.


Clopidogrel with aspirin in High-risk patients with Acute Non-disabling Cerebrovascular Events II (CHANCE-2): rationale and design of a multicentre randomised trial.
In patients with a minor ischaemic stroke or transient ischaemic attack (TIA), separate trials have shown that dual antiplatelet therapy with clopidogrel plus aspirin (clopidogrel-aspirin) or ticagrelor plus aspirin (ticagrelor-aspirin) are more effective than aspirin alone in stroke secondary prevention. However, these two sets of combination have not been directly compared. Since clopidogrel was less effective in stroke patients who were CYP2C19 loss-of-function (LOF) allele carriers, whether ticagrelor-aspirin is clinically superior to clopidogrel-aspirin in this subgroup of patients with stroke is unclear. To describe the rationale and design considerations of the Clopidogrel in High-risk patients with Acute Non-disabling Cerebrovascular Events (CHANCE-2) trial. CHANCE-2 is a randomised, double-blind, double-dummy, placebo-controlled, multicentre trial that compares two dual antiplatelet strategies for minor stroke or TIA patients who are CYP2C19 LOF allele carriers: ticagrelor (180 mg loading dose on day 1 followed by 90 mg twice daily on days 2-90) or clopidogrel (300 mg loading dose on day 1 followed by 75 mg daily on days 2-90), plus open-label aspirin with a dose of 75-300 mg on day 1 followed by 75 mg daily on day 2-21. All will be followed for 1 year. The primary efficacy outcome is any stroke (ischaemic or haemorrhagic) within 3 months and the primary safety outcome is any severe or moderate bleeding event within 3 months. The CHANCE-2 trial will evaluate whether ticagrelor-aspirin is superior to clopidogrel-aspirin for minor stroke or TIA patients who are CYP2C19 LOF allele carriers. NCT04078737.


Increasing omega-3 desaturase expression in tomato results in altered aroma profile and enhanced resistance to cold stress.
One of the drawbacks in improving the aroma properties of tomato (Solanum lycopersicum) fruit is the complexity of this organoleptic trait, with a great variety of volatiles contributing to determine specific quality features. It is well established that the oxylipins hexanal and (Z)-hex-3-enal, synthesized through the lipoxygenase pathway, are among the most important aroma compounds and impart in a correct proportion some of the unique fresh notes in tomato. Here, we confirm that all enzymes responsible for the synthesis of these C6 compounds are present and active in tomato fruit. Moreover, due to the low odor threshold of (Z)-hex-3-enal, small changes in the concentration of this compound could modify the properties of the tomato fruit aroma. To address this possibility, we have overexpressed the omega-3 fatty acid desaturases FAD3 and FAD7 that catalyze the conversion of linoleic acid (18:2) to linolenic acid (18:3), the precursor of hexenals and its derived alcohols. Transgenic OE-FAD tomato plants exhibit altered fatty acid composition, with an increase in the 18:3/18:2 ratio in leaves and fruits. These changes provoke a clear variation in the C6 content that results in a significant alteration of the (Z)-hex-3-enal/hexanal ratio that is particularly important in ripe OE-FAD3FAD7 fruits. In addition to this effect on tomato volatile profile, OE-FAD tomato plants are more tolerant to chilling. However, the different behaviors of OE-FAD plants underscore the existence of separate fatty acid fluxes to ensure plant survival under adverse conditions.


A specific mechanism for nonspecific activation in reporter-gene assays.
The importance of bioluminescence in enabling a broad range of high-throughput screening (HTS) assay formats is evidenced by widespread use in industry and academia. Therefore, understanding the mechanisms by which reporter enzyme activity can be modulated by small molecules is critical to the interpretation of HTS data. In this Perspective, we provide evidence for stabilization of luciferase by inhibitors in cell-based luciferase reporter-gene assays resulting in the counterintuitive phenomenon of signal activation. These data were derived from our analysis of luciferase inhibitor compound structures and their prevalence in the Molecular Libraries Small Molecule Repository using 100 HTS experiments available in PubChem. Accordingly, we found an enrichment of luciferase inhibitors in luciferase reporter-gene activation assays but not in assays using other reporters. In addition, for several luciferase inhibitor chemotypes, we measured reporter stabilization and signal activation in cells that paralleled the inhibition determined using purified luciferase to provide further experimental support for these contrasting effects.


Thirty-day perioperative outcomes in resection of cervical lymphatic malformations.
Limited information exists regarding short-term morbidity in the resection of lymphatic malformations. In order to make informed collaborative medical decisions, clinicians and families would benefit from information on 30-day outcomes and the expected course associated with surgical excision of lymphatic malformations. A retrospective chart review was conducted to develop a case series of patients who underwent resection of lymphatic malformation at a pediatric tertiary care center between June 1, 2007 and September 30, 2016. Demographic data, disease characteristics, operative details, post-operative care, and adverse events in the 30-day post-operative period were analyzed. Primary outcomes included facial nerve dysfunction, seroma formation, re-admission, and overall rate of any complications. Secondary outcomes included operative time, duration of stay, and duration of drain placement. Forty-nine excisions were performed in 46 patients (21 male, 25 female). Median age was 5 years. All but 7 cases were performed as the initial primary intervention. Median operative time was 96 min (range 22-224). Higher stage lesions (3-5) were associated with a longer operative time (p = .03). Median length of stay was 2 days (range 0-35). Higher stage lesions were associated with an increased length of stay (p = .0004). Median duration of drain placement was 2 days (range 0-14), and was longer in higher stage lesions (p = .0002). Higher stage lesions (p = .002) and cases ultimately found to have residual disease (p = .019) were associated with an increased overall rate of complications; there was no association between cyst type and rate of complications. Seroma formation (31%) and transient facial nerve weakness (26%) were the two most common complications observed. There was no association between stage or cyst type and likelihood of seroma formation. Seromas resolved after a median duration of 3 months and transient facial nerve weakness resolved after a median duration of 2 months. 3 patients required return to the OR and 1 patient was readmitted. The overall rate of adverse events after surgical excision of cervical lymphatic malformations is relatively low. Increased rates of complications can be expected with higher stage. Similarly, for these higher stage lesions, a longer operative time, hospital stay, and duration of drain placement can be expected.


How do friends influence smoking uptake? Findings from qualitative interviews with identical twins.
The smoking behavior of friends is a major risk factor for adolescent smoking uptake. To explore the social context of smoking experimentation and consolidation with a particular focus on friends, the authors interviewed both members of 14 young adult identical twin pairs who were discordant for smoking. The different smoking status of twins was connected to their different friendship groups and development of different identities. Smoking respondents gravitated to the behaviors and images of the peer group who smoked. Many nonsmokers felt strong pressure from their peers not to smoke and spoke about how the images conveyed by smoking were inconsistent with their peer group's image. Adolescents and young adults are aware of the messages that smoking can convey to others and exploit these images to construct a social identity.


Alternatively spliced tissue factor promotes breast cancer growth in a β1 integrin-dependent manner.
Full-length tissue factor (flTF), the coagulation initiator, is overexpressed in breast cancer (BrCa), but associations between flTF expression and clinical outcome remain controversial. It is currently not known whether the soluble alternatively spliced TF form (asTF) is expressed in BrCa or impacts BrCa progression. We are unique in reporting that asTF, but not flTF, strongly associates with both tumor size and grade, and induces BrCa cell proliferation by binding to β1 integrins. asTF promotes oncogenic gene expression, anchorage-independent growth, and strongly up-regulates tumor expansion in a luminal BrCa model. In basal BrCa cells that constitutively express both TF isoforms, asTF blockade reduces tumor growth and proliferation in vivo. We propose that asTF plays a major role in BrCa progression acting as an autocrine factor that promotes tumor progression. Targeting asTF may comprise a previously unexplored therapeutic strategy in BrCa that stems tumor growth, yet does not impair normal hemostasis.


Serum uric acid concentrations in horses heterozygous for combined immunodeficiency.
Serum uric acid concentrations were determined in horses known to be carriers of combined immunodeficiency gene(s) and in presumed noncarrier horses. Uric acid concentrations were significantly higher (P less than 0.005) in carrier horses than in presumed noncarrier horses. However, there was some overlap in serum uric acid concentrations between carrier and presumed noncarrier horses.


Clinical definition of respiratory viral infections in young children and potential bronchiolitis misclassification.
Viral respiratory infections are often grouped as a single respiratory syndrome named 'viral bronchiolitis', independently of the viral etiology or individual risk factors. Clinical trials and guidelines have used a more stringent definition of viral bronchiolitis, including only the first episode of wheezing in children less than 12 months of age without concomitant respiratory comorbidities. There is increasing evidence suggesting that this definition is not being followed by pediatric care providers, but it is unclear to what extent viral respiratory infections are currently misclassified as viral bronchiolitis using standard definitions. We conducted a retrospective analysis of hospitalized young children (≤3 years) due to viral respiratory infections. Bronchiolitis was defined as the first wheezing episode less than 12 months of age. Demographic variables and comorbidities were obtained by electronic medical record review. The study comprised a total of 513 hospitalizations (n=453). Viral bronchiolitis was diagnosed in 144 admissions (28.1%). Notably, we identified that the majority of children diagnosed with bronchiolitis (63%) were misclassified as they had prior episodes of wheezing. Many children with bronchiolitis misclassification had significant comorbidities, including prematurity (51%), neuromuscular conditions (9.8%), and congenital heart disease (9.8%). Misclassification of bronchiolitis is a common problem that may lead to inappropriate management of viral respiratory infections in young children. A comprehensive approach that takes into consideration viral etiology and individual risk factors may lead to a more accurate clinical assessment of this condition and would potentially prevent bronchiolitis misclassification.


Characteristics and Prognosis of Stroke in Living Donor Renal Transplant Recipients.
We aimed to determine the characteristics and vascular outcomes of stroke in renal transplant (RT) recipients and compare them with those in patients on hemodialysis (HD) and those with no renal replacement therapy (RRT). In this prospective observational study, 717 patients (mean age, 70.8 years; male, 60.5%) with acute ischemic stroke within one week of onset were consecutively enrolled and followed for one year. The patients were classified into three groups: (1) living donor RT recipients (n=27); (2) patients on maintenance HD before the index stroke (n=39); and (3) those with no history of RRT (n=651). The primary outcome was a composite of major adverse cardiovascular events (MACE). Diabetic nephropathy was the most common reason for RRT in both RT and HD patients. RT patients were more likely to have embolic stroke of undetermined source (33.3%) than others, whereas HD patients more often had cardioembolism (51.3%). No difference was observed in the MACE risk between the patients in RT and non-RRT groups (annual rate, 11.3% vs. 13.1%; log-rank P=0.82; hazard ratio [95% confidence interval], 0.92 [0.29-2.98]). In contrast, HD patients had a greater risk of MACE than those with no RRT (annual rate, 28.2% vs. 13.1%; log-rank P=0.019; hazard ratio [95% confidence interval], 2.24 [1.16-4.3]). The underlying etiologies of stroke differed in RT and HD patients. The one-year risk of MACE for stroke patients who had received an RT was lower than that for patients undergoing HD and comparable with that of patients with no RRT.


Abnormal cell responses and role of TNF-α in impaired diabetic wound healing.
Impaired diabetic wound healing constitutes a major health problem. The impaired healing is caused by complex factors such as abnormal keratinocyte and fibroblast migration, proliferation, differentiation, and apoptosis, abnormal macrophage polarization, impaired recruitment of mesenchymal stem cells (MSCs) and endothelial progenitor cells (EPCs), and decreased vascularization. Diabetes-enhanced and prolonged expression of TNF- α also contributes to impaired healing. In this paper, we discuss the abnormal cell responses in diabetic wound healing and the contribution of TNF- α .


Review: efficient rehabilitation trial designs using disease progress modeling: a pediatric traumatic brain injury example.
The identification of possible treatment effects against a background of spontaneous recovery is a major challenge to the successful completion of randomized clinical trials (RCTs) in rehabilitation research. Conventional trial outcomes such as the differences between group means of an outcome measure at a fixed time point are inefficient to an extent that is a major problem, particularly for exploratory studies seeking preliminary evidence of efficacy. To quantitate gains in study power over conventional fixed-end-point designs by using parametric end points derived from the modeling of the time course of recovery after brain injury. Nonlinear mixed effects (NLME) modeling of the recovery trajectories of 103 children rehabilitating after traumatic brain injury (TBI) as reflected in serial WeeFIM scores was performed. Pseudoreplicate data sets were generated replicating the statistical characteristics of the original data set, and these formed the basis of clinical trial simulations to derive robust estimates of study power. Parametric end points derived from modeling of recovery improve study power (and reduce necessary sample size) by up to 5 times in this example. Parametric end points derived from models of recovery trajectories offer an efficient alternative design for exploratory clinical studies of rehabilitation interventions.


Ascorbate accumulation during sulphur deprivation and its effects on photosystem II activity and H2 production of the green alga Chlamydomonas reinhardtii.
In nature, H2 production in Chlamydomonas reinhardtii serves as a safety valve during the induction of photosynthesis in anoxia, and it prevents the over-reduction of the photosynthetic electron transport chain. Sulphur deprivation of C. reinhardtii also triggers a complex metabolic response resulting in the induction of various stress-related genes, down-regulation of photosynthesis, the establishment of anaerobiosis and expression of active hydrogenase. Photosystem II (PSII) plays dual role in H2 production because it supplies electrons but the evolved O2 inhibits the hydrogenase. Here, we show that upon sulphur deprivation, the ascorbate content in C. reinhardtii increases about 50-fold, reaching the mM range; at this concentration, ascorbate inactivates the Mn-cluster of PSII, and afterwards, it can donate electrons to tyrozin Z(+) at a slow rate. This stage is followed by donor-side-induced photoinhibition, leading to the loss of charge separation activity in PSII and reaction centre degradation. The time point at which maximum ascorbate concentration is reached in the cell is critical for the establishment of anaerobiosis and initiation of H2 production. We also show that ascorbate influenced H2 evolution via altering the photosynthetic electron transport rather than hydrogenase activity and starch degradation.


Tomographic study of the arthroscopic approaches to the hip joint.
The anatomic depth of the hip joint has long been one of the limiting factors in the development of arthroscopy of this particular joint. A major factor would seem to be variation in body size. The main purpose of this study was to quantify the distance between the joint and the skin in usual arthroscopic approaches. In vivo radiologic study using computed tomography. We studied the distance from the center of the acetabulum to the skin. The lines that we studied correspond to the paratrochanteric, the anterolateral, and anterior arthroscopic entry points. Although notable differences exist from one individual to another, the average values of the aforementioned are 12.4 cm, 11.2 cm, and 9.8 cm, respectively. The distances of these portals are greater in women than in men (P <.05), and there is no statistically significant relationship to age. The results of the present work suggest that surgical tools needed for arthroscopy of the hip should be more than 16 cm long to guarantee performing hip arthroscopy comfortably in more that 95% of the population.


Prisoners' knowledge of HIV/AIDS and its prevention in Kerman, Islamic Republic of Iran.
Knowledge of prisoners regarding HIV/AIDS in Kerman was evaluated. Analysis indicated that the sample (n = 350) of prisoners had relatively high knowledge about HIV/AIDS and its modes of transmission. However, they had a lower level of knowledge about HIV/AIDS prevention. The overall knowledge of men about AIDS was significantly lower than women. Persons aged 46 years and older and illiterate inmates had the least knowledge about modes of transmission. In addition, the knowledge of illiterate prisoners about HIV/AIDS prevention was significantly lower than others. Evaluation of attitudes and practices of prisoners and implementation of educational programmes regarding HIV/AIDS are suggested.


Is shade beneficial for mediterranean shrubs experiencing periods of extreme drought and late-winter frosts?
Plants are naturally exposed to multiple, frequently interactive stress factors, most of which are becoming more severe due to global change. Established plants have been reported to facilitate the establishment of juvenile plants, but net effects of plant-plant interactions are difficult to assess due to complex interactions among environmental factors. An investigation was carried out in order to determine how two dominant evergreen shrubs (Quercus ilex and Arctostaphylos uva-ursi) co-occurring in continental, Mediterranean habitats respond to multiple abiotic stresses and whether the shaded understorey conditions ameliorate the negative effects of drought and winter frosts on the physiology of leaves. Microclimate and ecophysiology of sun and shade plants were studied at a continental plateau in central Spain during 2004-2005, with 2005 being one of the driest and hottest years on record; several late-winter frosts also occurred in 2005. Daytime air temperature and vapour pressure deficit were lower in the shade than in the sun, but soil moisture was also lower in the shade during the spring and summer of 2005, and night-time temperatures were higher in the shade. Water potential, photochemical efficiency, light-saturated photosynthesis, stomatal conductance and leaf 13C composition differed between sun and shade individuals throughout the seasons, but differences were species specific. Shade was beneficial for leaf-level physiology in Q. ilex during winter, detrimental during spring for both species, and of little consequence in summer. The results suggest that beneficial effects of shade can be eclipsed by reduced soil moisture during dry years, which are expected to be more frequent in the most likely climate change scenarios for the Mediterranean region.


Clinical features of liver involvement in adult patients with listeriosis. Review of the literature.
Clinical features of liver involvement due to Listeria monocytogenes infection in adults are rarely reported in literature. This is surprising, regarding the current opinion that the portal system is extensively involved in the first stages of pathogenesis in invasive L. monocytogenes disease. A literature search in the PubMed and Embase database revealed 34 cases with clinical features of hepatic involvement due to listeriosis. We systematically analyzed all case reports with respect to clinical manifestations, treatment and outcome. In addition, we added clinical information on a patient diagnosed with a solitary liver abscess due to L. monocytogenes, who was seen at our institution. This review describes the different presentations of liver-involvement reported in listeriosis; solitary liver abscess, multiple liver abscesses and diffuse or granulomatous hepatitis. Distinction between these different forms of liver involvement is clinically relevant as they have a different outcome. We delve into the different pathogenic events leading to different forms of liver involvement. In addition, diagnostic modalities and possible treatments are reviewed.


A contribution with review to the description of mineralization of bone and other calcified tissues in vivo.
This manuscript considers certain aspects of mineral deposition in bone and other vertebrate calcifying tissues in order to examine physical, chemical, and biological factors important in the mineralization process. The paper in a discussion format principally presents a new data and the formulation of concepts based on such data as well as a summary of background material as necessary review. Mineralization is found to occur at spatially independent sites throughout the organic extracellular tissue matrices. Matrix vesicles and collagen fibrils each may serve as independent nucleation centers for mineral with vesicle mineralization being local and collagen mineralization dominating the tissues as a whole. Collagen fibril organization is suggested to be such that hole zones are aligned in three dimensions, creating extensive channels for mineral accommodation. Nucleation occurs initially in hole zones and crystal growth leads to the development of plate-like mineral particles whose orientation, disposition, and sizes within fibrils are detailed. Effects of diffusion, crystallinity, and critical nucleation and growth events are described with respect to their influence on mineral deposition in bulk and local regions of tissue matrices.


Perceptual learning modules in mathematics: enhancing students' pattern recognition, structure extraction, and fluency.
Learning in educational settings emphasizes declarative and procedural knowledge. Studies of expertise, however, point to other crucial components of learning, especially improvements produced by experience in the extraction of information: perceptual learning (PL). We suggest that such improvements characterize both simple sensory and complex cognitive, even symbolic, tasks through common processes of discovery and selection. We apply these ideas in the form of perceptual learning modules (PLMs) to mathematics learning. We tested three PLMs, each emphasizing different aspects of complex task performance, in middle and high school mathematics. In the MultiRep PLM, practice in matching function information across multiple representations improved students' abilities to generate correct graphs and equations from word problems. In the Algebraic Transformations PLM, practice in seeing equation structure across transformations (but not solving equations) led to dramatic improvements in the speed of equation solving. In the Linear Measurement PLM, interactive trials involving extraction of information about units and lengths produced successful transfer to novel measurement problems and fraction problem solving. Taken together, these results suggest (a) that PL techniques have the potential to address crucial, neglected dimensions of learning, including discovery and fluent processing of relations; (b) PL effects apply even to complex tasks that involve symbolic processing; and (c) appropriately designed PL technology can produce rapid and enduring advances in learning.


Overlapping network meta-analyses on the same topic: survey of published studies.
To assess how common it is for a published network meta-analysis (NMA) to have other published overlapping NMAs, and to evaluate these overlaps. A total of 88 NMAs of randomized controlled trials evaluating the comparative effectiveness of health interventions were randomly selected. For each of these, we searched for NMAs on the same topic. A random sample of 40 pairs (an index NMA and one of its overlapping NMAs) was selected to assess the overlap in terms of nodes, treatments and references. The topic with the largest number of overlapping NMAs was described in depth. In all, 68 of the 88 index NMAs had at least one overlapping NMA: 77% [95% confidence interval (CI), 69-86%]. We identified 515 pairs of overlapping NMAs. Among the 40 randomly selected pairs, 73% (95% CI, 58-88%) of nodes, 79% (95% CI, 72-86%) of treatments and 48% (95% CI, 37-59%) of references included in the index NMAs were also found in the respective overlapping NMAs. Efficacy of biologics in rheumatoid arthritis had the largest number of overlapping NMAs, with 28 NMAs published between 2003 and 2014. Differences in selection and definition of nodes of treatments resulted in different network geometries. There were also differences in both the direction and the statistical significance of effects. Published NMAs exhibit extensive overlap and potential redundancy. Erratic retrieval of eligible trials, and lack of consensus on the range of interventions to be considered and how they might be merged or split in different nodes, may cause confusion.


The effects of Lysergic Acid Diethylamide (LSD) on the Positive Valence Systems: A Research Domain Criteria (RDoC)-Informed Systematic Review.
The renewed interest in psychedelic research provides growing evidence of potentially unique effects on various aspects of reward processing systems. Using the Research Domain Criteria (RDoC) framework, as proposed by the National Institute of Mental Health, we aim to synthesize the existing literature concerning the impact of lysergic acid diethylamide (LSD) on the RDoC's Positive Valence Systems (PVS) domain, and to identify potential avenues for further research. Two LSD-related terms (lysergic acid diethylamide and LSD) and 13 PVS-related terms (reward, happiness, bliss, motivation, reinforcement learning, operant, conditioning, satisfaction, decision making, habit, valence, affect, mood) were used to search electronic databases such as PubMed, Scopus, PsychINFO, and Web of Science for relevant articles. A manual search of the reference list resulted in nine additional articles. After screening, articles and data were evaluated and included based on their relevance to the objective of investigating the effects of LSD on the PVS. Articles and data were excluded if they did not provide information about the PVS, were observational in nature, lacked comparators or reference groups, or were duplicates. A risk of bias assessment was performed using the National Toxicology Program's Office of Health Assessment and Translation (NTP OHAT) risk of bias (RoB) tool. Data from the included articles were collected and structured based on the RDoC bio-behavioral matrix, specifically focusing on the PVS domain and its three constituent constructs: reward responsiveness, reward learning, and reward valuation. We reviewed 28 clinical studies with 477 participants. Lysergic acid diethylamide, assessed at self-report (23 studies), molecular (5 studies), circuit (4 studies), and paradigm (3 studies) levels, exhibited dose-dependent mood improvement (20 short-term and 3 long-term studies). The subjective and neural effects of LSD were linked to the 5-HT2A receptor (molecular). Animal studies (14 studies) suggested LSD could mildly reinforce conditioned place preference without aversion and reduce responsiveness to other rewards. Findings on reward learning were inconsistent but hinted at potential associative learning enhancements. Reward valuation measures indicated potential reductions in effort expenditure for other reinforcers. Our findings are consistent with our previous work, which indicated classical psychedelics, primarily serotonin 2A receptor agonists, enhanced reward responsiveness in healthy individuals and patient populations. Lysergic acid diethylamide exhibits a unique profile in the reward learning and valuation constructs. Using the RDoC-based framework, we identified areas for future research, enhancing our understanding of the impact of LSD on reward processing. However, applying RDoC to psychedelic research faces limitations due to diverse study designs that were not initially RDoC-oriented. Limitations include subjective outcome measure selection aligned with RDoC constructs and potential bias in synthesizing varied studies. Additionally, some human studies were open-label, introducing potential bias compared to randomized, blinded studies.


Novel polygonized single-wall carbon nanotube bundles.
We have synthesized novel crystalline ropes of "polygonized" single-wall carbon nanotubes (SWCNTs). The tubes exhibit rounded-hexagonal cross sections in contrast to the earlier observations of nearly circular tubes. To investigate the structural characteristics of the lattice of SWCNTs we have performed extensive molecular-dynamics simulations. We find several metastable structures of the lattice characterized by different tube cross sections, hexagonal, rounded-hexagonal, and circular, and increasing cell volume. The competition between different tube shapes as a function of tube diameter is analyzed and compared to experiments.


Role of skeletal muscle in motor neuron development.
The current paper is a continuation of our work most recently described in Kablar, 2011. Here, we show lists of up- and down-regulated genes obtained by a cDNA microarray analysis that compared developing mouse MyoD-/- limb musculature (MyoD-dependent, innervated by Lateral Motor Column motor neurons) and Myf5-/- back (epaxial) musculature (Myf5-dependent, innervated by Medial Motor Column motor neurons) to the control and to each other, at embryonic day 13.5 which coincides with the robust programmed cell death of motor neurons and the inability of myogenesis to undergo its normal progression in the absence of Myf5 and MyoD that at this embryonic day cannot substitute for each other. We wanted to see if/how the myogenic program couples with the neurotrophic one, and also to separate Lateral from Medial column trophic requirements, potentially relevant to Motor Neuron Diseases with the predilection for the Lateral column. Several follow-up steps revealed that Kif5c, Stxbp1 and Polb, differentially expressed in the MyoD-/- limb muscle, and Ppargc1a, Glrb and Hoxd10, differentially expressed in the Myf5-/- back muscle, are actually regulators of motor neuron numbers. We propose a series of follow-up experiments and various ways to consider our current data.


Antitumor Effect of Hyperoside Loaded in Charge Reversed and Mitochondria-Targeted Liposomes.
Hyperoside (HYP), a flavonol glycoside compound, has been shown to significantly inhibit the proliferation of malignant tumors. Mitochondria serve as both "energy factories" and "suicide weapon stores" of cells. Targeted delivery of cytotoxic drugs to the mitochondria of tumor cells and tumor vascular cells is a promising strategy to improve the efficacy of chemotherapy. We report a novel dual-functional liposome system possessing both extracellular charge reversal and mitochondrial targeting properties to enhance drug accumulation in mitochondria and trigger apoptosis of cancer cells. L-lysine was used as a linker to connect 2,3-dimethylmaleic anhydride (DMA) and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE) to yield a new compound, DSPE-Lys-DMA (DLD). Then, DLD was mixed with other commercially available lipids to form charge reversed and mitochondria-targeted liposomes (DLD-Lip). The size, morphology, zeta potential, serum stability, and protein adsorption of the HYP loaded DLD-Lip (HYP/DLD-Lip) were measured. The release profile, cellular uptake, in vitro and in vivo toxicity, and anticancer activity of HYP/DLD-Lip were investigated. The results showed that the mean diameter of the liposomes was less than 200 nm. The zeta potential of the liposomes was negative at pH 7.4. However, the zeta potential was positive at weak acidic pH values with the cleavage of the DMA amide. The charge reversion of HYP/DLD-Lip facilitated the cellular internalization and mitochondrial accumulation for enhanced antitumor effect. The strongest tumor growth inhibition (TGI 88.79%) without systemic toxicity was observed in DLD/HYP-Lips-treated CBRH-7919 tumor xenograft BALB/C mice. The charge reversed and mitochondria-targeted liposomes represented a promising anticancer drug delivery system for enhanced anticancer therapeutic efficacy.


Photoelectrochemical biosensor using enzyme-catalyzed in situ propagation of CdS quantum dots on graphene oxide.
An innovative photoelectrochemical (PEC) biosensor platform was designed based on the in situ generation of CdS quantum dots (QDs) on graphene oxide (GO) using an enzymatic reaction. Horseradish peroxidase catalyzed the reduction of sodium thiosulfate with hydrogen peroxide to generate H2S, which reacted with Cd(2+) to form CdS QDs. CdS QDs could be photoexcited to generate an elevated photocurrent as a readout signal. This strategy offered a "green" alternative to inconvenient presynthesis procedures for the fabrication of semiconducting nanoparticles. The nanomaterials and assembly procedures were characterized by microscopy and spectroscopy techniques. Combined with immune recognition and on the basis of the PEC activity of CdS QDs on GO, the strategy was successfully applied to a PEC assay to detect carcinoembryonic antigen and displayed a wide linear range from 2.5 ng mL(-1) to 50 μg mL(-1) and a detection limit of 0.72 ng mL(-1) at a signal-to-noise ratio of 3. The PEC biosensor showed satisfactory performance for clinical sample detection and was convenient for determining high concentrations of solute without dilution. This effort offers a new opportunity for the development of numerous rapid and convenient analytical techniques using the PEC method that may be applied in the design and preparation of various solar-energy-driven applications.


D4 Receptor Activation Differentially Modulates Hippocampal Basal and Apical Dendritic Synapses in Freely Moving Mice.
Activation of D4 receptors (D4Rs) has been shown to improve cognitive performance, potentially affecting synaptic strength. We investigated the D4R agonist PD 168077 (PD) in hippocampal CA1 of freely moving mice. We electrically stimulated in stratum oriens (OR) or radiatum (RAD) and evoked local field potentials (LFPs). Intraperitoneally injected PD dose-dependently and reversibly attenuated LFPs for longer time in basal (OR) than apical (RAD) dendrites. High-frequency stimulation induced LTP that was stronger and more stable in OR than RAD. LTP lasted at least 4 h during which the paired-pulse ratio remained reduced. A PD concentration not affecting synaptic transmission was sufficient to reduce LTP in OR but not in RAD. A PD concentration reducing synaptic transmission reduced the early phase LTP in OR additionally and the late phase LTP in RAD exclusively. Furthermore, cell type-specific expression of mCherry in DATCre mice generated fluorescence in dorsal CA1 that was highest in lacunosum moleculare and similar in OR/RAD, indicating that midbrain dopaminergic fibers distribute evenly in OR/RAD. Together, the D4R-mediated modulation of hippocampal synaptic transmission and plasticity is stronger in OR than RAD. This could affect information processing in CA1 neurons, since signals arriving via basal and apical afferents are distinct.


Towards the Identification and Characterization of Putative Adult Human Lens Epithelial Stem Cells.
The anterior lens epithelium has the ability to differentiate into lens fibres throughout its life. The present study aims to identify and functionally characterize the adult stem cells in the human lens epithelium. Whole mounts of lens epithelium from donor eyes (normal/cataract) were immunostained for SOX2, gap junction protein alpha 1 (GJA1), PAX6, α, β and γ-crystallins, followed by a confocal analysis. The functional property of adult stem cells was analysed by their sphere forming ability using cultured lens epithelial cells from different zones. Based on marker expression, the lens epithelium was divided into four zones: the central zone, characterized by a small population of PAX6+, GJA1-, β-crystallin- and γ-crystallin- cells; the germinative zone, characterized by PAX6+, GJA1+, β-crystallin- and γ-crystallin-; the transitional zone, characterized by PAX6+, GJA1+, β-crystallin+ and γ-crystallin-; and the equatorial zone, characterized by PAX6+/-, GJA1+, β-crystallin+, and γ-crystallin+ cells. The putative lens epithelial stem cells identified as SOX2+ and GJA1 membrane expression negative cells were located only in the central zone (1.89 ± 0.84%). Compared to the other zones, a significant percentage of spheres were identified in the central zone (1.68 ± 1.04%), consistent with the location of the putative adult lens epithelial stem cells. In the cataractous lens, an absence of SOX2 expression and a significant reduction in sphere forming ability (0.33 ± 0.11%) were observed in the central zone. The above findings confirmed the presence of putative stem cells in the central zone of the adult human lens epithelium and indicated their probable association with cataract development.


Metastatic Extramammary Paget's Disease of Scrotum Responds Completely to Single Agent Trastuzumab in a Hemodialysis Patient: Case Report, Molecular Profiling and Brief Review of the Literature.
Extramammary Paget's disease (EMPD) is a rare cancer. Although EMPD is usually noninvasive and treated with local therapy, once metastatic the prognosis of EMPD is poor and treatment options are limited. We report a case of a complete response to single agent trastuzumab in a hemodialysis patient with metastatic Her2/neu overexpressed EMPD of the scrotum. Molecular profiling of his case as well as 12 other EMPD and 8 mammary Paget disease (MPD) cases was completed and revealed multiple biomarker aberrations. Overexpression of Her2 was frequently noted (30%-40%) in both EMPD and MPD patients and when present can be effectively treated with Her2 targeted agents. Trastuzumab therapy can be safely utilized in a hemodialysis patient. In addition, multiple protein overexpression and loss were seen in EMPD including PD-1, PD-L1, PTEN, and AR as well as PIK3CA mutation. These findings may lead to possible therapeutic interventions targeting these pathways in a disease with few effective treatment options.


Pausing of flagellar rotation is a component of bacterial motility and chemotaxis.
When bacterial cells are tethered to glass by their flagella, many of them spin. On the basis of experiments with tethered cells it has generally been thought that the motor which drives the flagellum is a two-state device, existing in either a counterclockwise or a clockwise state. Here we show that a third state of the motor is that of pausing, the duration and frequency of which are affected by chemotactic stimuli. We have recorded on video tape the rotation of tethered Escherichia coli and Salmonella typhimurium cells and analyzed the recordings frame by frame and in slow motion. Most wild-type cells paused intermittently. The addition of repellents caused an increase in the frequency and duration of the pauses. The addition of attractants sharply reduced the number of pauses. A chemotaxis mutant which lacks a large part of the chemotaxis machinery owing to a deletion of the genes from cheA to cheZ did not pause at all and did not respond to repellents by pausing. A tumbly mutant of S. typhimurium responded to repellents by smooth swimming and to attractants by tumbling. When tethered, these cells exhibited a normal rotational response but an inverse pausing response to chemotactic stimuli: the frequency of pauses decreased in response to repellents and increased in response to attractants. It is suggested that (i) pausing is an integral part of bacterial motility and chemotaxis, (ii) pausing is independent of the direction of flagellar rotation, and (iii) pausing may be one of the causes of tumbling.


Understanding how health systems facilitate primary care redesign.
To understand how health systems are facilitating primary care redesign (PCR), examine the PCR initiatives taking place within systems, and identify barriers to this work. A purposive sample of 24 health systems in 4 states. Data were systematically reviewed to identify how system leaders define and implement initiatives to redesign primary care delivery and identify challenges. Researchers applied codes which were based on the theoretical PCR literature and created new codes to capture emerging themes. Investigators analyzed coded data then produced and applied a thematic analysis to examine how health systems facilitate PCR. Semi-structured telephone interviews with 162 system executives and physician organization leaders from 24 systems. Leaders at all 24 health systems described initiatives to redesign the delivery of primary care, but many were in the early stages. Respondents described the use of centralized health system resources to facilitate PCR initiatives, such as regionalized care coordinators, and integrated electronic health records. Team-based care, population management, and care coordination were the most commonly described initiatives to transform primary care delivery. Respondents most often cited improving efficiency and enhancing clinician job satisfaction, as motivating factors for team-based care. Changes in payment and risk assumption as well as community needs were commonly cited motivators for population health management and care coordination. Return on investment and the slower than anticipated rate in moving from fee-for-service to value-based payment were noted by multiple respondents as challenges health systems face in redesigning primary care. Given their expanding role in health care and the potential to leverage resources, health systems are promising entities to promote the advancement of PCR. Systems demonstrate interest and engagement in this work but face significant challenges in getting to scale until payment models are in alignment with these efforts.


Antibiotic Resistance Results of Helicobacter pylori in a University Hospital: Comparison of the Hybridization Test and Real-Time Polymerase Chain Reaction.
H. pylori is a bacterial pathogen in the human stomach which infects about 50% of the world population. Untreated infection can lead to various diseases leading to cancer. Some of the H. pylori strains are asymptomatic, but some of them cause more severe diseases. Standard treatment protocol used for the treatment of H. pylori infection is triple therapy, which includes omeprazole as a proton pump inhibitor (PPI) and two antibiotics usually consist of amoxicillin and clarithromycin or metronidazole. In the recent years, because of the increase in the rate of antibiotic resistance, the eradication rate has decreased. We evaluated 140 patients who applied to a university hospital gastroenterology department and underwent biopsy during endoscopy. In these patients, we analysed floroquinolone and clarithromycin resistance using the GenoType® HelicoDR (Hain Life Science, Germany). We also used the real-time method for clarithromycin resistance. We found the number and rate of floroquinolone resistance as 20 (25.6%) and clarithromycine resistance as 31 (39.7%). With the real-time PCR method, we detected clarithromycine resistance in 26 (33.3%) patients. These results were not statistically significant. Discussion and Conclusion. Our results show similarity to the other studies held in our country. There should be more studies for the policy of eradication through our country.


Sensitive detection of rare disease-associated cell subsets via representation learning.
Rare cell populations play a pivotal role in the initiation and progression of diseases such as cancer. However, the identification of such subpopulations remains a difficult task. This work describes CellCnn, a representation learning approach to detect rare cell subsets associated with disease using high-dimensional single-cell measurements. Using CellCnn, we identify paracrine signalling-, AIDS onset- and rare CMV infection-associated cell subsets in peripheral blood, and extremely rare leukaemic blast populations in minimal residual disease-like situations with frequencies as low as 0.01%.


Squamous cell carcinoma of the hallux.
This article reports on a case of malignant degeneration of a hallux nail bed ulcer of 30 years' duration. Histologically, this lesion was determined to be a squamous cell carcinoma, a type of lesion that is also known as Marjolin's ulcer. The diagnosis, histologic findings, and treatment of patients with cutaneous squamous cell carcinoma are discussed.


Establishing the stability and reversibility of protein pyrophosphorylation with synthetic peptides.
Protein pyrophosphorylation is emerging as a new post-translational modification, yet its role in cellular signaling remains poorly characterized. Important factors in determining the biological relevance of pyrophosphorylation include understanding the chemical and biochemical stability of the pyrophosphoryl group and elucidating the reversibility of modification in a cellular context. Towards this end, we prepared a series of synthetic pyrophosphopeptides, which were utilized to demonstrate that the modification is quite inert over a wide pH range but can be removed biochemically by alkaline phosphatases. Importantly, we observed enzyme-dependent removal of the pyrophosphate in mammalian and yeast cell lysates using the synthetic pyrophosphopeptides. The findings provide evidence for the reversibility of pyrophosphorylation and thereby highlight the potential impact of this modification on cellular signal transduction pathways.


Fermentation characteristics of Fusariumoxysporum grown on acetate.
In this study, the growth characteristics of Fusariumoxysporum were evaluated in minimal medium using acetate or different mixtures of acetate and glucose as carbon source. The minimum inhibitory concentration (MIC) of acetic acid that F.oxysporum cells could tolerate was 0.8%w/v while glucose was consumed preferentially to acetate. The activity of isocitrate lyase was high when cells were grown on acetate and acetate plus glucose indicating an activation of the glyoxylate cycle. Investigation of the metabolic fingerprinting and footprinting revealed higher levels of intracellular and extracellular TCA cycle intermediates when F.oxysporum cells were grown on mixtures of acetate and glucose compared to growth on only glucose. Our data support the hypothesis that a higher flux through TCA cycle during acetate consumption could significantly increase the pool of NADH, resulting in the activation of succinate-propionate pathway which consumes reducing power (NADH) via conversion of succinate to propionyl-CoA and produce propionate.


Environmental endocrine disruptors in farm animal reproduction: research and reality.
In this review, possible comparative advantages of studying endocrine disruption in farm animals vs laboratory rodents are discussed. First, using farm animals, the generality of findings in laboratory rodents are challenged. Farm animals may in certain aspects be better models for humans than laboratory rodents, and sometimes there might be methodological advantages in using farm animals. Second, there are several in vitro studies based on cell-culture systems from sows and cows where the effects of chemicals on sex steroid secretion can be measured and maturation and fertilization of oocytes may be assessed. These in vitro systems are powerful tools for dissecting the mechanisms of action for endocrine-disrupting chemicals. Third, in a set of recent in vivo studies using sheep, goats and pigs, in which very different exposure regimens to endocrine-disrupting chemicals have been used, a full panel of reproductive parameters pertinent to farm animals were assessed. Clinically, it is suggested that endocrine disruption in farm animals should be considered when impaired reproduction could be linked to change in source of feed or pasture. Finally, epigenetic and toxicogenomic approaches can be particularly rewarding in elucidating endocrine disruption in future farm animal studies.


[Endocrine studies on the Prader-Labhart-Willi syndrome: puberty induction in a 19-year-old boy after long-term treatment with an LHRH analog].
In two girls (14 and 16 years) and one boy (19 years) with PLW-syndrome and pronounced obesity (240, 210 and 77% overweight) endocrine function tests were carried out. Growth hormone secretion was decreased but normalized after reduction of weight. Thyroxin levels as well as basal and TRH stimulated TSH concentrations were normal. HCG application in the boy induced no rise of the normal basal testosterone levels. Oral glucose tolerance test demonstrated an increased stimulation of insulin in two cases, no other symptoms of diabetes mellitus were found. In the LHRH test an insufficient rise of gonadotropins was found. However, after two weeks of pernasal application of an LHRH analogue (D-Leu6-des-Gly10-EA) the gonadotropin stimulation was distinctly improved and onset of puberty was induced in the male patient. These results are indicative of a hypothalamic disturbance in patients with PLW-syndrome.


Impacts of phototherapy on DNA damage and total oxidant/ antioxidant status in jaundiced newborns.
Bulut Ö, Dürüyen S. Impacts of phototherapy on DNA damage and total oxidant/antioxidant status in jaundiced newborns. Turk J Pediatr 2019; 61: 697-703. Jaundice is common in newborn babies, and pathologic hyperbilirubinemia is frequently treated with phototherapy. Although it is considered to be safe, the side effects of phototherapy in newborns are still a matter of debate. In this study, the impacts of two types of phototherapy, conventional fluorescence and intensive light-emitting diodes (LED), on DNA damage and total oxidant/ antioxidant status in jaundiced newborns were assessed. The study group included 40 newborns (gestation age ≥ 37 weeks) on days 2-8 after birth. Newborns were divided into two groups on the basis of need for phototherapy: 20 newborns were exposed to conventional phototherapy (Group I); and 20 infants were exposed to intensive phototherapy (Group II). Blood samples were taken from all infants at admission and after phototherapy to determine plasma 8-hydroxy-2-deoxy-guanosine (8-OH-dG; a marker of DNA damage), total oxidant status (TOS), and total antioxidant capacity (TAC). The oxidative stress index (OSI) was also calculated. Demographic information was recorded, and DNA damage, TOS, and TAC were compared. There were no differences in demographic information between the two groups. There were no significant differences in DNA damage, TOS, TAC, and OSI between Groups I and II before phototherapy (p > 0.05) and no significant differences in DNA damage, TOS, TAC, and OSI between the two groups after phototherapy (p > 0.05). However, the TAC decreased significantly in both groups after phototherapy (p < 0.01). These findings suggest that conventional and intensive phototherapies do not affect DNA damage and oxidative stress, supporting the safety of its use as the preferred treatment for jaundiced newborns.


Inductively coupled plasma mass spectrometric analysis of calcium isotopes in human serum: a low-sample-volume acid-equilibration method.
Analytical methods for measuring the calcium isotope distribution in enriched human serum samples that use low blood volumes, simple preparation methods, and rapid analysis are important in clinical studies of calcium kinetics. Previously, sample preparation by oxalate precipitation typically required 500 micro L of serum. This method was time-consuming, and the blood volume required was limiting in circumstances when only a small amount of serum could be obtained. Serum was collected from humans who were administered (42)Ca, and 20 micro L of serum was mixed with 2 mL of 0.22-0.67 mol/L HNO(3) at room temperature for between 1 min and 16 h. The (42)Ca/(43)Ca ratio in the supernatant was measured by a magnetic sector inductively coupled plasma mass spectrometer (ICP-MS). Calcium isotope ratios from these equilibration solutions were compared with data from oxalate-precipitated serum samples to determine the optimum equilibrium time and the effect of acid concentration on equilibrium. Various amounts of aggregated particles developed in different acid-serum mixtures. These affected the time required for isotope equilibration in the mixture. The shortest equilibrium time needed for the calcium isotopes varied from 1 to 6 h for samples acidified with 0.22-0.45 mol/L HNO(3). Data obtained from these solutions were consistent with data from oxalate-precipitated calcium. The precision of (42)Ca/(43)Ca ratio measurements was better than 0.5%. We have developed a simple, rapid sample preparation technique for ICP-MS analysis in which 20 micro L of serum can be used for accurate measurement of the calcium isotope distribution in a sample with good precision and a rapid analysis time.


Keystone-designed buried de-epithelialized flap: A novel technique for obliterating small to moderately sized dead spaces.
Effective obliteration of dead space after reconstructive surgery facilitates a good cosmetic outcome and prevention of delayed wound healing and recurrent infection.We evaluated the efficacy of a keystone-designed buried de-epithelialized (KBD) flap for the obliteration of small to moderately sized surgical dead spaces.We reviewed the medical records of patients who received a KBD flap following removal of a mass or debridement of necrotic tissue from September 2015 to February 2016. The diagnosis, site, dead space dimensions, flap width, drain data, complications, and follow-up duration were recorded.Twenty-eight KBD flaps were evaluated, including 9 cases of fat necrosis, 7 cases of epidermal cyst, and 12 cases of lipoma. Dead space dimensions ranged from 2 × 1.5 × 1 cm to 10 × 5 × 3 cm, with a mean depth of 2.01 cm. Flap sizes ranged from 2.5 × 1 cm to 11 × 3 cm, with a mean flap width of 2.01 cm. No postoperative complications, such as seroma or hematoma, occurred. The cosmetic results were favorable, and all patients were satisfied with their final outcomes.The KBD flap is useful for the obliteration of small to moderately sized surgical dead spaces both spatially and physiologically and shows excellent cosmetic outcomes.