The N-butylcarbamate derivative of galantamine acts as an allosteric potentiating ligand on alpha7 nicotinic receptors in hippocampal neurons: clinical implications for treatment of Alzheimer's disease.
Alzheimer's disease (AD) is a progressive neurodegenerative disorder characterized by cognitive impairments that become severe enough to interfere with the daily activities of patients and eventually lead to death (Chung and Cummings, 2000). Arecent study reports that approx 24 million people suffer from dementia worldwide. If the mortality rate does not change and no curative or preventive treatment is developed, this number is expected to double every 20 yr worldwide (Ferri et al., 2005). Although the causes of AD remain obscure, it has been reported that incremental loss of cholinergic neurons and of nicotinic receptor (nAChR) function/expression in specific brain regions correlates well with the severity of the symptoms at early stages of the disease (Hellström-Lindahl et al., 1999; Nordberg, 2001; Perry et al., 2001; Wevers et al., 1999). In patients with more advanced stages of AD, such a correlation between the magnitude of nAChR loss and of cognitive decline does not appear to exist (Sabbagh et al., 2001). The nicotinic cholinergic system plays a central role in modulating different forms of associative learning known to be impaired in AD patients, including the eyeblink classical conditioning (Woodruff-Pak, 2001), and in maintaining neuronal viability. Neuroprotection and cognitive improvement result from increasing the activity of different nAChR subtypes, including those bearing the alpha7 subunit (Carlson et al., 1998; Hejmadi et al., 2003; Kihara et al., 1997; Levin et al., 2006). Thus, increasing nAChR activity in the brain was proposed as a mechanism to slow down the progression of the disease (Maelicke and Albuquerque, 1996).


Hand, foot, and mouth disease: current scenario and Indian perspective.
Hand, foot, and mouth disease (HFMD), first reported in New Zealand in 1957 is caused by Coxsackievirus A16 (CVA16) and human enterovirus 71 (HEV71) and occasionally by Coxsackievirus A4-A7, A9, A10, B1-B3, and B5. This is characterized by erythematous papulo vesicular eruptions over hand, feet, perioral area, knees, buttocks and also intraorally mostly in the children. HFMD has been known for its self limiting course. Only small scale outbreaks have been reported from United States, Europe, Australia, Japan and Brazil for the first few decades. However, since 1997 the disease has conspicuously changed its behavior as noted in different Southeast Asian countries. There was sharp rise in incidence, severity, complications and even fatal outcomes that were almost unseen before that period. Following the near complete eradication of poliovirus, HEV71, the non-polio enterovirus, may become the greatest threat to cause significant neurological complications. This adds to the fact that effective therapy or vaccine is still a far reaching goal. There are reports of disease activity in different corners of India since 2004. Although of milder degree, continuous progress to affect larger parts of the country may indicate vulnerability of India from possible future fatal outbreaks. Low level of awareness among the health care providers may prove critical.


Minimizing E-factor in the continuous-flow synthesis of diazepam and atropine.
Minimizing the waste stream associated with the synthesis of active pharmaceutical ingredients (APIs) and commodity chemicals is of high interest within the chemical industry from an economic and environmental perspective. In exploring solutions to this area, we herein report a highly optimized and environmentally conscious continuous-flow synthesis of two APIs identified as essential medicines by the World Health Organization, namely diazepam and atropine. Notably, these approaches significantly reduced the E-factor of previously published routes through the combination of continuous-flow chemistry techniques, computational calculations and solvent minimization. The E-factor associated with the synthesis of atropine was reduced by 94-fold (about two orders of magnitude), from 2245 to 24, while the E-factor for the synthesis of diazepam was reduced by 4-fold, from 36 to 9.


Cerebrospinal fluid biomarkers in clinically isolated syndromes and multiple sclerosis.
A panel of three cerebrospinal fluid (CSF) markers for clinically isolated syndromes (CIS) and multiple sclerosis (MS), based on SDS-PAGE, 2-D maps, and immunoblot results, is here proposed. No individual marker has any specificity, though, since they appear in a number of other neurological diseases. However the set of three, with the respective modulation sign (up-regulated or maintained at constant level), appears to be unique for MS. These proteins are: tau protein (levels remaining constant and undistinguishable from controls, contrary to up- and downregulation in other neurological disorders); 14-3-3 protein (strong upregulation of distinct isoforms) and cystatin C (changing in accordance to disease stage and progression). As an additional evidence, one can rely in the pattern of isoforms of 14-3-3, as obtained by 2-D maps and Western blot analysis: this pattern further distinguishes the variation of this protein from other neurological syndromes, notably sporadic Creutzfeldt-Jakob disease (sCJD), motor neuron diseases and other dementias. In contrast, a similar qualitative and quantitative upregulation of 14-3-3 is observed in Guillain-Barré syndrome (GBS), a demyelinating condition affecting the peripheral nervous system. To the best of our knowledge, this is the first time in which such a panel of biomarkers is reported in MS.


A second allosteric site in Escherichia coli aspartate transcarbamoylase.
Escherichia coli aspartate transcarbamoylase is feedback inhibited by CTP and UTP in the presence of CTP. Here, we show by X-ray crystallography that UTP binds to a unique site on each regulatory chain of the enzyme that is near but not overlapping with the known CTP site. These results bring into question all of the previously proposed mechanisms of allosteric regulation in aspartate transcarbamoylase.


Conformational dynamics of p53 N-terminal TAD2 region under different solvent conditions.
Although the mystery molecule p53 has been studied extensively, still several unknown mechanisms need to be elucidated. Being a central hub of cellular signaling pathways, the function of p53 is precisely executed synergistically by its intrinsically disordered and structural domains. The disordered N-terminal region further modulates p53 function by undergoing promiscuous binding and folding with several partners with the help of TAD1 and TAD2 motifs. Among these regions, a significant contribution is made by TAD2 in terms of binding affinities. This heterogeneity in p53 TAD region motivates to employ a reductionist approach to understand the folding behavior of TAD2 region independently under a broad range of different pH, temperature and solvent conditions. Since the intracellular environment is complex and crowded with a variety of molecules providing different type of surfaces from polar to hydrophobic, it is mandatory to characterize the conformational heterogeneity of disordered proteins to completely understand their function. Different types of alcohols were used to estimate the structure forming capabilities of the TAD2 peptides using circular dichroism, fluorescence and lifetime spectroscopy. The alcohols ethanol, TFE and HFIP were previously known to induce increasing levels of hydrophobic environments in water-alcohol mixtures respectively. Our results have shown that TAD2 peptide undergoes a dehydration dependent induction of hydrophobic interactions leading towards structural transitions in presence of organic solvents. This study is highlighting the importance of hydrophobic surfaces playing a crucial role in TAD2 interaction and conformational transitions.


Management of the early stage of Alzheimer's disease: a systematic review of literature over the past 10 years.
Alzheimer's disease is the most widespread senile dementia in the world and diagnoses are on the rise due to the aging of the world's population and this is causing a significant increase in health spending on the care of the disease. Current studies highlight the need for rapid diagnosis in order to prevent the therapeutic approach from being ineffective, and a comprehensive management of the disease, both with pharmacological and integrative therapies (safety of the environment, music therapy, diet, physical activity, recreational activities, etc.). Regarding the care provided by caregivers, the need to educate these figures is fundamental, to support them economically and psycho-socially (especially during the most difficult stages of the disease) and to provide them with technical skills to effectively and safely address the patient's care. Aim of this study was to investigate the current evidence from scientific literature about the early stage of Alzheimer's disease in order to educate and support caregivers with specific technical and practical skills. A systematic review of the scientific literature of the last 10 years, has been conducted in September 2019 through Medline (PubMed), Scopus and Cinahl databases, in accordance with the criteria of PRISMA Statement guideline (Preferred Reporting Items for Systematic reviews and Meta-Analyses). The included studies were related to Alzheimer's disease and patients' management. Thirty-six articles met the inclusion criteria but only for 26 of these it was possible to retrieve and access the full text. The evidences from this study provide information on how to effectively address Alzheimer's disease. The studies address different aspects of the disease, with different methodologies, thus providing heterogeneous evidence. Future studies with reproducible methodologies should be undertaken in order to achieve robustness in results.


Factors associated with glucose metabolism disorders after kidney transplantation.
Post-transplant diabetes mellitus (PTDM), pre-diabetes-impaired glucose tolerance (IGT) and impaired fasting glucose (IFG) are frequent complications after organ transplantation. The aim of this study was to assess the frequency of PTDM, IFG and IGT in a group of renal transplant recipients, to compare the frequency of glucose metabolism disorders in subjects treated with tacrolimus and with cyclosporine, and to establish the influence of different risk factors on the development of glucose metabolism disorders. We examined 206 non-diabetic kidney allograft recipients (age 46.4 ± 12.3 years, time since transplantation 45.5 ± ± 33.6 months, BMI 26.3 ± 4.5 kg/m2). Glucose metabolism disorders were diagnosed using an oral glucose tolerance test. Logistic regression was used to assess the influence of each risk factor (age, BMI, waist circumference, physical activity, the presence of cardiovascular disease, positive family history of diabetes, cholesterol and triglycerides concentration) on the development of glucose metabolism disorders. In 103 patients (50%), we diagnosed glucose metabolism disorders. 19% of patients had PTDM, 14% IFG, and 17% IGT. We did not find any differences in the frequency of glucose metabolism disorders between patients treated with tacrolimus and with cyclosporine. Multivariate analysis identified BMI and a family history of diabetes as independent risk factors of glucose metabolism disorders. We found a high prevalence of glucose metabolism disorders in the examined group. This suggests that kidney transplant recipients should be screened for these disturbances. Patients with higher BMI and with first-degree relatives with diabetes had an increased risk of glucose metabolism disorders after kidney transplantation.


HDNetDB: A Molecular Interaction Database for Network-Oriented Investigations into Huntington's Disease.
Huntington's disease (HD) is a progressive and fatal neurodegenerative disorder caused by an expanded CAG repeat in the huntingtin gene. Although HD is monogenic, its molecular manifestation appears highly complex and involves multiple cellular processes. The recent application of high throughput platforms such as microarrays and mass-spectrometry has indicated multiple pathogenic routes. The massive data generated by these techniques together with the complexity of the pathogenesis, however, pose considerable challenges to researchers. Network-based methods can provide valuable tools to consolidate newly generated data with existing knowledge, and to decipher the interwoven molecular mechanisms underlying HD. To facilitate research on HD in a network-oriented manner, we have developed HDNetDB, a database that integrates molecular interactions with many HD-relevant datasets. It allows users to obtain, visualize and prioritize molecular interaction networks using HD-relevant gene expression, phenotypic and other types of data obtained from human samples or model organisms. We illustrated several HDNetDB functionalities through a case study and identified proteins that constitute potential cross-talk between HD and the unfolded protein response (UPR). HDNetDB is publicly accessible at http://hdnetdb.sysbiolab.eu .


Prevention, treatment and outcomes of bacterial meningitis in childhood.
Meningitis is more likely to occur in childhood that at any other age. Preventive measures, in the form of vaccination programmes targeted at certain at-risk age groups, have drastically reduced the incidence in recent years. Early identification of symptoms remains vital in order to prevent mortality and reduce the likelihood of long-term effects.


Determination of Ten Corticosteroids in Illegal Cosmetic Products by a Simple, Rapid, and High-Performance LC-MS/MS Method.
The aim of our present work was the development of a rapid high-performance liquid chromatography method with electrospray ionization and tandem mass spectrometry detection (LC-ESI-MS/MS) for the determination of several corticosteroids in cosmetic products. Corticosteroids are suspected to be illegally added in cosmetic preparations in order to enhance the curative effect against some skin diseases. Sample preparation step consists in a single extraction with acetonitrile followed by centrifugation and filtration. The compounds were separated by reversed-phase chromatography with water and acetonitrile (both with 0.1% formic acid) gradient elution and detected by ESI-MS positive and negative ionization mode. The method was validated at the validation level of 0.1 mg kg-1. Linearity was studied in the 5-250 μg L-1 range and linear coefficients (r2) were all over 0.99. The accuracy and precision of the method were satisfactory. The LOD ranged from 0.085 to 0.109 mg kg-1 and the LOQ from 0.102 to 0.121 mg kg-1. Mean recoveries for all the analytes were within the range 91.9-99.2%. The developed method is sensitive and useful for detection, quantification, and confirmation of these corticosteroids in cosmetic preparations and can be applied in the analysis of the suspected samples under investigation.


Calcium and vitamin D have a synergistic role in a rat model of kidney stone disease.
Vitamin D supplementation in humans should be accompanied by calcium administration to avoid bone demineralization through vitamin D receptor signaling. Here we analyzed whether long-term exposure of rats to vitamin D supplementation, with or without a calcium-rich diet, would promote kidney stone formation. Four groups of rats received vitamin D alone (100,000 UI/kg/3 weeks), a calcium-enriched diet alone, both vitamin D supplementation and calcium-enriched diet, or a standard diet (controls) for 6 months. Serum and urine parameters and crystalluria were monitored. Kidney stones were assessed by 3-dimensional micro-computed tomography, infrared spectroscopy, von Kossa/Yasue staining, and field emission scanning electron microscopy. Although serum calcium levels were similar in the 4 groups, rats receiving vitamin D had a progressive increase in urinary calcium excretion over time, especially those receiving both calcium and vitamin D. However, oral calcium supplementation alone did not increase urinary calcium excretion. At 6 months, rats exposed to both calcium and vitamin D, but not rats exposed to calcium or vitamin D alone, developed significant apatite kidney calcifications (mean volume, 0.121 mm(3)). Thus, coadministration of vitamin D and increased calcium intake had a synergistic role in tubular calcifications or kidney stone formation in this rat model. Hence, one should be cautious about the cumulative risk of kidney stone formation in humans when exposed to both vitamin D supplementation and high calcium intake.


Nitrotyrosine and nitrate/nitrite levels in cardiac arrest survivors treated with endovascular hypothermia.
The protective effect of therapeutic hypothermia in cardiac arrest survivors (CAS) has been previously well documented. Animal studies have indicated that attenuation of tissue oxidative stress (OS) may be involved in the mechanisms that lead to the beneficial effect of hypothermia. The extent of OS and nitric oxide (NO) production in adult CAS treated with endovascular hypothermia is, however, unknown. A total of 11 adult patients who experienced cardiac arrest out of hospital were included in the present study, and all were treated with mild hypothermia using the Thermogard XP (Alsius, USA) endovascular system. A target core temperature of 33 °C was maintained for 24 hours, with a subsequent rewarming rate of 0.15 °C per hour, followed by normothermia at 36.8 °C. Blood samples for the measurement of nitrotyrosine and nitrate/nitrite levels were drawn at admission and every 6 hours thereafter for two days. During the hypothermic period, the levels of nitrotyrosine and nitrates/nitrites were comparable with baseline values. During the rewarming period, serum levels of both parameters gradually increased and, during the normothermic period, the levels were significantly higher compared with hypothermic levels (nitrotyrosine, P<0.001; nitrates/nitrites, P<0.05). In our study, significantly lower levels of nitrotyrosine and nitrates/nitrites were demonstrated during hypothermia compared with levels during the normothermic period in adult CAS. These data suggest that attenuation of OS and NO production may be involved in the protective effect of hypothermia in adult CAS.


Intranasal immunisation with recombinant Toxoplasma gondii actin partly protects mice against toxoplasmosis.
Toxoplasma gondii is a ubiquitous protozoan intracellular parasite, the causative agent of toxoplasmosis, and a worldwide zoonosis for which an effective vaccine is needed. Actin is a highly conserved microfilament protein that plays an important role in the invasion of host cells by T. gondii. This study investigated the immune responses elicited by BALB/c mice after nasal immunisation with a recombinant T. gondii actin (rTgACT) and the subsequent protection against chronic and lethal T. gondii infections. We evaluated the systemic response by proliferation, cytokine and antibody measurements, and we assessed the mucosal response by examining the levels of TgACT-specific secretory IgA (SIgA) in nasal, vaginal and intestinal washes. Parasite load was assessed in the liver and brain, and the survival of mice challenged with a virulent strain was determined. The results showed that the mice immunised with rTgACT developed high levels of specific anti-rTgACT IgG titres and a mixed IgG1/IgG2a response with a predominance of IgG2a. The systemic immune response was associated with increased production of Th1 (IFN-γ and IL-2), Th2 (IL-4) and Treg (IL-10) cytokines, indicating that not only Th1-type response was induced, but also Th2- and Treg-types responses were induced, and the splenocyte stimulation index (SI) was increased in the mice immunised with rTgACT. Nasal immunisation with rTgACT led to strong mucosal immune responses, as seen by the increased secretion of SIgA in nasal, vaginal and intestinal washes. The vaccinated mice displayed significant protection against lethal infection with the virulent RH strain (survival increased by 50%), while the mice chronically infected with RH exhibited lower liver and brain parasite loads (60.05% and 49.75%, respectively) than the controls. Our data demonstrate, for the first time, that actin triggers a strong systemic and mucosal response against T. gondii. Therefore, actin may be a promising vaccine candidate against toxoplasmosis.


Quality of life among people with epilepsy: a cross-sectional study from rural southern India.
Epilepsy can be associated with profound physical, social and psychological consequences and it has an impact on a person's quality of life. We assessed the quality of life and factors associated with a poor quality of life, among adults with epilepsy in a rural block of Tamil Nadu. We interviewed 91 epilepsy patients from 20 randomly selected villages using a structured questionnaire including World Health Organization Quality of Life BREF (WHOQOL-BREF), Patient Health Questionnaire 2 (PHQ-2) and Generalized Anxiety Disorder 7 (GAD-7) questionnaires. The mean (SD) total score of the quality of life scale was 61.49 (12.56). Those who were single, separated or widowed (t statistic= -2.71, p<0.01), had not completed primary education (t statistic=-2.308, p<0.05), not currently going for work (t statistic=-2.748, p<0.01), had seizure in the past one year (t statistic=-4.068, p<0.01),had depressive symptoms (t statistic=-3.207, p<0.01),had higher anxiety scores (t statistic=-2.727, p<0.01), had low scores in the quality of life questionnaire. Multivariate analysis showed increasing age, education less than grade V, being unmarried, widowed or separated, lower per capita income, a high anxiety score and experiencing a seizure episode in the past one year to be significantly associated with a low score in the WHOQOL-BREF questionnaire (adjusted R²=0.378, SE 9.90). The presence of anxiety, lack of primary education, being single, separated or widowed, increasing age, low per capita income and having a seizure episode in the past year are associated with lower quality of life among people with epilepsy.


Astrocytic GABA receptors.
GABA receptors are distributed widely throughout the central nervous system on a variety of cell types. It has become increasingly clear that astrocytes, both in cell culture and tissue slices, express abundant GABAA receptors. In astrocytes, GABA activates Cl(-)-specific channels that are modulated by barbiturates and benzodiazepines; however, the neuronal inverse agonist methyl-4-ethyl-6, 7-dimethoxy-beta-carboline-3-carboxylate enhances the current in a subpopulation of astrocytes. The properties of astrocytic GABAA receptors, therefore, are remarkably similar to their neuronal counterparts, with only a few pharmacological exceptions. In stellate glial cells of the pituitary pars intermedia, GABA released from neuronal terminals activates postsynaptic potentials directly. The physiological significance of astrocytic GABAA-receptor activation remains unknown, but it may be involved in extracellular ion homeostasis and pH regulation. At present, there is considerably less evidence for the presence of GABAB receptors on astrocytes. The data that have emerged, however, indicate a prominent role for second-messenger regulation by this receptor.


Awareness about pulse polio immunization among the general population in Delhi.
A study was conducted in three intensive pulse polio immunization (IPPI) centres in Delhi on 24th September, 2000 to assess the awareness about poliomyelitis and the IPPI programme. A structured questionnaire was used to elicit the information from 182 accompanying persons (41 males and 141 females) by qualified medical doctors. Majority (92.9%) of the respondents knew that the administered drops were polio vaccine. Major sources of information were television (32.9%), relatives or friends (24.2%), health staff (20.9%), poster or leaflets (15.9%). Only 18.1% knew the next IPPI day. Out of 43 claiming to know the number of subsequent IPPI days during 2000, only 22 could tell correctly. A large number of respondents (86.2%) knew that IPPI would help in preventing polio in children. Paralysis of limbs as a clinical feature of polio was known to 70.3%, 45.6% knew that polio cannot be cured and 21.9% perceived that polio could also lead to death. The nationwide intensive awareness campaign for polio eradication was found to be partially effective in disseminating the information.


The relationship of minority stress and flexible coping to psychological well being in lesbian and bisexual women.
This study explores minority stress among lesbian and bisexual women by examining the relationship of sexist and heterosexist events, self-concealment, and self-monitoring to positive psychological well being. The sample was made up of 373 lesbian and bisexual women, including 77 women of color. Results of a multiple regression analysis of the total sample indicated that, considered simultaneously, self-concealment and self-monitoring explained significant variance in positive psychological well being, while sexist and heterosexist events, though present, were not significantly related to positive psychological well being. Results were similar for the women of color alone. This research provides evidence for the resiliency of lesbian and bisexual women and offers some applicable concepts (e.g., self-concealment and self-monitoring) to consider in research and practice with lesbian and bisexual female clients.


Effects of a brief mindfulness-meditation intervention on neural measures of response inhibition in cigarette smokers.
Research suggests that mindfulness-practices may aid smoking cessation. Yet, the neural mechanisms underlying the effects of mindfulness-practices on smoking are unclear. Response inhibition is a main deficit in addiction, is associated with relapse, and could therefore be a candidate target for mindfulness-based practices. The current study hence investigated the effects of a brief mindfulness-practice on response inhibition in smokers using behavioral and electroencephalography (EEG) measures. Fifty participants (33 females, mean age 20 years old) underwent a protocol of cigarette exposure to induce craving (cue-exposure) and were then randomly assigned to a group receiving mindfulness-instructions or control-instructions (for 15 minutes approximately). Immediately after this, they performed a smoking Go/NoGo task, while their brain activity was recorded. At the behavioral level, no group differences were observed. However, EEG analyses revealed a decrease in P3 amplitude during NoGo vs. Go trials in the mindfulness versus control group. The lower P3 amplitude might indicate less-effortful response inhibition after the mindfulness-practice, and suggest that enhanced response inhibition underlies observed positive effects of mindfulness on smoking behavior.


Shear stress regulates the endothelial Kir2.1 ion channel.
Endothelial cells (ECs) line the mammalian vascular system and respond to the hemodynamic stimulus of fluid shear stress, the frictional force produced by blood flow. When ECs are exposed to shear stress, one of the fastest responses is an increase of K(+) conductance, which suggests that ion channels are involved in the early shear stress response. Here we show that an applied shear stress induces a K(+) ion current in cells expressing the endothelial Kir2.1 channel. This ion current shares the properties of the shear-induced current found in ECs. In addition, the shear current induction can be specifically prevented by tyrosine kinase inhibition. Our findings identify the Kir2.1 channel as an early component of the endothelial shear response mechanism.


The Feasibility of a Fast Liver MRI Protocol for Lesion Detection of Adults at 3.0-T.
To investigate the feasibility of a fast liver magnetic resonance imaging (MRI) protocol for lesion detection in adults using 3.0-T MRI. A fast liver MRI exam protocol was proposed. The protocol included motion-resistant coronal T2-w sequence, axial T2-w fast spin echo sequence with fat suppression, axial in-op phase gradient recalled echo (GRE) T1, axial diffusion weighted imaging (DWI), and axial contrast-enhanced T1 sequences. To evaluate the diagnostic capacity of the proposed protocol, 31 consecutive patients (20 males and 11 females; mean age, 53.2 years) underwent a liver MRI exam with conventional sequences, including the proposed protocol as a subset. Images from the conventional protocol and extracted abbreviated protocol were independently read, and the diagnostic concordance rate was assessed for each patient. The concordance analysis is presented as the proportion of concordant cases between the two protocols. The net measurement time of the fast liver MRI protocol without adjustment and waiting time were 4 min and 28 s. In the 31 patients included in this study, 139 suspicious findings were found from both the conventional liver MR protocol and the fast liver MRI protocol. The diagnostic concordance rate was 96.4%. The fast liver MRI protocol is feasible at 3.0-T, with a shorter exam time and high diagnostic concordance compared to the conventional liver MRI workflow.


Inhibition of cyclin dependent kinase 9 by dinaciclib suppresses cyclin B1 expression and tumor growth in triple negative breast cancer.
Cyclin-dependent kinases (CDKs) are potential cancer therapeutic targets because of their critical role in promoting cell growth. Dinaciclib is a novel CDK inhibitor currently under clinical evaluation for the treatment of advanced malignancies. In this study, we demonstrated the anti-tumor activity of dinaciclib in triple negative breast cancer (TNBC) patient derived xenograft (PDX) and cell lines in vitro and in vivo. Treatment with dinaciclib induced cell cycle arrest at G2/M phase and marked apoptosis. These changes were accompanied by reduced phosphorylation of CDK1 and retinoblastoma (Rb) protein and decreased protein levels of cyclin B1, cMYC and survivin. We further demonstrated that siRNA knockdown of CDK9, the kinase subunit of positive transcription elongation factor b (P-TEFb), instead of CDK1 or CDK2, reduced the levels of cyclin B1 and MYC in TNBC cell lines. These data support the importance of CDK9, in addition to CDK1, in mediating the growth inhibitory effect of dinaciclib in TNBC. Further investigation of CDK9 as a therapeutic target in TNBC is needed.


Validation of the prognostic value of NF-κB p65 in prostate cancer: A retrospective study using a large multi-institutional cohort of the Canadian Prostate Cancer Biomarker Network.
The identification of patients with high-risk prostate cancer (PC) is a major challenge for clinicians, and the improvement of current prognostic parameters is an unmet clinical need. We and others have identified an association between the nuclear localization of NF-κB p65 and biochemical recurrence (BCR) in PC in small and/or single-centre cohorts of patients. In this study, we accessed 2 different multi-centre tissue microarrays (TMAs) representing cohorts of patients (Test-TMA and Validation-TMA series) of the Canadian Prostate Cancer Biomarker Network (CPCBN) to validate the association between p65 nuclear frequency and PC outcomes. Immunohistochemical staining of p65 was performed on the Test-TMA and Validation-TMA series, which include PC tissues from patients treated by first-line radical prostatectomy (n = 250 and n = 1,262, respectively). Two independent observers evaluated the p65 nuclear frequency in digital images of cancer tissue and benign adjacent gland tissue. Kaplan-Meier curves coupled with a log-rank test and univariate and multivariate Cox regression models were used for statistical analyses of continuous values and dichotomized data (cutoff of 3%). Multivariate analysis of the Validation-TMA cohort showed that p65 nuclear frequency in cancer cells was an independent predictor of BCR using continuous (hazard ratio [HR] 1.02 [95% CI 1.00-1.03], p = 0.004) and dichotomized data (HR 1.33 [95% CI 1.09-1.62], p = 0.005). Using a cutoff of 3%, we found that this biomarker was also associated with the development of bone metastases (HR 1.82 [95% CI 1.05-3.16], p = 0.033) and PC-specific mortality (HR 2.63 [95% CI 1.30-5.31], p = 0.004), independent of clinical parameters. BCR-free survival, bone-metastasis-free survival, and PC-specific survival were shorter for patients with higher p65 nuclear frequency (p < 0.005). As the small cores on TMAs are a limitation of the study, a backward validation of whole PC tissue section will be necessary for the implementation of p65 nuclear frequency as a PC biomarker in the clinical workflow. We report the first study using the pan-Canadian multi-centre cohorts of CPCBN and validate the association between increased frequency of nuclear p65 frequency and a risk of disease progression.


[Progressive myelopathy caused by human T-cell lymphotropic virus type I (HTLV-I) in 3 patients in The Netherlands].
One man and two women (aged 30, 44 and 46, respectively) were seen between 1987 and 1992 with a myelopathy caused by the human T-cell lymphotropic virus type I (HTLV-I). The first symptoms were impaired gait in the man and micturition disorders in the women. Diagnosing took 2 to 4 years, possibly due to the fact that many Dutch physicians are unfamiliar with the disease. The diagnosis was based on originating from an endemic area (i.e. Surinam and the Caribbean basin), the clinical picture and the presence of antibodies against HTLV-I in blood and CSF. The disease in a number of years leads to spastic paraparesis, incontinence for urine and dependence on a wheelchair.


[Treatment of acute lung injury due to sea water aspiration in dogs by early massive bronchoalveolar lavage: an experimental study].
To investigate the therapeutic effects of massive bronchoalveolar lavage at early stage of acute lung injury induced by sea water aspiration. Twelve mongrel dogs were subjected to sea water infusion of the entire lungs and were subsequently randomized into 2 groups. The dogs in group A received intravenous saline transfusion 24 h after injury, while those in group B underwent bronchoscopic lavage of injured lungs with 450 ml 0.45% saline 15 min after injury. Unassisted breathing was maintained in both groups, and the gas exchange function of the bilateral lungs and the levels of lactate dehydrogenase, alkaline phosphatase in the bronchoalveolar fluid were observed, with the hemodynamics indices monitored continuously. The lungs of the dogs were finally removed for morphological study under both light and electron microscopes, and the water content and wet- to dry-weight ratio of the lung tissues were determined. A total lavage volume of approximately 30 ml/kg x b.w. was used, and the liquid volume remaining in the lungs after operation ranged from 80 to 120 ml. All the dogs manifested significant declination in pulmonary function after the drowning. PaO2 in group B decreased significantly soon after the lavage but then increased gradually to a higher levels than group A. The dogs in group B had smaller lung water content and lower levels of lactate dehydrogenase and alkaline phosphatase in the bronchoalveolar fluid, showing improved oxygenation in comparison with group A. The inflammatory reaction and edema in lungs were also markedly alleviated in group B. Treatment with massive lung lavage in early stage after sea water aspiration can effectively protect the lung tissues from damage and alleviate the inflammatory reaction and pulmonary edema, helping to improve the lung function.


Andrographolide inhibits infectious bronchitis virus-induced apoptosis, pyroptosis, and inflammation.
Avian infectious bronchitis virus (IBV), a coronavirus, causes a huge economic loss to the poultry industry. Andrographolide (APL) is a compound with a variety of pharmacological properties, including antiviral and anti-inflammatory effects. In this study, APL was evaluated for antiviral activity by its anti-apoptotic, anti-pyroptosis, and anti-inflammatory effects. The cytotoxicity of APL was determined by the MTT method. We investigated the therapeutic impact of APL on IBV through a plate assay. We explored that APL inhibited IBV-induced apoptosis, pyroptosis, and inflammation in HD11 cells by RT-qPCR and immunofluorescence. Also, it was verified in the clinical chicken embryo trial. We found that APL down-regulated apoptosis-related genes Caspase-3, Caspase-8, Caspase-9, Bax, Bid, and Bak, down-regulated pyroptosis gene DFNA5, and down-regulated inflammation-related genes (NF-κB, NLRP3, iNOS, TNF-α, and IL-1β). In addition, APL reduced the reactive oxygen species (ROS) production in cells. Finally, clinical trials showed that APL inhibited IBV-induced apoptosis, pyroptosis, and inflammation, as well as reduced the mortality and malformation of chicken embryos. In this study, we delved into the antiviral properties of APL in the context of chicken macrophage (HD11) infection with IBV. Our findings confirm that andrographolide effectively inhibits apoptosis, pyroptosis, and inflammation by IBV infection. Furthermore, this inhibition was verified on chicken embryos in vivo. This inhibition suggests a substantial potential for APL as a therapeutic agent to mitigate the harmful effects of IBV on host cells.


Development of a rheological analogue to periprosthetic fluid.
The purpose of this work is to design a rheological analogue to periprosthetic fluid, for potential use in wear testing of orthopaedic implants. Polymer solutions of sodium carboxymethyl cellulose (CMC), xanthan and mixtures thereof were prepared, and the experimentally determined viscosity-shear rate dependence was analyzed using the three-parameter modified Cross model. A mixture containing 0.185 wt% CMC and 0.075 wt% xanthan was identified as being the closest rheological match for periprosthetic fluid in both steady shear and oscillatory modes of deformation.


[Surgical treatment of lymphatic malformations].
Lymphatic malformations remain a therapeutic challenge. Many treatments by the past led to poor success. The wide variety of clinical presentations makes it difficult to outline specific management programs. Often, diagnosis or complication circumstances as infection, bleeding, airway obstruction, or handicap force acting. Thus, a specific follow-up joined to a multidisciplinary analysis are the key of well-planned surgery. Through our experience and a review of literature, we will describe principles of lymphatic malformations surgery and specific technics for specific locations.


Arabidopsis mTERF15 is required for mitochondrial nad2 intron 3 splicing and functional complex I activity.
Mitochondria play a pivotal role in most eukaryotic cells, as they are responsible for the generation of energy and diverse metabolic intermediates for many cellular events. During endosymbiosis, approximately 99% of the genes encoded by the mitochondrial genome were transferred into the host nucleus, and mitochondria import more than 1000 nuclear-encoded proteins from the cytosol to maintain structural integrity and fundamental functions, including DNA replication, mRNA transcription and RNA metabolism of dozens of mitochondrial genes. In metazoans, a family of nuclear-encoded proteins called the mitochondrial transcription termination factors (mTERFs) regulates mitochondrial transcription, including transcriptional termination and initiation, via their DNA-binding activities, and the dysfunction of individual mTERF members causes severe developmental defects. Arabidopsis thaliana and Oryza sativa contain 35 and 48 mTERFs, respectively, but the biological functions of only a few of these proteins have been explored. Here, we investigated the biological role and molecular mechanism of Arabidopsis mTERF15 in plant organelle metabolism using molecular genetics, cytological and biochemical approaches. The null homozygous T-DNA mutant of mTERF15, mterf15, was found to result in substantial retardation of both vegetative and reproductive development, which was fully complemented by the wild-type genomic sequence. Surprisingly, mitochondria-localized mTERF15 lacks obvious DNA-binding activity but processes mitochondrial nad2 intron 3 splicing through its RNA-binding ability. Impairment of this splicing event not only disrupted mitochondrial structure but also abolished the activity of mitochondrial respiratory chain complex I. These effects are in agreement with the severe phenotype of the mterf15 homozygous mutant. Our study suggests that Arabidopsis mTERF15 functions as a splicing factor for nad2 intron 3 splicing in mitochondria, which is essential for normal plant growth and development.


Analysis of Occupational Safety and Health Administration (OSHA) noise standard violations over 50 years: 1972 to 2019.
Noise exposure has long been an occupational health concern and has been an important area of focus of the Occupational Safety and Health Administration (OSHA) since its founding. Nevertheless, it remains unclear what effects OSHA's noise standards have had on employer efforts to reduce risks. Consequently, a review of OSHA noise standard violations was performed to clarify the violation trends between 1972 and 2019. Using the OSHA Information System, researchers identified 119 305 violations involving four noise standards between 1972 and 2019: 29 CFR 1910.95, occupational noise exposure in general industry; 1926.52, occupational noise exposure in construction; 1926.101, hearing protection in construction, and 1904.10, recording criteria for cases involving occupational hearing loss. Violation frequencies of noise standard subparagraphs and relationships to factors such as industry differences were analyzed using descriptive statistics and t tests. The most commonly violated noise standard was 1910.95 in manufacturing. Such violations rose between 1972 and 1985 and then declined steadily. Whether in general industry or construction, four noise standards were most-frequently cited: lack of feasible administrative or engineering controls (1910.95[b] and 1926.52[d]) and inadequate hearing conservation program (1910.95[c] and 1926.52[b]). These violations were more highly penalized (mean = $1036.50) than other subparagraph violations (mean = $915.80). Programmed and unprogrammed inspections generated similar violation quantities except between 1980 and 1985, when programmed inspections exhibited a sharp spike in violations. The study identified trends in OSHA noise standard violations and possible explanations for those trends. The study findings can support development of more practical noise-exposure protection policy.


Dark-rearing uncovers novel gene expression patterns in an obligate cave-dwelling fish.
Extreme environments often result in the evolution of dramatic adaptive features. The Mexican tetra, Astyanax mexicanus, includes 30 different populations of cave-dwelling forms that live in perpetual darkness. As a consequence, many populations have evolved eye loss, reduced pigmentation, and amplification of nonvisual sensory systems. Closely-related surface-dwelling morphs demonstrate typical vision, pigmentation, and sensation. Transcriptomic assessments in this system have revealed important developmental changes associated with the cave morph, however, they have not accounted for photic rearing conditions. Prior studies reared individuals under a 12:12 hr light/dark (LD) cycle. Here, we reared cavefish under constant darkness (DD) for 5+ years. From these experimental individuals, we performed mRNA sequencing and compared gene expression of surface fish reared under LD conditions to cavefish reared under DD conditions to identify photic-dependent gene expression differences. Gene Ontology enrichment analyses revealed a number of previously underappreciated cave-associated changes impacting blood physiology and olfaction. We further evaluated the position of differentially expressed genes relative to QTL positions from prior studies and found several candidate genes associated with these ecologically relevant lighting conditions. In sum, this work highlights photic conditions as a key environmental factor impacting gene expression patterns in blind cave-dwelling fish.


Coincidence bell inequality for three three-dimensional systems.
We construct a Bell inequality for coincidence probabilities on a three three-dimensional (qutrit) system. We show that this inequality is violated when each observer measures two noncommuting observables, defined by the so-called unbiased six-port beam splitter, on a maximally entangled state of two qutrits. The strength of the violation agrees with the numerical results presented by Kaszlikowski et al, quant-ph/0202019. It is proven that the inequality defines facets of the polytope of local variable models.


Effect of the secretome of mesenchymal stem cells overexpressing BMP-9 on osteoblast differentiation and bone repair.
The secretome present in the conditioned medium (CM) of mesenchymal stem cells (MSCs) is a promising tool to be used in therapies to promote bone regeneration. Considering the high osteogenic potential of the bone morphogenetic protein 9 (BMP-9), we hypothesized that the secretome of MSCs overexpressing BMP-9 (MSCsBMP-9 ) enhances the osteoblast differentiation of MSCs and the bone formation in calvarial defects. CM of either MSCsBMP-9 (CM-MSCsBMP-9 ) or MSCs without BMP-9 overexpression (CM-MSCsVPR ) were obtained at different periods. As the CM-MSCsBMP-9 generated after 1 h presented the highest BMP-9 concentration, CM-MSCsBMP-9 and CM-MSCsVPR were collected at this time point and used to culture MSCs and to be injected into mouse calvarial defects. The CM-MSCsBMP-9 enhanced the osteoblast differentiation of MSC by upregulating RUNX2, alkaline phosphatase (ALP) and osteopontin protein expression, and ALP activity, compared with CM-MSCsVPR . The CM-MSCsBMP-9 also enhanced the bone repair of mouse calvarial defects, increasing bone volume, bone volume/total volume, bone surface, and trabecular number compared with untreated defects and defects treated with CM-MSCsVPR or even with MSCsBMP-9 themselves. In conclusion, the potential of the MSCBMP-9 -secretome to induce osteoblast differentiation and bone formation shed lights on novel cell-free-based therapies to promote bone regeneration of challenging defects.


Circulating Exosomal miRNA as Diagnostic Biomarkers of Neurodegenerative Diseases.
Neurodegenerative diseases (NDDs) are a group of diseases caused by chronic and progressive degeneration of neural tissue. The main pathological manifestations are neuronal degeneration and loss in the brain and/or spinal cord. Common NDDs include Alzheimer disease (AD), Parkinson disease (PD), Huntington disease (HD), and amyotrophic lateral sclerosis (ALS). The complicated pathological characteristics and different clinical manifestations of NDDs result in a lack of sensitive and efficient diagnostic methods. In addition, no sensitive biomarkers are available to monitor the course of NDDs, predict their prognosis, and monitor the therapeutic response. Despite extensive research in recent years, analysis of amyloid β (Aβ) and α-synuclein has failed to effectively improve NDD diagnosis. Although recent studies have indicated circulating miRNAs as promising diagnostic biomarkers of NDDs, the miRNA in the peripheral circulation is susceptible to interference by other components, making circulating miRNA results less consistent. Exosomes are small membrane vesicles with a diameter of approximately 30-100 nm that transport proteins, lipids, mRNA, and miRNA. Because recent studies have shown that exosomes have a double-membrane structure that can resist ribonuclease in the blood, giving exosomal miRNA high stability and making them resistant to degradation, they may become an ideal biomarker of circulating fluids. In this review, we discuss the applicability of circulating exosomal miRNAs as biomarkers, highlight the technical aspects of exosomal miRNA analysis, and review studies that have used circulating exosomal miRNAs as candidate diagnostic biomarkers of NDDs.


Involvement of Arabidopsis Acyl Carrier Protein 1 in PAMP-Triggered Immunity.
Plant fatty acids (FAs) and lipids are essential in storing energy and act as structural components for cell membranes and signaling molecules for plant growth and stress responses. Acyl carrier proteins (ACPs) are small acidic proteins that covalently bind the fatty acyl intermediates during the elongation of FAs. The Arabidopsis thaliana ACP family has eight members. Through reverse genetic, molecular, and biochemical approaches, we have discovered that ACP1 localizes to the chloroplast and limits the magnitude of pattern-triggered immunity (PTI) against the bacterial pathogen Pseudomonas syringae pv. tomato. Mutant acp1 plants have reduced levels of linolenic acid (18:3), which is the primary precursor for biosynthesis of the phytohormone jasmonic acid (JA), and a corresponding decrease in the abundance of JA. Consistent with the known antagonistic relationship between JA and salicylic acid (SA), acp1 mutant plants also accumulate a higher level of SA and display corresponding shifts in JA- and SA-regulated transcriptional outputs. Moreover, methyl JA and linolenic acid treatments cause an apparently enhanced decrease of resistance against P. syringae pv. tomato in acp1 mutants than that in WT plants. The ability of ACP1 to prevent this hormone imbalance likely underlies its negative impact on PTI in plant defense. Thus, ACP1 links FA metabolism to stress hormone homeostasis to be negatively involved in PTI in Arabidopsis plant defense. [Formula: see text] Copyright © 2022 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.


First Insight into a Nationwide Genotypic Diversity of Mycobacterium tuberculosis among Previously Treated Pulmonary Tuberculosis Cases in Benin, West Africa.
Molecular studies on tuberculosis (TB) are rare in low-resource countries like Benin, where data on molecular study on previously treated TB cases is unavailable. From January to December 2014, all smear- and culture-positive previously treated pulmonary TB patients from all TB clinics were systematically recruited. Drug susceptibility testing and spoligotyping were performed on all isolates. Of the 100 patients recruited, 71 (71.0%) were relapse cases and 24 (24.0%) were failure cases, while 5 (5.0%) were default cases. Resistance rate to any first-line drug was 40.0%, while 12.0% of strains were multidrug-resistant (MDR) and no strain was extensively drug-resistant (XDR). A total of 40 distinct spoligotypes were found to be corresponding to a genotypic diversity of 40.0%. ST61 was the most predominant spoligotype with prevalence of 33.0%. In all, 31 single spoligotypes and nine clusters were observed with 2 to 33 strains per cluster giving a clustering rate of 69.0%. Euro-American (Lineage 4) was the most prevalent lineage (74.0%) and Lineage 2 was associated with resistance to streptomycin. This first insight into genetic diversity of previously treated pulmonary TB patients in Benin showed a relatively high genetic diversity of Mycobacterium tuberculosis.


An education program which prepares nurses to teach breast self-examination.
The importance of competent and regular breast self-examination in the early detection and improved outcome of breast cancer cannot be emphasized enough. In their contacts with women, nurses are in an opportune position to promote BSE through education and demonstration. Breast self-examination is an area where many nurses are not comfortable. However, with planned and organized instruction, professional nurses can assume the role of teaching this important skill. Breast self-examination can then be incorporated into any clinical setting where professional nursing is practiced.


Polyelectrolyte multilayer-assisted immobilization of zero-valent iron nanoparticles onto polymer nanofibers for potential environmental applications.
We report a facile approach to synthesizing and immobilizing zero-valent iron nanoparticles (ZVI NPs) onto polyelectrolyte (PE) multilayer-assembled electrospun polymer nanofibers for potential environmental applications. In this approach, negatively charged cellulose acetate (CA) nanofibers fabricated by electrospinning were assembled with multilayers of poly(diallyldimethylammonium chloride) (PDADMAC) and polyacrylic acid (PAA) through electrostatic layer-by-layer assembly. The formed PAA/PDADMAC multilayers onto CA nanofibers were then used as a nanoreactor to complex Fe(II) ions through the binding with the free carboxyl groups of PAA for subsequent reductive formation of ZVI NPs. Combined scanning electron microscopy, transmission electron microscopy, energy dispersive spectroscopy, Fourier transform infrared spectroscopy, and thermogravimetry analysis studies demonstrate that the ZVI NPs are successfully synthesized and uniformly distributed into the PE multilayers assembled onto the CA nanofibers. The produced hybrid nanofibrous mats containing ZVI NPs were found to exhibit superior capability to decolorize acid fuchsin, an organic dye in dyeing wastewater. We show that the loading capacity of ZVI NPs can be tuned by changing the number of PE layers and the cycles of binding/reduction process. Increasing the number of the binding/reduction cycles leads to a slight bigger size of the ZVI NPs, which is not beneficial for improving the reactivity of ZVI NPs. The present approach to synthesizing and immobilizing ZVI NPs onto polymer nanofibers opens a new avenue to fabricating various fiber-based composite materials with a high surface area to volume ratio for environmental, catalytic, and sensing applications.


Cigarette Smoking and Pancreatic Cancer Survival.
Purpose Cigarette smoking is associated with increased incidence of pancreatic cancer. However, few studies have prospectively evaluated the association of smoking with patient survival. Patients and Methods We analyzed survival by smoking status among 1,037 patients from two large US prospective cohort studies diagnosed from 1986 to 2013. Among 485 patients from four prospective US cohorts, we also evaluated survival by prediagnostic circulating levels of cotinine, a metabolite of nicotine that is proportional to tobacco smoke exposure. On the basis of prediagnosis cotinine levels, we classified patients as nonsmokers (< 3.1 ng/mL), light smokers (3.1-20.9 ng/mL), or heavy smokers (≥ 21.0 ng/mL). We estimated hazard ratios (HRs) for death by using Cox proportional hazards models, with adjustment for age, sex, race/ethnicity, body mass index, diabetes status, diagnosis year, and cancer stage. Results The multivariable-adjusted HR for death was 1.37 (95% CI, 1.11 to 1.69) comparing current smokers with never smokers ( P = .003). A statistically significant negative trend in survival was observed for increasing pack-years of smoking ( Ptrend = .008), with HR for death of 1.49 (95% CI, 1.05 to 2.10) for > 60 pack-years of smoking versus never smoking. Survival among former smokers was similar to that for never smokers, regardless of time since quitting. Heavy smokers defined by prediagnostic circulating cotinine levels had a multivariable-adjusted HR for death of 1.76 (95% CI, 1.23 to 2.51) compared with nonsmokers. Among patients with circulating cotinine levels measured within 5 years before diagnosis, heavy smokers had a multivariable-adjusted HR for death of 2.47 (95% CI, 1.24 to 4.92) compared with nonsmokers. Conclusion Cigarette smoking was associated with a reduction in survival among patients with pancreatic cancer.


Willingness to pay and accept risks to cure chronic disease.
Measurements of disease burden focus most often on economic outputs--neglecting effects on quality of life. More comprehensive quantification is based on what people would pay or risk to avoid illness. Many, however, find it difficult to respond thoughtfully to hypothetical questions about what they would pay or risk. With response rates frequently under 50 per cent, the practicality of these methods has been of concern. In this study, specially trained interviewers asked 247 subjects with rheumatoid arthritis how much of their income they would pay and how large a mortal risk they would accept to achieve a hypothetical cure. Ninety-eight per cent of the subjects estimated their maximum acceptable risk (MAR) at an average 27 per cent chance of immediate death. Eighty-four per cent gave plausible responses to the willingness-to-pay (WTP) questions, with a mean WTP of 22 per cent of household income. The aspect of disease most strongly associated with WTP was impairment in activities of daily living; measured pain was most associated with MAR. The response rates achieved indicate the overall feasibility of these methods; the associations of WTP and MAR with other variables suggest systematic consideration of personal circumstances.


Hypoxia induces differentiation of pulmonary artery adventitial fibroblasts into myofibroblasts.
Activation of the alpha-smooth muscle actin (alpha-SMA) gene during the conversion of fibroblasts into myofibroblasts is an essential feature of various fibrotic conditions. Microvascular compromise and thus local environmental hypoxia are important components of the fibrotic response. The present study was thus undertaken to test the hypothesis that hypoxia can induce transdifferentiation of vascular fibroblasts into myofibroblasts and also to evaluate potential signaling mechanisms governing this process. We found that hypoxia significantly upregulates alpha-SMA protein levels in bovine pulmonary artery adventitial fibroblasts. Increased alpha-SMA expression is controlled at the transcriptional level because the alpha-SMA gene promoter activity, assayed via a luciferase reporter, was markedly increased in transfected fibroblasts exposed to hypoxia. Hypoxic induction of the alpha-SMA gene was mimicked by overexpression of constitutively active Galphai2 (alphai2Q205L) but not Galpha16 (alpha-16Q212L). Blockade of hypoxia-induced alpha-SMA expression with pertussis toxin, a Galphai antagonist, confirmed a role for Galphai in the hypoxia-induced transdifferentiation process. c-Jun NH2-terminal kinase (JNK) inhibitor II and SB202190, but not U0126, also attenuated alpha-SMA expression in hypoxic fibroblasts, suggesting the importance of JNK in the differentiation process. Hypoxia-induced increase in bromodeoxyuridine incorporation, which occurred concomitantly with hypoxia-induced differentiation, was blocked by U0126, suggesting that DNA synthesis and alpha-SMA expression take place through simultaneously activated parallel signaling pathways. Neutralizing antibody against transforming growth factor-beta1 blocked only 30% of the hypoxia-induced alpha-SMA promoter activity. Taken together, our results suggest that hypoxia induces differentiation of vascular fibroblasts into myofibroblasts by upregulating the expression of alpha-SMA, and this increase in alpha-SMA level occurs through Galphai- and JNK-dependent signaling pathways.


Noninvasive mechanical body contouring: a preliminary clinical outcome study.
L.P.G.'s Endermologie is a massage method consisting of positive pressure rolling, in conjunction with applied negative pressure to both the, skin and subcutaneous tissues (L.P.G. Endermologie U.S.A., 3101 North Federal Highway, Suite 301 Fort Lauderdale, Florida 33306, U.S.A., (800) 222-3911). Devised in France during the 1970s, L.P.G.'s original purpose was to soften scars and standardize physical therapy; however, patients treated with the L.P.G. machine also showed an improvement in body contour and skin texture. Since then, several thousand machines have been in use in France as an alternative method for altering fat distribution in the subcutaneous plane. The authors began a study to determine the safety and efficacy of this machine. This study is composed of 22 women between the ages of 24 and 48. All 22 women completed at least seven sessions of treatments. Six of these 22 women completed all 14 sessions of the prescribed treatments. The study group exhibited a wide range of body habitus, initial weights, and final results. Of the 22 women who completed seven sessions of treatment, three had an increase in body weight and a mean index (see Materials and Methods) reduction in body diameter of 1.38 cm (0.5 in). Three of the six patients who completed all 14 treatment sessions had an increase in body weight and a mean index reduction in body diameter of 2.85 cm (1.12 in). All but one of the patients had a decrease in their mean body diameter index, regardless of their loss or gain in weight.


Elevation of ceruloplasmin activity involved in changes of hepatic metal concentration in primary biliary cirrhosis.
Primary biliary cirrhosis (PBC) patients showed increased ceruloplasmin activity and Cu content in the serum as compared to the control group. Moreover, Cu, Fe, and Zn contents were measured in the livers of autopsied cases. The PBC patients showed increased Cu content and reduced Fe and Zn levels. In PBC cases, Cu transportation via bile is disturbed due to collapse of the interlobular bile ducts and consequently Cu accumulates in the liver. Thus, it is likely that serum ceruloplasmin activity increases to metabolize the accumulated hepatic Cu and that Cu content increases in the serum as a ceruloplasmin-bound form. Furthermore, reduction of Fe and Zn contents in the liver of the PBC patients indicated the possible relationship of ceruloplasmin to Fe and Zn metabolism as well as Cu metabolism.


[The role of anatomical hepatectomy in the treatment of intrahepatic cholangiocarcinoma].
Objective: To evaluate the role of anatomical hepatectomy in the treatment of intrahepatic cholangiocarcinoma. Methods: The cases of intrahepatic cholangiocarcinoma who received curative surgery in two hospitals from 2010 to 2015 were analyzed retrospectively. Among the 98 patients enrolled in this study, 55 were male and 43 were female. The median age was 61 years. According to receiving anatomical hepatectomy or not, the 98 cases were divided into two groups: non-anatomical hepatectomy(n=30) and anatomical hepatectomy(n=68). The surgical results were compared between the two groups.Survival curves were plotted by the Kaplan-Meier method and compared by the log-rank test. The influence of each prognostic factor identified by univariate analysis was multivariate analysis by Cox's proportional hazard regression. Results: The duration of surgery was significantly prolonged in the anatomical hepatectomy group((196.4±94.9)minutes vs. (166.2±65.7)minutes, P=0.027), while there was no significant difference in terms of other surgical results such as intraoperative blood transfusion, postoperative morbidity and mortality rate. Compared to non-anatomical hepatectomy, anatomical hepatectomy significantly improved long-term survival results(14 months vs. 11 months)(χ2=4.641, P=0.031). Single variable analysis indicated that tumor differentiation, tumor numbers, T stage, N stage, anatomical hepatectomy and adjuvant therapy significantly affected overall survival. Multivariate analysis demonstrated that tumor numbers(HR=0.522, 95% CI: 0.259-0.974, P=0.042) and anatomical hepatectomy(HR=1.858, 95%CI: 1.092-3.161, P=0.022) were two independent prognostic factors for overall survival. Conclusion: Compared to non-anatomical hepatectomy, anatomical hepatectomy performed for intrahepatic cholangiocarcinoma is not only safe but also beneficial for long-term survival.


Molecular cloning of porcine Siglec-3, Siglec-5 and Siglec-10, and identification of Siglec-10 as an alternative receptor for porcine reproductive and respiratory syndrome virus (PRRSV).
In recent years, several entry mediators have been characterized for porcine reproductive and respiratory syndrome virus (PRRSV). Porcine sialoadhesin [pSn, also known as sialic acid-binding immunoglobulin-type lectin (Siglec-1)] and porcine CD163 (pCD163) have been identified as the most important host entry mediators that can fully coordinate PRRSV infection into macrophages. However, recent isolates have not only shown a tropism for sialoadhesin-positive cells, but also for sialoadhesin-negative cells. This observation might be partly explained by the existence of additional receptors that can support PRRSV binding and entry. In the search for new receptors, recently identified porcine Siglecs (Siglec-3, Siglec-5 and Siglec-10), members of the same family as sialoadhesin, were cloned and characterized. Only Siglec-10 was able to significantly improve PRRSV infection and production in a CD163-transfected cell line. Compared with sialoadhesin, Siglec-10 performed equally effectively as a receptor for PRRSV type 2 strain MN-184, but it was less capable of supporting infection with PRRSV type 1 strain LV (Lelystad virus). Siglec-10 was demonstrated to be involved in the endocytosis of PRRSV, confirming the important role of Siglec-10 in the entry process of PRRSV. In conclusion, it can be stated that PRRSV may use several Siglecs to enter macrophages, which may explain the strain differences in the pathogenesis.


COL6A1, the candidate gene for ossification of the posterior longitudinal ligament, is associated with diffuse idiopathic skeletal hyperostosis in Japanese.
Genetic screening of collagen 6A1 gene (COL6A1) in patients with diffuse idiopathic skeletal hyperostosis (DISH) recruited in Japan and the Czech Republic. To investigate allelic associations between DISH and nucleotide variants of COL6A1. DISH is a skeletal hyperostotic disease characterized by ligamentous ossification of the anterolateral side of the spine. Ossification of the posterior longitudinal ligament (OPLL) is a related disorder with DISH, and COL6A1 was identified as a susceptibility gene to OPLL. COL6A1 was examined for susceptibility in DISH patients from Japan and the Czech Republic. Seven single nucleotide polymorphisms of COL6A1 were genotyped by direct sequencing. The allele frequencies were compared between 97 Japanese DISH patients and 298 Japanese controls, and between 96 Czech DISH patients and 96 Czech controls by chi2 test. The intron 32 (-29) single nucleotide polymorphisms of COL6A1 was significantly associated with the Japanese DISH patients (chi2 = 9.33; P = 0.0022), but not with the Czech DISH patients. Because COL6A1 could be a susceptibility to the occurrence of DISH and OPLL in the Japanese population, we consider that COL6A1 could be responsible for the hyperostotic state, leading to ectopic bone formation in the spinal ligament.


Interactions of law and ethics affecting reproductive choice.
Controversies affecting reproductive choice can often be resolved within interactions of legal and ethical decision-making. This paper addresses three topics, following the methodology presented in Reproductive Health and Human Rights: Integrating Medicine, Ethics, and Law, by R.J. Cook, B.M. Dickens and M.F. Fathalla (Oxford University Press, 2003). The book's 15 case studies each addresses medical, ethical, legal and human rights aspects, and structural approaches at clinical, healthcare system and societal levels. STERILIZATION: Individual self-determination supports legal and ethical rights of intellectually competent persons to sterilization. Sterilization of intellectually compromised persons was historically abused, causing reactions of excessively protective prohibition. ABORTION: Most developed countries have liberalized abortion legislation, thereby reducing abortion-related mortality and morbidity, but many developing countries retain repressive colonial laws. Over 95% of the estimated 20 million unsafe abortions annually occur in developing countries. COURT-ORDERED CAESAREAN DELIVERIES: A concern in developed countries is the willingness of some courts to order Caesarean procedures over competent women's objections.


Pilot alcohol violations reported in U.S. newspapers, 1990-2006.
Alcohol violations by airline pilots are rare yet remain a public concern. Such incidents often generate widespread news coverage. This study examines the frequency and characteristics of alcohol violation incidents involving airline pilots reported in U.S. newspapers. The database of Lexis-Nexis, which contains full-text articles for over 350 newspapers, was searched to identify alcohol violation incidents involving airline pilots in the U.S. between January 1990 and June 2006. Information pertaining to the pilot, flight, blood alcohol concentration (BAC), and consequence was ascertained for each incident based on the newspaper coverage. During the study period, newspapers reported on a total of 13 incidents of alcohol violations involving 17 pilots. All but two of the incidents occurred during January 2002 through June 2006. The majority (85%) of the incidents were first identified by airport personnel, such as security screeners, based on suspicion of alcohol use by the pilot. Subsequent alcohol testing revealed a mean BAC of 90 mg/dL (ranging from 10 mg x dL(-1) to 182 mg x dL(-1)). Of the 17 pilots, 6 were known to be prosecuted criminally, including 5 who were sentenced to jail terms. Incidents of alcohol violations by airline pilots reported in U.S. newspapers have increased in recent years. This increase is likely due in part to increased detection resulting from enhanced aviation security and enforcement following the September 2001 terrorist attacks.


Maintenance of geometric alterations associated with percutaneous mitral valve repair: real-time three-dimensional echocardiographic assessment in an ovine model.
Percutaneous catheter-based mitral annuloplasty (PTMA) exploits the anatomic proximity of the coronary sinus (CS) to the mitral valve apparatus. Acute results of PTMA have been favorable, but the durability of the geometric alterations associated with PTMA has not been reported. The study aim was to assess the three-dimensional (3D) geometry of the mitral annulus (MA) in normal sheep at 20 weeks after PTMA implantation. A PTMA device was implanted percutaneously in the CS of 10 normal sheep without mitral regurgitation. All animals were followed for 20 weeks with real-time 3D echocardiography (RT3DE). The MA area, the diagonal diameters in four directions, and the angle alpha, representing the degree of the saddle shape of MA, were determined. No significant hemodynamic, pathologic or mechanical complications were observed during implantation or follow up. Both, the MA area (from 4.8 +/- 0.9 cm2 to 3.7 +/- 0.9 cm2) and anterior-posterior (A-P) diameter (from 21.4 +/- 3.0 mm to 17.6 +/- 2.4 mm) were reduced immediately after the procedure (both p <0.05). The angle alpha decreased after the procedure (from 142.0 +/- 11.5 degrees to 128.3 +/- 15.6 degrees; p <0.05). These changes remained stable over the 20-week follow up period. RT3DE demonstrates that PTMA reduces the MA area and A-P diameter and maintains the physiologic curved or saddle shape of the MA. These changes remained stable for 20 weeks after device implantation.


Double-antibody radioimmunoassay of serum insulin: effect of use of hormone-depleted human serum.
Human serum can be depleted of insulin and growth hormone by treatment with dextran-coated charcoal or by dialysis, or both. Inclusion of such hormone-depleted serum in the standard curve of a double-antibody radioimmunoassay for immunoreactive insulin more nearly mimics the conditions under which an unknown human serum sample is assayed. Compared to the usual standard curve in which serum albumin is the only protein, the addition of hormone-depleted serum can cause an increase (by an average 64% under the conditions we used) in the absolute value for apparent insulin in serum. The effect of hormone-depleted serum should be tested in the standard curves of double-antibody radioimmunoassays and included routinely in these standard curves if it changes the results. When used in conjunction with an eventual reference standard for human insulin, this modification of the insulin assay may make the measured values of human serum insulin from different laboratories more comparable.


The clinician-scientist in neuropsychiatry: a position statement from the Committee on Research of the American Neuropsychiatric Association.
Neuropsychiatric research seeks to improve the lives of patients with brain-based behavioral disturbances. There has been dramatic progress in diagnosis and treatment of neuropsychiatric disorders, and progress in neuroscience and biotechnology promises further success. Paradoxically, recent trends threaten to erode this progress. In this environment, neuropsychiatric clinician-scientists must advocate for the importance of research. This position statement defines neuropsychiatric research, describes current challenges to the neuropsychiatric clinician-scientist, summarizes research opportunities, describes how future neuropsychiatric clinician-investigators should be trained, and makes recommendations for promoting neuropsychiatric research.


Reduced mortality rates in a cohort of long-term underground iron-ore miners.
Historically, working in iron-ore mines has been associated with an increased risk of lung cancer and silicosis. However, studies on other causes of mortality are inconsistent and in the case of cancer incidence, sparse. The aim of this study was to examine the association between iron-ore mining, mortality and cancer incidence. A 54-year cohort study on iron-ore miners from mines in northern Sweden was carried out comprising 13,000 workers. Standardized rate ratios were calculated comparing the disease frequency, mortality, and cancer incidence with that of the general population of northern Sweden. Poisson regression was used to evaluate the association between the durations of employment and underground work, and outcome. Underground mining was associated with a significant decrease in adjusted mortality rate ratios for cerebrovascular and digestive system diseases, and stroke. For several outcomes, elevated standardized rate ratios were observed among blue-collar workers relative to the reference population. However, only the incidence of lung cancer increased with employment time underground (P < 0.001). Long-term iron-ore mining underground was associated with lower rates regarding several health outcomes. This is possibly explained by factors related to actual job activities, environmental exposure, or the selection of healthier workers for long-term underground employment.


The lncRNA ANRIL Gene rs2151280 GG Genotype is Associated with Increased Susceptibility to Recurrent Miscarriage in a Southern Chinese Population.
Genetic factors may play an important role in susceptibility to recurrent miscarriage. Some cardiovascular disease-related candidate genes have been shown to be associated with recurrent miscarriage. Long noncoding RNA ANRIL has been confirmed to be associated with susceptibility to various diseases, such as cardiovascular disease. However, it remains unclear whether the ANRIL gene polymorphism is related to recurrent miscarriage susceptibility. Three ANRIL gene polymorphisms (rs2151280, rs1063192 and rs564398) were genotyped in 819 controls and 610 recurrent miscarriage patients through TaqMan real-time polymerase chain reaction. The odds ratios and 95% confidence intervals (CIs) were used to assess the strength of each association. Our results showed that the ANRIL rs2151280 GG genotype was associated with increased susceptibility to recurrent miscarriage (GG vs AA: adjusted OR=1.527, 95% CI=1.051-2.218, p=0.0262; GG vs AG/AA adjusted OR=1.460, 95% CI=1.021-2.089, p=0.0381). By combining the analysis of the risk genotypes in the three SNPs, we found that individuals with 2-3 risk genotypes had a significantly increased risk of recurrent miscarriage compared with those with a 0-1 risk genotype (adjusted OR=1.728, 95% CI=1.112-2.683, p=0.0149). This risk was more significant in subgroups of women less than 35-40 years of age and women with 2-3 miscarriages. These results suggested that a specific SNP in the ANRIL gene may be associated with increased susceptibility to recurrent miscarriage in a southern Chinese population.


Identification of a novel spinal nociceptive-motor gate control for Aδ pain stimuli in rats.
Physiological responses to nociceptive stimuli are initiated within tens of milliseconds, but the corresponding sub-second behavioral responses have not been adequately explored in awake, unrestrained animals. A detailed understanding of these responses is crucial for progress in pain neurobiology. Here, high-speed videography during nociceptive Aδ fiber stimulation demonstrated engagement of a multi-segmental motor program coincident with, or even preceding, withdrawal of the stimulated paw. The motor program included early head orientation and adjustments of the torso and un-stimulated paws. Moreover, we observed a remarkably potent gating mechanism when the animal was standing on its hindlimbs and which was partially dependent on the endogenous opioid system. These data reveal a profound, immediate and precise integration of nociceptive inputs with ongoing motor activities leading to the initiation of complex, yet behaviorally appropriate, response patterns and the mobilization of a new type of analgesic mechanism within this early temporal nociceptive window.


Community orthodontists and craniofacial care: results of a Washington state survey.
Little is known about community orthodontists' previous training in, experience with, or receptivity to caring for children with craniofacial disorders. (1) To characterize the current level of participation by Washington state orthodontists in craniofacial care; and (2) to identify factors that promote or impede community orthodontists' involvement in caring for children with craniofacial conditions. Mail survey. A 26-item questionnaire was designed and mailed to all active orthodontists in Washington state (N = 230). Question topics included practice characteristics, training and experience with craniofacial conditions, concerns related to public and private insurance, and communication with craniofacial teams. Of eligible respondents, 68% completed the survey. Most orthodontists' patient panels were made up of patients who either have private insurance or pay cash for services. On average, 2% of respondents' patients were Medicaid beneficiaries. Only 20% of respondents had seen more than three patients with cleft lip and/or palate in the past 3 years. Although a minority of orthodontists receive referrals from (27%) or are affiliated with (11%) craniofacial teams, most orthodontists perceived craniofacial care positively and were interested to learn more about craniofacial care and to accept additional patients with these conditions. Results of this survey can inform potential strategies to increase access to orthodontic care for children with craniofacial disorders. These would include developing an organized training, referral, and communication system between community orthodontists and state craniofacial teams and considering a case-management approach to facilitate this process.


The aroma of goat milk: seasonal effects and changes through heat treatment.
Goat milk was characterized and analyzed by human sensory evaluation and gas chromatography/olfactometry (GC/O). Most potent odor-active compounds were determined in (a) raw goat's milk from two different seasons and (b) heated goat's milk after different treatment intensities. A trained panel found sensorial differences between winter and summer milks (seasonal effect) and milks from different farms (farm-specific effect). A total of 54 odor-active compounds with flavor dilution (FD) factors ≥8 were detected of which 42 odorants were identified. 4-Ethyloctanoic acid, 3-methylindole (skatol) and one unknown compound (RI 2715) showed highest intensities in all raw milks. With heat treatment, goat-like, stable-like, and (cooked) milk-like odor characteristics decreased while caramel-like or vanilla-like notes increased. In total, 66 odor-active compounds were detected in heated goat milks (FD ≥ 8). To the best of our knowledge, only 16 of the 42 identified odorants were reported before in raw goat's milk. Additionally, for the first time the presence of 1-benzopyran-2-one (coumarin) could be confirmed in ruminant milk.


The Importance of the Study of Cognitive Performance Enhancement for U.S. National Security.
The American military is embarking on the 'Third Offset'-a strategy designed to produce seismic shifts in the future of warfare. Central to the approach is the conjoining of humans, technology, and machines to deliver a decisive advantage on the battlefield. Because technology will spread rapidly and globally, tactical overmatch will occur when American operators possess a competitive edge in cognition. Investigation of cognitive enhancing therapeutics is not widely articulated as an adjunct to the Third Offset, yet failure to study promising agents could represent a strategic vulnerability. Because of its legacy of research into therapeutic agents to enhance human-machine interplay, the aerospace medical community represents a front-running candidate to perform this work. Notably, there are strong signals emanating from gambling, academic, and video-gaming enterprises that already-developed stimulants and other agents provide cognitive benefits. These agents should be studied not only for reasons of national security, but also because cognitive enhancement may be a necessary step in the evolution of humankind. To illustrate these points, this article will assert that: 1) the need to preserve and enhance physical and cognitive health will become more and more important over the next century; 2) aeromedical specialists are in a position to take the lead in the endeavor to enhance cognition; 3) signals of enhancement of the type useful to both military and medical efforts exist aplenty in today's society; and 4) the aeromedical community should approach human enhancement research deliberately but carefully.Malish RG. The importance of the study of cognitive performance enhancement for U.S. national security. Aerosp Med Hum Perform. 2017; 88(8):773-778.


Sleep disturbances in drug naïve Parkinson's disease (PD) patients and effect of levodopa on sleep.
Parkinson's disease (PD) is associated with sleep disturbances, attributed to the neurodegenerative process and therapeutic drugs. Studies have found levodopa to increase wakefulness in some patients while increasing sleepiness in others. To confirm sleep disturbances in drug naïve PD patients and understand the impact of levodopa on their sleep. Twenty-three drug naïve PD patients and 31 age-gender matched controls were compared using the Parkinson's Disease Sleep Scale (PDSS) and Epworth Sleepiness Scale (ESS). A polysomnogram objectively compared sleep quality. Of the 23 patients, the 12 initiated on levodopa were reassessed subjectively and through polysomnography after 2 months of therapy. Data was expressed as mean ± standard deviation, median, and range. Continuous variables were analyzed by Student's T test for normally distributed data and Mann-Whitney U test for skewed data. Discrete variables were compared by Chi Square tests (Pearson Chi square Test or Fisher's Exact Test). Wilcoxon signed ranks test was applied in the analysis of paired data pre- and post-levodopa. A P value < 0.05 was considered as statistically significant. Statistical analysis of the data was done using the Statistical Package for the Social Sciences (SPSS) version 12. Drug naïve PD patients had lower PDSS scores than controls. The sleep architecture changes observed on polysomnogram were reduced NREM Stage III and REM sleep and increased sleep latency and wake after sleep onset time. Following levodopa, improved sleep efficiency with reduced sleep latency and wake after sleep onset time was noted, coupled with improved PDSS scores. However, NREM Stage III and REM sleep duration did not increase. PD patients take longer to fall asleep and have difficulty in sleep maintenance. Sleep maintenance is affected by nocturia, REM behavioral disorder, nocturnal cramps, akinesia, and tremors, as observed in PDSS scores. Levodopa improves sleep efficiency by improving motor scores without altering sleep architecture. Poor sleep quality and sleep architecture changes occur secondary to the neurodegenerative process in PD patients. Though levodopa improves sleep quality by reducing rigidity and tremor, it does not reverse sleep architecture changes.


[Treatment of arrhythmia in coronary patients and hypertensives with beta blockers and Depasan retard].
In the ambulatory of an internal specialist a number of patients suffering from angina pectoris or hypertonia together with arrhythmic troubles received an initial treatment with beta-blockers over a period of 21 days. In cases in which arrhythmia persisted after this initial period the treatment was continued for another 21 days with the addition of Depasan retard in function of a second medicament. This combined treatment in form of an open study was extended to a total number of 50 patients presenting ventricular extrasystolia in 45 cases and in 5 cases absolute arrhythmia in addition to the main disease. Treatment with Depasan retard showed good or satisfactory results in 39 out of 45 patients suffering from ventricular extrasystoles, whilst no effect could be obtained in the 5 cases with absolute arrhythmia. No signs of incompatibility or unwanted interactions were observed in the course of this open study. Based on these results it can be concluded that Depasan retard should be recommended in those cases where on account of persistent arrhythmical troubles and especially extrasystoles, during a treatment with beta-blockers in patients suffering from angina pectoris or hypertonia, and additional treatment with anti-arrhythmical medicaments appears to be indicated.


ABSENCE OF POLLEN DISCOUNTING IN A GENOTYPE OF IPOMOEA PURPUREA EXHIBITING INCREASED SELFING.
Throughout southeastern North America, the annual morning glory Ipomoea purpurea exhibits a polymorphism at a locus that influences the intensity of floral pigmentation. Previous studies have shown that when rare, the homozygous white genotype has a greater selfing rate than the homozygous dark genotype. In the absence of pollen discounting (a reduction in transmission of pollen to other plants by genotypes that exhibit increased selfing) and inbreeding depression, this increased selfing rate should favor the white allele. Experiments reported here confirm that the white genotype has elevated selfing rates when rare but indicate pollen discounting is not associated with elevated selfing. Rather, white genotypes contribute more pollen to the outcross pollen pool. The disparity between genotypes in both selfing rates and success at pollen contribution to other plants disappears at intermediate to high frequencies of the white allele. Pollinator movements are consistent with the pattern of selfing. These results suggest that elevated selfing and enhanced success at pollen donation contribute to maintenance of the white allele in natural populations of morning glories.


[Pilonidal sinus - diagnosis at the intersection of general and plastic surgery].
Pilonidal sinus predominantly affects young patients. Improper treatment results in long-term restrictions in everyday life and incapacity for work. The aim of the study was to find out what the results and current treatment options of pilonidal sinus are. This is a retrospective analysis of 67 patients treated at the Department of Surgery of the Second Medical Faculty of Charles University and Motol Hospital in the period 20102013. We evaluated the type of surgery and infectious complications in the wound, as well as age, sex, BMI, smoking, employment of the patients, duration of wound drainage, length of hospital stay, time required to complete healing of the surgical wound, the surgeons erudition and disease recurrence. 50 (75%) patients underwent primary closure in the midline, Limberg flap was used in 15 (22%) patients. In 2 (3%) patients, the wound was left without suture. In the group of patients who had not undergone flap reconstruction, secondary wound healing occurred in 20 (40%) patients. In the group of patients where flap reconstruction was used, secondary healing occurred in 3 patients (20%). Relapse of the disease within one year occurred in the group of patients with primary suture in the midline in 4 (8%) patients; other patients had no recurrence. According to our experience as well as literary data, the treatment of choice is the extirpation of the sinus with primary suture beyond the midline using a flap reconstruction technique.


Antimicrobial Efficacy of a Novel Automated Irrigation Device As Compared to Conventional Needle Irrigation Against Enterococcus faecalis: An In Vitro Study.
Aim The present study aims to compare the antibacterial efficacy of a novel automated endodontic irrigation device with that of a manual irrigation technique. Materials and methods The present study considered 45 extracted single-rooted teeth. After sectioning the teeth, the specimens were inoculated with Enterococcus faecalis (E. faecalis) bacteria for three weeks. Instrumentation was done using ProTaper Gold rotary files to size 50 and 5% taper. Based on the irrigation protocol, the experimental samples were divided into Group I: control (normal saline irrigation) (n = 15); Group II: conventional needle irrigation (n = 15); and Group III: automated irrigation (n = 15). The irrigation in Group III was done using the automated irrigation device. After the experimental irrigation, the pre-weighted dentinal shaving was collected in Eppendorf tubes, diluted 10 times, and incubated in the Petri dish with tryptone soy agar (TSA) for 48 hours. Finally, the colony-forming unit (CFU) counts were assessed. IBM SPSS Statistics for Windows version 23.0 (Armonk, NY, USA, IBM Corp.) was used for data analysis. Intergroup comparisons were made using the non-parametric Kruskal-Wallis test. Results The mean CFU count (CFU/ml) for the groups was: Group 1 (normal saline irrigation): 3.67x101; Group 2 (manual irrigation): 2.69× 101; Group 3 (automated irrigation): 1.57× 101. A statistically significant reduction in E. faecalis CFU count was seen among the groups assessed (p<0.01). The automated irrigation group exhibited the most substantial reduction in E. faecalis CFU count. Group 2 showed a significant difference compared to Group 1 (p<0.05). Conclusions The novel automated endodontic irrigation device was superior to manual irrigation in reducing E. faecalis CFU counts. This indicates that the automated irrigation device has the potential to enhance bacterial elimination efficacy during endodontic treatment.


A Case of Parvovirus-Related Haemophagocytic Lymphohistiocytosis in a Patient with HbH Disease.
Hemophagocytic lymphohistiocytosis (HLH) is rare and life-threatening medical emergency. Parvovirus infection is rarely associated with HLH. We report a case of parvovirus-related HLH in a patient with alpha thalassaemia (HbH disease). The patient responded well to a course of dexamethasone without the need of etoposide. Based on our literature search, this is the first case of parvovirus related HLH in a patient with HbH disease in the medical literature.


Inheritance of random amplified polymorphic DNA (RAPD-PCR) markers and their use in population genetic studies of Calomys musculinus (Rodentia, Muridae), the reservoir of Argentine hemorrhagic fever.
In order to assess the reliability of RAPD markers in the estimation of the genetic structure of natural populations of the murid rodent Calomys musculinus (reservoir of Junin virus, ethiological agent of Argentine Hemorrhagic Fever), we have analyzed the heritability of RAPD bands in 10 parents and their offspring (33 individuals). Fourteen out of a total of 119 bands obtained were absent in the parental patterns, but consistently amplified in offspring from some families. These bands can be eliminated from analyses. Overall degree of band sharing between individuals, including non-parental bands, correctly grouped members of a family in the same cluster in a UPGMA tree, with a high bootstrap percentage. Results support the usefulness of RAPDs as hereditable markers. One hundred polymorphic RAPD loci were identified in three natural populations of C. musculinus. Mean expected heterozygosity in three natural populations ranged from 0.206 to 0.220. Allele frequency based and phenotype based measures of genetic differentiation among natural populations of C. musculinus gave similar results (Weir and Cockerham's theta = 0.133; Excoffier et al.'s phi = 0.127). These values were considerably higher than those found previously using allozymes as genetic markers, and are compatible with moderate to low levels of gene flow among populations.


Nurses' knowledge of error in blood pressure measurement technique.
Errors in measuring blood pressure may have significant impact on the investigation and treatment of patients. Errors arise from faults in measurement technique or the equipment used. In Australia, blood pressure measurement technique is taught to nurses during their undergraduate education and may not be reviewed again. This observational, descriptive study surveyed clinical nurses at a metropolitan teaching hospital at shift hand-over time. Participation was voluntary and anonymous by 78 nurses who answered a questionnaire to determine the need and focus for updating blood pressure measurement technique. Sixty-one per cent of participants conformed to currently accepted practice in identifying systolic blood pressure, and 71% diastolic blood pressure; 54% correctly interpreted a description of blood pressure sounds containing an auscultatory gap. Correct answers for assessment of faulty equipment were given by 58%, assessing cuff size by 57%, arm position for seated measurement by 14%, determination of inflation pressure by 29% and deflation rate 62%. Incidence of terminal digit preference was 32%. These findings indicate that knowledge of participants was inadequate to perform blood pressure measurement in a standardized manner, and prevent introduced error.


Feulgen DNA cytophotometry in histologic sections of mammary neoplasms.
Because of a lack of suitable archival material, it is rarely possible to make retrospective studies of the correlation between the prognosis for a patient with mammary carcinoma and the distribution of nuclear DNA in the cells of the neoplasm. An investigation of the possibility of using sections cut from paraffin-embedded specimens showed that such sections are not suitable for use in retrospective studies of breast carcinoma. Because of such factors as the heterogeneity in size and shape of the nuclei from neoplastic cells and their tendency to mold around each other, determinations of DNA content of cells in sections were extremely difficult; in this particular carcinoma it was found that the distribution of nuclear DNA as obtained from a Feulgen-stained histologic section was not the same as that obtained from a Feulgen-stained imprint smear, and some polyploid tumors were erroneously classified as aneuploid.


Comparison of S-T segment/heart rate slope with exercise thallium imaging and conventional S-T segment criteria in detecting coronary artery disease: effect of exercise level on accuracy.
The S-T segment/heart rate (ST/HR) slope has been proposed as a more accurate electrocardiographic criterion for the diagnosis of coronary artery disease, but studies comparing the diagnostic value of the ST/HR slope with exercise thallium imaging are scant. The aims of this study were to assess the diagnostic accuracy of the ST/HR slope, conventional S-T segment criteria and exercise thallium imaging in detecting coronary artery disease and to evaluate the effect of exercise level on the sensitivity and specificity of these three techniques. Ninety consecutive patients underwent treadmill testing and exercise thallium single-photon emission computed tomography (SPECT) simultaneously. All 90 patients also underwent coronary angiography within 1 month of the exercise test. We found that exercise thallium SPECT had a significantly higher diagnostic accuracy than either the ST/HT slope or conventional S-T segment criteria (thallium imaging 82%, ST/HR slope 67%, conventional S-T segment criteria 63%). The overall accuracy of the ST/HR slope was slightly but insignificantly greater than conventional S-T segment criteria (ST/HR slope 67%, conventional S-T segment criteria 63%; p = 0.639). In 50 patients with a lower exercise level, defined as not achieving 85% of their maximal predicted heart rate, the accuracy of the ST/HR slope was insignificantly greater (ST/HR slope 72%, conventional S-T segment criteria 66%; p = 0.517). In 40 patients with adequate exercise, the accuracy rate was the same (60%) for both the ST/HR slope and conventional S-T segment criteria. We conclude that exercise thallium imaging is more accurate than the ST/HR slope in diagnosing coronary artery disease and that the accuracy of the ST/HR slope is marginally better than conventional S-T segment criteria only in patients with a lower exercise level, and not in those with adequate exercise.


The effects of Lactobacillus reuteri microcapsules on radiation-induced brain injury by regulating the gut microenvironment.
A radiation-induced brain injury (RIBI) is a major adverse event following radiotherapy of malignant tumors. RIBI would affect cognitive function, leading to a series of complications and even death. However, the pathogenesis of RIBI is still unclear, and it still lacks specific therapeutic drugs. The gut-brain bidirectional communication may be mediated by various microbiota and metabolites in the gastrointestinal tract. Probiotics are closely related to physiological health. The theory of the gut-brain axis provides us with a new idea to improve the gut microenvironment by supplementing probiotics against RIBI. Here, Lactobacillus reuteri microcapsules (LMCs) were prepared, which were predominantly irregular spheres with a rough surface under a scanning electron microscope and a narrow size distribution ranging from 20 to 700 μm. The transmission electron microscopy images showed that the structure of microcapsules containing Lactobacillus reuteri (L. reuteri) was a core and shell structure. The survival of L. reuteri in microcapsules was significantly more than that of free L. reuteri in the simulated stomach environment of pH 1.2. 16S rDNA sequencing showed that LMCs observably increased the relative abundance of Lactobacillus in RIBI mice. More importantly, compared with the RIBI model mice, the behavior of RIBI mice treated with LMCs was significantly improved. In addition, LMCs greatly alleviated the pathological damage of the hippocampus and intestines in the mice after irradiation and reduced the level of TNF-α and IL-6 in vivo. Generally, LMCs are a promising oral preparation, which provide new ideas and methods for the treatment of RIBI.