Differences in COVID-19-Related Testing and Healthcare Utilization by Race and Ethnicity in the Veterans Health Administration.
Recent reports indicate differences in COVID-19-related care and outcomes between Black and White Americans. We examine the COVID-19-related healthcare utilization and mortality by race and ethnicity of patients tested for SARS-CoV-2 in the Veterans Health Administration (VHA). A retrospective cohort study. We used the VHA COVID-19 shared data resources between February 1 and June 30, 2020. Veterans tested for SARS-CoV-2 virus by VHA. Three racial-ethnicity groups of Black, Hispanic, and White (as reference) veterans. Main outcomes are testing rate, positivity rate, hospitalization rate, ICU admission rate, and in-hospital mortality. Controlling for sex, age, and Elixhauser comorbidity index, we report adjusted odds ratios (aOR) and 95% confidence intervals (95% CI) from logistic regression models. Of the 8,667,996 active veteran enrollees, 252,702 were tested by VHA from February to June, 2020, with 20,500 positive results and 4,790 hospitalizations. The testing rate was 4.4% among Black and 4.7% among Hispanic veterans compared to White veterans, 2.8%. The testing positivity rate was similarly elevated among Black (12.2%) and Hispanic (11.6%) veterans compared to White veterans (6.0%). The aORs of hospitalization in Black veterans (1.88; 95% CI 1.74, 2.03) and Hispanic veterans (1.41; 95% CI 1.25, 1.60) were higher compared to White veterans. No significant differences by race and ethnicity were observed in OR or aOR of ICU admission and in-hospital death among hospitalized patients. On a national level, the VHA was more likely to test and hospitalize Black and Hispanic veterans compared to White veterans, but there were no significant differences in ICU admission or in-hospital mortality among those hospitalized. This pattern of differences may relate to social determinants of health, factors affecting access to non-VHA care, or preferences for VHA care affecting initial care seeking, but not in-hospital outcomes.


Inhibition of sphingomyelin hydrolysis: targeting the lipid mediator ceramide as a key regulator of cellular fate.
Ceramide, an intracellular lipid mediator, is generated by transient hydrolysis of sphingomyelin in response to agonists inducing inflammation and apoptosis, ionizing radiation, chemotherapeutics or ischaemia/reperfusion. An elevated intracellular ceramide production is predominantly induced by an elevated hydrolytic activity of sphingomyelinases or by the activity of enzymes controlling de novo synthesis, such as ceramide synthase. Ceramide is implicated in various cellular responses, acting as an autonomous intracellular effector in cell cycle regulation, differentiation, senescence, and apoptosis. Furthermore, by changing membrane properties it contributes to the assembly and interactions of various signal transduction molecules. The lipid mediator is subsequently metabolised by ceramidase and sphingosine kinase, and other key players, which determine the dynamic balance between the intracellular levels of ceramide and its breakdown products (the ceramide/S1P rheostat). Together with sphingomyelinases, they are crucial for cell signalling, cellular survival or initiation of apoptosis. Sphingomyelinases are regarded as key enzymes in the regulated activation of the sphingomyelin cycle. Up to now, five isoforms of sphingomyelinases are known, differentiated by agonists, intracellular localization, pH optimum and essential co-factors. Herein, we focus on the biochemical background and the action of pharmacologically interesting compounds capable of interfering with sphingomyelinases and outline their potential implications for medicinal chemistry.


A computational and experimental investigation of the mechanical properties of single ZnTe nanowires.
One-dimensional nanostructures such as ZnTe, CdTe, Bi(2)Te(3) and others have attracted much attention in recent years for their potential in thermoelectric devices among other applications. A better understanding of their mechanical properties is important for the design of devices. A combined experimental and computational approach has been used here to investigate the size effects on the Young's modulus of ZnTe nanowires (NWs). The mechanical properties of individual ZnTe nanowires in a wide diameter range (50-230 nm) were experimentally measured inside a high resolution transmission electron microscope using an atomic force microscope probe with the ability to record in situ continuous force-displacement curves. The in situ observations showed that ZnTe NWs are flexible nanostructures with the ability to withstand relatively high buckling forces without becoming fractured. The Young's modulus is found to be independent of nanowire diameter in the investigated range, in contrast to reported results for ZnO NWs and carbon nanotubes where the modulus increases with a decrease in diameter. Molecular dynamics simulations performed for nanowires with diameters less than 20 nm show limited size dependence for diameters smaller than 5 nm. The surface atoms present lower Young's modulus according to the simulations and the limited size dependency of the cylindrical ZnTe NWs is attributed to the short range covalent interactions.


Management of acute pancreatitis.
Acute pancreatitis (AP) is a common medical condition with extensive morbidity and mortality. Approximately 210,000 Americans are hospitalized each year; and 5% of patients with AP will die. It is also an expensive condition, costing 2.6 billion dollars (United States) in 2009 alone. Moreover, the incidence is increasing - the National Hospital Discharge Survey showed hospitalizations increased from 78 per 100,000 in 2007 to 90 per 100,000 just three years later in 2010. There is no proven pharmacologic entity to treat the inflammatory response associated with acute pancreatitis; supportive care with IV fluids, bowel rest and pain control are the mainstays of therapy. Recently, new developments to help increase survival and minimize morbidity with several key interventions have been investigated. This summary highlights new studies and meta-analyses to provide current opinion on treatment of this morbid condition.


Insight does not come at random: Individual gray matter networks relate to clinical and cognitive insight in schizophrenia.
Impaired clinical and cognitive insight are prevalent in schizophrenia and relate to poorer outcome. Good insight has been suggested to depend on social cognitive and metacognitive abilities requiring global integration of brain signals. Impaired insight has been related to numerous focal gray matter (GM) abnormalities distributed across the brain suggesting dysconnectivity at the global level. In this study, we test whether global integration deficiencies reflected in gray matter network connectivity underlie individual variations in insight. We used graph theory to examine whether individual GM-network metrics relate to insight in patients with a psychotic disorder (n = 114). Clinical insight was measured with the Schedule for the Assessment of Insight-Expanded and item G12 of the Positive and Negative Syndrome Scale, and cognitive insight with the Beck Cognitive Insight Scale. Individual GM-similarity networks were created from GM-segmentations of T1-weighted MRI-scans. Graph metrics were calculated using the Brain Connectivity Toolbox. Networks of schizophrenia patients with poorer clinical insight showed less segregation (i.e. clustering coefficient) into specialized subnetworks at the global level. Schizophrenia patients with poorer cognitive insight showed both less segregation and higher connectedness (i.e. lower path length) of their brain networks, making their network topology more "random". Our findings suggest less segregated processing of information in patients with poorer cognitive and clinical insight, in addition to higher connectedness in patients with poorer cognitive insight. The ability to take a critical perspective on one's symptoms (clinical insight) or views (cognitive insight) might depend especially on segregated specialized processing within distinct subnetworks.


Obtaining data on patient race, ethnicity, and primary language in health care organizations: current challenges and proposed solutions.
To provide an overview of why health care organizations (HCOs) should collect race, ethnicity, and language data, review current practices, discuss the rationale for collecting this information directly from patients, and describe barriers and solutions. Hospitals and HCOs with data from their own institutions may be more likely to look at disparities in care, design targeted programs to improve quality of care, and provide patient-centered care. Yet data collection is fragmented and incomplete within and across organizations. A major factor affecting the quality of data is the lack of understanding about how best to collect this information from patients. If HCOs make a commitment to systematically collect race/ethnicity and language data from patients, it would be a major step in enhancing the ability of HCOs to monitor health care processes and outcomes for different population groups, target quality initiatives more efficiently and effectively, and provide patient-centered care.


Acromioclavicular septic arthritis and sternoclavicular septic arthritis with contiguous pyomyositis.
Acromioclavicular (AC) and sternoclavicular (SC) septic arthritis with contiguous pyomyositis are rare, especially in immunocompetent individuals. We report a case of septic AC joint with pyomyositis of the deltoid and supraspinatus muscles and a separate case with septic SC joint with pyomysitis of the sternocleidomastoid muscle. Both patients had similar presentations of infections with Staphylococcus aureus and were successfully treated with surgical incision and drainage followed by prolonged antibiotic therapy.


Chromosome 1 rearrangements involving the genes TPR and NTRK1 produce structurally different thyroid-specific TRK oncogenes.
The NTRK1 gene in the q arm of chromosome 1 encodes one of the receptors for the nerve growth factor and is frequently activated as an oncogene in papillary thyroid carcinomas. The activation is due to chromosomal rearrangements juxtaposing the NTRK1 tyrosine kinase domain to 5'-end sequences from different genes. The thyroid TRK oncogenes are activated by recombination with at least three different genes: the gene coding for tropomyosin and TPR, both on chromosome 1,and TFG on chromosome 3. In a previous study, we showed that two tumors carrying the TPR/NTRK1 rearrangement contained structurally different oncogenes named TRK-T1 and TRK-T2. In this paper, we report (1) the cDNA structure of TRK-T2, (2) evidence that TRK-T2 is generated by different rearrangements in two thyroid tumors, and (3) a detailed analysis of the three different TPR/NTRK1 rearrangements. With molecular studies based on Southern blot hybridization, cloning, and sequencing, we show that all the rearrangements are nearly balanced, involving deletion, insertion, or duplication of only few nucleotides. In one case, an additional rearrangement involving sequences derived from chromosome 17 was detected.


Postactivation potentiation of force is independent of h-reflex excitability.
The contractile history of muscle can potentiate electrically evoked force production. A link to voluntary force production, related in part to an increase in reflex excitability, has been suggested. Our purpose was to quantify the effect of postactivation potentiation on voluntary force production and spinal H-reflex excitability during explosive plantar flexion actions. Plantar flexor twitch torque, soleus H-reflex amplitudes, and the rate of force development of explosive plantar flexion were measured before and after 4 separate conditioning trials (3 x 5 s maximal contractions). Twitch torque and rate of force production during voluntary explosive plantar flexion were significantly increased (P < .05) while H-reflex amplitudes remained unchanged. Although twitch torque was significantly higher after conditioning, leading to a small increase in the rate of voluntary force production, this was unrelated to changes in reflex excitability. We conclude that postactivation potentiation may result in a minor increase in the rate of voluntary isometric force production that is unrelated to neural excitability.


Semisynthetic lectin-4-dimethylaminopyridine conjugates for labeling and profiling glycoproteins on live cell surfaces.
Glycoproteins on cell surfaces play important roles in biological processes, including cell-cell interaction/signaling, immune response, and cell differentiation. Given the diversity of the structure of glycans, labeling and imaging of selected glycoproteins are challenging, although several promising strategies have been developed recently. Here, we design and construct semisynthetic reactive lectins (sugar-binding proteins) that are able to selectively label glycoproteins. Congerin II, an animal galectin, and wheat germ agglutinin are conjugated with 4-dimethylaminopyridine (DMAP), a well-known acyl transfer catalyst by our affinity-guided DMAP method and Cu(I)-assisted click chemistry. Selective labeling of glycoproteins is facilitated by the DMAP-tethered lectin catalysts both in vitro and on living cells. Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) analysis enabled us to isolate labeled glycoproteins that are uniquely exposed on distinct cell lines. Furthermore, the combination of immunoprecipitation with mass spectrometry (MS)-fingerprinting techniques allowed us to characterize 48 glycoproteins endogenously expressed on HeLa cells, and some low-abundant glycoproteins, such as epidermal growth factor receptor (EGFR) and neuropilin-1, were successfully identified. Our results demonstrate that semisynthetic DMAP-tethered lectins provide a new tool for labeling and profiling glycoproteins on living cells.


Pulmonary and systemic effects of the phosphodiesterase inhibitor dipyridamole in newborn lambs with persistent pulmonary hypertension.
Nitric oxide (NO) relaxes vascular smooth muscle by increasing the intracellular concentration of cGMP. In the pulmonary circulation, cGMP is inactivated by specific phosphodiesterases (PDE5). Dipyridamole, a clinically approved drug, has inhibitory activity against PDE5 and has been reported to augment the response to inhaled NO in persistent pulmonary hypertension of the newborn (PPHN). We wished to determine whether dipyridamole alone, or in combination with NO, can be used to treat a newborn lamb model of PPHN. In newborn lambs with PPHN, dipyridamole infused at 0.02 mg/kg/min for 45 min alone, or in combination with 5 ppm of inhaled NO for the final 15 min, significantly decreased pulmonary and systemic blood pressure, decreased pulmonary vascular resistance, and increased pulmonary blood flow. There was no significant difference between the pulmonary vascular effects of 5 ppm NO alone compared with the effects of NO combined with dipyridamole. In control lambs, the 45-min infusion of dipyridamole did not change pulmonary pressure whereas systemic pressure decreased by 28 +/- 3%. These systemic effects in control lambs persisted 90 min after discontinuing the dipyridamole infusion. Systemic arteries isolated from both control and PPHN lambs were significantly more sensitive to dipyridamole than pulmonary arteries. We conclude that dipyridamole has significant hemodynamic effects in both the pulmonary and systemic circulations of newborn lambs with pulmonary hypertension as well as in the systemic circulation of newborn control lambs. The pronounced effects of dipyridamole on the systemic circulation limits its utility as an adjunct to inhaled NO in the treatment of PPHN.


N-Acetyl Cysteine-Mediated Improvements in Dental Restorative Material Biocompatibility.
The fibroblast-rich gingival tissue is usually in contact with or adjacent to cytotoxic polymer-based dental restoration materials. The objective of this study was to determine whether the antioxidant amino acid, N-acetyl cysteine (NAC), reduces the toxicity of dental restorative materials. Human oral fibroblasts were cultured with bis-acrylic, flowable composite, bulk-fill composite, self-curing acrylic, and titanium alloy test specimens. Cellular behavior and function were analyzed on and around the materials. Impregnation of the bulk-fill composite and self-curing acrylic with NAC reduced their toxicity, improving the attachment, growth, and function of human oral fibroblasts on and around the materials. These mitigating effects were NAC dose dependent. However, NAC impregnation of the bis-acrylic and flowable composite was ineffective, with no cells attaching to nor around the materials. Although supplementing the culture medium with NAC also effectively improved fibroblast behaviors, direct impregnation of materials with NAC was more effective than supplementing the cultures. NAC-mediated improvements in fibroblast behavior were associated with reduced production of reactive oxygen species and oxidized glutathione together with increased glutathione reserves, indicating that NAC effectively directly scavenged ROS from materials and reinforced the cellular antioxidant defense system. These results establish a proof of concept of NAC-mediated improvements in biocompatibility in the selected dental restorative materials.


[Role of prostaglandins in regulation of kidney function in heart failure].
Conditions like heart failure that augment the activity of neurohumoral mechanisms i.e. the renin-angiotensin systems, sympathetic nerve activity and vasopressin secretion are commonly associated with a decreased effective blood volume and a reduced renal perfusion. This leads to an increased dependence of renal hemodynamics on endogenous renal prostaglandin synthesis as a vasodilator and natriuretic counter-regulating system. We investigated the role of prostaglandins in renal functional control in an experimental setting of congestive heart failure by chronic inhibition of cyclooxygenase by indomethacin. In chronic moderate heart failure plasma levels of prostaglandin E2 and prostacyclin were unchanged whereas the urinary excretion of prostaglandin E2 was significantly increased, indicating an augmented synthesis within the kidney (Figures 1 to 3). After inhibition of prostaglandin synthesis we observed a profound increase of renal vascular resistance associated with a reduction of effective renal plasma flow and renal blood flow. This was mainly due to a constriction of the vas afferens of the glomerulum. This led to an impairment of renal function indicated by an increase of serum creatinine and blood urea nitrogen associated with a reduction of urinary flow and fluid retention (Figures 4 and 5). We also studied in a randomized, double-blind, placebo-controlled, parallel-group trial in 40 patients with congestive heart failure effects of acetylsalicylic acid (500 mg t.i.d.) on renal functional parameters. In patients with normal sodium intake acetylsalicylic acid reduced urinary prostaglandin E2 concentration by 37% which led to a reduction of daily urinary sodium excretion by 29% in comparison to placebo (Figure 6). These results clearly show the importance of vasodilator prostaglandins in the regulation of kidney function in heart failure where inhibition of cyclooxygenase results in profound deterioration of renal perfusion and kidney function and retention of fluid and sodium.


Long term prognosis of fatty liver: risk of chronic liver disease and death.
Fatty liver is a common histological finding in human liver biopsy specimens. It affects 10-24% of the general population and is believed to be a marker of risk of later chronic liver disease. The present study examined the risk of development of cirrhotic liver disease and the risk of death in a cohort diagnosed with pure fatty liver without inflammation. A total of 215 patients who had a liver biopsy performed during the period 1976-1987 were included in the study. The population consisted of 109 non-alcoholic and 106 alcoholic fatty liver patients. Median follow up time was 16.7 (0.2-21.9) years in the non-alcoholic and 9.2 (0.6-23.1) years in the alcoholic group. Systematic data collection was carried out by review of all medical records. All members of the study cohort were linked through their unique personal identification number to the National Registry of Patients and the nationwide Registry of Causes of Death, and all admissions, discharge diagnoses, and causes of death were obtained. In the non-alcoholic fatty liver group, one patient developed cirrhosis during the follow up period compared with 22 patients in the alcoholic group. Survival estimates were significantly (p<0.01) different between the two groups, for men as well as for women, with a higher death rate in the alcoholic fatty liver group. Survival estimates in the non-alcoholic fatty liver group were not different from the Danish population. This study revealed that patients with type 1 non-alcoholic fatty liver disease have a benign clinical course without excess mortality.


Temporal reproducibility of IgG and IgM autoantibodies in serum from healthy women.
Autoantibodies are present in healthy individuals and altered in chronic diseases. We used repeated samples collected from participants in the NYU Women's Health Study to assess autoantibody reproducibility and repertoire stability over a one-year period using the HuProt array. We included two samples collected one year apart from each of 46 healthy women (92 samples). We also included eight blinded replicate samples to assess laboratory reproducibility. A total of 21,211 IgG and IgM autoantibodies were interrogated. Of those, 86% of IgG (n = 18,303) and 34% of IgM (n = 7,242) autoantibodies showed adequate lab reproducibility (coefficient of variation [CV] < 20%). Intraclass correlation coefficients (ICCs) were estimated to assess temporal reproducibility. A high proportion of both IgG and IgM autoantibodies with CV < 20% (76% and 98%, respectively) showed excellent temporal reproducibility (ICC > 0.8). Temporal reproducibility was lower after using quantile normalization suggesting that batch variability was not an important source of error, and that normalization removed some informative biological information. To our knowledge this study is the largest in terms of sample size and autoantibody numbers to assess autoantibody reproducibility in healthy women. The results suggest that for many autoantibodies a single measurement may be used to rank individuals in studies of autoantibodies as etiologic markers of disease.


Evaluation of a multiplex panel for the diagnosis of acute infectious diarrhea in immunocompromised hematologic patients.
Diarrhea is a frequent complication in hematologic patients, being an infectious cause frequently suspected. Rapid and accurate detection of gastrointestinal pathogens is vital in immunocompromised hosts. The aim of this study was to compare routine diagnostic methods versus a multiplex polymerase chain reaction (PCR) assay for the diagnosis of infectious diarrhea in immunocompromised hematologic patients. We conducted a prospective observational study from March 2015 to January 2016 to compare conventional methods for the diagnosis of infectious diarrhea with FIlmArray GI Panel (BioFire-bioMérieux, France). Samples from adult immunocompromised hematologic patients with acute diarrhea were collected. In cases with discordant results, a second multiplex assay was performed (Allplex, Seegene, Korea). The result was considered positive or negative when the same result was obtained by at least two of the methods. A total of 95 samples were obtained from 95 patients (median age of 52 years (46-64)). Sixty-one (64%) episodes were hospital-acquired and 34 (36%) were community-acquired diarrhea. Twenty-five (26%) patients had a positive microbiological result, being Clostridium difficile the most frequent pathogen, followed by Campylobacter spp and norovirus. The concordance between FilmArray methods was good (k = 0.79). The FilmArray GI panel showed a sensitivity of 95%, a specificity of 100% for positive results. The time required to obtain results was markedly reduced with the use of multiplex PCR methods. Multiplex molecular panels provide a rapid and sensitive tool for the diagnosis of infectious diarrhea, thereby allowing more timely clinical decisions in immunocompromised hematologic patients.


Coamplification of prostate stem cell antigen (PSCA) and MYC in locally advanced prostate cancer.
Gain of sequences on chromosome arm 8q is a common feature of prostate cancer that may correlate with metastatic and androgen-independent progression. The target gene(s) for this gain is not known, although MYC is amplified in a subset of advanced tumors and is one potential candidate. Prostate stem cell antigen (PSCA) is a prostate-specific cell surface protein that maps to chromosome region 8q24.2 and is overexpressed in prostate cancer. Our aim in this study was to test the hypothesis that PSCA overexpression may result from overrepresentation of chromosome arm 8q. Twenty locally advanced prostate cancers were analyzed by dual-probe fluorescence in situ hybridization (FISH) for alterations of MYC and PSCA. Extra copies of MYC were found in 12/20 (60%) tumors, including 5 (25%) with simple gain (no increase in MYC copy number relative to the chromosome 8 centromere) and 7 (35%) with an additional increase (AI or overrepresentation) in MYC copy number relative to the centromere. In the five cases with simple gain of MYC, there was a concomitant gain of PSCA. PSCA was overrepresented in 5/7 (71%) cases with AI of MYC. Immunohistochemical staining of the 20 tumors with monoclonal antibodies specific for PSCA showed a high degree of correlation between PSCA gene overrepresentation and protein overexpression. Four of 5 tumors with AI of PSCA overexpressed PSCA protein, compared with only 2/15 tumors with a normal PSCA copy number or simple gain of PSCA (P = 0.014). These results demonstrate that PSCA is co-overrepresented with MYC in a majority of cases, but may not be a necessary part of the 8q amplicon. PSCA protein overexpression can result from AI of PSCA and might be useful as a cell surface marker on prostate cancer cells with 8q overrepresentation. Genes Chromosomes Cancer 27:95-103, 2000.


Automated software for counting and measuring Hyalella genus using artificial intelligence.
Amphipods belonging to the Hyalella genus are macroinvertebrates that inhabit aquatic environments. They are of particular interest in areas such as limnology and ecotoxicology, where data on the number of Hyalella individuals and their allometric measurements are used to assess the environmental dynamics of aquatic ecosystems. In this study, we introduce HyACS, a software tool that uses a model developed with the YOLOv3's architecture to detect individuals, and digital image processing techniques to extract morphological metrics of the Hyalella genus. The software detects body metrics of length, arc length, maximum width, eccentricity, perimeter, and area of Hyalella individuals, using basic imaging capture equipment. The performance metrics indicate that the model developed can achieve high prediction levels, with an accuracy above 90% for the correct identification of individuals. It can perform up to four times faster than traditional visual counting methods and provide precise morphological measurements of Hyalella individuals, which may improve further studies of the species populations and enhance their use as bioindicators of water quality.


The effect of protocatechuic acid on ovarian histopathology and reserve in rat ovarian torsion model.
The aim of the study is to investigate the effects of Protocatechuic Acid (PCA), which is an antioxidant, anti-inflammatory and anti-apoptotic agent, on ovarian tissue and ovarian reserve against ischemia-reperfusion (IR) injury in a rat ovarian torsion model. Reactive oxygen radicals cause histopathological changes in the ovarian tissue during the reperfusion phase. PCA may have protective effects on ovarian tissue and reserve due to its antioxidant and antiapoptotic properties. A total of 24 Wistar adult female rats were divided into 3 groups as the control (sham operation, n = 8), IR (Ischemia-Reperfusion, n = 8), and IR+PCA (Ischemia-Reperfusion + 80 mg/kg protocatechuic acid, n = 8). The IR and IR + PCA groups underwent 3 hours of ischemia followed by 3 hours of ovarian reperfusion. Protocatechuic acid (80 mg/kg) was administered to the IR+PCA group 30 minutes before reperfusion. After reperfusion, the ovaries were removed for histopathological and biochemical examination. Histopathological score and TUNEL+ cell count were significantly lower and AMH expression level was significantly higher in the IR+PCA group when compared to the IR group (p <0.05). However, in the comparison of the follicle counts, there was no statistically significant difference between all groups. Due to the increase in antioxidant activity, the MDA levels were found to be significantly lower in the IR+PCA group compared to the IR group (p < 0.05). Protocatechuic acid may be an effective antioxidant in protecting ovarian tissue and follicle reserve against IR injury of the ovary (Tab. 1, Fig. 4, Ref. 36).


[Cavernous angioma. Clinical observations and prognosis of 133 patients].
Cavernous angiomas represent 9% of the vascular malformations that affect nervous system. The principal mode of onset is cerebral hemorrhage and epilepsy, and can be sporadic of multiple. In the last, there is a familiar factor specially found in Mexician-American. In our Country there is no data of the clinical or demographic characteristics of the disease. The aim of this study is to describe the clinical characteristics and prognosis of 133 patients with cavernous angiomas consecutively attended in the National Institute of Neurology of Mexico City. Since 1988 we evaluated a total of 146 cases of cavernous angiomas, 133 were confirmed by MRI and included in the analysis of this study. In every case we registered the demographic characteristic, neurological manifestations, and prognosis divided in good o bad outcome, according to the Glasgow outcome scale (1 and > or = 2 respectively). Data were analyzed with descriptive statistics with the Chi square test, and p was set at 0.05 level. The mean age was 34.3 +/- 14.6 years; 50.4% in male. Eighty seven percent were unique, and the principal manifestation was supratentorial in 65%, infratentorial in 24.8%, supra-infratentorial in 6.8%, and spinal in 3.8%. The clinical manifestations were intracerebral hemorrhage in 58.7%, epilepsy in 48.1%, headache in 37.6%, neurological focalization not secondary to hemorrhage in 8.3%, and incidental in 2.3%. The 6 month outcome was good in 80% of patient assessed by the Glasgow outcome scale. In our serie the mean age or presentation was 34 years old, the mean neurological manifestations were cerebral hemorrhage and epilepsy, and the outcome was good in the majority of the cases.


Effect of ozone on DNA-repair deficient mutants of Bacillus subtilis.
The wild type strains and DNA-repair deficient mutant strains of B. subtilis were exposed to ozone. The results show that the lethal effect of ozone is greatly influenced by factors such as ozone concentration, treatment time, growth phase, and treatment of medium. The recA and recC loci seem to be most important and perhaps the polA and uvrA loci may also be required, but the recB and recD loci are not required for the repair of DNA-damase induced by ozone in B. subtilis.


[The age-dependent effect of copper ions on different strains of the wine yeast (Saccharomyces [corrected] cerevisiae var vini)].
The chronological ageing of wine yeast strains isolated from natural populations and the resistance of cells of different age to copper ions have been studied. In total, 25 wine yeast strains derived from the "Tsolikauri" population of Sviri village (Imereti, West Georgia) were tested. The strains were transferred to the standard (control samples) and the copper sulphate-containing complete media and stored at 2 degrees C temperature. 10, 20 and 30 days after setting up the cultures their resistance to copper ions were examined. Methylene blue dye was used for identification of survived cells. Young strain cultures have revealed different resistance to copper ions. The strains under investigation may be conditionally divided into three morphs, such as: "sensitive" (capable of growing on 1-2 mM medium. 6 strains belong to this group), "mild" (grow on 3-4 mM medium--17 strains and "resistant" (can grow on 5 mM medium, includes 3 strains). The population was found to be polymorphic by the resistance to copper ions. The "sensitive" and "resistant" strains in spite of ageing (10 or 20 days after the onset of culturing) retain their resistance to copper ions, while the "mild" morphs exhibited some degree of variability. In the "aged" (30-day long) cultures strong inhibitory activity of copper ions was apparently observed. In contrast to the others, the resistance was sustained in the "resistant" morphs. The frequency of living cells was defined in the strains grown on the control and copper ion-containing substrates. Survived and died cells were counted. The strains belonged to the sensitive morphs contained high number of died cells--26-30%. The same index was rather low in the other morphs with no difference between them. Further ageing of the cultures (30 days and more) caused so sharp decrease in the liability of the strains, that they completely lose their resistance. Thus, the examined populations of the wine yeast were found to be polymorphic by their lifespan. They differ from each other by the speed of chronological ageing, that should be attributed to their genetic structure.


[Immunogenic properties of brucella protective antigen upon revaccination].
The authors generalized the materials on the experimental study of the immunological efficacy of brucella protective antigen in revaccination. Revaccination 12 months after the immunization of guinea pigs with an antigen in a dose equal to the primary immunization dose (0.6 mg) reproduced the protection from the infection with a virulent Br. melitensis culture in 78% of the animals. The antigen proved to be effective only at the remote period between the revaccination and the primary antigen vaccination (12 months). The data obtained pointed to a high protective action of the antigen in the guinea pig revaccination 9 to 12 months after their immunization with live brucella Br. abortus 19-BA vaccine.


Metal salts and complexes of 1,1'-dinitramino-5,5'-bitetrazole.
Potassium 1,1'-dinitramino-5,5'-bitetrazolate is one of the most promising primary explosives which is currently under investigation for different applications. This is due to its high initiation power and the exclusion of heavy metals. To close the gap, the remaining alkali metal salts such as the lithium 6, sodium 7, rubidium 8 and cesium 9 salts were synthesized by reaction of the highly soluble ammonium salt 5 with its corresponding metal hydroxide solutions. In addition, the highly explosive silver salt 10 as well as several other transition metal(ii) amine complexes with nickel(ii) 11, copper(ii) 12 and zinc(ii) 13 were prepared in a similar manner. The structure of all compounds was determined by X-ray diffraction. The sensitivities toward impact, friction, heat and electrostatic discharge as well as their behavior on laser irradiation of the transition metal complexes were explored.


Quantitative analysis of the selectivity of radioligands for subtypes of beta adrenergic receptors.
In tissues with two classes of binding sites for a drug, it is common to estimate the proportion of each class of binding site by inhibiting the binding of a radioligand with a selective unlabeled ligand. Accurate estimates of the density of each class of binding site, however, will be obtained only if the radioligand is nonselective or used at a concentration that saturates both classes of binding sites. A method of simultaneous regression analysis of multiple inhibition curves, using the program MLAB on the PROPHET system, was used to quantify the selectivity of radioligands for beta-1 or beta-2 adrenergic receptors. The selectivity of [125I]iodopindolol, [125I]iodocyanopindolol, [125I]iodohydroxybenzylpindolol and [3H]dihydroalprenolol for beta-1 and beta-2 adrenergic receptors was assessed by inhibiting the binding of each radioligand with the beta-1-selective unlabeled ligand ICI 89,406 at increasing concentrations of the radioligand, using membranes prepared from C6 glioma cells, which have both beta-1 and beta-2 adrenergic receptors. Scatchard plots for all four radioligands were linear, with correlation coefficients greater than 0.95. [125I]Iodopindolol and [125I]iodocyanopindolol were 3.2- and 2-fold selective, respectively, and [125I]iodohydroxybenzylpindolol and [3H]dihydroalprenolol were 5.8- and 2.3-fold selective, respectively, for beta-2 adrenergic receptors. Values obtained for the densities of beta-1 and beta-2 adrenergic receptors and the affinities of the receptors for ICI 89,406 were independent of the radioligand used.(ABSTRACT TRUNCATED AT 250 WORDS)


Increased DNA methyltransferase 1 protein expression in human transitional cell carcinoma of the bladder.
DNA methylation is a key regulator of gene transcription and genomic stability, and alteration of DNA methylation is one of the most consistent epigenetic changes in human cancers. We elucidated the significance of aberrant protein expression of DNA methyltransferase (DNMT) 1, a major enzyme involved in the determination of genomic methylation patterns, during human urothelial carcinogenesis. A total of 61 samples of normal urothelium, 89 noncancerous urothelium samples showing no remarkable histological changes from patients with bladder cancer (NBCs), 78 dysplastic urothelium samples and 174 transitional cell carcinoma samples (TCCs) were subjected to immunohistochemical analysis for DNMT1. The incidence of nuclear DNMT1 immunoreactivity in NBCs (65%) was significantly higher than in normal urothelium (20%, p <0.0001) and the incidence was even higher in dysplastic urothelium samples (84%, p = 0.0017). The incidence of nuclear DNMT1 immunoreactivity was 87% in TCCs and the intensity of nuclear immunoreactivity was markedly increased in TCCs compared with that in dysplastic urothelium samples. DNMT1 expression levels had already increased in NBCs in which the proliferating cell nuclear antigen labeling index had not yet increased. Increased DNMT1 protein expression correlated significantly with histological grade (p <0.0001). DNMT1 protein expression was higher in nonpapillary tumors (p = 0.0001), especially flat carcinoma in situ, than in papillary tumors. Progressively increasing expression of DNMT1 protein is not entirely a secondary result of increased cell proliferative activity, but rather it is associated with urothelial carcinogenesis even during the precancerous stages. In particular, it is associated with the development of flat carcinoma in situ, which is considered to be a precursor of nodular invasive carcinoma of the bladder.


Pressure flow analysis may aid in identifying women with outflow obstruction.
We refined recently developed pressure flow cutoff values for female bladder outlet obstruction and applied these values in a consecutive group of women undergoing urodynamic testing for various lower urinary tract symptoms. A total of 87 women with clinical obstruction determined by history and presenting complaint were enrolled in our prospective evaluation of pressure flow studies. We identified 3 groups of participants according to the suspected cause of obstruction, including prolapse in 33, previous incontinence surgery in 25, and no likely source of obstruction identified from history and physical examination only in 29. An additional 124 patients presenting for evaluation of stress urinary incontinence served as controls. Optimal cutoff values for determining obstruction were developed using receiver operating characteristic curves. To determine the prevalence of bladder outlet obstruction these values were prospectively applied to 106 women undergoing urodynamics for various voiding complaints. In controls the average maximum flow rate was 23 cc per second and average detrusor pressure was 21.9 cm. water, whereas the corresponding values in those with clinical obstruction were 10.7 cc per second and 40.8 cm. water (p <0.001). No differences were noted in the various obstruction groups. Receiver operating characteristics analysis revealed that cutoff values of 11 cc per second or less and 21 cm. water or more optimized the selection of patients with bladder outlet obstruction. Using these values we noted a bladder outlet obstruction prevalence of 20% in a consecutive cohort of women undergoing urodynamic studies at our center. We propose cutoff pressure flow values for identifying women with bladder outlet obstruction although they should be used only in conjunction with the overall clinical situation. Neither pressure flow data only nor clinical symptoms alone may be sufficient for diagnosing obstruction in women.


Predictive potential role of glutathione S-transferases polymorphisms on prognosis of breast cancer.
The aim of this study was to evaluate the clinical response to chemotherapy and treatment outcome of breast cancers patients in the presence of the GSTM1 null/present, GSTT1 null/present, and GSTP1 IIe105Val polymorphisms. Genotyping of GSTP1 rs1695, GSTT1 deletion and GSTM1 deletion was carried out on a 384-well plate format on the Sequenom MassARRAY platform. Of 382 patients, 202 patients showed good response to chemotherapy, 51 died, and 155 showed progression at the end of the study. Patients carrying GG genotype and G allele of GSTP1 rs1695 were associated with poor response to chemotherapy. In the Cox proportional hazards model, after adjusting for potential confounding factors, patients carrying GG genotype and G allele of GSTP1 rs1695 were correlated with a shorter overall survival (OS). Variants of GSTP1 rs1695 are associated with response to chemotherapy and PFS and OS of breast cancer patients, and this gene polymorphism could help in the design of individualized therapy.


Summary measures were a useful alternative for analyzing therapeutic clinical trial data.
To compare use of both a pooled index and area under the curve (AUC) to the standard analysis of a randomized controlled trial of antibiotics for patients with Alzheimer's disease. Using data from a randomized, placebo-controlled trial of antibiotics for patients with Alzheimer's disease, a pooled index of six outcome measures was constructed. Each change score was standardized by dividing by the standard deviation, and the six standardized change scores were averaged. The Standardized Alzheimer's Disease Assessment Scale cognitive subscale and the pooled index were plotted against time and the AUC was calculated. The AUC analysis of the pooled index showed significant treatment effect over the 12-month period. In contrast, none of the individual measures showed an effect at more than one time point. The AUC analyses of the Standardized Alzheimer's Disease Assessment Scale cognitive subscale, the primary outcome of the trial, showed no significant difference over the entire 12-month period, although the original individual time-point analysis showed a difference at 6 months alone. Pooled indices and AUC analyses may provide important insight into therapeutic effect of agents tested in randomized clinical trials of patients with Alzheimer's disease.


Confirmation of the R653Q polymorphism of the trifunctional C1-synthase enzyme as a maternal risk for neural tube defects in the Irish population.
The risk of neural tube defects (NTDs) is known to have a significant genetic component that could act through either the NTD patient and/or maternal genotype. The success of folic acid supplementation in NTD prevention has focused attention on polymorphisms within folate-related genes. We previously identified the 1958G>A (R653Q) polymorphism of the trifunctional enzyme MTHFD1 (methylenetetrahydrofolate-dehydrogenase, methenyltetrahydrofolate-cyclohydrolase, formyltetrahydrofolate synthetase; often referred to as 'C1 synthase') as a maternal risk for NTDs, but this association remains to be verified in a separate study to rule out a chance finding. To exclude this possibility, we genotyped an independent sample of mothers with a history of an NTD-affected pregnancy derived from the same Irish population. In this sample there was a significant excess of 1958AA homozygote mothers of NTD cases (n=245) compared to controls (n=770). The direction and magnitude of risk (odds ratio 1.49 (1.07-2.09), P=0.019) is consistent with our earlier finding. Sequencing of the MTHFD1 gene revealed that this association is not being driven by another common variant within the coding region. We have established that the MTHFD1 1958G>A polymorphism has a significant role in influencing a mother's risk of having an NTD-affected pregnancy in the Irish population.


iTRAQ experimental design for plasma biomarker discovery.
There is considerable interest in using mass spectrometry for biomarker discovery in human blood plasma. We investigated aspects of experimental design for large studies that require analysis of multiple sample sets using iTRAQ reagents for sample multiplexing and quantitation. Immunodepleted plasma samples from healthy volunteers were compared to immunodepleted plasma from patients with osteoarthritis in eight separate iTRAQ experiments. Our analyses utilizing ProteinPilot software for peptide identification and quantitation showed that the methodology afforded excellent reproducibility from run to run for determining protein level ratios (coefficient of variation 11.7%), in spite of considerable quantitative variances observed between different peptides for a given protein. Peptides with high variances were associated with lower intensity iTRAQ reporter ions, while immunodepletion prior to sample analysis had a negligible affect on quantitative variance. We examined the influence of different reference samples, such as pooled samples or individual samples on calculating quantitative ratios. Our findings are discussed in the context of optimizing iTRAQ experimental design for robust plasma-based biomarker discovery.


The role of growth factors and ammonia in the genesis of hypokalemic nephropathy.
Hypokalemia is a common electrolyte abnormality encountered in clinical practice. It can be identified in an asymptomatic patient undergoing routine electrolyte screening or can manifest itself as part of a number of functional abnormalities in a variety of organs and systems. Among the most commonly recognized complications are profound effects on the cardiovascular and neuromuscular systems, together with abnormalities in acid-base regulation. In humans, hypokalemia contributes to the development of hypertension and predisposes patients to a variety of ventricular arrhythmias, including ventricular fibrillation. Commonly recognized neuromuscular complications include weakness, cramping, and myalgia. Hypokalemia also affects systemic acid-base homeostasis by interfering with multiple components of the renal acid-base regulation and is a frequent cause of metabolic alkalosis. Less known, however, is the role of potassium deficiency in causing progressive renal failure. In animals, potassium deficiency stimulates renal enlargement because of cellular hypertrophy and hyperplasia. If potassium deficiency persists, interstitial infiltrates appear in the renal interstitial compartment, and eventually tubulointerstitial fibrosis develops. In humans, longstanding hypokalemia is associated with the development of renal cysts, chronic interstitial nephritis, and progressive loss of renal function, the so-called hypokalemic nephropathy. This review focuses on the potential mechanisms involved in the development of the hypokalemic nephropathy with emphasis on the role of ammonia and growth factors in its pathogenesis.


Measurement models for sexual orientation in a community twin sample.
Multivariate structural equation modeling techniques have been applied to examine the causes of individual differences in responses to several items concerning sexual orientation. To minimize potential ascertainment and response biases, the study sample involved a large (N = 4901) community-based cohort of Australian twins aged 18-52 who answered an anonymous questionnaire on sexual behavior and attitudes. The statistical power of the analysis was increased by the availability of multiple measures of sexual orientation (behaviors, attitudes and feelings), providing stronger evidence for the existence of additive genetic influences on this phenotype than in a previous analysis (Bailey et al., 2000). Estimates of the heritability of homosexuality in this sample ranged between 50 and 60% in females but were significantly lower (heritability of approximately 30%) in males.


Progress in the performance of HIV early infant diagnosis services in Zambia using routinely collected data from 2006 to 2016.
Early diagnosis and treatment initiation of HIV-infected infants can greatly reduce the risk of infant mortality. The WHO recommends testing HIV-exposed infants at 6 weeks of age and immediate initiation of antiretroviral therapy if positive. This study aimed to determine the feasibility of using an electronic health records system to evaluate the performance of Zambia's HIV Early Infant Diagnosis services. A retrospective analysis of routinely collected data from the Zambian SmartCare database was performed for the period January 2006 to December 2016. The study population includes all HIV-infected infants (n = 32,593) registered during this period on treatment for HIV. Univariable logistic regression was conducted to identify factors associated with later infant testing and treatment initiation. The mean age at infant HIV test decreased from 10.10 months in 2006 to 3.49 months in 2016. Infants born in 2015 were almost 4 times more likely to be tested under 2 months of age compared to infants born in 2006 (OR: 3.72, p-value: < 0.001). The mean time from diagnosis to treatment initiation decreased from 220 days in 2006 to 9 days in 2015. There was substantial regional variability with infants in the provinces of Copperbelt, Luapula and Southern performing best in outcomes and Eastern, Lusaka and Western performing the worst. HIV-exposed infants born more recently have significantly better outcomes than infants born a decade ago in Zambia, which could be as a result of increased attention and funding for HIV programmes.


Dynamic mechanisms of blood vessel growth.
The formation of a polygonal configuration of proto-blood-vessels from initially dispersed cells is the first step in the development of the circulatory system in vertebrates. This initial vascular network later expands to form new blood vessels, primarily via a sprouting mechanism. We review a range of recent results obtained with a Monte Carlo model of chemotactically migrating cells which can explain both de novo blood vessel growth and aspects of blood vessel sprouting. We propose that the initial network forms via a percolation-like instability depending on cell shape, or through an alternative contact-inhibition of motility mechanism which also reproduces aspects of sprouting blood vessel growth.


COX-2 from the injury milieu is critical for the initiation of periosteal progenitor cell mediated bone healing.
Although a critical role of COX-2 in bone repair has been established, the mechanism involved remains unclear. During early inflammatory phase of bone healing, COX-2 is produced by the surrounding inflammatory cells as well as bone/cartilage progenitors. Based on the temporal and spatial expression of COX-2 during the early phase of fracture healing, we hypothesize that COX-2 from both sources is critical for progenitor cell activation, proliferation and differentiation. To directly test this we utilized a murine femoral grafting model, in which live segmental grafts from the same strains were transplanted and donor versus host cell involvement in healing was assessed. Specifically, fresh femur cortical bone grafts of 4 mm in length from COX-2(-/-) (KO) mice were transplanted into wild type (WT) mice with the same sized segmental defect in femurs. Similarly, grafts from WT were transplanted into the defects in KO mice. As controls, transplantations between wild types, and transplantations between KO were also performed. Histologic analyses showed that WT-to-WT transplantation resulted in normal endochondral bone healing as evidenced by markedly induction of neovascularization and periosteal bone formation on donor graft. In contrast, transplantation of KO graft into KO host led to 96% reduction of bone formation and near elimination of donor cell-initiated periosteal bone formation. Similarly, transplantation of WT graft into a KO host resulted in 87% reduction of bone formation (n=8, p>0.05), indicating that KO host impaired WT donor progenitor cell expansion and differentiation. When a KO graft was transplanted into WT host, KO donor periosteal cell-initiated endochondral bone formation was restored. Histomorphometric analyses demonstrated 10-fold increase in bone formation and 3-fold increase in cartilage formation compared to KO-to-KO transplantation (n=8, p<0.05), suggesting that COX-2 deficient donor cells were capable to differentiate and form bone when placed in a WT host. Taken together, our data strongly suggest that COX-2 is critical for initiation of periosteal cortical bone healing. The early induction of COX-2 constitutes a crucial host-healing environment for activation and differentiation of donor periosteal progenitors. Elimination of COX-2 at the early stage of healing could lead to detrimental effects on periosteal progenitor cell-initiated cortical bone repair.


Ultrasensitive detection of tumor-specific mutations in saliva of patients with oral cavity squamous cell carcinoma.
Oral cavity squamous cell carcinoma (OCSCC) is the most common head and neck malignancy. Although the survival rate of patients with advanced-stage disease remains approximately 20% to 60%, when detected at an early stage, the survival rate approaches 80%, posing a pressing need for a well validated profiling method to assess patients who have a high risk of developing OCSCC. Tumor DNA detection in saliva may provide a robust biomarker platform that overcomes the limitations of current diagnostic tests. However, there is no routine saliva-based screening method for patients with OCSCC. The authors designed a custom next-generation sequencing panel with unique molecular identifiers that covers coding regions of 7 frequently mutated genes in OCSCC and applied it on DNA extracted from 121 treatment-naive OCSCC tumors and matched preoperative saliva specimens. By using stringent variant-calling criteria, mutations were detected in 106 tumors, consistent with a predicted detection rate ≥88%. Moreover, mutations identified in primary malignancies were also detected in 93% of saliva samples. To ensure that variants are not errors resulting in false-positive calls, a multistep analytical validation of this approach was performed: 1) re-sequencing of 46 saliva samples confirmed 88% of somatic variants; 2) no functionally relevant mutations were detected in saliva samples from 11 healthy individuals without a history of tobacco or alcohol; and 3) using a panel of 7 synthetic loci across 8 sequencing runs, it was confirmed that the platform developed is reproducible and provides sensitivity on par with droplet digital polymerase chain reaction. The current data highlight the feasibility of somatic mutation identification in driver genes in saliva collected at the time of OCSCC diagnosis.


The Society for Vascular Surgery lower extremity threatened limb classification system based on Wound, Ischemia, and foot Infection (WIfI) correlates with risk of major amputation and time to wound healing.
The purpose of this study was to evaluate whether the new Society for Vascular Surgery (SVS) Wound, Ischemia, and foot Infection (WIfI) classification system correlates with important clinical outcomes for limb salvage and wound healing. A total of 201 consecutive patients with threatened limbs treated from 2010 to 2011 in an academic medical center were analyzed. These patients were stratified into clinical stages 1 to 4 on the basis of the SVS WIfI classification. The SVS objective performance goals of major amputation, 1-year amputation-free survival (AFS) rate, and wound healing time (WHT) according to WIfI clinical stages were compared. The mean age was 58 years (79% male, 93% with diabetes). Forty-two patients required major amputation (21%); 159 (78%) had limb salvage. The amputation group had a significantly higher prevalence of advanced stage 4 patients (P < .001), whereas the limb salvage group presented predominantly as stages 1 to 3. Patients in clinical stages 3 and 4 had a significantly higher incidence of amputation (P < .001), decreased AFS (P < .001), and delayed WHT (P < .002) compared with those in stages 1 and 2. Among patients presenting with stage 3, primarily as a result of wound and ischemia grades, revascularization resulted in accelerated WHT (P = .008). These data support the underlying concept of the SVS WIfI, that an appropriate classification system correlates with important clinical outcomes for limb salvage and wound healing. As the clinical stage progresses, the risk of major amputation increases, 1-year AFS declines, and WHT is prolonged. We further demonstrated benefit of revascularization to improve WHT in selected patients, especially those in stage 3. Future efforts are warranted to incorporate the SVS WIfI classification into clinical decision-making algorithms in conjunction with a comorbidity index and anatomic classification.


In silico and in vitro evaluation of tetrahydropyridine compounds as efflux inhibitors in Mycobacterium abscessus.
Herein, we evaluated tetrahydropyridine (THP) compounds (NUNM) as antimicrobials and inhibitors of the efflux mechanism in M. abscessus. subsp. abscessus. The modulation factor (MF) of efflux inhibitors was calculated from the minimum inhibitory concentrations (MICs) of amikacin (AMI), ciprofloxacin (CIP) and clarithromycin (CLA) in the absence and presence of subinhibitory concentrations of the NUNM compounds and canonical inhibitors carbonyl cyanide m-chlorophenyl hydrazone (CCCP) and verapamil (VP). The kinetics of the intracellular accumulation of the fluorimetric substrate ethidium bromide (EtBr) was evaluated and calculated by the relative final fluorescence (RFF). In addition, molecular modeling simulations for the MmpL5 and Tap efflux transporters with ligands (CLA, NUNM, CCCP, VP and EtBr) were performed to better understand the efflux mechanism. We highlight the NUNM01 compound because it reduced the MICs of AMI, CIP and CLA by 4-, 4- and 16-fold, respectively, had the highest effect on EtBr accumulation (RFF = 3.1) and showed a significant in silico affinity for the evaluated proteins in docking simulations. Based on the analyses performed in vitro and in silico, we propose that NUNM01 is a potential pharmacophore candidate for the development of a therapeutic adjuvant for M. abscessus infections.


A novel SMOC1 pathogenic homozygous variant in a fetus with mesomelia of the lower limbs, micrognathia and hypertelorism and an incidental finding of CYP21A2-related congenital adrenal hyperplasia.
Trio exome sequencing was performed on a fetus with bilateral mesomelia of the lower limbs with significant angulation of the tibial bones, micrognathia and hypertelorism detected on ultrasound scan at 19 + 0 weeks gestation. The couple is consanguineous. A homozygous pathogenic frameshift variant in the SMOC1 gene (c.339_340del p.(Phe114Cysfs*40)) was detected and both parents were shown to be heterozygous. Pathogenic variants in the SMOC1 gene are associated with microphthalmia with limb anomalies which multidisciplinary team discussion determined to be causal of the scan anomalies detected. The fetus was also a compound heterozygote for CYP21A2 pathogenic variants, confirming a second diagnosis of non-classical congenital adrenal hyperplasia, which was felt incidental to the scan findings. The risk that this couple's next pregnancy would be affected by either of these disorders is 1 in 4 (25%) and demonstrates the importance of genetic diagnoses for the family and implications for future pregnancies.


Women's experiences and expectations after disclosure of intimate partner abuse to a healthcare provider: A qualitative meta-synthesis.
To identify and synthesise the experiences and expectations of women victim/survivors of intimate partner abuse (IPA) following disclosure to a healthcare provider (HCP). The databases MEDLINE, Embase, CINAHL, PsychINFO, SocINDEX, ASSIA and the Cochrane Library were searched in February 2020. Included studies needed to focus on women's experiences with and expectations of HCPs after disclosure of IPA. We considered primary studies using qualitative methods for both data collection and analysis published since 2004. Studies conducted in any country, in any type of healthcare setting, were included. The quality of individual studies was assessed using an adaptation of the Critical Appraisal Skills Programme checklist for qualitative studies. The confidence in the overall evidence base was determined using Grading of Recommendations, Assessment, Development and Evaluations (GRADE)-Confidence in the Evidence from Reviews of Qualitative Research methods. Thematic synthesis was used for analysis. Thirty-one papers describing 30 studies were included in the final review. These were conducted in a range of health settings, predominantly in the USA and other high-income countries. All studies were in English. Four main themes were developed through the analysis, describing women's experiences and expectations of HCPs: (1) connection through kindness and care; (2) see the evil, hear the evil, speak the evil; (3) do more than just listen; and (4) plant the right seed. If these key expectations were absent from care, it resulted in a range of negative emotional impacts for women. Our findings strongly align with the principles of woman-centred care, indicating that women value emotional connection, practical support through action and advocacy and an approach that recognises their autonomy and is tailored to their individual needs. Drawing on the evidence, we have developed a best practice model to guide practitioners in how to deliver woman-centred care. This review has critical implications for practice, highlighting the simplicity of what HCPs can do to support women experiencing IPA, although its applicability to low-income and-middle income settings remains to be explored.


Place of dexfenfluramine in the management of obesity.
The treatment of obesity remains a puzzling challenge because long-term maintenance of weight loss--one of the most suitable goals--is rarely achieved with conventional methods. Among the theoretical measures able to maintain a permanent and bearable constraint to obtain the maintenance of weight loss is long-term (life-long?) pharmacotherapy. Dexfenfluramine is a good candidate for such a use; the demonstration of its long-term efficacy, i.e., maintenance of the initial weight loss with no escape with time, is in progress (ISIS study). Although the correct use of dexfenfluramine in obese patients remains to be more precisely defined, its pharmacological profile leads us to consider that it could be useful for those with normo or hyperphagia and stress induced compulsive feeding behavior, and/or for avoiding relapse in restrained patients. The use of dexfenfluramine must not excuse other types of interventions when needed.


Patient experiences of closed-incision negative pressure therapy on groin incisions after discharge following peripheral arterial surgery: A qualitative study.
Closed-incision negative pressure therapy may lower the risk of surgical site infections in patients after peripheral arterial surgery. To explore patient experience of negative pressure therapy applied to groin incisions after discharge following peripheral arterial surgery, and to study their perception and attitudes toward the self-care information sheet they received at the vascular department. A qualitative study underpinned by Gadamer's philosophical hermeneutics was conducted semi-structured interviews by telephone around day seven after therapy ended with ten participants. All had received self-care information sheet at the discharge and been home with closed-incision negative pressure therapy for 3-6 days. The participants had open peripheral arterial surgery in the groin in form of femoral thromboendarterectomy. Kvale and Brinkmann's research guided the data collection, analysis, and interpretation. Patients found themselves coping with an unfamiliar situation after peripheral arterial surgery and the need arose to conceal the pump and tubing that were part of their incision treatment to protect their self-image. Their treatment became a constant companion, with some patients viewing the equipment as an extension of their bodies and others feeling its impact on activities of daily living. Patients perceived the treatment as providing reassurance, albeit with constraints, leading to feelings of manageability and an increasing sense of control. They viewed the written information as informative but with room for improvement. Patient experiences of closed-incision negative pressure therapy on groin incisions after discharge following peripheral arterial surgery showed that they perceived it as safe and manageable. Patients need support, however, in learning how to hide the treatment and to expand their own involvement and improve self-care. The study found that patient involvement and individually tailored information is essential to facilitating a healthy transition from hospital to self-care at home and that written information must be improved further.


Stress and symptoms in patients with interstitial cystitis: a life stress model.
Objectives. Stress-related exacerbation of interstitial cystitis (IC) symptoms has frequently been reported. Previous research has found stress-related IC symptom exacerbation in an experimental model. However, this relationship has not been objectively examined with daily life stressors. We used a prospective daily symptom diary method to investigate the relationships among stress and bladder symptoms in patients with IC and age-matched healthy controls.Methods. Forty-five previously diagnosed female patients with IC completed a bladder symptom and stress diary nightly for 1 month; 31 female age-matched healthy controls completed a similar diary for 7 days. The symptom questions were modified from the Interstitial Cystitis Data Base study.Results. Patients reported greater mean daily stress, bladder pain, urgency, and daytime and nocturnal frequency than controls (all P values less than 0.001). Among all patients, a significant relationship between stress and urgency was observed. In addition, a significant relationship between stress and pain was observed among patients with moderate and severe disease. As the disease severity increased, more pronounced relationships between stress and the symptoms of urgency and pain were evidenced. Greater stress was associated with greater nocturnal frequency among patients with more severe disease. These stress-symptom relationships were not observed among the controls.Conclusions. Higher levels of stress were related to greater pain and urgency in patients with IC but not in the controls. In addition, the relationship of stress and these IC symptoms was stronger among patients with more severe disease. The results indicate that life stress is associated with greater IC symptoms, particularly among patients whose disease is not well controlled.


[Postoperative anxiety and its relationship with life quality in patients with Ménière's diseases].
To investigate the postoperative anxiety and its relationship with life quality in patients with Ménière's diseases. Methods: A total of 68 patients with Ménière's disease, who received the treatment of endolymphatic sac decompression from 2010 to 2016, were enrolled. They finished two scales (the self-rating anxiety scale and the quality of life questionnaire for endolymphatic sac decompression of Ménière's disease) by snail mail. The patients were divided into different groups based on their preoperative course, postoperative follow-up time, clinic stage and quality of life after endolymphatic sac decompression surgery, and their anxiety was analyzed. In addition, 109 gender and age-matched patients with other diseases of otorhinolaryngology served as the control group, and their anxiety was also analyzed. Results: There was no statistical difference in the postoperative anxiety among the patients with preoperative course for less than 1 year, 1 to 5 years or more than 5 years (all P>0.05). There was no statistical difference in the patients with different duration of follow-up (P>0.05). There was no statistical difference in the patients with the different clinic stages (all P>0.05). There was no statistical difference in the patients with the improved life or non-improved life after surgery (P>0.05). The degree of the anxiety in the Ménière's group was more severe than that in the control group (P<0.01). Conclusion: It needs to keep in mind that a certain degree of postoperative anxiety still keeps in patients with Ménière's disease.


A meta-analysis of randomized clinical trials assessing hemodialysis access thrombosis based on access flow monitoring: where do we stand?
The National Kidney Foundation Kidney Disease Outcomes Quality Initiative recommends the routine use of hemodialysis arteriovenous (AV) access surveillance to detect hemodynamically significant stenoses and appropriately correct them to reduce the incidence of thrombosis and to improve accesses patency rates. Access blood flow monitoring is considered as one of the preferred surveillance method for both AV fistulas (AVF) and AV grafts (AVG); however, published studies have reported conflicting results of its utility that led healthcare professionals to doubt the benefits of this surveillance method. We performed a meta-analysis of the published randomized controlled trials (RCTs) of AV access surveillance using access blood flow monitoring. Our hypothesis was that access blood flow monitoring lowers the risk of AV access thrombosis and that the outcome differs between AVF and AVG. The estimated overall pooled risk ratio (RR) of thrombosis was 0.87 (95% confidence interval [CI], 0.67-1.13) favoring access blood flow monitoring. The pooled RR of thrombosis were 0.64 (95% CI, 0.41-1.01) and 1.06 (95% CI, 0.77-1.46) in the subgroups of only AVF and only AVG, respectively. Our results added to the uncertainty of access blood flow monitoring as a surveillance method of hemodialysis accesses.


Perspectives on domestication: the history of our relationship with man's best friend.
We are a nation of dog lovers. Never before in our history have we spent more time, money, and emotional energy on a group of animals that are kept solely for companionship. Pet food sales are a multi-billion dollar industry, and pet owners are spending more than 11 billion dollars each year on veterinary care. This devotion is further illustrated by the exponential growth of the pet supply industry, including increasing numbers of pet superstores, play-parks, training centers, and doggie day care centers. During the 1980s, recognition of the human-animal bond led to serious study of the roles that dogs play in our lives. These studies have shown that pets provide significant benefits to our emotional, physical, and social well being. It is ironic then, that at a time when we recognize and appreciate our bonds with animal companions, dark elements of this relationship are equally pervasive. Animal shelters in the United States kill between 3 and 4 million dogs and cats annually. Dog fighting, although outlawed, has reached epidemic proportions in some areas of the country. Episodes of animal cruelty and neglect are reported with alarming frequency in the media; so frequently that discussions of the connection between animal cruelty and human violence have become daily parlance. How then did we come to have such paradoxical perceptions and treatment of our canine companions? This question is explored through an examination of the ancestry of the dog and the prevailing myths and facts about domestication. Historical and present-day perceptions of the wolf and the impact that these attitudes may have upon perceptions of dogs are examined.


Baby care products: possible sources of infant phthalate exposure.
Phthalates are man-made chemicals found in personal care and other products. Recent studies suggest that some phthalates can alter human male reproductive development, but sources of infant exposure have not been well characterized. We investigated the relationship between phthalate metabolite concentrations in infant urine and maternal reported use of dermally applied infant care products. We measured 9 phthalate metabolites in 163 infants who were born in 2000-2005. An infant was considered to have been exposed to any infant care product that the mother reported using on her infant within 24 hours of urine collection. Results of multiple linear regression analyses are reported as the ratio of metabolite concentrations (with 95% confidence intervals) in exposed and unexposed infants. We standardized concentrations by forming z scores and examined combined exposure to multiple metabolites. In most (81%) infants, > or = 7 phthalate metabolites were above the limit of detection. Exposure to lotion was predictive of monoethyl phthalate and monomethyl phthalate concentrations, powder of monoisobutyl phthalate, and shampoo of monomethyl phthalate. Z scores increased with number of products used. Most associations were stronger in younger infants. Phthalate exposure is widespread and variable in infants. Infant exposure to lotion, powder, and shampoo were significantly associated with increased urinary concentrations of monoethyl phthalate, monomethyl phthalate, and monoisobutyl phthalate, and associations increased with the number of products used. This association was strongest in young infants, who may be more vulnerable to developmental and reproductive toxicity of phthalates given their immature metabolic system capability and increased dosage per unit body surface area.


Effect of basic and nonbasic amino acid substitutions on transport induced by simian virus 40 T-antigen synthetic peptide nuclear transport signals.
A previous study demonstrated the ability of a synthetic peptide homologous to the simian virus 40 T-antigen nuclear transport signal to induce the nuclear transport of carrier proteins and the dependence of peptide-induced transport on a positive charge at the lysine corresponding to amino acid 128 of T antigen. In this investigation synthetic peptides were utilized to examine the effect on transport of amino acid substitutions within the T-antigen nuclear transport signal. Nuclear transport was evaluated by immunofluorescence after microinjection of protein-peptide conjugates into the cytoplasm of mammalian cells. Substitution of other basic amino acids at position 128 revealed a hierarchy for nuclear transport. The rate of nuclear transport was most rapid when a lysine was at position 128 followed in descending order by arginine, D-lysine, ornithine, and p-aminophenylalanine. Peptide-induced nuclear transport was dependent upon a positively charged amino acid at positions 128 and 129, since substitutions of neutral asparagines at these positions abolished transport. However, partial transport was observed with the peptide having an asparagine at position 128 when a high number of peptides were conjugated to the carrier protein.


[Isolation and immunoenzyme determination of human beta 2-microglobulin].
Two procedures for isolation of homogeneous beta 2-microglobulin from urine of patients with systemic lupus erythematosus were developed: a procedure for isolation of beta 2-microglobulin with two-stage gel chromatography on Sephacryl S-200 and a procedure for isolation of homogeneous beta 2-microglobulin with affinity chromatography on rabbit polyclonal antibodies. The microglobulins isolated with the two procedures showed identical physicochemical properties and were used in development of a competitive immunoenzymatic assay method for determination of beta 2-microglobulin in the blood. The method was approved for determination of beta 2-microglobulin in blood serum of patients.


Demographic, Clinical and Management Characteristics of Newly Diagnosed COPD Patients in Turkey: A Real-Life Study.
In order to determine the clinical and sociodemographic characteristics of newly diagnosed treatment-naïve asthma and COPD patients in Turkey, a multicenter study in 2012 was initiated . We aimed to investigate the characteristics and therapies of COPD patients in the original study in more detail. This nation-wide, multicentric, non-interventional, prospective, real-life observational cohort study was conducted in 122 centers. The newly diagnosed patients were not receiving any treatment before the recruitment. Their general characteristics, the combined GOLD 2011 COPD categories and exacerbation histories were noted. The patients were followed up with 3 voluntary visits for 1 year. Their adherence to the inhaled treatment according to GOLD 2011 was evaluated during follow-up visits. The study included 776 COPD patients. Their mean age was 59.4±9.1 years, and 11.9% of the patients were female. 35.1% of the patients were in the GOLD 2011 C and D category. 12.6% are frequent exacerbators, and 52.8% had at least one comorbid condition. 71.8% overtreatment rate was detected. Their attendance rates for three follow-up visits became 55.9%, 32.9% and 18.7%, respectively. The adherence rate to the treatment was measured as 81.9%. Although these patients were diagnosed for the first time, the GOLD C and D categories and frequent exacerbator phenotype were found at a high rate. They were usually prescribed an overtreatment regimen. We think that newly diagnosed COPD patients should be evaluated carefully, and best effort should be made to treat these patients in accordance with the recommendations of the major COPD guidelines.


Mode of Action of the Catalytic Site in the N-Terminal Ribosome-Inactivating Domain of JIP60.
Jasmonate-induced protein 60 (JIP60) is a ribosome-inactivating protein (RIP) from barley (Hordeum vulgare) and is involved in the plant immune response dependent on jasmonate hormones. Here, we demonstrate in Nicotiana benthamiana that transient expression of the N-terminal domain of JIP60, from which the inhibitor domain (amino acids 163-185) is removed, initiates cell death, leading to extensive necrosis of leaf tissues. We used structure prediction of JIP60 to identify potential catalytic amino acids in the active site and tested these by mutagenesis and in planta assays of necrosis induction by expression in N. benthamiana, as well as through an in vitro translation-inactivation assay. We found that Tyr 96, Glu 201, Arg 204, and Trp 234 in the presumptive active site of JIP60 are conserved in 815 plant RIPs in the Pfam database that were identified by HUMMR as containing a RIP domain. When these amino acid residues are individually mutated, the necrosis-inducing activity is completely abolished. We therefore propose that the role of these amino acids in JIP60 activity is to depurinate adenosine in ribosomes. This study provides insight into the catalytic mechanism of JIP60.


Photoinduced formation mechanism of the thymine-thymine (6-4) adduct.
The photoinduced mechanism leading to the formation of the thymine-thymine (6-4) photolesion has been studied by using the CASPT2//CASSCF approach over a dinucleotide model in vacuo. Following light absorption, localization of the excitation on a single thymine leads to fast singlet-triplet crossing that populates the triplet (3)(nπ*) state of thymine. This state, displaying an elongated C(4)═O bond, triggers (6-4) dimer formation by reaction with the C(5)═C(6) double bond of the adjacent thymine, followed by a second intersystem crossing, which acts as a gate between the excited state of the reactant and the ground state of the photoproduct. The requirement of localized excitation on just one thymine, whose main decay channel (by radiationless repopulation of its ground state) is nonphotochemical, can rationalize the experimentally observed low quantum yield of formation for the thymine-thymine (6-4) adduct.


Submucosal tunneling endoscopic resection for the treatment of rectal submucosal tumors originating from the muscular propria layer.
The objective was to evaluate the clinical value of submucosal tunneling endoscopic resection (STER) for the treatment of submucosal tumors (SMTs) originating from the muscular propria (MP) in the rectum. The clinicopathological data of 12 cases with rectal SMTs originating from the MP layer performed STER in our center from January 2012 to June 2014 were analyzed retrospectively. Three males and nine females (M/F, 1/3) were studied in this series. The median age of the patients was 53.5 (range, 41-84) years. The tumors located 5-10 cm from the anal verge. En bloc STER was performed successfully in all of the 12 cases. The median size of resected specimens was 1.4 cm (range 1.0-3.0 cm). The median procedure time was 49.5 min (range 40-70 min). Three patients developed low fever after the operation and were all recovered after receiving intravenous antibiotics. One of these three cases developed mucosa perforation, which was closed immediately with metal clips. One patient developed subcutaneous emphysema in one lower limb, which disappeared with conservative treatments 2 weeks after the STER procedure. The median hospital stays were 3.1 (range 2-8) days. Postoperative pathological outcomes revealed schwannoma in 3 cases, leiomyoma in 2 cases, stromal tumor in 5 cases, and proliferation of collagen fibers nodular degeneration in 2 cases. No lesion residual or recurrence was found during postoperative follow-up of 4-33 months. STER is a feasible, safe, and effective method for treating SMTs originating from the MP layer in the rectum.


Reliability and validity of the subjective component of the American Orthopaedic Foot and Ankle Society clinical rating scales.
This study evaluates the criterion validity of the subjective component of the American Orthopaedic Foot and Ankle Society (AOFAS) clinical rating scales by correlating scores obtained with these rating scales to scores obtained with the Foot Function Index (FFI) in patients with foot and ankle conditions. To date, the AOFAS scoring scales have not been shown to provide valid information despite their popularity. The FFI, on the other hand, has previously been shown to provide valid information in regard to conditions affecting the foot and ankle. A moderately strong inverse criterion validity correlation (Pearson correlation coefficient = -0.68) was shown when preoperative patients were administered both the AOFAS and FFI questionnaires, and the resultant scores were compared. Test-retest reliability measurements showed no significant difference (P = .27) between preoperative AOFAS scale scores measured at least 2 weeks apart. Construct validity was shown (P = .006) when dependent preoperative and postoperative (at least 3 months) AOFAS scale scores were compared, indicative of the clinical rating scales' ability to discriminate and predict quality of life related to foot and ankle conditions. The moderate level of correlation, satisfactory degree of reliability, and responsiveness (ability to distinguish differences between preoperative and postoperative conditions in the same patient) observed in this study suggest that the subjective component of the AOFAS clinical rating scales provides quality-of-life information that conveys acceptable validity regarding conditions affecting the foot and ankle.


Obeticholic acid-a new therapy in PBC and NASH.
Obeticholic acid (OCA) is a semi-synthetic hydrophobic bile acid (BA) analogue that is highly selective agonist of farnesoid X receptor (FXR), a key nuclear BA receptor, which induces expression of gut-derived hormones, in particular fibroblast growth factor 19. The resulting beneficial effects of OCA on glucose and lipid metabolism and particularly hepatic inflammation make it a candidate for the treatment of a variety of conditions including primary biliary cholangitis (PBC) and nonalcoholic steatohepatitis (NASH). In PBC patients who have not initially responded to ursodeoxycholic acid, OCA has been shown in double-blind controlled clinical trials to significantly reduce serum alkaline phosphatase. To date, OCA is the only therapy licensed by the FDA, EMA and endorsed by NICE as second line therapy for PBC.No medications are currently approved in Europe or the USA for the treatment of NASH.In recent clinical trials, OCA has been shown encouraging results by improving liver blood tests and reducing liver fibrosis with no worsening of NASH. OCA is the established second line therapy for PBC in those patients who fail to adequately respond to ursodeoxycholic acid. The main side effects of OCA treatment in both PBC and NASH is that of dose-dependent pruritis which can lead to treatment discontinuation in ~1-10% of patients. In addition, OCA-treated patients may also exhibit (reversible) alterations in serum lipid levels; most notably a small decrease in high density lipoprotein cholesterol. It is not yet known whether these changes carry a long-term cardiovascular risk in NASH.In addition, the relatively high cost of OCA may limit its use in cash-limited health systems. Additional clinical trials are in progress to ascertain the long-term effects of OCA on survival in PBC and NASH. New FXR agonists with a lower rate of side effects are being developed and trialed. Combination therapy with other agents may offer increased efficacy.


CD200 included in a 4-marker modified Matutes score provides optimal sensitivity and specificity for the diagnosis of chronic lymphocytic leukaemia.
CD200, a transmembrane type Ia glycoprotein belonging to the immunoglobulin superfamily, has been shown to have a differential expression in B-cell neoplasms. Here, we retrospectively assessed the diagnostic relevance of CD200 on 427 patients with B-cell chronic neoplasms in leukemic phase (median age, 69 y; range, 35-97 y). The final diagnosis based on the investigator's assessment was chronic lymphocytic leukaemia (CLL) in 75% of cases and non-CLL in 25% of cases. Sensitivity and specificity for the diagnosis of CLL (vs non-CLL) were calculated for the following markers: CD200, CD5, CD22, CD23, CD79b, FMC7, and SmIg. CD23 was the only marker without a statistically significant difference between the investigator assessment and the flowcytometric analysis. The other markers were unable-when individually evaluated-to discriminate between CLL and non-CLL, requiring the integration into a scoring system. The modified score no. 1 (addition of CD200) showed superimposable sensitivity and specificity compared with the Matutes score. The substitution of CD79b (modified score no. 2), surface membrane immunoglobulins (SmIg) (modified score no. 3), and CD79b and FMC7 (modified score no. 4) with CD200 showed that only the modified score no. 4 had both higher sensitivity and higher specificity compared with standard Matutes score. In conclusion, this work defines a simplified score, compared with the classical Matutes score, for the differential diagnosis of chronic B-cell leukaemia-which only requires 4 markers instead of 5 (CD5, CD23, CD200, and SmIg).


Problem based review: the patient presenting with hyponatraemia.
Hyponatraemia is the most common electrolyte anomaly and is associated with significant morbidity and mortality. Patients with severe hyponatraemia often present to acute medical units with non-specific symptoms which can progress to overt neurological manifestations. There are many causes of hyponatriaemia, the most common being drug-induced causes, particularly thiazide diuretics, and the Syndrome of Inappropriate ADH Secretion (SIADH). Initial assessment should include a careful evaluation of the patient's volume status, which helps to identify the most likely cause. This article utilises a recent case which presented to our AMU to illustrate the importance of a careful and systematic assessment of patients presenting to hospital with hyponatraemia. The new vasopressin receptor antagonists are explored as an option for the management of severe hyponatraemia.


[Social network and health promotion of homeless population].
One of the outcomes of globalization has been a constant increase of the number of people living in the streets of metropolis such as São Paulo: those are not only homeless, but also people who live in the streets and earn their living from it. We have surveyed the institutions working with adult homeless population in the central region of Sao Paulo. Their goals and strategies were described, as well as how they associate with each other. Our main hypothesis is that our reality is still formed by isolated actions, based on mere assistance policies that help to maintain the problem of homelessness and turn it into a chronicle situation. Although, the participative methodology we chose has shown a small change from this model towards the idea of the emancipation of the subject. Even so, the idea of a technical knowledge still looms large and keeps the voice of the population in the background. The constitution of a network based on inter-sectorial actions (according to the perspective of health promotion) indicates the possibility of building innovatory public policies.


Signal transduction events in Chinese hamster ovary cells expressing human CD14; effect of endotoxin desensitization.
Previous studies suggest that endotoxin (LPS) stimulation of CD14 receptors may be coupled to heterotrimeric G proteins. However, characterization of the G protein-coupled signaling pathways is incomplete. Also, specific changes in the transduction pathways occur in a phenomenon known as LPS tolerance or desensitization induced by prior exposure to LPS. In the present study, we examined potential CD14-dependent G protein-coupled signaling events in response to LPS, and changes in signaling in these pathways during LPS desensitization in Chinese Hamster Ovary (CHO) cells. LPS stimulated inhibitory kappa B alpha (IkappaB alpha) degradation and p38 phosphorylation in CHO cells transfected with human CD14 receptor (CHO-CD14), but not in CHO cells transfected with vector only. However, activation of these signaling events diverged early in the signal transduction pathways. Pretreatment with pertussis toxin, which inactivates inhibitor G protein (G alpha i) function, significantly inhibited LPS-induced p38 phosphorylation, but not LPS-induced IkappaB alpha degradation. Mastoparan, a putative G alpha i agonist, synergized with LPS to induce p38 phosphorylation. Thus, LPS stimulation of p38 phosphorylation is, in part, G alpha i coupled, whereas IkappaB alpha degradation is not. In subsequent studies, CHO-CD14 cells were desensitized by prior LPS exposure. LPS-desensitized cells exhibited augmented IkappaB alpha content and were refractory to LPS-induced IkappaB alpha degradation and p38 phosphorylation. Pretreatment with cycloheximide, a protein synthesis inhibitor, prevented the effect of LPS desensitization on augmenting cellular IkappaB alpha content and its refractoriness to LPS-induced degradation. However, cycloheximide pretreatment did not prevent impaired p38 phosphorylation in desensitized cells. IkappaB alpha upregulation in LPS tolerance may occur through increased synthesis and/or induction of protein that suppress IkappaB alpha degradation. The latter protein synthesis-dependent mechanisms may be distinct from mechanismis inhibiting p38 phosphorylation in tolerance. These findings suggest that LPS tolerance induces CD14-dependent signaling alterations in G alpha i-coupled pathways leading to mitogen-activated (MAP) kinase activation as well as G alpha i-independent pathways inducing IkappaB alpha degradation.


Precision Medicine in Internal Medicine.
Medicine has long sought to match diagnostic and treatment approaches to the particular needs and risks of individual patients. The decreasing cost and increasing ease of genetic sequencing have propelled the rise of precision medicine. Precision medicine aims to use genetic and other information to provide care tailored to the individual patient, with the goal of improving clinical outcomes and minimizing unnecessary diagnostic and therapeutic interventions. Although developments in genetic sequencing have the potential to transform clinical care, there are important limitations, including uncertainty in the clinical interpretation of many genetic variants and concerns about privacy, discrimination, and cost. To help clinicians understand the basics of genetic sequencing and how to apply it in clinical practice, Annals of Internal Medicine is launching a new "Precision Medicine" series. This introduction provides a general overview of clinical sequencing, with a focus on germline variation. Subsequent articles will use a case-based format to provide concise summaries of specific clinical precision medicine scenarios that are relevant to the practice of internal medicine. These cases will highlight specific clinical indications; interpretation of genetic test results; and ethical, legal, cost, and privacy issues related to genetic testing. The goal is to provide practical information on the appropriate application and interpretation of genomics in routine clinical practice.


Loss of hepatic AMP-activated protein kinase impedes the rate of glycogenolysis but not gluconeogenic fluxes in exercising mice.
Pathologies including diabetes and conditions such as exercise place an unusual demand on liver energy metabolism, and this demand induces a state of energy discharge. Hepatic AMP-activated protein kinase (AMPK) has been proposed to inhibit anabolic processes such as gluconeogenesis in response to cellular energy stress. However, both AMPK activation and glucose release from the liver are increased during exercise. Here, we sought to test the role of hepatic AMPK in the regulation of in vivo glucose-producing and citric acid cycle-related fluxes during an acute bout of muscular work. We used 2H/13C metabolic flux analysis to quantify intermediary metabolism fluxes in both sedentary and treadmill-running mice. Additionally, liver-specific AMPK α1 and α2 subunit KO and WT mice were utilized. Exercise caused an increase in endogenous glucose production, glycogenolysis, and gluconeogenesis from phosphoenolpyruvate. Citric acid cycle fluxes, pyruvate cycling, anaplerosis, and cataplerosis were also elevated during this exercise. Sedentary nutrient fluxes in the postabsorptive state were comparable for the WT and KO mice. However, the increment in the endogenous rate of glucose appearance during exercise was blunted in the KO mice because of a diminished glycogenolytic flux. This lower rate of glycogenolysis was associated with lower hepatic glycogen content before the onset of exercise and prompted a reduction in arterial glucose during exercise. These results indicate that liver AMPKα1α2 is required for maintaining glucose homeostasis during an acute bout of exercise.


Mapping elements distribution in carapace of Caretta caretta: A strategy for biomonitoring contamination in sea turtles?
This study analyzed the carapace distribution of Ca, Cd, Cr, Cu, Mg, Mn, Pb, Sb, U, V and Zn by GF-AAS and ICP-AES in one specimen of Caretta caretta from Mediterranean Sea. Calcium, Mg, Mn, Pb, U, Zn were mainly distributed in the central area while Cd, Cr, Cu, Sb, V in lateral areas. Cadmium, Cr, Mg, Mn, Sb, U and V were different between lateral areas. The different distribution may be related to several exposures during lifetime and/or the shell ossification during growth. Carapace may be a suitable matrix for metal biomonitoring, however, further studies are required to confirm these findings.


Host-directed therapy to combat mycobacterial infections.
Upon infection, mycobacteria, such as Mycobacterium tuberculosis (Mtb) and nontuberculous mycobacteria (NTM), are recognized by host innate immune cells, triggering a series of intracellular processes that promote mycobacterial killing. Mycobacteria, however, have developed multiple counter-strategies to persist and survive inside host cells. By manipulating host effector mechanisms, including phagosome maturation, vacuolar escape, autophagy, antigen presentation, and metabolic pathways, pathogenic mycobacteria are able to establish long-lasting infection. Counteracting these mycobacteria-induced host modifying mechanisms can be accomplished by host-directed therapeutic (HDT) strategies. HDTs offer several major advantages compared to conventional antibiotics: (a) HDTs can be effective against both drug-resistant and drug-susceptible bacteria, as well as potentially dormant mycobacteria; (b) HDTs are less likely to induce bacterial drug resistance; and (c) HDTs could synergize with, or shorten antibiotic treatment by targeting different pathways. In this review, we will explore host-pathogen interactions that have been identified for Mtb for which potential HDTs impacting both innate and adaptive immunity are available, and outline those worthy of future research. We will also discuss possibilities to target NTM infection by HDT, although current knowledge regarding host-pathogen interactions for NTM is limited compared to Mtb. Finally, we speculate that combinatorial HDT strategies can potentially synergize to achieve optimal mycobacterial host immune control.