Delta Checks in the clinical laboratory.
International standards and practice guidelines recommend the use of delta check alerts for laboratory test result interpretation and quality control. The value of contemporary applications of simple univariate delta checks determined as an absolute change, percentage change, or rate of change to recognize specimen misidentification or other laboratory errors has not received much study. This review addresses these three modes of calculation, but in line with the majority of published work, most attention is focused on the identification of specimen misidentification errors. Investigation of delta check alerts are time-consuming and the yield of identified errors is usually small compared to the number of delta check alerts; however, measured analytes with low indices of individuality frequently perform better. While multivariate approaches to delta checks suggest improved usefulness over simple univariate delta check strategies, some of these are complex and not easily applied in contemporary laboratory information systems and middleware. Nevertheless, a simple application of delta checks may hold value in identifying clinically significant changes in several clinical situations: for acute kidney injury using changes in serum creatinine, for risk of osmotic demyelination syndrome using rapid acute changes in serum sodium levels, or for early triage of chest pain patients using high sensitivity troponin assays. A careful and highly selective approach to identifying delta check analytes, calculation modes, and thresholds before putting them into practice is warranted; then follow-up with careful monitoring of performance and balancing true positives, false negatives, and false positives among delta check alerts is needed.


Expansion of CMV-mediated NKG2C+ NK cells associates with the development of specific de novo malignancies in liver-transplanted patients.
Solid cancers are a major adverse outcome of orthotopic liver transplantation (OLT). Although the use of chronic immunosuppression is known to play a role in T cell impairment, recent insights into the specificities of NK cells led us to reassess the potential modulation of this innate immune cell compartment after transplantation. Our extensive phenotypic and functional study reveals that the development of specific de novo noncutaneous tumors post-OLT is linked to unusual NK cell subsets with maturation defects and to uncommon cytokine production associated with the development of specific cancers. Remarkably, in CMV(+) patients, the development de novo head/neck or colorectal tumors is linked to an aberrant expansion of NK cells expressing NKG2C and a high level of intracellular TNF-α, which impact on their polyfunctional capacities. In contrast, NK cells from patients diagnosed with genitourinary tumors possessed a standard immature signature, including high expression of NKG2A and a robust production of IFN-γ. Taken together, our results suggest that under an immunosuppressive environment, the interplay between the modulation of NK repertoire and CMV status may greatly hamper the spectrum of immune surveillance and thus favor outgrowth and the development of specific de novo tumors after OLT.


Pyrimidone inhibitors targeting Chikungunya Virus nsP3 macrodomain by fragment-based drug design.
The macrodomain of nsP3 (nsP3MD) is highly conserved among the alphaviruses and ADP-ribosylhydrolase activity of Chikungunya Virus (CHIKV) nsP3MD is critical for CHIKV viral replication and virulence. No small molecule drugs targeting CHIKV nsP3 have been identified to date. Here we report small fragments that bind to nsP3MD which were discovered by virtually screening a fragment library and X-ray crystallography. These identified fragments share a similar scaffold, 2-pyrimidone-4-carboxylic acid, and are specifically bound to the ADP-ribose binding site of nsP3MD. Among the fragments, 2-oxo-5,6-benzopyrimidine-4-carboxylic acid showed anti-CHIKV activity with an IC50 of 23 μM. Our fragment-based drug discovery approach provides valuable information to further develop a specific and potent nsP3 inhibitor of CHIKV viral replication based on the 2-pyrimidone-4-carboxylic acid scaffold. In silico studies suggest this pyrimidone scaffold could also bind to the macrodomains of other alphaviruses and coronaviruses and thus, have potential pan-antiviral activity.


[Awareness, treatment and control of hypertension and related factors in the jurisdictional areas of primary health care posts in a rural community of Korea].
This study aimed to identify and assess the factors related to the awareness, treatment, and control of hypertension based on jurisdictional areas of primary health care posts in a rural community of Korea. This study was performed on 4598 adults aged over 30 years in a rural community and we measured their blood pressure (BP) from October. 2007 to August. 2009. Hypertension is defined as a condition characterized by a systolic BP ≥ 140 mmHg, a diastolic BP ≥ 90 mmHg or reported treatment with antihypertensive medications. We analyzed the factors related with the prevalence, awareness, treatment and control of hypertension using chi-square test and multivariate logistic regression analysis. The age-adjusted prevalence of hypertension was 34.7%. The age-adjusted rates of hypertension awareness, treatment and control were 50.6%, 93.9% and 64.1%, respectively. Awareness of hypertension was related with increasing age. Higher awareness was found among men who were felt more stress, were obese and had hypercholesterolemia, and among women who were regularly taking medicine for hypertension, were obese and had diabetes mellitus. In women, the hypertension treatment was related a Medical aid and education for hypertension management. Controlled hypertension was more common among men who were educated about the management of hypertension and among women who had hypercholesterolemia. The awareness of hypertension was low and the control of hypertension was high compared with the nationwide data (KNHANES 2005). The results suggest that understanding the characteristics of hypertension in a community is important to perform a community based hypertension control program.


[Risk and risk perception].
After having delineated the concept of risk and having described the type of risks that are relevant to dental practice, an overview of empirical and theoretical work on risk perception is given. The perception of risks is determined both by the actual magnitude and by perceptual bias, originating from either general perceptual phenomena such as (cognitive) availability or the ego-relevance of risks: persons in general underestimate risks for themselves as compared to the risks for others (unrealistic optimism).


Case studies of the attainment of insight in dream sessions: replication and extension.
To replicate and extend the Hill, Knox, et al. (2007) case study of a client who attained insight in one session of dream work, the authors examined two additional single-session cases: one in which a client gained insight and another in which a client did not. The observations across all three cases suggest that the two clients who acquired insight had positive attitudes toward dreams; were motivated and involved in session; and were nonresistant, trusting, and affectively present but not overwhelmed. The client who did not gain insight questioned the value of dreams and was resistant, untrusting, and emotionally overwhelmed. Therapist adherence and competence using the dream model, ability to manage countertransference, and effective use of probes for insight distinguished the therapists whose clients gained insight from the therapist whose client did not.


Employment status influences the weekly patterns of suicide among alcohol misusers.
As part of the National Suicide Prevention Project in Finland, a nationwide psychological autopsy study, all suicide victims (n = 1397) over a 12-month period were investigated concerning factors associated with any variation in suicide frequency between weekdays and weekends. In particular, employment status was expected to have influenced the weekly pattern of alcohol misuse, and thereby to have caused clustering of suicides at weekends among the employed. Among suicide victims who had misused alcohol, those in employment were significantly more likely to have committed suicide during the weekend that those without work (52% vs. 34%, p < 0.001). In logistic regression analysis, employment was the only independent variable significantly associated with suicide at the weekend. According to forensic chemical analysis, those classified as misusers had frequently used alcohol at the time of suicide, regardless of the weekday, and slightly more often if employed. The clustering of suicides at weekends among employed alcohol misusers is probably explained by a weekly pattern in the use of alcohol, which suggests that besides the established risk factors for suicide among alcohol misusers, the act of using alcohol per se also contributes to the suicidal act.


Exposure to electrical contact currents and the risk of childhood leukemia.
The objectives of this study were to examine the association between contact current exposure and the risk of childhood leukemia and to investigate the relationship between residential contact currents and magnetic fields. Indoor and outdoor contact voltage and magnetic-field measurements were collected for the diagnosis residence of 245 cases and 269 controls recruited in the Northern California Childhood Leukemia Study (2000-2007). Logistic regression techniques produced odds ratios (OR) adjusted for age, sex, Hispanic ethnicity, mother's race and household income. No statistically significant associations were seen between childhood leukemia and indoor contact voltage level [exposure ≥90th percentile (10.5 mV): OR = 0.83, 95% confidence interval (CI): 0.45, 1.54], outdoor contact voltage level [exposure ≥90th percentile (291.2 mV): OR = 0.89, 95% CI: 0.48, 1.63], or indoor magnetic-field levels (>0.20 µT: OR = 0.76, 95% CI: 0.30, 1.93). Contact voltage was weakly correlated with magnetic field; correlation coefficients were r = 0.10 (P = 0.02) for indoor contact voltage and r = 0.15 (P = 0.001) for outdoor contact voltage. In conclusion, in this California population, there was no evidence of an association between childhood leukemia and exposure to contact currents or magnetic fields and a weak correlation between measures of contact current and magnetic fields.


Echinacea-associated anaphylaxis.
A woman with atopy experienced anaphylaxis after taking, among other dietary supplements, a commercial extract of echinacea. Hypersensitivity was confirmed by skinprick and RAST testing. Regular ingestion of echinacea by up to 5% of surveyed patients with atopy, combined with detection of echinacea-binding IgE in atopic subjects (19% by skin testing; 20% with moderate to strong reactivity by RAST testing), raises the possibility of severe allergic reactions, even with first-time use, due to cross-reactivity with other structurally similar allergens. Patients with atopy should be cautioned about the risk of developing life-threatening reactions to complementary medicines, including echinacea.


Changes in function and ultrastructure of striatal dopaminergic terminals that regenerate following partial lesions of the SNpc.
Following partial substantia nigra lesions, remaining dopaminergic neurones sprout, returning terminal density in the dorsal striatum to normal by 16 weeks. This suggests regeneration and maintenance of terminal density is regulated to release appropriate levels of dopamine. This study examined the structure and function of these reinnervated terminals, defining characteristics of dopamine uptake and release, density and affinity of the dopamine transporter (DAT) and ultrastructural morphology of dopamine terminals in the reinnervated dorsal striatum. Finally, rotational behaviour of animals in response to amphetamine was examined 4 and 16 weeks after substantia nigra pars compacta (SNpc) lesions. Dopamine transport was markedly reduced 16 weeks after lesioning along with reduced density and affinity of DAT. Rate of dopamine release and peak concentration, measured electrochemically, was similar in lesioned and control animals, while clearance was prolonged after lesioning. Ultrastructurally, terminals after lesioning were morphologically distinct, having increased bouton size, vesicle number and mitochondria, and more proximal contacts on post-synaptic cells. After 4 weeks, tendency to rotate in response to amphetamine was proportional to lesion size. By 16 weeks, rotational behaviour returned to near normal in animals where lesions were less than 70%, although some animals demonstrated unusual rotational patterns at the beginning and end of the amphetamine effect. Together, these changes indicate that sprouted terminals are well compensated for dopamine release but that transport mechanisms are functionally impaired. We discuss these results in terms of implications for dyskinesia and other behavioural states.


Berberine inhibits myofibroblast differentiation in nasal polyp-derived fibroblasts via the p38 pathway.
The purposes of this study were to determine whether berberine has any effect on phenotype changes and extracellular matrix (ECM) production in nasal polyp-derived fibroblasts (NPDFs) and to investigate the underlying molecular mechanism. NPDFs were pre-treated with berberine prior to induction by transforming growth factor (TGF)-β1. The expression of α-smooth muscle actin (SMA) and collagen type I mRNA was determined by a reverse transcription-polymerase chain reaction, and the expression of α-SMA protein and collagen type I was determined by western blotting and/or immunofluorescent staining. The total soluble collagen production was analysed by the SirCol collagen assay. The expression of several signaling molecules of the TGF-β1 pathway was evaluated by western blot analysis. In TGF-β1-induced NPDFs, berberine significantly inhibited the expression of α-SMA and collagen type I mRNA and reduced α-SMA and collagen protein levels. Berberine only suppressed the expression of pp38 among the evaluated signaling molecules. SB203580 (a specific inhibitor of p38 kinase) markedly suppressed the increased expression of collagen type I and α-SMA in TGF-β1-induced NPDFs. Berberine exerts suppressive effects on phenotype changes and ECM production in NPDFs via p38 signaling pathway interference. The findings provide new therapeutic options for ECM production in nasal polyps.


Replacement of short segments within transmembrane domains of MC2R disrupts retention signal.
The proteolysis of the pro-opiomelanocortin precursor results in the formation of melanocortins (MCs), a group of peptides that share the conserved -H-F-R-W- sequence, which acts as a pharmacophore for five subtypes of MC receptors (MCRs). MC type 2 receptor (MC2R; also known as ACTHR) is the most specialized of all the MCRs. It is predominantly expressed in the adrenal cortex and specifically binds ACTH. Unlike other MCRs, it requires melanocortin receptor accessory protein 1 (MRAP) for formation of active receptor and for its transport to the cell membrane. The molecular mechanisms underlying this specificity remain poorly understood. In this study, we used directed mutagenesis to investigate the role of various short MC2R sequence segments in receptor membrane trafficking and specific activation upon stimulation with ligands. The strategy of the study was to replace two to five amino acid residues within one MC2R segment with the corresponding residues of MC4R. In total, 20 recombinant receptors C-terminally fused to enhanced green fluorescent protein were generated and their membrane trafficking efficiencies and cAMP response upon stimulation with α-MSH and ACTH(1-24) were estimated during their stand-alone expression and coexpression with MRAP. Our results indicate that both the motif that determines the ligand-recognition specificity and the intracellular retention signal are formed by a specific extracellular structure, which is supported by the correct alignment of the transmembrane domains. Our results also indicate that the aromatic-residue-rich segment of the second extracellular loop is involved in the effects mediated by the second ACTH pharmacophore (-K-K-R-R-).


[Curve of labeled mitoses in different kinetic states of cell proliferation. III. Experiments with a mathematical model].
By means of computer simulation, the dynamic structure of labeled mitoses curve (LMC) has been investigated. Experiments using computer demonstrated the dependence of LMC form not only on the values of temporal parameters of the mitotic cycle, but also on the state of cell proliferation kinetics before and after pulse labeling. Of particular interest was a case of a temporal block of cell entry into the S period which has been studied in a greater detail. The results of modelling experiments have shown that the universally accepted graphic procedures for the evaluation of mean durations of S- and G2-periods (using 50%-level of LMC) yield the satisfactory estimates only for systems with stationary age distribution for cells in the mitotic cycle (steady, in the strict sense, and exponential states of cell proliferation kinetics). The paper presents also the kinetic interpretation of certain abnormalities in LMC form arising from some peculiarities of cell population kinetics in the real biological experiment.


Assessing congenital heart defects in the cocaine-exposed neonate.
An estimated 100,000 "crack babies" are born each year in the United States. Because in addition to a variety of other congenital anomalies, neonatal cardiac defects have recently been linked to maternal cocaine use, the critical care nurse needs to conduct a detailed cardiac status assessment on all cocaine-exposed infants.


[Brucellosis in pregnancy].
Brucellosis is rare in pregnancy. Recently, an increase in the incidence of this disease has been observed in our area. We present 7 cases of brucellosis in pregnancy and review the literature on the effects of brucellosis on the outcome of pregnancy. Brucellosis is rare in the Middle East and Africa and the most common source of infection is unpasteurized milk products. Brucella is a coccobacillus, gram-negative bacterium, whose hosts are mostly animals. There is controversy about the relationship between brucellosis and the outcome of pregnancy. There is some evidence that there is a higher rate of complications such as abortion, premature rupture of membranes and preterm delivery in infected animals. Reasons for this difference in the impact of brucella in animals and man include the absence of the carbohydrate erythritol in the human placenta, which appears to be a preferential medium and growth factor for brucella in the placentas of animals. There is uncertainty regarding effects of brucella in early pregnancy and no evidence of its transplacental passage in later pregnancy, causing adverse obstetrical outcome, although recently there has been a single report of Brucella abortus (biotype 2). We present 7 cases of brucellosis in pregnant women found between 1977-1988. Its incidence among the women who delivered here is 0.007% (7/92, 768 deliveries). Our first case was complicated by preterm premature rupture of membranes and preterm delivery in the 20th week of gestation. In 2 other cases there was preterm delivery with 1 developing clinical chorioamnionitis. The 4 remaining women delivered at term, although 1 had preterm premature rupture of membranes and intra-uterine growth retardation, and 2 had postpartum endometritis.


Diagnosis of carcinoma of the pancreas by assay of immunosuppressive acidic protein.
To establish a procedure for screening of pancreatic cancer, we assayed the serum level of immunosuppressive acidic protein (IAP) in patients with malignant and benign pancreatic diseases and compared it with the serum level of carcinoembryonic antigen (CEA). In 96% of 46 pancreatic cancer patients, the serum IAP level averaged 928 micrograms/ml; while 500 micrograms/ml or below was obtained as the normal value in 211 healthy adults. The serum IAP level may be elevated in the presence of non-malignant, inflammatory pancreatic diseases. For the differential diagnosis of pancreatic cancer, therefore, other laboratory data need to be taken into account. The serum IAP level was correlated with the progression of pancreatic cancer and may provide a means to determine whether tumor resection was radical or admitting of recurrence. The serum IAP level in samples obtained from different sites of the portal system did not reflect tumor location in patients with pancreatic cancer.


Clock Gene Expression in the Suprachiasmatic Nucleus of Hibernating Arctic Ground Squirrels.
Most organisms have a circadian system, entrained to daily light-dark cycles, that regulates 24-h rhythms of physiology and behavior. It is unclear, however, how circadian systems function in animals that exhibit seasonal metabolic suppression, particularly when this coincides with the long-term absence of a day-night cycle. The arctic ground squirrel, Urocytellus parryii, is a medium-sized, semi-fossorial rodent that appears above-ground daily during its short active season in spring and summer before re-entering a constantly dark burrow for 6 to 9 months of hibernation. This hibernation consists of multiple week-long torpor bouts interrupted by short (< 20 h) arousal intervals when metabolism and body temperature (Tb) return to normal levels. Here, we used immunohistochemistry to measure the expression of daily or circadian rhythms of the protein products of 3 circadian clock genes, PER1, PER2, BMAL1, and the neural activity marker c-FOS in the suprachiasmatic nucleus (SCN) of arctic ground squirrels before, during, and after the first torpor bout of hibernation. Before torpor, while under 12:12-h light:dark conditions, animals showed significant daily rhythms in their Tb, as well as in protein expression levels of PER1 and PER2, but not BMAL1. Upon entering first torpor (Tb < 30°C), animals were moved into constant darkness. When sampled at 6-h intervals-beginning 24 h after the last light out, with Tb 3°C to 4°C-there were no circadian oscillations in PER1, PER2, or c-FOS expression. Sampling across 24 h during the first spontaneous arousal interval, c-FOS expression was elevated only when Tb reached 20°C and PER1 and PER2 expression did not show any Tb- or time-dependent changes. These results suggest that the central circadian clock might have stopped functioning during hibernation in this species, and the timing of arousal from torpor in arctic ground squirrels is unlikely to be controlled by the circadian clock within the SCN.


Continuous Glucose Monitoring Time-in-Range and HbA1c Targets in Pregnant Women with Type 1 Diabetes.
The CONCEPTT trial compared real-time Continuous Glucose Monitoring (RT-CGM) to capillary glucose monitoring in pregnant women with type 1 diabetes. We analyzed CGM and glycated hemoglobin (HbA1c) measures in first (n = 221), second (n = 197), and third (n = 172) trimesters, aiming to examine target glucose attainment and associations with pregnancy outcomes. CGM targets were Time-in-range (TIR) > 70%, Time-above-range (TAR) <25%, and Time-below-range (TBR) < 4%, and HbA1c targets < 6.5% (National Institute for Health and Care Excellence [NICE]) and HbA1c < 6.0% in second and third trimesters (American Diabetes Association [ADA]). TIR/TAR/TBR targets were achieved by 7.7/14.5/30.3% participants in first, 10.2/14.2/52.8% in second, and 35.5/37.2/52.9% in third trimesters. CGM target attainment was low but increased during pregnancy and with RT-CGM use. In the adjusted analyses, achieving TBR target was associated with a higher risk of pre-eclampsia and neonatal hypoglycemia. ADA HbA1c target attainment was low and unchanged during pregnancy (23.5/27.9/23.8%) but increased with RT-CGM use. In the adjusted analyses, HbA1c target attainment was associated with a lower risk of preterm birth, large-for-gestational age and neonatal hypoglycemia. We conclude that CONCEPTT trial participants had a low rate of CGM and of HbA1c target attainment. Attainment of CGM and NICE HbA1c targets increased throughout gestation and all targets (both NICE/ADA HbA1c and CGM) were more likely to be achieved by RT-CGM users, at 34 weeks' gestation. ADA HbA1c target achievement was independently associated with better perinatal outcomes, while the independent association of TBR target achievement with increased risk warrants further study. ClinicalTrials.gov Registration Identifier NCT01788527.


[The inhibition of lidocaine metabolism by various barbiturates in rat hepatic microsome].
To evaluate the effects of various barbiturates on lidocaine metabolism by cytochrome P-450 (P-450), enzyme kinetics were analyzed in an in vitro study using rat hepatic microsomes. Phenobarbital, amobarbital, hexobarbital, pentobarbital, and thiamylal showed the mixed type inhibition of lidocaine metabolism with inhibition constants being 4.89, 1.08, 2.76, 0.77 and 0.65 mM, respectively. Same as lidocaine, all barbiturates used in the present study, corresponding to binding with P-450, induced the I type of spectral change of P-450. Since these did not affect cytochrome C reductase activity, it was suggested that this inhibition of lidocaine metabolism in hepatic microsomes may have been caused by the reduction of activity on P-450 by the barbiturates.


Review article: Novel technologies in the treatment and monitoring of advanced and relapsed epithelial ovarian cancer.
Epithelial Ovarian cancer (EOC) is the fifth most common cause of cancer death in females in the UK. It has long been recognized to be a set of heterogeneous diseases, with high grade serous being the most common subtype. The majority of patients with EOC present at an advanced stage (FIGO III-IV), and have the largest risk for disease recurrence from which a high percentage will develop resistance to chemotherapy. Despite continual advances in diagnostics, imaging, surgery and treatment of EOC, there has been little variation in the survival rates for patients with EOC. In this review we will introduce novel bioengineering advances in modelling the lymphatic system and real-time tissue monitoring to improve the clinical and therapeutic outcome for patients with EOC. We discuss the advent of the non-invasive "liquid biopsy" in the surveillance of patients undergoing treatment and follow-up. Finally, we present new bioengineering advances for palliative care of patients to lessen symptoms of patients with ascites and improve quality of life.


Association of Helicobacter pylori infection with the Lewis and ABO blood groups in dyspeptic patients.
Helicobacter pylori infection is a basic risk factor for chronic gastritis, and gastric carcinoma. Based on some studies, the reason is binding of H. pylori to H and Le(b) antigens in gastric mucosa. However, some other findings have not determined any association between the infection and these antigens. Because of this controversy and the fact that H. pylori infection and gastric adenocarcinoma are common diseases in Iran, the assessment of the association of H. pylori infection with these blood groups could be valuable. In a cross sectional study on 135 adult dyspeptic patients in Mashhad, Iran, from 2009 to 2010, H. pylori infection was evaluated by using the Heliprobe (14)C-urea breath test and the ABO and Lewis blood group antigens were determined by the tube method. Association between the Lewis and ABO phenotypes with H. pylori infection were analysed by Fisher's exact test. A P ≤ 0.05 was considered to be significant. 68 (50.4%) patients were positive for H. pylori infection. The frequencies of the ABO, Lewis and secretion phenotypes were not significant in the infected and non-infected patients. We also did not find a significant association between Le(a) and Le(b) antigens and this infection. We could not establish a significant association between the Lewis, ABO and secretion phenotypes with H. pylori infection. Diversity of sequences of blood group antigen b-binding adhesion (babA gene) of H. pylori may be a reason why our findings are different from other studies in other geographic areas.


Cervical lymphadenopathy in childhood: nasopharyngeal carcinoma as a challenging diagnosis.
Nasopharyngeal carcinoma (NPC) is a carcinoma that arises from the nasopharyngeal mucosa and differs from other head and neck carcinomas by its unique histologic, epidemiologic, and biologic characteristics. NPC is rare in most countries, especially Europe and North America. However, it has a high incidence in several regions of South China. The incidence variability of NPC, among different geographical and ethnic groups, indicates a combination of genetic susceptibility, infection by Epstein-Barr virus and environmental factors. NPC is classified into three histological subtypes according to the 1991 World Health Organization classification: squamous cell carcinoma, nonkeratinizing carcinoma, and basaloid squamous cell carcinoma. The symptoms of patients with NPC are related to the primary tumor site and the degree of dissemination. Therefore, patients can remain asymptomatic during a long period of time. Imaging exams and biopsy of the tumor mass generally are sufficient to establish the diagnosis. NPC is a rare disease among children. The authors report a case of a 12-year-old boy who sought medical attention complaining of a progressive growing tumoral mass on the right side of the neck. The computed tomography images of the head and neck and the histological examination of a cervical lymph node biopsy diagnosed a metastatic NPC.


Treatment of GM2 Gangliosidosis in Adult Sandhoff Mice Using an Intravenous Self-Complementary Hexosaminidase Vector.
GM2 gangliosidosis is a neurodegenerative, lysosomal storage disease caused by the deficiency of β-hexosaminidase A enzyme (Hex A), an α/β-subunit heterodimer. A novel variant of the human hexosaminidase α-subunit, coded by HEX M, has previously been shown to form a stable homodimer, Hex M, that hydrolyzes GM2 gangliosides (GM2) in vivo. The current study assessed the efficacy of intravenous (IV) delivery of a self-complementary adeno-associated virus serotype 9 (scAAV9) vector incorporating the HEXM transgene, scAAV9/HEXM, including the outcomes based on the dosages provided to the Sandhoff (SD) mice. Six-week-old SD mice were injected with either 2.5E+12 vector genomes (low dose, LD) or 1.0E+13 vg (high dose, HD). We hypothesized that when examining the dosage comparison for scAAV9/HEXM in adult SD mice, the HD group would have more beneficial outcomes than the LD cohort. Assessments included survival, behavioral outcomes, vector biodistribution, and enzyme activity within the central nervous system. Toxicity was observed in the HD cohort, with 8 of 14 mice dying within one month of the injection. As compared to untreated SD mice, which have typical survival of 16 weeks, the LD cohort and the remaining HD mice had a significant survival benefit with an average/median survival of 40.6/34.5 and 55.9/56.7 weeks, respectively. Significant behavioral, biochemical and molecular benefits were also observed. The second aim of the study was to investigate the effects of IV mannitol infusions on the biodistribution of the LD scAAV9/HEXM vector and the survival of the SD mice. Increases in both the biodistribution of the vector as well as the survival benefit (average/median of 41.6/49.3 weeks) were observed. These results demonstrate the potential benefit and critical limitations of the treatment of GM2 gangliosidosis using IV delivered AAV vectors.


Evaluation of early-onset cardiotoxic effects of anthracyclines used during the treatment of childhood acute lymphoblastic leukemia by speckle-tracking echocardiography.
Anthracyclines are widely used in the treatment of acute lymphoblastic leukemia (ALL). However, cardiotoxicity is the most critical side effect that requires dose limitation. It is thought to occur at first exposure, but the clinical presentation may occur years later. In this study, we aimed to determine the time of initial damage and cardiotoxicity that develops in children with ALL. In this prospective study, 13 patients with newly diagnosed intermediate-risk precursor B cell ALL treated with the ALL-IC BFM 2009 protocol were included. Conventional echocardiography, tissue Doppler imaging (TDI), and speckle-tracking echocardiography (STE) were performed in all the patients before chemotherapy, after completing the induction phase, and at the end of the reinduction phase. The mean age of the patients was 7.8±4.6 (3.1-16.3) years. Myocardial velocity during systole (Sm) determined by TDI at the interventricular septum significantly decreased during the induction phase. Despite a decrease in STE parameters, a statistically significant reduction was determined in the global longitudinal strain rate at both left and right ventricles at the end of the induction. Nevertheless, a statistically significant increase was observed among the conventional echocardiographic findings in the left ventricular end-diastolic diameter at the end of the reinduction. During the treatment of ALL, subclinical anthracycline-associated cardiotoxicity develops in the early stages of treatment. The findings detected by TDI and STE could be missed by conventional echocardiography. We recommend evaluating patients with these newly developed techniques to detect subclinical cardiotoxicity at an early stage and starting appropriate therapy on time.


Tumor necrosis factor-alpha-stimulated membrane type 1-matrix metalloproteinase production is modulated by epidermal growth factor receptor signaling in human gingival fibroblasts.
Membrane type 1-matrix metalloproteinase (MT1-MMP) is a collagenolytic enzyme involved in connective tissue remodeling. In periodontal tissues, either cytokines or growth factors regulate the production of proteolytic enzymes. Mice deficient in epidermal growth factor receptor (EGFR) show a reduced expression of MT1-MMP, suggesting that this receptor may play an important role in MT1-MMP production. The present study evaluated the role of the inflammatory cytokine tumor necrosis factor-alpha (TNF-alpha) and EGFR in the production of MT1-MMP in gingival fibroblasts. Primary cultures of human gingival fibroblasts were cultured over plastic or a type I collagen matrix and stimulated with TNF-alpha and EGF. A selective EGFR inhibitor (AG1478) was used to interfere with this signaling pathway. Production of MT1-MMP and activation of proMMP-2 were studied using Western blot and gelatin zymography, respectively. Activation of EGFR signaling was assessed through immunoprecipitation and Western blot. Expression of EGFR ligands was determined through reverse transcriptase-polymerase chain reaction. Treatment of gingival fibroblasts cultured over a collagen matrix with TNF-alpha stimulated proMMP-2 activation and MT1-MMP production. However, after using AG1478, both responses were inhibited. Tumor necrosis factor-alpha induced EGFR transactivation and stimulated the expression of the mRNA for the EGFR ligands heparin binding-epidermal growth factor (HB-EGF) and transforming growth factor-alpha (TGF-alpha). The present study shows that TNF-alpha may stimulate MT1-MMP production through transactivation of EGFR. Tumor necrosis factor-alpha may also modulate the expression of the EGFR ligands TGF-alpha and HB-EGF. Production of MT1-MMP by TNF-alpha requires interaction with EGFR, suggesting that tissue remodeling is controlled by cross-communication between diverse signaling pathways in gingival fibroblasts.


Exceptionally high but still growing predatory reef fish biomass after 23 years of protection in a Marine Protected Area.
Marine Protected Areas (MPAs) help replenish fish assemblages, though different trophic levels may show diverse recovery patterns. Long-term protection is required to achieve total recovery but poaching events may prevent the achievement of full carrying capacity. Here, we have analysed the effect of long-term protection on the entire reef fish community and the different trophic levels in the Cabo de Palos-Islas Hormigas MPA (SE Spain; SW Mediterranean Sea) in order to assess their recovery patterns after 23 years of protection. We compared the values for carrying capacity obtained with the maximum values achieved at regional scale, and we assessed the effect of a reduction in the surveillance over a few years, during which poaching events increased, on the recovery patterns. We found that, overall, biomass of fishes increased with time while density diminished. In particular, piscivorous and macro-invertivore fish increased while the other trophic groups remained constant or declined, suggesting top-down processes. For the entire study period, those trophic groups were approaching carrying capacity; however, when accounting only for the period in which enforcement was high and constant, they grew exponentially, indicating that full carrying capacity may have not been achieved yet. When compared to other Mediterranean MPAs, the Cabo de Palos-Islas Hormigas MPA showed values for biomass that were disproportionately higher, suggesting that local factors, such as habitat structure and associated oceanographic processes, may be responsible for the dynamics found. Our results help to understand the potential trajectories of fish assemblages over a consolidated MPA and highlight empirically how the reduction of surveillance in a period may change the recovery patterns.


Anti-inflammatory and anti-arthritic effects of taraxasterol on adjuvant-induced arthritis in rats.
Taraxasterol was isolated from the traditional Chinese medicinal herb Taraxacum which has been frequently used as a remedy for inflammatory diseases. In the present study, we determined the in vivo anti-arthritic effect of taraxasterol on arthritis induced by Freund's complete adjuvant (FCA) in rats. Rats were immunized with FCA by intradermal injection into the right hind metatarsal footpad, and were orally treated daily with taraxasterol at 2, 4 and 8mg/kg from day 2-28 after immunization. Paw swelling, arthritis index, body weight, spleen index and thymus index were evaluated. The levels of TNF-α, IL-1β, PGE2, OPG and RANKL in sera were measured using ELISA. Histopathological changes in joint tissues were examined using hematoxylin and eosin (H&E). Taraxasterol significantly suppressed paw swelling and arthritis index, attenuated body weight loss, decreased the spleen index and thymus index induced by FCA. Furthermore, taraxasterol significantly inhibited the overproduction of serum TNF-α, IL-1β, PGE2 and RANKL, and increased serum OPG production in FCA-induced rats. Histopathological examination indicated that taraxasterol attenuated synovial hyperplasia, bone and cartilage damage, and inflammatory cell infiltration. These results suggest that taraxasterol has the potential protective effect against FCA-induced arthritis in rats.


Knockdown of lncRNA ZEB2NAT suppresses epithelial mesenchymal transition, metastasis and proliferation in breast cancer cells.
Breast cancer is the second most common cause of cancer-related mortality in women. Breast cancer metastasis which usually is observed at the last stage is the major cause of breast cancer-related death. Long non-coding RNAs (lncRNAs) are member of the non-coding RNA family. It is known that lncRNAs have important functions in the genes regulation of different processes and pathways such as EMT (Epithelial mesenchymal transition), metastasis and apoptosis. Therefore, it is inevitable that lncRNAs have potential contribution for the understanding of cancer pathogenesis. lncRNA-ZEB2NAT is the natural antisense transcript of ZEB2. Herein, we investigated the effects of lncRNA-ZEB2NAT on process of EMT, metastasis and apoptosis in MCF7 and MDA-MB-231 breast cancer cells. The effect of ZEB2NAT on the expression of important genes in EMT, metastasis and apoptosis, and some protein levels was determined by qRT-PCR and western blot analysis, respectively. The effects of ZEB2NAT on cell proliferation, apoptosis, invasion and colony formation were evaluated using XTT, annexin V, invasion and colony assays, respectively. The ZEB2NAT knockdown caused anti-metastatic and apoptotic effects. The ZEB2NAT knockdown resulted in a decrease in ZEB2 and N-cadherin but an increase in E-cadherin protein levels. In addition, it was determined that ZEB2NAT knockdown significantly decreased cell proliferation and stimulated apoptosis in both cells. It was found that ZEB2NAT knockdown significantly decreased invasion and colony formation in both cells. ZEB2NAT knockdown showed anti-metastatic and apoptotic effect by affecting the important genes in both cells. These results have suggested that ZEB2NAT has an important role in EMT, metastasis and apoptosis in breast cancer and ZEB2NAT knockdown caused significant anti-cancer activities.


Specific labelling of the active site of T7 RNA polymerase.
We describe a method for specifically labelling T7 RNA polymerase at (or near) the active site. Enzyme molecules that have been modified by covalent attachment of a benzaldehyde nucleotide derivative in the presence of template DNA are subsequently incubated with radioactively labelled nucleoside triphosphates. Labelling of the enzyme occurs as a result of the formation of the first phosphodiester bond. The labelling is template-directed and the expected specificity of initiation at individual T7 promoters is observed. The label has been localized to an 80 kd tryptic fragment that contains the carboxy-terminal portion of the enzyme.


Methionine Induces LAT1 Expression in Dairy Cow Mammary Gland by Activating the mTORC1 Signaling Pathway.
Methionine is the limiting amino acid for milk protein synthesis in dairy cows. The effect of methionine availability on milk protein synthesis is dependent on its active transport into cells through amino acid transporters. L-type amino acid transporter 1 (LAT1), which induces the transport of neutral amino acids, is highly expressed in lactating mammary gland. However, the effect of methionine on LAT1 expression and the mechanism governing this process in dairy cow mammary gland are poorly understood. In this study, we show that treatment of dairy cow mammary epithelial cells with increasing concentrations of methionine for 24 h resulted in increased expression of LAT1 and its associated protein 4F2 heavy chain (4F2hc). Maximal expression levels occurred after treatment with 0.6 mM methionine. Methionine treatment also increased cell viability and β-casein synthesis. Western blots showed that methionine induced LAT1 and 4F2hc expression by activating mammalian target of rapamycin complex 1 (mTORC1) signaling. Inhibition of mTORC1 signaling by rapamycin or raptor siRNA prevented the upregulation of LAT1 and 4F2hc. These results indicate that methionine may activate the mTORC1 signaling pathway and further increase LAT1 and 4F2hc expression in dairy cow mammary gland, thus affecting milk protein synthesis.


DAF-16 and SMK-1 Contribute to Innate Immunity During Adulthood in Caenorhabditis elegans.
Aging is accompanied by a progressive decline in immune function termed "immunosenescence". Deficient surveillance coupled with the impaired function of immune cells compromises host defense in older animals. The dynamic activity of regulatory modules that control immunity appears to underlie age-dependent modifications to the immune system. In the roundworm Caenorhabditis elegans levels of PMK-1 p38 MAP kinase diminish over time, reducing the expression of immune effectors that clear bacterial pathogens. Along with the PMK-1 pathway, innate immunity in C. elegans is regulated by the insulin signaling pathway. Here we asked whether DAF-16, a Forkhead box (FOXO) transcription factor whose activity is inhibited by insulin signaling, plays a role in host defense later in life. While in younger C. elegans DAF-16 is inactive unless stimulated by environmental insults, we found that even in the absence of acute stress the transcriptional activity of DAF-16 increases in an age-dependent manner. Beginning in the reproductive phase of adulthood, DAF-16 upregulates a subset of its transcriptional targets, including genes required to kill ingested microbes. Accordingly, DAF-16 has little to no role in larval immunity, but functions specifically during adulthood to confer resistance to bacterial pathogens. We found that DAF-16-mediated immunity in adults requires SMK-1, a regulatory subunit of the PP4 protein phosphatase complex. Our data suggest that as the function of one branch of the innate immune system of C. elegans (PMK-1) declines over time, DAF-16-mediated immunity ramps up to become the predominant means of protecting adults from infection, thus reconfiguring immunity later in life.


The virulence factor AvrXa7 of Xanthomonas oryzae pv. oryzae is a type III secretion pathway-dependent nuclear-localized double-stranded DNA-binding protein.
AvrXa7 is a member of the avrBs3 avirulence gene family, which encodes proteins targeted to plant cells by a type III secretion apparatus. AvrXa7, the product of avrXa7, is also a virulence factor in strain PXO86 of Xanthomonas oryzae pv. oryzae. Avirulence and virulence specificities are associated with the central repeat domain, which, in avrXa7, consists of 25.5 direct repeat units. Mutations in three C-terminal nuclear localization signal motifs eliminated avirulence and virulence activities in rice and severely reduced nuclear localization in a yeast assay system. Both pathogenicity functions and nuclear localization were restored on the addition of the sequence for the nuclear localization signal motif from SV40 T-antigen. The loss of avirulence activity because of mutations in the acidic transcriptional activation domain was restored by addition of the activation domain from the herpes simplex viral protein VP16. The activation domain was also required for virulence activity. However, the VP16 domain could not substitute for the endogenous domain in virulence assays. In gel shift assays, AvrXa7 bound double-stranded DNA with a preference for dA/dT rich sequences. The results indicate that products of the avrBs3-related genes are virulence factors targeted to host cell nuclei and have the potential to interact with the host DNA and transcriptional machinery as part of their mode of action. The results also suggest that the host defensive recognition mechanisms are targeted to the virulence factor site of action.


Mechanisms Explaining Muscle Fatigue and Muscle Pain in Patients with Myalgic Encephalomyelitis/Chronic Fatigue Syndrome (ME/CFS): a Review of Recent Findings.
Here, we review potential causes of muscle dysfunction seen in many patients with myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) such as the effects of oxidative and nitrosative stress (O&NS) and mitochondrial impairments together with reduced heat shock protein production and a range of metabolic abnormalities. Several studies published in the last few years have highlighted the existence of chronic O&NS, inflammation, impaired mitochondrial function and reduced heat shock protein production in many patients with ME/CFS. These studies have also highlighted the detrimental effects of chronically elevated O&NS on muscle functions such as reducing the time to muscle fatigue during exercise and impairing muscle contractility. Mechanisms have also been revealed by which chronic O&NS and or impaired heat shock production may impair muscle repair following exercise and indeed the adaptive responses in the striated muscle to acute and chronic increases in physical activity. The presence of chronic O&NS, low-grade inflammation and impaired heat shock protein production may well explain the objective findings of increased muscle fatigue, impaired contractility and multiple dimensions of exercise intolerance in many patients with ME/CFS.


Current ophthalmology practice patterns for syphilitic uveitis.
Syphilitic uveitis is re-emerging alongside the systemic infection. In July 2017, an international group of uveitis-specialised ophthalmologists formed the International Ocular Syphilis Study Group to define current practice patterns. 103 Study Group members based in 35 countries completed a 25-item questionnaire focused on case load, clinical presentations, use and interpretation of investigations, treatment and clinical indicators of poor prognosis. Members managed a mean of 6.1 patients with syphilitic uveitis in clinics that averaged 707 annual cases of uveitis (0.9%); 53.2% reported increasing numbers over the past decade. Patients presented to more members (40.2%) during secondary syphilis. Uveitis was usually posterior (60.8%) or pan (22.5%); complications included optic neuropathy, macular oedema and posterior synechiae. All members diagnosed syphilitic uveitis using serological tests (simultaneous or sequential testing algorithms), and 97.0% routinely checked for HIV co-infection. Cerebrospinal fluid (CSF) analysis was ordered by 90.2% of members, and 92.7% took uveitis plus Venereal Disease Research Laboratory test (VDRL) or fluorescent treponemal antibody absorption test (FTA-ABS) to indicate neurosyphilis. Patients were commonly co-managed with infectious disease physicians, and treated with penicillin for at least 10-14 days, plus corticosteroid. Features predicting poor outcome included optic neuropathy (86.3%) and initial misdiagnosis (63.7%). Reasons for delayed diagnosis were often practitioner-related. 82.5% of members tested every patient they managed with uveitis for syphilis. This comprehensive report by an international group of uveitis-specialised ophthalmologists provides a current approach for the management of syphilitic uveitis.


[Induction of paradoxical sleep (PS) by infusion of cerebrospinal fluid from PS-deprived rats].
In the rat, alpha-Methyldopa (alpha-MD, 50 mg/kg i.p.) induced during 8 hrs. an important reduction of paradoxical sleep (PS) and an increase of light slow waves sleep (SWS1). These effects were reversed by intraventricular infusion of cerebrospinal fluid (CSF) from PS-deprived donor rats, and PS restauration depended directly on the duration of the deprivation in the donor. It is probable that hypnogenic substances accumulate in the CSF during PS-deprivation, and that these factors can by-pass the noradrenergic step in the chain of biochemical events normally leading to the appearance of PS.


Therapy-induced clearance of HCV core antigen from plasma predicts an end of treatment viral response.
During viral assembly, viral proteins are released into plasma and can be used to infer viral load. The Architect hepatitis C virus (HCV) core antigen (Ag) assay is a potential alternative to HCV RNA quantification for measuring response to therapy and predicting an end of treatment viral response (EOTR). The HCVp22Ag assay was used to infer viral load in 68 window RNA-containing samples and in 284 samples from baseline to week 14 of ribavirin/interferon treatment in 23 patients with EOTR including three who relapsed, 20 not achieving EOTR and 11 controls. HCV Ag and RNA correlated well (r = 0.86) with linear dose responses on dilution. In patients on therapy and control patients, plasma HCV antigen was detected in 51 of 54 with an interpolated LOD cut off between 10(3) and 10(4) RNA IU/mL. Plasma HCV antigenaemia and plasma RNA levels were significantly different in EOTR from non-EOTR patients at 3 days after treatment start and all times thereafter. Positive and negative EOTR predictive values for HCV RNA >2 log drop and HCV Ag loss at 12 weeks were 70% and 74%, 85% and 93% respectively. HCV Ag reactivity has a linear dose response independent of genotype and correlates well with HCV RNA. The failure to clear HCV Ag is as accurate as the failure to clear HCV RNA at twelve weeks into therapy in predicting the likelihood of failure to achieve EOTR. HCV Ag potentially offers a convenient alternative to RNA measurement for defining a futility flag in HCV therapy.


Quantitative analysis of mouse mammary tumor virus in milk in two sublines of RIII/AnOk mice with low and high mammary tumor incidence.
Mammary tumors have ceased to develop in descendants of one female of the RIII/AnOk mouse strain which was brought from the Mouse Colony of Okayama University Medical School in 1975, while descendants of another female of this strain have maintained a high tumor incidence. The former and latter descendants were separated out as RIII/AnOk/2 and RIII/AnOk/1 sublines, respectively. The amount of mouse mammary tumor virus (MMTV) in the milk of individual females in these two sublines was determined by means of a sensitive enzyme immunoassay. No MMTV antigens were detected in 59 milk samples of RIII/AnOk/2 females collected during the 1st to 7th lactation periods. On the other hand, all the milk samples of RIII/AnOk/1 females contained MMTV antigens with concentrations ranging from 9 to 400 micrograms/ml. When RIII/AnOk/2 mice were foster-nursed by RIII/AnOk/1 mothers, the infected RIII/AnOk/2 females produced MMTV in the milk to almost the same degree as did the RIII/AnOk/1 females. No genetic differences between the two sublines were observed by the use of reciprocal skin grafting, tumor transplantation, and analysis of biochemical genetic markers. These results indicate that the arrest of mammary tumor development in the RIII/AnOk/2 subline was due to the disappearance of MMTV in the milk.


Adjusting for spatial variation when assessing individual-level risk: A case-study in the epidemiology of snake-bite in Sri Lanka.
Health outcomes and causality are usually assessed with individual level sociodemographic variables. Studies that consider only individual-level variables can suffer from residual confounding. This can result in individual variables that are unrelated to risk behaving as proxies for uncaptured information. There is a scarcity of literature on risk factors for snakebite. In this study, we evaluate the individual-level risk factors of snakebite in Sri Lanka and highlight the impact of spatial confounding on determining the individual-level risk effects. Data was obtained from the National Snakebite Survey of Sri Lanka. This was an Island-wide community-based survey. The survey sampled 165,665 individuals from all 25 districts of the country. We used generalized linear models to identify individual-level factors that contribute to an individual's risk of experiencing a snakebite event. We fitted separate models to assess risk factors with and without considering spatial variation in snakebite incidence in the country. Both spatially adjusted and non-adjusted models revealed that middle-aged people, males, field workers and individuals with low level of education have high risk of snakebites. The model without spatial adjustment showed an interaction between ethnicity and income levels. When the model included a spatial adjustment for the overall snakebite incidence, this interaction disappeared and income level appeared as an independent risk factor. Both models showed similar effect sizes for gender and age. HEmployment and education showed lower effect sizes in the spatially adjusted model. Both individual-level characteristics and local snakebite incidence are important to determine snakebite risk at a given location. Individual level variables could act as proxies for underling residual spatial variation when environmental information is not considered. This can lead to misinterpretation of risk factors and biased estimates of effect sizes. Both individual-level and environmental variables are important in assessing causality in epidemiological studies.


Total glossectomy with laryngeal preservation.
To evaluate the results of a planned combined therapy with surgery and postoperative radiotherapy in the management of large (T3 to T4) squamous cell carcinomas of the tongue-tongue base in properly selected patients. Twenty-one patients underwent this procedure in a 4-year period. The selection criteria were directed to identify those patients in whom laryngeal preservation was feasible and who were motivated. Five categories of patients were included: those with (1) large primary tumors of the tongue-tongue base; (2) recurrence after initial radiotherapy, brachytherapy, chemotherapy, or a combination of these modalities; (3) recurrence after initial surgery and radiotherapy; (4) double primary tumors; and (5) second or third primary tumors arising in the tongue or base of the tongue. Preoperative investigation included examination under anesthetic and magnetic resonance imaging. Depending on the proximity of the primary tumor to the mandible, a mandibular split, a marginal resection, or a segmental resection was carried out. Soft-tissue replacement was achieved by a myocutaneous or a muscle flap of the pectoralis muscle with split skin. Whenever possible tissues of the contralateral floor of the mouth were also used. Laryngeal suspension was performed in all cases. Postoperative radiotherapy consisted of megavolt therapy, 66 Gy in 6 weeks to the primary site and both sides of the neck. Resumption of swallowing and speech was achieved in all patients. External deformity was slight. Patients were able to return to their families. Total glossectomy with laryngeal preservation in properly selected patients provides local and regional control and preserves quality of life.


Phantom Study of Arterial Localization using Tactile Sensor Array and a Normal Vs. Shear Pulse Pressure Propagation Method.
Locating the radial artery reliably is a key challenge in reducing patient risks from complications in Trans-Radial Access, which is an important clinical method for catheterization, cardiac monitoring, and neuroendovascular procedures. New tactile sensing technology is being developed to bridge the skill, cost, and performance gap between ultrasonic needle guidance, and manual palpation, for use in developing countries. This paper further develops tactile artery localization with a novel algorithm for arterial localization based on the properties of a curved tactile sensor array. Using tactile sensor insensitivity to shear loading, coupled with a radial pulse wave propagation path, the position of the artery can be found at the intersection of a normal and tangential vector from the array corresponding to maximum and minimum pulse pressure measurement locations respectively. This was validated in a simple silicone phantom study Results: The proposed method measured with MAE= 0.58±0.25mm whilst the artery is within range of the tactile array, compared with 0.81±0.57mm for a comparative method of simple pulse localization. This showed improvement in arterial localization and repeatability, and was within 1 arterial radius, expected to reduce the risk of missing the artery, or perforating the side wall.Clinical Relevance- Robust and repeatable arterial localization is important for reducing the failure rate of trans-radial (and other arterial) procedures, and thus reducing the risk of harmful complications.


The "3 in 1" Study: Pooling Self-Taken Pharyngeal, Urethral, and Rectal Samples into a Single Sample for Analysis for Detection of Neisseria gonorrhoeae and Chlamydia trachomatis in Men Who Have Sex with Men.
Triple-site testing (using pharyngeal, rectal, and urethral/first-void urine samples) for Neisseria gonorrhoeae and Chlamydia trachomatis using nucleic acid amplification tests detects greater numbers of infections among men who have sex with men (MSM). However, triple-site testing represents a cost pressure for services. MSM over 18 years of age were eligible if they requested testing for sexually transmitted infections (STIs), reported recent sexual contact with either C. trachomatis or N. gonorrhoeae, or had symptoms of an STI. Each patient underwent standard-of-care (SOC) triple-site testing, and swabs were taken to form a pooled sample (PS) (pharyngeal, rectal, and urine specimens). The PS was created using two methods during different periods at one clinic, but we analyzed the data in combination because the sensitivity of the two methods did not differ significantly for C. trachomatis (P = 0.774) or N. gonorrhoeae (P = 0.163). The sensitivity of PS testing (92%) was slightly lower than that of SOC testing (96%) for detecting C. trachomatis (P = 0.167). For N. gonorrhoeae, the sensitivity of PS testing (90%) was significantly lower than that of SOC testing (99%) (P < 0.001). When pharynx-only infections were excluded, the sensitivity of PS testing to detect N. gonorrhoeae infections increased to 94%. Our findings show that pooling of self-taken samples could be an effective and cost-saving method, with high negative predictive values. (Interim results of this study were presented at the BASHH 2013 summer meeting.).


Hand infections.
Special approaches are required for management of infected wounds involving the hand. Care must be taken to consider involvement of joint, bone, and tendon, or tendon sheath as well as deep fascial spaces. The hand is also prone to infections with unusual microbiology that can result from animal and human bites as well as soil, plant, and water exposures. The special anatomic and microbiologic considerations are critical for selection of proper surgical and medical therapy to preserve function of this vital structure.


Sex differences in aripiprazole sensitization from adolescence to adulthood.
The present study investigated the potential sex differences in repeated aripiprazole (ARI) treatment-induced behavioral sensitization from adolescence to adulthood, and to determine whether ARI sensitization can be transferred to olanzapine (OLZ) and/or clozapine (CLZ) using the conditioned avoidance response (CAR) and phencyclidine-induced (PCP) hyperlocomotion tests of antipsychotic activity. Male and female Sprague-Dawley adolescence rats (P46) were first treated with ARI (10mg/kg) for 5 consecutive days (P46-50) and tested for avoidance response and ARI-induced inhibition of PCP-induced hyperlocomotion. After they became adults (>P68), rats were challenged with ARI (1.5mg/kg, sc) (P70), OLZ (0.5mg/kg, sc; P73), CLZ (5mg/kg, sc; P76) and again with ARI (1.5mg/kg, sc; P84) and tested for avoidance response and ARI-induced inhibition of PCP-induced hyperlocomotion again. During the drug treatment period in adolescence, repeated ARI treatment suppressed avoidance response, inhibited the PCP-induced hyperlocomotion, and these effects were progressively increased across the 5-day period in both males and females, confirming the induction of ARI sensitization. On the challenge days, rats previously treated with ARI in adolescence also had significantly lower avoidance and lower PCP-induced hyperlocomotion than the previous vehicle rats, confirming the expression of ARI sensitization and its persistence into adulthood. More importantly, female rats made significantly more avoidances than males in both ARI and vehicle groups, indicating higher sensitivity to the acute and long-term effects of ARI. Further, on the OLZ and CLZ challenge days, prior ARI treatment seemed to increase sensitivity to OLZ exposure, however, this increase was not significant. Similarly, rats also showed an ARI sensitization to OLZ and CLZ on challenge days. Collectively, results from this experiment demonstrated a sex difference in response to ARI and enhanced inhibition of PCP-induced hyperlocomotion in animals that were pretreated with ARI as compared to controls.


Eat walk engage: an interdisciplinary collaborative model to improve care of hospitalized elders.
High-quality, efficient health care for older patients is a priority for health care systems. Acute Care for Elders units improve outcomes but there is a need for generalizable models of care that adopt the principles pioneered in these units. This report describes Eat Walk Engage, a collaborative care model on a general medical ward in Brisbane, Australia. The model focused on early mobilization, feeding assistance, and cognitive stimulation. Using the Promoting Action on Research Implementation in Health Services implementation framework, the facilitation team enabled the clinical team to recognize barriers and develop solutions. Challenges included unclear responsibility, workload concerns, and risk aversion. Implementation strategies included engaging champions, education, audit and feedback, task delineation and delegation, improving physical resources, and workforce redesign. During the first 18 months, audits showed improved nursing documentation in targeted domains and improved performance of mobilizing and cognitive strategies; length of stay for older inpatients fell by 3 days on the intervention ward.


Selection and characterization of interferon-sensitive cells derived from an interferon-resistant NIH 3T3 line.
We have developed a selection protocol to isolate interferon (IFN)-sensitive subclones directly from an IFN-resistant cell population. The protocol uses encephalomyocarditis virus (EMCV) as a selection agent in combination with pretreatment with low doses of IFN and subsequent growth in the presence of virus-neutralizing antiserum. We have applied this protocol to the partially IFN-resistant NIH 3T3 clone 1 line and have obtained a number of IFN-sensitive subclones. Sensitivity to IFN was restricted to protection against EMCV. Replication of vesicular stomatitis virus as well as cell growth were resistant to IFN treatment as in the original clone 1 line. We have compared levels of 2',5'-oligoadenylate (2-5A) synthetase, dsRNA-activated protein kinase and 2-5A-dependent RNase in some IFN-sensitive subclones and found no difference from the resistant clone 1 cells. Markedly decreased levels of 2-5A-dependent RNase and thus a defective 2-5A pathway have been implicated as a possible cause for the partial resistance of clone 1 cells to IFN. Since the selected IFN-sensitive subclones are of the same phenotype in this respect as the clone 1 line we suggest that inhibition of EMCV in these lines occurs through a mechanism independent of the 2-5A system.


Structure and analysis of the human dimethylglycine dehydrogenase gene.
Dimethylglycine dehydrogenase (DMGDH; E.C. 220.127.116.11) is an enzyme involved in the catabolism of choline, catalyzing the oxidative demethylation of dimethylglycine (DMG) to form sarcosine. Subsequently, sarcosine dehydrogenase (SDH; E.C. 18.104.22.168) converts sarcosine to glycine via a similar reaction. Both enzymes are found as monomers in the mitochondrial matrix, and both contain 1 mol of covalently bound flavin adenine dinucleotide. DMGDH and SDH also utilize a noncovalently bound folate coenzyme that receives the "1-carbon" groups that are removed by DMGDH and SDH, forming "active formaldehyde." We have recently described a new inborn error of metabolism of DMGDH characterized by an unusual fish-like body odor. To augment our study of this new disorder, we have isolated two human genomic clones that together contain 16 exons of coding sequence for the hDMGDH gene. Fluorescent in situ hybridization analysis of the hDMGDH gene indicates that it is found on chromosome 5q12.2-q12.3. In addition, several polymorphisms have been identified in the hDMGDH cDNA sequence. Population analysis of two Ser/Pro polymorphisms found 367 amino acids apart reveals a skew of alleles, with the haplotypes Ser/Pro or Pro/Ser (79%) overrepresented compared to the number of Ser/Ser or Pro/Pro alleles observed. Possible functional consequences of these findings are discussed. Characterization of the gene structure for hDMGDH will aid in the study of patients with inherited defects of this enzyme.


Charged Carbon Nanomaterials: Redox Chemistries of Fullerenes, Carbon Nanotubes, and Graphenes.
Since the discovery of buckminsterfullerene over 30 years ago, sp2-hybridised carbon nanomaterials (including fullerenes, carbon nanotubes, and graphene) have stimulated new science and technology across a huge range of fields. Despite the impressive intrinsic properties, challenges in processing and chemical modification continue to hinder applications. Charged carbon nanomaterials (CCNs), formed via the reduction or oxidation of these carbon nanomaterials, facilitate dissolution, purification, separation, chemical modification, and assembly. This approach provides a compelling alternative to traditional damaging and restrictive liquid phase exfoliation routes. The broad chemistry of CCNs not only provides a versatile and potent means to modify the properties of the parent nanomaterial but also raises interesting scientific issues. This review focuses on the fundamental structural forms: buckminsterfullerene, single-walled carbon nanotubes, and single-layer graphene, describing the generation of their respective charged nanocarbon species, their interactions with solvents, chemical reactivity, specific (opto)electronic properties, and emerging applications.


Insight into catalysis of a unique GTPase reaction by a combined biochemical and FTIR approach.
Rap1 and Rap2 are the only small guanine nucleotide-binding proteins of the Ras superfamily that do not use glutamine for GTP hydrolysis. Moreover, Rap1GAP, which stimulates the GTPase reaction of Rap1 10(5)-fold, does not have the classical "arginine finger" like RasGAP but presumably, introduces an asparagine residue into the active site. Here, we address the requirements of this unique reaction in detail by combining various biochemical methods, such as fluorescence spectroscopy, stopped-flow and time-resolved Fourier transform infrared spectroscopy (FTIR). The fluorescence spectroscopic assay monitors primarily protein-protein interaction steps, while FTIR resolves simultaneously the elementary steps of functional groups labor-free, but it is less sensitive and needs higher concentrations. Combining both methods allows us to distinguish weather mechanistic defects caused by mutation are due to affinity or due to functionality. We show that several mutations of Asn290 block catalysis. Some of the mutants, however, still form a complex with Rap1*GDP in the presence of BeF(x) but not AlF(x), supporting the notion that fluoride complexes are indicators of the ground versus transition state. Mutational analysis also shows that Thr61 is not required for catalysis. While replacement of Thr61 of Rap1 by Leu eliminates GTPase activation by Rap1GAP, the T61A and T61Q mutants have only a minor effect on catalysis, but change the relative rates of cleavage and (P(i)(-)) release. While Rap1GAP(N290A) is completely inactive on wild-type Rap1, it can act on Rap1(T61Q), arguing that Asn290 in trans has a role in catalysis similar to that of the intrinsic Gln in Ras and Rho. Finally, since FTIR works at high, and thus mostly saturating, concentrations, it can clearly separate effects on affinity from purely catalytic modifications, showing that Arg388, conserved between RapGAPs and mutated in the homologous RheBGAP Tuberin, affects binding affinity severely but has no effect on the cleavage reaction itself.


Business and continuity of operations: care of the critically ill and injured during pandemics and disasters: CHEST consensus statement.
During disasters, supply chain vulnerabilities, such as power, transportation, and communication, may affect the delivery of medications and medical supplies and hamper the ability to deliver critical care services. Disasters also have the potential to disrupt information technology (IT) in health-care systems, resulting in interruptions in patient care, particularly critical care, and other health-care business functions. The suggestions in this article are important for all of those involved in a large-scale pandemic or disaster with multiple critically ill or injured patients, including front-line clinicians, hospital administrators, and public health or government officials. The Business and Continuity of Operations Panel followed the American College of Chest Physicians (CHEST) Guidelines Oversight Committee's methodology in developing key questions regarding medication and supply shortages and the impact disasters may have on healthcare IT. Task force members met in person to develop the 13 key questions believed to be most relevant for Business and Continuity of Operations. A systematic literature review was then performed for relevant articles and documents, reports, and gray literature reported since 2007. No studies of sufficient quality were identified upon which to make evidence-based recommendations. Therefore, the panel developed expert opinion-based suggestions using a modified Delphi process. Eighteen suggestions addressing mitigation strategies for supply chain vulnerabilities including medications and IT were generated. Suggestions offered to hospitals and health system leadership regarding medication and supply shortages include: (1) purchase key medications and supplies from more than one supplier, (2) substituted medications or supplies should ideally be similar to those already used by an institution's providers, (3) inventories should be tracked electronically to monitor medication/supply levels, (4) consider higher inventories of medications and supplies known or projected to be in short supply, (5) institute alternate use protocols when a (potential) shortage is identified, and 6) support government and nongovernmental organizations in efforts to address supply chain vulnerability. Health-care IT can be damaged in a disaster, and hospitals and health system leadership should have plans for urgently reestablishing local area networks. Planning should include using portable technology, plans for providing power, maintenance of a patient database that can accompany each patient, and protection of patient privacy. Additionally, long-term planning should include prioritizing servers and memory disk drives and possibly increasing inventory of critical IT supplies in preparedness planning. The provision of care to the critically ill or injured during a pandemic or disaster is dependent on key processes, such as the supply chain, and infrastructure, such as IT systems. Hospitals and health systems will help minimize the impact of medication and supply shortages with a focused strategy using the steps suggested. IT preparedness for maintaining local area networks, functioning clinical information systems, and adequate server and memory storage capacity will greatly enhance preparedness for hospital and health system clinical and business operations.


Change in energy expenditure and physical activity in response to aerobic and resistance exercise programs.
Exercise is considered an important component of a healthy lifestyle but there remains controversy on effects of exercise on non-exercise physical activity (PA). The present study examined the prospective association of aerobic and resistance exercise with total daily energy expenditure and PA in previously sedentary, young men. Nine men (27.0 ± 3.3 years) completed two 16-week exercise programs (3 exercise sessions per week) of aerobic and resistance exercise separated by a minimum of 6 weeks in random order. Energy expenditure and PA were measured with the SenseWear Mini Armband prior to each intervention as well as during week 1, week 8 and week 16 of the aerobic and resistance exercise program. Body composition was measured via dual x-ray absorptiometry. Body composition did not change in response to either exercise intervention. Total daily energy expenditure on exercise days increased by 443 ± 126 kcal/d and 239 ± 152 kcal/d for aerobic and resistance exercise, respectively (p < 0.01). Non-exercise moderate-to-vigorous PA, however, decreased on aerobic exercise days (-148 ± 161 kcal/d; p = 0.03). There was no change in total daily energy expenditure and PA on non-exercise days with aerobic exercise while resistance exercise was associated with an increase in moderate-to-vigorous PA during non-exercise days (216 ± 178 kcal/d, p = 0.01). Results of the present study suggest a compensatory reduction in PA in response to aerobic exercise. Resistance exercise, on the other hand, appears to facilitate non-exercise PA, particularly on non-exercise days, which may lead to more sustainable adaptations in response to an exercise program.


Chinese version of the Cancer Support Person's Unmet Needs Survey-Sort Form: A psychometric study.
Psychometrically rigorous and comprehensive needs assessment measures for Chinese general cancer caregivers are relatively scarce. We described the development and psychometric evaluation of the Chinese version of the Cancer Support Person's Unmet Needs Survey-Short Form (SPUNS-SFC). A forward-backward translation method was utilised to develop the SPUNS-SFC. The construct validity, internal consistency, convergent validity and discriminant validity of the SPUNS-SFC were evaluated. A total of 1,026 Chinese cancer caregivers completed the SPUNS-SFC. Items 10, 11, 13, 24 and 26 were removed as more than 80% of respondents reported having no unmet needs on these items. Exploratory factor analysis revealed a five-factor structure, which accounted for 78.47% of the total variance, and consisted of the following domains: information, healthcare access and continuity, personal and emotional needs, worries about the future, and financial needs. Internal consistency of the measure was high, with Cronbach's alpha coefficients ranging from 0.87 to 0.95 for the five domains. The measure illustrated adequate evidence of convergent validity, demonstrated by significant correlations with multiple measures of psychological well-being. Known-groups validity was established, as 87.5% of the hypotheses were supported. This study indicates the SPUNS-SFC is a reliable and valid measure of the unmet needs of Chinese cancer caregivers.


The unintended consequences of New Zealand's primary health care reforms.
In 2001, the New Zealand government commenced a program to reform the organization of publicly funded primary care services. While there have been several positive results of this reform, including the reduction of patient co-payments and the extension of the range of primary care services, the government's program was a hastily implemented attempt to place primary care, the delivery of which is dominated by private doctors, under firm state control. It was also an attempt to override preexisting arrangements. As such, the government succeeded in its goal of establishing new primary health organizations (PHOs), but there were also significant unintended consequences. As detailed in this article, these consequences include (1) the creation of a labyrinthine funding and organizational system with a variable capacity to deliver on the government's reform objectives, (2) an increase in the power and scope of preexisting doctor organizations combined with a government unable to wrest control over the setting of patient co-payment levels, and (3) an emerging lack of clarity about future directions for the primary health care sector.


Feasibility and Accuracy of a Nonmedical Research Person in Assimilation and Calculation of Acute Physiologic Assessment and Chronic Health Evaluation Scores in an Indian Intensive Care Unit.
The Physiologic Assessment and Chronic Health Evaluation (APACHE) score assimilation and calculation, as well as other demographic data collection, is inherent to research and nonresearch related needs of intensive care. There may be a role for well-trained nonmedical personnel to collect this vital material to enhance research and the standard of care in the Intensive Care Units (ICUs) in countries that are poorly funded and resourced in terms of medical personnel. The aim of this study is to verify the interrater reliability of a trained nonmedical personnel and ICU trainee in the collection and calculation APACHE scores. In a prospective study, two raters who were blinded, one a trained nonmedical ward clerk and another an ICU trainee, assimilated data and calculated APACHE scores for 60 consecutive patients admitted to two tertiary mixed ICUs (with a total of 19 beds). Primary outcomes were to assess interrater and interclass correlation as well as the agreement of scores between the two raters. There was an excellent correlation of APACHE scores (Kappa coefficient of 0.92) and Bland-Altman plot depicted overall good agreement with low bias between raters. A well-trained and supervised nonmedical research person can assimilate and calculate APACHE II scores with good agreement with an ICU trainee. This may help in deriving data from medically understaffed ICUs in India, thus promoting much-needed research from such ICUs.


IDENTIFICATION OF CONOIDIN A AS A COVALENT INHIBITOR OF PEROXIREDOXIN II.
Conoidin A (1) is an inhibitor of host cell invasion by the protozoan parasite Toxoplasma gondii. In the course of studies aimed at identifying potential targets of this compound, we determined that it binds to the T. gondii enzyme peroxiredoxin II (TgPrxII). Peroxiredoxins are a widely conserved family of enzymes that function in antioxidant defense and signal transduction, and changes in PrxII expression are associated with a variety of human diseases, including cancer. Disruption of the TgPrxII gene by homologous recombination had no effect on the sensitivity of the parasites to 1, suggesting that TgPrxII is not the invasion-relevant target of 1. However, we showed that 1 binds covalently to the peroxidatic cysteine of TgPrxII, inhibiting its enzymatic activity in vitro. Studies with human epithelial cells showed that 1 also inhibits hyperoxidation of human PrxII. These data identify Conoidin A as a novel inhibitor of this important class of antioxidant and redox signaling enzymes.


Origin of Coding RNA from Random-Sequence RNA.
D-ribose and D-arabinose differ only by the steric orientation of their C2-OH groups. The initial reactions and emergence of RNA depended on the position, reactivity, and flexibility of the C2-OH moiety in the ribose molecule. The steric relationship of the C2- and C3-OH groups favored the selection of ribose, ribonucleotide, and RNA synthesis and excluded the possibility of xenonucleic acid-based life on Earth. This brief review provides a hypothesis based on the absence of nucleotides and enzymes under prebiotic conditions and on the polymerization of ribose 5-phosphate units leading to the polarized formation of the ribose-phosphate backbone. The strong covalent bond formation in the sugar-phosphate backbone was followed by the somewhat less reactive interaction between ribose and nucleobase and supplemented by even weaker hydrogen-bonded and stacking interactions. This hypothesis proposes a scheme how prebiotic random-sequence RNA was formed under abiotic conditions and hydrolyzed to oligomers and nucleotides. The term random-sequence prebiotic RNA refers to nucleobases attached randomly to the ribose-phosphate backbone and not to cellular RNA sequences as proteins and cells did not probably exist at the time of abiotic RNA formation. It is hypothesized that RNA generated under abiotic conditions containing random nucleobases was hydrolyzed to nucleotides that served as a pool for the selected synthesis of genetic RNA.


Cuproptosis-related molecular patterns and gene (ATP7A) in hepatocellular carcinoma and their relationships with tumor immune microenvironment and clinical features.
Cuproptosis has been studied in various aspects as a new form of cell death. We hope to explore the molecular patterns and genes related to cuproptosis in evaluating and predicting the prognosis of hepatocellular carcinoma (HCC), as well as the impact of tumor immune microenvironment. Sixteen cuproptosis related gene (CRGs) and cuproptosis related molecular and gene characteristics were comprehensively analyzed from 492 HCC samples. Cuproptosis related molecular patterns were generated by consensus clustering algorithm, including cuproptosis clusters, cuproptosis gene clusters (CGC) and cuproptosis score (CS). The characteristics of tumor microenvironment (TME) and tumor immune cells were described by the ssGSEA and ESTIMATE algorithms. Cuproptosis score was established to assess the clinical characteristics, prognostic and immunotherapy. The role and mechanism of CRG (ATP7A) in HCC, as well as its relationship with TME and immune checkpoints, have been further explored. The results of somatic mutation, copy number variations (CNV), and CRGs expression in HCC suggested the CRGs might participate in the HCC oncogenesis. The cuproptosis clusters were closely related to the clinical pathological characteristics, biological processes, and prognosis of HCC. The three CGC was revealed to be consistent with the three immune infiltration characterizations, including immune-high, immune-mid, and immune-low subtypes. Higher CS was characterized by decreased TMB, activated immunity, higher immune cell proportion score (IPS) and better overall survival (OS), which indicated higher CS was immune-high type and with better treatment effect and prognosis. The ATP7A had the highest hazard ratio (HR = 1.465, p < .001), was high expression in HCC tissues and with a shorter 5-year OS. Knocking down ATP7A could enhance intracellular copper concentration, cause a decrease in DLAT expression, and induce cuproptosis and inhibit cell proliferation and migration. ATP7A was also positively correlated with most cancer immune cells and immune checkpoints. Taken together, this research revealed the cuproptosis related molecular patterns and genes associated with the clinical pathological characteristics, TME phenotype and prognosis of HCC. The CS will further deepen our understanding of the TME characteristics of HCC, and the involvement of ATP7A in cuproptosis will provide new ideas for predicting HCC prognosis and immunotherapy.


Motor nerve conduction velocity in spinal muscular atrophy of childhood.
The ulnar and posterior tibial conduction velocities were measured in 29 children with spinal muscular atrophy, 14 of whom had the servere form of the disease. The ulnar nerve velocity was slow in 12 of the 14 severely affected infants, but normal or fast in 11 of 14 children less severely affected. The corresponding results for the posterior tibial nerve were slow velocities in 11 of 12 infants in the severe group and normal or fast in all 11 infants less severely affected. The difficulty in distinguishing infantile spinal muscular atrophy from peripheral neuropathy is emphasized.


Comparative Autonomic Responses to Diagnostic Interviewing between Individuals with GAD, MDD, SAD and Healthy Controls.
Dysregulation of the autonomic nervous system (ANS) has been well documented in individuals diagnosed with a range of psychological disorders, including generalized anxiety disorder (GAD) and major depressive disorder (MDD). Moreover, these disorders both confer an increased risk of cardiovascular disease-which may relate to increased sympathetic and decreased parasympathetic tone. Extant research has indicated a reduction in autonomic flexibility in GAD, and while reduced flexibility has also been seen in MDD, the specific physiological alterations have been more difficult to categorize due to methodological limitations, including high co-morbidity rates with anxiety disorders. Prior studies have largely assessed autonomic functioning in stress paradigms or at the trait level, yet to date, no research has investigated the ANS during a diagnostic interview, a ubiquitous task employed in both research and clinical settings. In this study we sought to identify physiological differences in both branches of the ANS across diagnostic categories in the context of a diagnostic interview. Participants (n = 82) were administered a structured clinical interview, during which heart rate (HR), respiratory sinus arrhythmia (RSA) and pre-ejection period (PEP) were recorded in participants carrying a diagnosis of GAD (n = 34), MDD (n = 22), Social Anxiety Disorder (SAD; n = 15) and healthy controls (n = 27). Person-specific linear regression models were employed to assess the level and slope for HR, RSA and PEP throughout the course of the interview. A multivariate analysis of variance (MANOVA) model was conducted to baseline differences in HR, RSA and PEP between diagnostic groups. Multiple regression models were then conducted to differences in slope of HR, RSA and PEP throughout the course of the interview amongst diagnostic groups, including both suppression and worry as moderators. Results indicated significant increases in RSA throughout the interview in MDD (p = 0.01) compared to healthy controls. Worry itself was found to be a more significant predictor of both decreased PEP (p = 0.02) and increased HR (p = 0.05). Suppression exhibited a dampening effect on individuals with worry and GAD, whereby those who suppressed had dampened HR responsiveness compared to those who did not suppress. These findings are consistent with existing literature supporting a decreased autonomic flexibility in certain psychological disorders, as well as indicate distinct physiological differences across certain transdiagnostic features of mood and anxiety disorders.


Scavenging versus predation: shark-bite injuries in stranded sea turtles in the southeastern USA.
Injuries inflicted by sharks are a frequent observation in stranded sea turtles. Sharks prey on live turtles and scavenge carcasses, which can create uncertainty as to the cause of stranding when sea turtles are found dead with shark-bite wounds. Consequently, attributing the cause of stranding to a shark attack based purely on the presence of the characteristic wounds can overestimate predation by sharks as a cause of mortality. To better characterize the timing of shark-bite wounds relative to death of sea turtles in the southeastern USA, we performed necropsies on 70 stranded turtles that were found dead in which the predominant observation was bite wounds without any grossly evident vital responses (inflammation or healing). Postmortem examination included assessment for evidence of exsanguination and histopathological evaluation of skeletal muscle comprising wound margins. We characterized wounds as antemortem, perimortem, or postmortem based on specific criteria related to the presence or absence of supravital and intravital responses. Most (80%) shark-bite wounds were postmortem, 10% were antemortem, and 10% were perimortem. We found that antemortem and postmortem wounds were similar in extent and location except for wounds that primarily involved the shell, which were never found in cases of scavenging. For sea turtles found dead in the southeastern USA, our findings suggest that most shark-bite wounds without externally evident vital responses are due to scavenging. Additionally, this scavenging can significantly damage a carcass, potentially obscuring the detection of other causes of mortality. These findings should be considered when using data derived from stranded sea turtles to conduct mortality assessments.


[Characteristics of the protein spectra of individual representatives of mycobacteria].
A study of proteinograms of some strains of mycobacteria with the use of the method of electrophoresis in polyacrylamide gel showed that individual strains had characteristic protein spectra which could serve as one of the possible taxonomic characteristics. Analysis of proteinograms of three groups of mycobacteria (tuberculosis, atypical and saprophytes) by electrophoretic mobility of the fractions, with consideration to their homologous character and by the intensity of protein fractions in the mobility zones pointed to heterogeneity of the group of atypical strains and their greater similarity to tuberculosis mycobacteria than to acid-fast saprophytes. Proteinograms of strains H37 Rv and Bov-8 proved to be identical; a marked similarity between the protein spectra of M. avium and M. battey was revealed.


Stereotactic radiosurgery for arteriovenous malformations, Part 4: management of basal ganglia and thalamus arteriovenous malformations.
The authors conducted a study to define the long-term outcomes and risks of stereotactic radiosurgery (SRS) for arteriovenous malformations (AVMs) of the basal ganglia and thalamus. Between 1987 and 2006, the authors performed Gamma Knife surgery in 996 patients with brain AVMs; 56 patients had AVMs of the basal ganglia and 77 had AVMs of the thalamus. In this series, 113 (85%) of 133 patients had a prior hemorrhage. The median target volume was 2.7 cm(3) (range 0.1-20.7 cm(3)) and the median margin dose was 20 Gy (range 15-25 Gy). Obliteration of the AVM eventually was documented on MR imaging in 78 patients and on angiography in 63 patients in a median follow-up period of 61 months (range 2-265 months). The actuarial rates documenting total obliteration after radiosurgery were 57%, 70%, 72%, and 72% at 3, 4, 5, and 10 years, respectively. Factors associated with a higher rate of AVM obliteration included AVMs located in the basal ganglia, a smaller target volume, a smaller maximum diameter, and a higher margin dose. Fifteen (11%) of 133 patients suffered a hemorrhage during the latency period and 7 patients died. The rate of post-SRS AVM hemorrhage was 4.5%, 6.2%, 9.0%, 11.2%, and 15.4% at 1, 2, 3, 5, and 10 years, respectively. The overall annual hemorrhage rate was 4.7%. When 5 patients with 7 hemorrhages occurring earlier than 6 months after SRS were removed from this analysis, the annual hemorrhage rate decreased to 2.7%. Larger volume AVMs had a higher risk of hemorrhage after SRS. Permanent neurological deficits due to adverse radiation effects (AREs) developed in 6 patients (4.5%), and in 1 patient a delayed cyst developed 56 months after SRS. No patient died of AREs. Factors associated with a higher risk of symptomatic AREs were larger target volume, larger maximum diameter, lower margin dose, and a higher Pollock-Flickinger score. Stereotactic radiosurgery is a gradually effective and relatively safe management option for deep-seated AVMs in the basal ganglia and thalamus. Although hemorrhage after obliteration did not occur in the present series, patients remain at risk during the latency interval between SRS and obliteration. The best candidates for SRS are patients with smaller volume AVMs located in the basal ganglia.


Major concerns in developing countries: applications of the Precautionary Principle in Ecuador.
Ecuador is a Latin American country with one of the biggest biodiversities. At the same time, social and environmental problems are also big. Poverty, political and social problems as well as questions like old transport systems, imported hazards from industrialized countries and lack of information and weak health care systems are the framework of this situation. The most common problems are the use of heavy metals in many activities without safety and health protection, a low technological oil production during two decades, intensive use of pesticides in agriculture, and some other chemical risks. A limited capacity to develop prevention strategies, reduced technical and scientific skills, and the absence of a reliable information and control system, lead to a weak response mechanism. The Precautionary Principle could help to stimulate prevention, protection and to have a new tool to improve the interest in environment and health problems. Reinforcing the presence of international organizations like the World Health Organization or the International Labour Organization, establishing bridges among scientific organizations from developed and developing countries and introducing the Precautionary Principle in the legislation and daily practices of industry and agriculture could lead to an improvement in our environment and health.


Internal laryngocoele and referred otalgia.
A 74-year-old woman presented with severe right ear pain associated with bleeding of 2 months' duration and vertigo. She was otherwise fit and well apart from arthritis. On examination there was dried blood overlying the right ear drum and very mild inflammation of the canal but no obvious acute infection. The dried blood was removed in subsequent clinic visits. The ear drum was intact and there was no evidence of infection. However, the patient still had persistent pain in the right ear. A diagnosis of referred otalgia was made. Nose, throat, neck, temporo-mandibular joint, oral examination and flexible nasoendoscopic examination were normal. She underwent MRI of her neck which revealed a right supraglottic mass. Laryngoscopy was performed. Intra-operative findings revealed a smooth right supraglottic mass, which was thought to be an internal laryngocoele. The laryngocoele was de-roofed. The histology specimen showed no evidence of neoplasia.


New information about resynchronisation.
Chronic atrio-biventricular pacing appears to become a very attractive and promising additive therapy for congestive heart failure with systolic left ventricular impairment. The best results can only be achieved by an optimal interaction between the cardiologists involved. Until now no sort of cardiac intervention has ever required such close and ongoing cooperation to serve the cardiac patient. These efforts are welcomed because congestive heart failure strongly reduces the quality of mental health and physical functioning. By means of questions and answers on new information about cardiac resynchronisation, this special article aims to provide more insight into the role of this complex pacing therapy.


Transforming Growth Factor-Beta (TGFβ) Signaling Pathway in Cholangiocarcinoma.
Cholangiocarcinoma is a deadly cancer worldwide, associated with a poor prognosis and limited therapeutic options. Although cholangiocarcinoma accounts for less than 15% of liver primary cancer, its silent nature restricts early diagnosis and prevents efficient treatment. Therefore, it is of clinical relevance to better understand the molecular basis of cholangiocarcinoma, including the signaling pathways that contribute to tumor onset and progression. In this review, we discuss the genetic, molecular, and environmental factors that promote cholangiocarcinoma, emphasizing the role of the transforming growth factor β (TGFβ) signaling pathway in the progression of this cancer. We provide an overview of the physiological functions of TGFβ signaling in preserving liver homeostasis and describe how advanced cholangiocarcinoma benefits from the tumor-promoting effects of TGFβ. Moreover, we report the importance of noncoding RNAs as effector molecules downstream of TGFβ during cholangiocarcinoma progression, and conclude by highlighting the need for identifying novel and clinically relevant biomarkers for a better management of patients with cholangiocarcinoma.