Methionine metabolism in human pregnancy.
Hyperhomocysteinemia during pregnancy, which is a consequence of perturbations in methionine and/or folate metabolism, has been implicated in adverse outcomes such as neural tube defects, preeclampsia, spontaneous abortion, and premature delivery. The adaptive changes in methionine metabolism during pregnancy in humans have not been determined. Our objective was to examine the kinetics of methionine and its rate of transsulfuration and transmethylation in healthy women with advancing gestation. The whole-body rate of appearance (Ra) of methionine and phenylalanine was measured in healthy pregnant women during the first (n = 10), second (n = 5), and third (n = 10) trimesters of pregnancy. These data were compared with those for nonpregnant women (n = 8). Tracers [1-(13)C]methionine, [C(2)H(3)]methionine, and [(2)H(5)]phenylalanine were administered as prime-constant rate infusions. The effect of enteral high-protein, mixed-nutrient load on tracer-determined variables was also examined. In pregnant women, the Ra of phenylalanine was significantly (P < 0.05) lower in the first trimester than in the second and third trimesters and was significantly lower than that in nonpregnant women. A linear positive correlation was evident between gestational age and phenylalanine Ra. The fractional rate and total rate of transsulfuration of methionine was significantly (P < 0.05) higher during the first trimester, whereas the rate of transmethylation was higher during the third trimester. Plasma concentrations of total cysteine and homocysteine were lower during pregnancy. Uncomplicated pregnancy in humans is associated with a higher rate of transsulfuration early in gestation and a higher rate of transmethylation of methionine in late gestation. These data may have implications for understanding the role of methionine and homocysteine in complications of pregnancy and for the nutritional care of pregnant women.


Religious orientation and mental health: a study with Turkish university students.
The present study examined the relationship between religious orientation and mental health symptoms among Turkish students. A total of 341 undergraduate students between the ages of 18 and 26 completed the Brief Symptom Inventory and Age Universal Intrinsic-Extrinsic Scale. Extrinsic orientation was found to be correlated positively with symptoms of anxiety and depression. There were also significant correlations between hostility scores and both intrinsic and extrinsic religious orientations. Patterns were similar to those reported previously, but correlations were mostly in the .10-.14 range. In multiple-regression models, extrinsic orientation emerged as the only significant predictor for hostility, anxiety, and depression; however, only 2% of the variance was explained.


Can working memory capacity be expanded by boosting working memory updating efficiency in older adults?
Working memory updating (updating) and working memory capacity (WMC) have been assumed to share a common mechanism. However, it is unclear whether WMC can be expanded by boosting the efficiency of updating, particularly during late adulthood. In this randomized controlled study, 33 older adults (aged 60 years and above, M = 69.53, SD = 5.21) were assigned to updating training (n = 17) and contact control (n = 16) groups. In the training group, updating was targeted by a running memory task and a chess game in each training session; whereas in the control group, motivational effects were estimated by their attendance to a series of mental health-related lectures. The boosted updating efficiency was demonstrated by direct training gains over sessions on updating criterion/training tasks, and significant frontocentral inflation at the N2 time range. Near transfer effects were revealed on untrained similar measures immediately after training and partly maintained at a 3-month follow-up. Far transfer effects to WMC were revealed by a significant increase in contralateral delay activity (CDA) ERP measures, although these effects did not manifest on WMC span tasks and other far transfer measures. Note that increased CDA was significantly correlated with the extent of direct gains in updating training. Our findings suggest that updating efficiency can be boosted in older adults; although it has not yet brought about an explicit expansion of WMC in our behavioral measures (i.e., on span tasks), capacity-related neural changes (i.e., on CDA) have nevertheless, started to show up in our short-term updating training program. (PsycINFO Database Record (c) 2018 APA, all rights reserved).


The Impact of Cannabis Decriminalization and Legalization on Road Safety Outcomes: A Systematic Review.
There is substantial debate concerning the impact of cannabis decriminalization and legalization on road safety outcomes. Seven databases were systematically searched: Embase, MEDLINE, and PsycINFO through Ovid as well as Web of Science Core Collection, SafetyLit, Criminal Justice Database (ProQuest), and Transport Research International Documentation (from inception to June 16, 2021). Eligible primary studies examined group-level cannabis decriminalization or legalization and a road safety outcome in any population. A total of 65 reports of 64 observational studies were eligible, including 39 that applied a quasi-experimental design. Studies examined recreational cannabis legalization (n=50), medical cannabis legalization (n=22), and cannabis decriminalization (n=5). All studies except 1 used data from the U.S. or Canada. Studies found mixed impacts of legalization on attitudes, beliefs, and self-reported driving under the influence. Medical legalization, recreational legalization, and decriminalization were associated with increases in positive cannabis tests among drivers. Few studies examined impacts on alcohol or other drug use, although findings suggested a decrease in positive alcohol tests among drivers associated with medical legalization. Medical legalization was associated with reductions in fatal motor-vehicle collisions, whereas recreational legalization was conversely associated with increases in fatal collisions. Increased cannabis positivity may reflect changes in cannabis use; however, it does not in itself indicate increased impaired driving. Subgroups impacted by medical and recreational legalization, respectively, likely explain opposing findings for fatal collisions. More research is needed concerning cannabis decriminalization; the impacts of decriminalization and legalization on nonfatal injuries, alcohol and other drugs; and the mechanisms by which legalization impacts road safety outcomes.


[Long-term results of preoperative hyperthermo-chemo-radiotherapy (HCR) for patients with esophageal carcinoma invading neighboring structures (T4)].
A total of 180 patients with esophageal carcinoma invading the neighboring structures (T4) were surgically treated by esophagectomy and reconstruction in the Department of Surgery II, Kyushu University from January 1965 to April 1997. Any of these cases with distant node metastasis and demonstrating organ metastasis or a combined resection of adjacent structures were excluded from this study. As a result, twenty-six patients treated with preoperative hyperthermo-chemo-radiotherapy (HCR Group), 39 treated either with preoperative radiotherapy or preoperative chemo-radiotherapy (R or CR Group) and 23 non-treated patients (Non-tx Group) were thus entered in this study. The 3-year survival rates after esophagectomy in HCR Group, R or CR Group and Non-tx Group were 26.5%, 0% and 9%, respectively, while the 5-year survival rate of the HCR group was 15.9%. The group with preoperative HCR thus showed a significantly more favorable outcome than R or CR Group and Non-tx Group. (p < 0.05). The significant difference observed in the prognosis was thought to be due to the reinforced effect of local regulation due to hyperthermia. Our data thus suggest that preoperative HCR contributes to the prolonged post-operative survival for carcinoma of the esophagus invading the neighboring structures.


New Developments in Hypertrophic Cardiomyopathy.
Hypertrophic cardiomyopathy is the leading cause of sudden death in young individuals and an important cause of heart failure at any age. In this review we discuss advances in investigation and management of this heterogenous disease. Improved cardiac imaging has allowed us to detail many of the structural abnormalities whereas the use of new techniques, predominantly in cardiac magnetic resonance imaging, has given us a greater insight in to tissue architecture, mechanism of contractile abnormalities, and function. Risk stratification remains challenging because of the low event rate in clinical studies. Multicentre registries have improved risk stratification for sudden cardiac death and multiple models can be used to aid decision-making for implantable defibrillator therapy. We discuss the current state of nonsurgical septal reduction therapy and results of multicentre registries. New approaches to septal reduction therapy including refinement of alcohol ablation and noncoronary interventions such as radiofrequency ablation of the septum show great promise. Surgical myectomy remains a major part of treatment; a greater recognition of abnormalities of the mitral valve apparatus can allow improved surgical options. Myocardial perfusion abnormalities are known to predict adverse outcome in hypertrophic cardiomyopathy and we discuss underlying mechanisms and relevance to management. The off-label use of currently licensed medicines such as ranolazine, perhexiline, calcium channel blockers, and renin-angiotensin system antagonists are discussed. A novel approach to medical treatment of the underlying sarcomeric disorder has been investigated and shows great potential.


Photoluminescence properties of multiple stacked planes of GaN/AlN quantum dots studied by near-field optical microscopy.
We have studied the photoluminescence properties of GaN quantum dots with submicrometre lateral resolution by means of near-field scanning optical microscopy. The instrument operated at room temperature and was implemented for near-ultra-violet spectroscopy in the illumination-mode configuration. The analysed sample consisted of several stacked planes of GaN/AlN quantum dots grown by molecular beam epitaxy on Si(111) substrate. The photoluminescence maps showed islands in the micrometre range emitting at different wavelengths, confirming the atomic force microscopy studies on the morphology of similar uncapped samples.


Synergistic effects of baicalein with gemcitabine or docetaxel on the proliferation, migration and apoptosis of pancreatic cancer cells.
Baicalein, a type of flavonoids extracted from Scutellaria baicalensis Georgi, has been reported to be a very promising drug for pancreatic cancer. However, it is unclear whether combination of baicalein with gemcitabine or docetaxel is synergistic to the treatment for pancreatic cancer (PC). We investigated the combinational effects of baicalein with gemcitabine or docetaxel on proliferation, cell cycle, migration and apoptosis of human PC cells. Administration of baicalein alone significantly inhibit the proliferation of PC cells. Notably, when it is combined with gemcitabine or docetaxel, combination index (CI) values calculated by Calcusyn software are smaller than 1, indicating the synergism of baicalein with gemcitabine or docetaxel for the treatment of PC cells. Consistently, EdU assay showed that administration of baicalein significantly enhanced the capacity of gemicitabine to inhibit proliferation of PC cells. Cell cycle analysis showed that high-concentration of baicalein was able to arrest PC cells in the S phase. Furthermore, low concentration of baicalein in combination with either gemcitabine or docetaxel exhibited strong suppression on the migration of PC cells. A further study using transmission electron microscope (TEM), DAPI staining and western blot showed that baicalein induced-apoptosis of PC cells might be via caspase-3/PARP signaling pathway. Notably, combination treatment was able to induce more severe cell apoptosis of PC cells. In conclusion, baicalein exhibited synergistic effects with gemcitabine or docetaxel on the treatment of PC cells.


Tobacco coverage in print media: the use of timing and themes by tobacco control supporters and opposition before a failed tobacco tax initiative.
Tobacco control policies gained ground nationwide in 2006, with voters in nine states approving legislation to strengthen clean indoor air policies and increase tobacco excise taxes. Despite having the second lowest cigarette tax rate in the nation, Missouri was unsuccessful in passing its 2006 ballot initiative to raise the tax. An important way to encourage health-related policy change such as increasing tobacco taxes is through media coverage of tobacco issues. We examined how tobacco issues were presented in Missouri's print media leading up to the 2006 election. This study analysed 1263 articles with tobacco content published in 187 Missouri newspapers in the year before the election. Articles were coded for general and tobacco-related characteristics including article type (news story, editorial, letter to the editor), tobacco control position (pro, neutral, anti) and article theme (economic, health, political). Most articles were news stories (73.6%) and pro-tobacco control (63.8%). The proportion of anti-tobacco control articles increased significantly (chi(2)=104.9, p<0.001) the month before the election, driven by an increase in economically themed articles. Economic articles were published more often in counties with less voter support for the tax (F=5.68, p<0.01). Finally, tobacco control position varied significantly across article types (chi(2)=148.3, p<0.01), with letters to the editor being anti-tobacco control most often. The media have a critical role in promoting public health goals and presenting health issues which influences formation of health policies. Tobacco control advocates must consider public opinion, opposition pressure, timing and themes in tobacco-related media coverage when promoting policy change.


Directional and spectral reflectance of the rat retinal nerve fiber layer.
To measure and describe the reflectance properties of a mammalian retinal nerve fiber layer (RNFL) and to determine the mechanisms responsible for the RNFL reflectance. An isolated rat retina suspended across a slit in a black membrane and mounted in a black perfusion chamber provided high quality images of the RNFL. Imaging microreflectometry was used to measure RNFL reflectance at wavelengths from 400 nm to 830 nm and as a function of illumination angle. The directional reflectance of rat RNFL at all wavelengths was consistent with the theory of light scattering by cylinders; each nerve fiber bundle scattered light into a conical sheet coaxial with the bundle. There was no evidence of a noncylindrical component at any wavelength. Measured reflectance spectra were consistent between animals, similar to ones previously measured in macaque, and varied with scattering angle. All spectra could be described by a two-mechanism cylindrical scattering model with three free parameters. At all wavelengths the reflectance of rat RNFL arises from light scattering by cylindrical structures. The highly directional nature of this reflectance can be an important source of measurement variability in clinical assessment of the RNFL. The reflectance spectra reveal a combination of mechanisms: At wavelengths shorter than approximately 570 nm the reflectance comes from cylinders with diameters much smaller than the wavelength, but at wavelengths longer than approximately 680 nm the reflectance comes from cylinders with effective diameters of 350 nm to 900 nm.


Transfer ribonucleic acid methylases of bone. Studies on vitamin A and D deficiency.
Methods were devised for the assay of tRNA methylases of rat bone. The activities of bone tRNA methylases are similar to those from other mammalian tissues. However, unlike reports on liver methylases, no inhibitors were found in the supernatant fraction from pH5 precipitate of bone extracts. The effects of vitamins A and D on the methylation of tRNA by cell-free extracts of rat bone were studied. Deficiency of either vitamin resulted in a decrease in the rate and extent of tRNA methylation, whereas the administration of vitamin A to hypovitaminotic-A rats and vitamin D to hypovitaminotic-D rats increased the rate and extent of tRNA methylation. These effects appear to be apart from changes in ribonuclease activity or in concentrations of calcium or magnesium. No evidence of inhibitors of tRNA methylases was found in bone extracts from vitamin-deficient rats nor of activators in bone extracts from deficient rats given vitamin A or D. The pattern of tRNA methylation under conditions of vitamin A or D deficiency was not changed, suggesting a generalized cellular deficiency. It was of significance to find that the specificity for methylation of specific bases in tRNA was different after the administration of vitamin A as contrasted with the effects of vitamin D. The possible significance of tRNA methylation to the biochemical action of the vitamins on bone is discussed.


[Floppy baby with macrocytic anemia and vegan mother].
We report the case of a 7 month-old girl that presented with acute anemia, generalized muscular hypotonia and failure to thrive. Laboratory evaluation revealed cobalamin deficiency, due to a vegan diet of the mother. The clinical triad of an acquired floppy baby syndrome with megaloblastic anemia and failure to thrive is pathognomic for infantile cobalamin deficiency. Neurological abnormalities are often irreversible and may be associated with delayed myelinization in the MRI. A normal cobalamin level in maternal serum and absence of anemia do not exclude subclinical deficiency. If cobalamin deficiency is suspected, e.g. in pregnant women on vegan diet, urinary methylmalonic acid excretion and plasma homocysteine levels should be determined and cobalamin substitution should be started at an early stage to avoid potentially irreversible damage of the fetus.


Intestinal control of human postprandial gastric function: the role of components of jejunoileal chyme in regulating gastric secretion and gastric emptying.
Intestinal control of gastric secretion and emptying were studied in healthy humans by using intubation-marker-perfusion techniques. An occluding balloon was used to separate the jejunoileal test segment from the more proximal gut. Mixed-nutrient, semielemental meals (300 and 600 calories) were administered while varying the jejunal perfusate: normal saline, jejunal chyme or its protein (casein hydrolysate), lipid (oleic acid), or carbohydrate (maltose) components. The meal in contact with the stomach and duodenum increased gastric secretion above fasting levels. This was partially inhibited by whole chyme in contact with the jejunum, the lipid and carbohydrate components of chyme mediating this inhibition. Jejunoileal chyme inhibited gastric emptying, with all three nutrient components acting as inhibitors. Effects on gastric secretion and emptying were dose-dependent, being greater after the larger meal and most prominent in the early postprandial period; these effects on gastric emptying were more prominent than those on gastric secretion. These results suggest that the intestinal phase of gastric function is directed mainly toward regulation of the reservoir function of the stomach: to adapt the gastric delivery of nutrients and fluid to the capability for efficient digestion and absorption.


One-Year Mortality, Causes of Death, and Cardiac Interventions in Patients with Postoperative Myocardial Injury.
To evaluate the role of routine troponin surveillance in patients undergoing major noncardiac surgery, unblinded screening with cardiac consultation per protocol was implemented at a tertiary care center. In this study, we evaluated 1-year mortality, causes of death, and consequences of cardiac consultation of this protocol. This observational cohort included 3224 patients ≥60 years old undergoing major noncardiac surgery. Troponin I was measured routinely on the first 3 postoperative days. Myocardial injury was defined as troponin I >0.06 μg/L. Regression analysis was used to determine the association between myocardial injury and 1-year mortality. The causes of death, the diagnoses of the cardiologists, and interventions were determined for different levels of troponin elevation. Postoperative myocardial injury was detected in 715 patients (22%) and was associated with 1-year all-cause mortality (relative risk [RR] 1.4, P = 0.004; RR 1.6, P < 0.001; and RR 2.2, P < 0.001 for minor, moderate, and major troponin elevation, respectively). Cardiac death within 1 year occurred in 3%, 5%, and 11% of patients, respectively, in comparison with 3% of the patients without myocardial injury (P = 0.059). A cardiac consultation was obtained in 290 of the 715 patients (41%). In 119 (41%) of these patients, the myocardial injury was considered to be attributable to a predisposing cardiac condition, and in 111 patients (38%), an intervention was initiated. Postoperative myocardial injury was associated with an increased risk of 1-year all-cause but not cardiac mortality. A cardiac consultation with intervention was performed in less than half of these patients. The small number of interventions may be explained by a low suspicion of a cardiac etiology in most patients and lack of consensus for standardized treatment in these patients.


Optimized labeling of bone marrow mesenchymal cells with superparamagnetic iron oxide nanoparticles and in vivo visualization by magnetic resonance imaging.
Stem cell therapy has emerged as a promising addition to traditional treatments for a number of diseases. However, harnessing the therapeutic potential of stem cells requires an understanding of their fate in vivo. Non-invasive cell tracking can provide knowledge about mechanisms responsible for functional improvement of host tissue. Superparamagnetic iron oxide nanoparticles (SPIONs) have been used to label and visualize various cell types with magnetic resonance imaging (MRI). In this study we performed experiments designed to investigate the biological properties, including proliferation, viability and differentiation capacity of mesenchymal cells (MSCs) labeled with clinically approved SPIONs. Rat and mouse MSCs were isolated, cultured, and incubated with dextran-covered SPIONs (ferumoxide) alone or with poly-L-lysine (PLL) or protamine chlorhydrate for 4 or 24 hrs. Labeling efficiency was evaluated by dextran immunocytochemistry and MRI. Cell proliferation and viability were evaluated in vitro with Ki67 immunocytochemistry and live/dead assays. Ferumoxide-labeled MSCs could be induced to differentiate to adipocytes, osteocytes and chondrocytes. We analyzed ferumoxide retention in MSCs with or without mitomycin C pretreatment. Approximately 95% MSCs were labeled when incubated with ferumoxide for 4 or 24 hrs in the presence of PLL or protamine, whereas labeling of MSCs incubated with ferumoxide alone was poor. Proliferative capacity was maintained in MSCs incubated with ferumoxide and PLL for 4 hrs, however, after 24 hrs it was reduced. MSCs incubated with ferumoxide and protamine were efficiently visualized by MRI; they maintained proliferation and viability for up to 7 days and remained competent to differentiate. After 21 days MSCs pretreated with mitomycin C still showed a large number of ferumoxide-labeled cells. The efficient and long lasting uptake and retention of SPIONs by MSCs using a protocol employing ferumoxide and protamine may be applicable to patients, since both ferumoxides and protamine are approved for human use.


Massive Ascites and Pleural Effusion in Preeclampsia.
Preeclampsia is defined as new onset hypertension and proteinuria after 20 weeks of gestation and complicates approximately 2-8% of all pregnancies. Release of vasoconstrictive agents, endothelial damage, hyperpermeability of the capillaries and microangiopathic haemolysis involves the basic pathophysiology. It has variable clinical presentation. Here, we report a case of severe preeclampsia who developed postpartum massive ascites and pleural effusion. Primigravid patient was admitted to our clinic at 35 weeks of gestation with very high blood pressure. In biochemical analysis, Alanine aminotransferase (ALT) was 401 U/L, Aspartate aminotransferase (AST) was 292 U/L. An emergency caesarean section was performed because of fetal distress. On the 2nd post-operative day, abdominal distension and severe abdominal pain occurred. On the 3rd post-operative day, her abdominal distension increased and Ultrasonography (USG) revealed massive ascites. Abdominal drainage was performed and albumin infusion was administered. On postoperative day 4, she still had abdominal distension and concomitant respiratory distress. Computed Tomography (CT) showed ascites and bilateral pleural effusion. Her complaint regressed on the following days.


Effects of Alkaline-Reduced Drinking Water on Irritable Bowel Syndrome with Diarrhea: A Randomized Double-Blind, Placebo-Controlled Pilot Study.
The purpose of this study was to investigate whether the ingestion of alkaline-reduced water (ARW) is helpful in improving the symptoms of diarrhea-predominant irritable bowel syndrome (IBS). Twenty-seven patients (male, 25.9%; mean, 41.7 years old) with diarrhea-predominant IBS were randomly allocated to two groups. For eight weeks, the ARW group (n = 13) ingested at least 2 liters/day of ARW, while the control group (n = 14) ingested placebo water. IBS symptom scores (quality-of-life, abdominal pain/discomfort), stool form, and frequency were assessed before and after treatment via questionnaires. Eight patients (61.5%) in the ARW group and six patients (42.9%) in the control group indicated that their symptoms had improved in more than four out of the eight weeks of treatment (p = 0.449). The IBS quality-of-life score significantly improved from 57.2 to 30.8 in the ARW group; this improvement was significantly greater than the slight improvement from 48.7 to 42.2 observed in the control group (p = 0.029). The abdominal pain score improved from 1.8 to 0.9 in the ARW group and from 1.8 to 1.1 in the control group, with no significant group difference (p = 0.232). Drinking ARW for eight weeks improves the quality of life in patients with diarrhea-predominant IBS.


Synaptonemal complexes during premeiotic DNA synthesis in oocytes of Drosophila melanogaster.
Well-synchronized populations of oocytes obtained by means of the "pupal system" (Grell, 1973a) have been examined to determine the time of appearance of the synaptonemal complex. The complex first appears in the most advanced oocytes between 132 and 138 hr of female development. Between 138 and 156 hr the complex apparently undergoes a fourfold increase in length. At 150 and 156 hr the complex system is extensive and present in virtually all oocytes. Previous studies using the pupal system have placed the period of premeiotic DNA synthesis between 132 and 162 hr. Thus, indirect evidence indicates that a significant portion of synaptonemal complex formation is coextensive with the main DNA replication in the oocyte. Direct evidence that DNA synthesis and complex formation occur simultaneously in oocytes has been obtained by electron microscope autoradiography. By definition, then, the stage of synaptonemal complex formation in Drosophila must include premeiotic interphase.


EvgS/EvgA, the unorthodox two-component system regulating bacterial multiple resistance.
EvgS/EvgA, one of the five unorthodox two-component systems in Escherichia coli, plays an essential role in adjusting bacterial behaviors to adapt to the changing environment. Multiple resistance regulated by EvgS/EvgA endows bacteria to survive in adverse conditions such as acidic pH, multidrug, and heat. In this minireview, we summarize the specific structures and regulation mechanisms of EvgS/EvgA and its multiple resistance. By discussing several unresolved issues and proposing our speculations, this review will be helpful and enlightening for future directions about EvgS/EvgA.


The X-ray crystal structure of full-length human plasminogen.
Plasminogen is the proenzyme precursor of the primary fibrinolytic protease plasmin. Circulating plasminogen, which comprises a Pan-apple (PAp) domain, five kringle domains (KR1-5), and a serine protease (SP) domain, adopts a closed, activation-resistant conformation. The kringle domains mediate interactions with fibrin clots and cell-surface receptors. These interactions trigger plasminogen to adopt an open form that can be cleaved and converted to plasmin by tissue-type and urokinase-type plasminogen activators. Here, the structure of closed plasminogen reveals that the PAp and SP domains, together with chloride ions, maintain the closed conformation through interactions with the kringle array. Differences in glycosylation alter the position of KR3, although in all structures the loop cleaved by plasminogen activators is inaccessible. The ligand-binding site of KR1 is exposed and likely governs proenzyme recruitment to targets. Furthermore, analysis of our structure suggests that KR5 peeling away from the PAp domain may initiate plasminogen conformational change.


Attentional retraining can reduce chocolate consumption.
There is emerging evidence that attentional biases are related to the consumption of substances such as alcohol and tobacco, and that attentional bias modification can reduce unwanted consumption of these substances. We present evidence for the first time to our knowledge that the same logical argument applies in the food and eating domain. We conducted two experiments that used a modified dot probe paradigm to train undergraduate women to direct their attention toward ("attend") or away from ("avoid") food cues (i.e., pictures of chocolate). In Experiment 1, attentional bias for chocolate cues increased in the "attend" group, and decreased in the "avoid" group. Experiment 2 showed that these training effects generalized to novel, previously unseen chocolate pictures. Importantly, attentional retraining affected chocolate consumption and craving. In both experiments, participants in the "avoid" group ate less chocolate in a so-called taste test than did those in the "attend" group. In addition, in Experiment 2, but not in Experiment 1, the "attend" group reported stronger chocolate cravings following training, whereas the "avoid" group reported less intense cravings. The results support predictions of cognitive-motivational models of craving and consumption that attentional biases play a causal role in consumption behavior. Furthermore, they present a promising avenue for tackling unwanted food cravings and (over)eating.


Immune system stimulation by the native gut microbiota of honey bees.
Gut microbial communities can greatly affect host health by modulating the host's immune system. For many important insects, however, the relationship between the gut microbiota and immune function remains poorly understood. Here, we test whether the gut microbial symbionts of the honey bee can induce expression of antimicrobial peptides (AMPs), a crucial component of insect innate immunity. We find that bees up-regulate gene expression of the AMPs apidaecin and hymenoptaecin in gut tissue when the microbiota is present. Using targeted proteomics, we detected apidaecin in both the gut lumen and the haemolymph; higher apidaecin concentrations were found in bees harbouring the normal gut microbiota than in bees lacking gut microbiota. In in vitro assays, cultured strains of the microbiota showed variable susceptibility to honey bee AMPs, although many seem to possess elevated resistance compared to Escherichia coli. In some trials, colonization by normal gut symbionts resulted in improved survivorship following injection with E. coli. Our results show that the native, non-pathogenic gut flora induces immune responses in the bee host. Such responses might be a host mechanism to regulate the microbiota, and could potentially benefit host health by priming the immune system against future pathogenic infections.


Knowledge and attitudes toward COVID-19 vaccination in Sudan: A cross-sectional study.
Vaccines are an essential part of public health interventions to mitigate the devastating health and non-health impacts of COVID-19 pandemic. Despite the fact that Sudan launched the COVID-19 vaccination program in March 2021, only 10% of the population received their two primary doses of vaccines by the end of May 2022. This delayed uptake of vaccines obviously warrants investigation. Therefore, we have conducted this study to evaluate the knowledge, attitude and acceptance of the general population in Sudan toward COVID-19 vaccines. A descriptive cross-sectional community-based study. The data were collected using an electronic questionnaire from 403 individuals living in Khartoum, Sudan. The data were processed using the Statistical Package for Social Sciences (SPSS), and data analysis was performed using appropriate tests. 51% of the participants were found to have sufficient knowledge about the COVID-19 vaccine, and the knowledge level is higher among those educated beyond the secondary school and those who were employed. Among those unvaccinated, only 47% of the participants expressed their intention to take the vaccine when offered to them. The major reason for not trusting the vaccine is safety concerns expressed by 65.5% of the unvaccinated. Higher education levels and employment were associated with an increase in sufficient knowledge about the vaccine in around half of the participants. However, most of participants had not taken the vaccine at the time of the study, and the trust in vaccines is not high. Effective interventions by the health authorities are needed to address these issues in order to accelerate the COVID-19 vaccination program in Sudan.


Fine tuning of hormonal signaling is linked to dormancy status in sweet cherry flower buds.
In temperate trees, optimal timing and quality of flowering directly depend on adequate winter dormancy progression, regulated by a combination of chilling and warm temperatures. Physiological, genetic and functional genomic studies have shown that hormones play a key role in bud dormancy establishment, maintenance and release. We combined physiological and transcriptional analyses, quantification of abscisic acid (ABA) and gibberellins (GAs), and modeling to further investigate how these signaling pathways are associated with dormancy progression in the flower buds of two sweet cherry cultivars. Our results demonstrated that GA-associated pathways have distinct functions and may be differentially related with dormancy. In addition, ABA levels rise at the onset of dormancy, associated with enhanced expression of ABA biosynthesis PavNCED genes, and decreased prior to dormancy release. Following the observations that ABA levels are correlated with dormancy depth, we identified PavUG71B6, a sweet cherry UDP-GLYCOSYLTRANSFERASE gene that up-regulates active catabolism of ABA to ABA glucosyl ester (ABA-GE) and may be associated with low ABA content in the early cultivar. Subsequently, we modeled ABA content and dormancy behavior in three cultivars based on the expression of a small set of genes regulating ABA levels. These results strongly suggest the central role of ABA pathway in the control of dormancy progression and open up new perspectives for the development of molecular-based phenological modeling.


Post-genomics nanotechnology is gaining momentum: nanoproteomics and applications in life sciences.
The post-genomics era has brought about new Omics biotechnologies, such as proteomics and metabolomics, as well as their novel applications to personal genomics and the quantified self. These advances are now also catalyzing other and newer post-genomics innovations, leading to convergences between Omics and nanotechnology. In this work, we systematically contextualize and exemplify an emerging strand of post-genomics life sciences, namely, nanoproteomics and its applications in health and integrative biological systems. Nanotechnology has been utilized as a complementary component to revolutionize proteomics through different kinds of nanotechnology applications, including nanoporous structures, functionalized nanoparticles, quantum dots, and polymeric nanostructures. Those applications, though still in their infancy, have led to several highly sensitive diagnostics and new methods of drug delivery and targeted therapy for clinical use. The present article differs from previous analyses of nanoproteomics in that it offers an in-depth and comparative evaluation of the attendant biotechnology portfolio and their applications as seen through the lens of post-genomics life sciences and biomedicine. These include: (1) immunosensors for inflammatory, pathogenic, and autoimmune markers for infectious and autoimmune diseases, (2) amplified immunoassays for detection of cancer biomarkers, and (3) methods for targeted therapy and automatically adjusted drug delivery such as in experimental stroke and brain injury studies. As nanoproteomics becomes available both to the clinician at the bedside and the citizens who are increasingly interested in access to novel post-genomics diagnostics through initiatives such as the quantified self, we anticipate further breakthroughs in personalized and targeted medicine.


Clustered Conserved Cysteines in Hyaluronan Synthase Mediate Cooperative Activation by Mg2+ Ions and Severe Inhibitory Effects of Divalent Cations.
Hyaluronan synthase (HAS) uses UDP-GlcUA and UDP-GlcNAc to make hyaluronan (HA). Streptococcus equisimilis HAS (SeHAS) contains four conserved cysteines clustered near the membrane, and requires phospholipids and Mg2+ for activity. Activity of membrane-bound or purified enzyme displayed a sigmoidal saturation profile for Mg2+ with a Hill coefficient of 2. To assess if Cys residues are important for cooperativity we examined the Mg2+ dependence of mutants with various combinations of Cys-to-Ala mutations. All Cys-mutants lost the cooperative response to Mg2+. In the presence of Mg2+, other divalent cations inhibited SeHAS with different potencies (Cu2+~Zn2+ >Co2+ >Ni2+ >Mn2+ >Ba2+ Sr2+ Ca2+). Some divalent metal ions likely inhibit by displacement of Mg2+-UDP-Sugar complexes (e.g. Ca2+, Sr2+ and Ba2+ had apparent Ki values of 2-5 mM). In contrast, Zn2+ and Cu2+ inhibited more potently (apparent Ki ≤ 0.2 mM). Inhibition of Cys-null SeHAS by Cu2+, but not Zn2+, was greatly attenuated compared to wildtype. Double and triple Cys-mutants showed differing sensitivities to Zn2+ or Cu2+. Wildtype SeHAS allowed to make HA prior to exposure to Zn2+ or Cu2+ was protected from inhibition, indicating that access of metal ions to sensitive functional groups was hindered in processively acting HA•HAS complexes. We conclude that clustered Cys residues mediate cooperative interactions with Mg2+ and that transition metal ions inhibit SeHAS very potently by interacting with one or more of these -SH groups.


Bioabsorbable pins for treatment of osteochondral fractures of the knee after acute patella dislocation in children and young adolescents.
A retrospective study was performed on the use of bioabsorbable pins in the fixation of osteochondral fractures (OCFs) after traumatic patellar dislocation in children. Eighteen children (13 females, 5 males) aged 11 to 15 years (mean age 13.1 years) with osteochondral fracture (OCF) of the knee joint were treated at the authors' institution. Followup ranged from 22 months to 5 years. Diagnosis was verified by X-ray and magnetic resonance imaging (MRI) of the knee and patella. In seven patients the osteochondral fragment was detached from the patella and in 11 it was detached from the lateral femoral condyle. All patients were subjected to open reduction and fixation of the lesion with bioabsorbable pins. Postoperatively, the knee was immobilized in a cast and all patients were mobilized applying a standardized protocol. Bone consolidation was successful in 17 of the 18 patients. Bioabsorbable pins reliably fix OCF in children and adolescents, demonstrating a high incidence of consolidation of the detached osteochondral fragment in short- and middle-term followup without requiring further operative procedures.


A super-SILAC based proteomics analysis of diffuse large B-cell lymphoma-NOS patient samples to identify new proteins that discriminate GCB and non-GCB lymphomas.
Diffuse large B-cell lymphoma-not otherwise specified (DLBCL-NOS) is a large and heterogeneous subgroup of non-Hodgkin lymphoma. DLBCL can be subdivided into germinal centre B-cell like (GCB) and activated B-cell like (ABC or non-GCB) using a gene-expression based or an immunohistochemical approach. In this study we aimed to identify additional proteins that are differentially expressed between GCB and non-GCB DLBCL. A reference super-SILAC mix, including proteins of eight B-cell lymphoma cell lines, was mixed with proteins isolated from seven non-GCB DLBCL and five GCB DLBCL patient tissue samples to quantify protein levels. Protein identification and quantification was performed by LC-MS. We identified a total of 4289 proteins, with a four-fold significant difference in expression between non-GCB and GCB DLBCL for 37 proteins. Four proteins were selected for validation in the same cases and replication in an independent cohort of 47 DLBCL patients by immunohistochemistry. In the validation cohort, we observed a non-significant trend towards the same differential expression pattern as observed in the proteomics. The replication study showed significant and consistent differences for two of the proteins: expression of glomulin (GLMN) was higher in GCB DLBCL, while expression of ribosomal protein L23 (RPL23) was higher in non-GCB DLBCL. These proteins are functionally linked to important pathways involving MYC, p53 and angiogenesis. In summary, we showed increased expression of RPL23 and decreased expression of GLMN in non-GCB compared to GCB DLBCL on purified primary DLBCL patient samples and replicated these results in an independent patient cohort.


Nipple shield use in the maternity ward increases risk of exclusive breastfeeding interruption in the first six months of infant's life.
The objective of this study was to investigate the association between use of silicone nipple shields in the maternity ward and exclusive breastfeeding interruption in the first 6 months of the infant's life. Cohort study. Interviews were conducted personally at the mother's home at 1 and 6 months postpartum and by telephone at 2 and 4 months. 287 mother-infant dyads. Lactating mothers were randomly selected at two maternity wards (one public, one private) in Porto Alegre, Brazil. Data were analyzed using Kaplan-Meier survival curves and Cox multivariate regression. The outcome of interest was exclusive breastfeeding interruption before 6 months of the infant's life. Nipple shields were used by 6.2 % of the women in the public maternity ward and by 25.8 % of those in the private setting. Median duration of exclusive breastfeeding was 11 days (95 %CI 0.0-36.9) among women who used the accessory vs. 89 days (95 %CI 60.8-117.2) among those who did not. Nipple shield use in the maternity ward was associated with exclusive breastfeeding interruption before 6 months of infant's life (adjusted risk ratio = 1.47; 95 %CI 1.01-2.15). The risk was higher in the first months of breastfeeding, ranging from 2.0 to 1.47 in the first and sixth months, respectively. The use of silicone nipple shields in the maternity ward increased the risk of exclusive breastfeeding interruption before 6 months of the infant's life, especially in the first months. These findings suggest caution in recommending this accessory to new mothers.


[Obscure gastrointestinal bleeding due to duodenal infiltration of a non-functioning pancreatic neuroendocrine tumor diagnosed by capsule endoscopy: a case report].
Non-functioning pancreatic neuroendocrine tumors (PNETs) are infrequent slow-growing, clinically-silent tumors. They are incidentally detected and some of them may present in advanced stages with local involvement of surrounding structures. The diagnostic accuracy of endoscopio ultrasound (EUS) and fine needle aspiration (FNA) biopsy is significantly lower in neuroendocrine tumors (46.7%) compared with adenocarcinoma (81.4%) and other histologies (75%). Therefore, preoperative diagnosis is very difficult. Exceptionally, hey present with gastrointestinal bleeding. We present a case of a non-functioning PNET initially diagnosed as cystic serous tumor of pancreas with EUS and FNA biopsy. Two years later patient presented obscure gastrointestinal bleeding due to duodenal infiltration. Diagnosis was made by capsule endoscopy.


A surprising range of modified-methionyl S-adenosylmethionine analogues support bacterial growth.
S-Adenosyl-l-methionine (AdoMet) is an essential metabolite, serving in a very wide variety of metabolic reactions. The enzyme that produces AdoMet from l-methionine and ATP (methionine adenosyltransferase, MAT) is thus an attractive target for antimicrobial agents. We previously showed that a variety of methionine analogues are MAT substrates, yielding AdoMet analogues that function in specific methyltransfer reactions. However, this left open the question of whether the modified AdoMet molecules could support bacterial growth, meaning that they functioned in the full range of essential AdoMet-dependent reactions. The answer matters both for insight into the functional flexibility of key metabolic enzymes, and for drug design strategies for both MAT inhibitors and selectively toxic MAT substrates. In this study, methionine analogues were converted in vitro into AdoMet analogues, and tested with an Escherichia coli strain lacking MAT (ΔmetK) but that produces a heterologous AdoMet transporter. Growth that yields viable, morphologically normal cells provides exceptionally robust evidence that the analogue functions in every essential reaction in which AdoMet participates. Overall, the S-adenosylated derivatives of all tested l-methionine analogues modified at the carboxyl moiety, and some others as well, showed in vivo functionality sufficient to allow good growth in both rich and minimal media, with high viability and morphological normality. As the analogues were chosen based on incompatibility with the reactions via which AdoMet is used to produce acylhomoserine lactones (AHLs) for quorum sensing, these results support the possibility of using this route to selectively interfere with AHL biosynthesis without inhibiting bacterial growth.


Host-pathogen coevolution increases genetic variation in susceptibility to infection.
It is common to find considerable genetic variation in susceptibility to infection in natural populations. We have investigated whether natural selection increases this variation by testing whether host populations show more genetic variation in susceptibility to pathogens that they naturally encounter than novel pathogens. In a large cross-infection experiment involving four species of Drosophila and four host-specific viruses, we always found greater genetic variation in susceptibility to viruses that had coevolved with their host. We went on to examine the genetic architecture of resistance in one host species, finding that there are more major-effect genetic variants in coevolved host-pathogen interactions. We conclude that selection by pathogens has increased genetic variation in host susceptibility, and much of this effect is caused by the occurrence of major-effect resistance polymorphisms within populations.


p90 ribosomal S6 kinase regulates activity of the renin-angiotensin system: a pathogenic mechanism for ischemia-reperfusion injury.
Increasing evidence suggests that local renin-angiotensin system (RAS) plays an important role in cardiac diseases. Elevated p90 ribosomal S6 kinase (RSK) activity has been observed in diabetic animal, as well as in human failing hearts. We hypothesize that RSK mediates cardiac dysfunction by up regulating local RAS signaling. In the present study, we show that the prorenin mRNA level was significantly increased (~5.6-fold) in transgenic mouse hearts with cardiac specific expression of RSK (RSK-Tg). The RSK-Tg mice were more vulnerable to ischemia/reperfusion (I/R) injury than non-transgenic littermate controls (NLC). To further understand the direct contribution of cardiac renin to I/R injury, we used a Langendorff system to evaluate the effect of renin inhibition by aliskiren in RSK-Tg mouse hearts. In the vehicle-perfused group, I/R significantly decreased left ventricular developed pressure (LVDP) in RSK-Tg hearts compared to NLC (7% versus 60% of the baseline). However, aliskiren perfusion significantly increased LVDP in RSK-Tg (7% to 61%, p<0.01) but not in NLC hearts (60% to 62%, n.s.). The protective effect of aliskiren in RSK-Tg hearts was further demonstrated with positive (contraction) dp/dt (6.5% to 63%, p<0.01) and rate pressure product (RPP) (5% to 51%, p<0.01). Moreover, aliskiren significantly decreased I/R induced infarction in RSK-Tg (60% to 32%, p<0.01), compared to NLC hearts (37% to 32%, n.s.). These results suggest that RSK plays a crucial role in regulating local cardiac renin, which contributes to I/R induced cardiac injury and dysfunction. Thus, renin inhibition may provide an alternative therapeutic strategy under conditions of increased RAS.


Plasma membrane charging of Jurkat cells by nanosecond pulsed electric fields.
The initial effect of nanosecond pulsed electric fields (nsPEFs) on cells is a change of charge distributions along membranes. This first response is observed as a sudden shift in the plasma transmembrane potential that is faster than can be attributed to any physiological event. These immediate, yet transient, effects are only measurable if the diagnostic is faster than the exposure, i.e., on a nanosecond time scale. In this study, we monitored changes in the plasma transmembrane potential of Jurkat cells exposed to nsPEFs of 60 ns and amplitudes from 5 to 90 kV/cm with a temporal resolution of 5 ns by means of the fast voltage-sensitive dye Annine-6. The measurements suggest the contribution of both dipole effects and asymmetric conduction currents across opposite sides of the cell to the charging. With the application of higher field strengths the membrane charges until a threshold voltage value of 1.4-1.6 V is attained at the anodic pole. This indicates when the ion exchange rates exceed charging currents, thus providing strong evidence for pore formation. Prior to reaching this threshold, the time for the charging of the membrane by conductive currents is qualitatively in agreement with accepted models of membrane charging, which predict longer charging times for lower field strengths. The comparison of the data with previous studies suggests that the sub-physiological induced ionic imbalances may trigger other intracellular signaling events leading to dramatic outcomes, such as apoptosis.


Mutation spectral changes in spermatogenic cells obtained from old mice.
Male reproductive health is compromised with increased paternal age, due at least in part, to an increased frequency of de novo germline mutations. Because of technical and sample limitations, there is a dearth of empirical information on the mechanism(s) that mediate this age-related increase in mutant frequency. To study this phenomenon, investigators have used as a model system a transgenic mouse strain that carries a lacI mutagenesis reporter transgene. This transgene displays a paternal age effect and overcomes many of the technical difficulties that have inhibited experimental analyses of age-related changes in the male germline. In this study, approximately 300 mutant lacI transgenes were recovered from defined spermatogenic cell types obtained from various aged lacI transgenic mice and sequenced. The spectrum representing mutations from spermatogenic cells of old mice revealed an increased prevalence of transversions compared to spectra for young and middle-aged mice. Five mutation hotspots were identified in spectra for spermatogenic cells from young and middle-aged mice, but no hotspots were identified in the spectrum for spermatogenic cells from old mice. These results suggest that the challenges to germline DNA change as the animal ages and that the increased mutant frequency observed with increased paternal age is not simply a greater accumulation of mutagenic events characteristic of spermatogenic cells from the young animal.


The adaptability of Escherichia coli thioredoxin to non-conservative amino acid substitutions.
The adaptability of Escherichia coli thioredoxin to the substitution of a series of non-natural amino acids has been investigated. Different thiosulfonated alkyl groups were inserted into the hydrophobic core of the protein in position 78 via disulfide bonding with a buried cysteine residue as previously described (Wynn R, Richards FM. 1993. Unnatural amino acid packing mutants of Escherichia coli thioredoxin produced by combined mutagenesis/chemical modification techniques. Protein Sci 2:395-403). The side chains added to the cysteine included methyl, ethyl, n-propyl, n-butyl, n-pentyl, and cyclo-pentyl derivatives. The side chains appear to exploit the presence of the large cavities to incorporate these variant forms, enabling the protein to fold and have some activity. Solution structural and kinetic data suggested that these substitutions had little effect on the overall fold of the protein. Thermodynamic data revealed that the entropic effect of restricting the side chains in the folded protein has an effect on the stability. The variant forms of thioredoxin have different propensities to form dimers despite the limited structural perturbations. Molecular modeling studies allow the conformation of the side chains to be assessed.


Imaging Features of Pediatric Sarcoidosis.
Sarcoidosis is a granulomatous inflammatory disease of uncertain cause. It occurs most commonly in young and middle-aged adults and less frequently in children; therefore, few data on pediatric sarcoidosis exist in the literature. The diagnosis and management of sarcoidosis remain challenging because of diverse and often nonspecific clinical and imaging findings. In addition, the clinical picture varies widely by age. Prepubertal and adolescent patients often present with adult-like pulmonary disease; however, early-onset sarcoidosis is typically characterized by the triad of arthritis, uveitis, and skin rash. Sarcoidosis is mostly a diagnosis of exclusion made by demonstrating noncaseating granulomas at histopathologic examination in patients with compatible clinical and radiologic findings. Although sarcoidosis often affects the lungs and thoracic lymph nodes, it can involve almost any organ in the body. The most common radiologic manifestation is pulmonary involvement, characterized by mediastinal and bilateral symmetric hilar lymphadenopathies with perilymphatic micronodules. Abdominal involvement is also common in children and often manifests as hepatomegaly, splenomegaly, and abdominal lymph node enlargement. Although neurosarcoidosis and cardiac sarcoidosis are rare, imaging is essential to the diagnosis of central nervous system and cardiac involvement because of the risky biopsy procedure and its low diagnostic yield due to focal involvement. Being familiar with the spectrum of imaging findings of sarcoidosis may aid in appropriate diagnosis and management. ©RSNA, 2023 Test Your Knowledge questions are available in the supplemental material.


Tomentosin induces apoptosis in pancreatic cancer cells through increasing reactive oxygen species and decreasing mitochondrial membrane potential.
The aim of this study was to determine possible anticancer effect of tomentosin, a natural sesquiterpene lactone, on pancreatic cancer cells. The cytotoxic effect of tomentosin was determined by XTT analysis. Colony formation and apoptosis analyzes were performed, Reactive oxygen species (ROS) level and change in mitochondrial membrane potential (MMP) were evaluated in control and tomentosin-treated cells. The effect of tomentosin on expression levels of apoptosis-related genes was determined by qRT-PCR and Caspase-3 and Caspase-9 proteins were analyzed by western blot. And, the effect of tomentosin on migration and invasion of cells were evaluated. The IC50 dose of tomentosin was found to be 31.11 μM in PANC-1 cells and 33.93 μM in MIA PaCa-2 cells for 48 h. And, treatment of tomentosin at IC50 dose suppressed the colony forming capacity of cells. While tomentosin increased apoptosis rate and ROS production, an decrease was observed in MMP. Tomentosin affected expression level of apoptosis-related genes and increased Caspase-3 and Caspase-9 protein levels. After tomentosin treatment, cell migration and invasion were suppressed. As a result, this study reveals that tomentosin has anticancer effects on pancreatic cancer cells, and therefore it predicts that tomentosin can be evaluated as an effective agent against pancreatic cancer.


Innovative shortcuts and initiatives in primary health care for rural/remote localities: a scoping review on how to overcome the COVID-19 pandemic.
The COVID-19 pandemic has emerged as one of the greatest challenges to societies, world health systems and science in the past century, making it imperative to restructure care networks. Therefore, it is essential to discuss the role and initiatives of primary health care (PHC) to deal with it. However, regarding the response to the pandemic, including the current global effort against COVID-19, the nuances of the rural/remote PHC context in the pandemic is barely visible. Rural and remote communities have differentiated health risks, such as socioeconomic disadvantages, difficulties in mobility and access to health services, in addition to linguistic and cultural barriers. This scoping review aimed to analyze the set of individual and collective initiatives and innovations developed to face the COVID-19 pandemic, within the PHC scope, in rural and remote areas. A scoping review methodology was applied to peer-reviewed articles. Eight databases were searched to identify scientific articles published in English, Spanish and Portuguese, initially from January 2020 to July 2021, complemented by a rapid review of articles published from January 2022 to April 2023. The main focus sought in the literature was the set of initiatives and innovations carried out within the PHC scope in rural and remote locations during the pandemic, as well as the comparison with pre-pandemic situations and between different countries. The bibliographic information of each search result was imported into Rayyan (Intelligent Systematic Review), followed by the screening and eligibility stages, performed independently by two reviewers, with a third reviewer being accessed in case of conflicts. This review included 54 studies, with publications mostly from Australia, Canada, the US and India. The main PHC initiatives were related to access; to the roles of community health workers and health surveillance; and to the importance of placing, retaining and valuing human resources in health. Cultural, equity and vulnerability issues occupy a major place among the initiatives. Regarding the innovations, telehealth and customized communication are highlighted. From an organizational point of view, rural and remote locations showed enormous flexibility to deal with the pandemic and to improve intersectoral activities at the local level. The description of rurality and remoteness is practically coincident with that of the specific populations, present in geographic areas of difficult sociospatial and cultural access. Rarely, there is an index to measure rurality, or its description deals with the need to overcome distances and obstacles. The findings highlight and summarize knowledge about initiatives and innovations developed to face the COVID-19 pandemic, within the PHC scope in rural and remote areas in the world. This review has identified collective, clinical, intersectoral and, mainly, organizational health initiatives. An articulation between different government levels would be paramount in evaluating the implementation of policies and protocols in rural and remote locations for future sanitary crises. Innovations and lessons learned are equally relevant in strengthening health services and systems. This issue calls for considerable further exploration by new reviews and empirical research that seek evidence to assess the sustainability and effectiveness of the implemented measures to face post-pandemic difficulties and other adversities.


Anti colon cancer components from Lebanese sage (Salvia libanotica) essential oil: Mechanistic basis.
Lebanese sage essential oil possesses antitumor properties, however, the bioactive components and antitumor mechanisms are not known. Here we show that combining the three sage bioactive compounds, Linalyl acetate (Ly), Terpeniol (Te) and Camphor (Ca), caused synergistic inhibition of the growth of two isogenic human colon cancer cell lines HCT-116 (p53(+/+) and p53(-/-)) and had no effect on growth of FHs74Int normal human intestinal cell line. In p53(+/+) cells, the combination of Ly + Te + Ca (10(-3) M of each) caused significant accumulation of cells in PreG(1) (64% at 48 hours); less preG(1) increase was observed in response to Ly + Te (25%) or Ly + Ca (14%). In p53(-/-) cells, Ly + Te + Ca caused cell accumulation in PreG(1) and G(2)/M phases. In response to the three components, 58% apoptosis occurred in p53(+/+) cells and 38% in p53(-/-) cells. Apoptosis by Ly + Te + Ca, in p53+/+ cells, was associated with increased Bax/Bcl-2 ratio and pp53/p53 ratio, cleavage and activation of caspase-3, loss of mitochondrial membrane potential and cytochrome c release. In p53(-/-) cells, the disruption of mitochondrial membrane potential was observed but to a lesser extent than in p53(+/+) cells and caspase activation or cleavage did not appear to be involved in drug-induced apoptosis. Sage components induced poly(ADP-ribose)-polymerase (PARP) cleavage in both p53(+/+) and p53(-/-) cell lines. Pretreatment with the caspase-3 inhibitor and pan caspase inhibitor abrogated drug-mediated apoptosis and blocked procaspase-3 activation and partially blocked PARP cleavage in p53(+/+) cells. Conversely, in p53(-/-) cells, pre-incubation with caspase inhibitors potentiated drug-induced cell death. It appears that apoptosis in p53(+/+) cells is through the mitochondrial-mediated, caspase-dependent pathway, while in p53(-/-) cells apoptosis is mostly caspase independent despite the presence and features indicating caspase-dependent cell death, such as cytochrome c release and PARP cleavage. Our findings encourage further studies of sage oil components as promising chemotherapeutic agents against colon cancer.


Reconstruction of complex cranial wounds with demineralized bone matrix and bilayer artificial skin.
Complex wounds involving loss of both cranium and scalp are difficult to reconstruct. Current solutions include both autogenous and alloplastic materials. Tissue engineering provides interesting alternatives for reconstruction of missing bone and soft tissue. Demineralized bone matrix and Integra, artificial bilayer skin substitute, have been used successfully to reconstruct bone and skin as isolated defects, but never in combination. This study investigates the possibility of using demineralized bone matrix in a gel carrier and Integra for combined cranial and scalp defects. The study was divided into two parts. In the first, demineralized bone matrix in the forms of Grafton Flex (n = 12) and Grafton putty (n = 12) was used to reconstruct 15-mm critical size cranial defects in female adult New Zealand White rabbits. In the control group (n = 6), the defect was left empty. The second part of the study investigated the use of Integra and Grafton Flex to reconstruct a 15-mm cranial defect with an overlying full-thickness scalp defect (n = 6). The first study revealed bony healing of the critical-size cranial defect with demineralized bone matrix. The second study demonstrated successful reconstitution of scalp and cranium with both demineralized bone matrix and Integra. We conclude that complex cranial defects involving bone and soft tissue may be successfully reconstructed with their tissue-engineered substrates, demineralized bone matrix (Grafton) and Integra artificial skin.


Diesel exhaust particulate--exposed macrophages cause marked endothelial cell activation.
Exposure to air pollution containing diesel exhaust particulate (DEP) is linked to adverse cardiovascular events. This study tested the hypothesis that DEP not only causes direct endothelial cell injury, but also induces indirect endothelial cell activation via the release of soluble proinflammatory cytokines from macrophages. Human umbilical vein endothelial cells (HUVECs) and monocyte-derived macrophages (MDMs) were incubated with DEP (1-100 μg/ml; 24 h). Supernatants were analyzed for monocyte chemotactic protein (MCP)-1, IL6, IL8, and TNF-α. Indirect actions of DEP were investigated by incubating HUVECs with conditioned media from DEP-exposed MDMs in the presence and absence of the TNF-α inhibitor, etanercept. A modified Boyden chamber assay was used to determine whether HUVECs treated in this manner induced monocyte chemotaxis. Direct incubation with DEP induced a modest increase in MCP-1 concentration, but had no effect on IL-6 or IL-8 release from HUVECs. In contrast, direct treatment of MDMs with DEP had no effect on MCP-1, but elevated IL-8 and TNF-α concentrations. Incubation with conditioned media from DEP-exposed MDMs caused a dramatic amplification in MCP-1 and IL-6, but not IL-8, release from HUVECs. The potentiation of HUVEC activation was suppressed by TNF-α inhibition. MCP-1- and IL-6-containing HUVEC supernatants caused increased monocyte chemotaxis that was not inhibited by anti-MCP-1 antibodies. We conclude that DEP has only modest direct endothelial effects. In contrast, proinflammatory cytokines released from particle-laden MDMs appear to exacerbate endothelial activation after DEP exposure.


Retinoic acid modulation of mRNA levels in malignant, nontransformed, and immortalized osteoblasts.
Clonal cell lines presumably "arrested" at a particular stage of differentiation are useful models to study the processes of differentiation in osteoblasts. UMR-201 is a presumptive preosteoblastic nontransformed rat clonal cell line with a limited life span in culture. Two immortalized cell lines, UMR-201-10A (10A) and UMR-201-10B (10B), were derived from UMR-201 by stable transfection with simian virus (SV) 40 large T antigen. This study compares the growth and profile of gene expression of the immortalized cell lines with those of UMR-201 and UMR-106-06, a rat clonal cell line with well-defined osteoblast-like phenotypic characteristics. All four cell lines constitutively expressed the mRNA for the gamma, alpha, and beta receptors for retinoic acid (RA), the growth hormone receptor, pro-alpha 1(I) collagen, osteonectin, bone proteoglycan I, and bone morphogenetic proteins (BMP) 1 and 2A. Alkaline phosphatase mRNA was absent in the preosteoblast cell lines but was induced by treatment with 10(-6) M RA, which also increased the steady-state levels of mRNA for osteopontin and BMP1. mRNA for matrix gla protein was constitutively present and further induced by RA in UMR-201 and 10B only. Messenger RNA for bone sialoprotein and bone morphogenetic protein 3 were constitutively expressed in UMR-106-06 and UMR-201 but absent in the immortalized cell lines. None of the cell lines expressed measurable mRNA for bone gla protein or bone proteoglycan II. 10B grew more rapidly than UMR-201, but unlike UMR-201, it was also able to proliferate in serum-free medium and exhibit anchorage-independent growth. In summary, this study identifies novel retinoic acid effects on gene expression in these cells. Differences noted in the expression of mRNAs between UMR-106-06 and the other cell lines may provide some insight into the sequence of expression of these phenotypic characteristics as osteoblasts differentiate.


Evaluation of antibiotic susceptibility testing by agar dilution and the Micro Media system (Fox Panel).
The susceptibilities of 350 gram-positive cocci and 638 gram-negative bacilli to various antimicrobial agents were compared by using the Micro-Media system (MMS) (Fox Panel) (Micro-Media Systems, Inc., Potomac, Md.) and a standard agar dilution procedure. Major discrepancies occurred with enterococci, among which 48 of 53 isolates (91%) were found to be resistant to penicillin G by agar dilution and reported as susceptible by the MMS. Other large discrepancies occurred with Staphylococcus aureus and Acinetobacter calcoaceticus subsp. anitratus, among which more than 40% of the isolates were judged to be resistant to ampicillin by agar dilution and susceptible by the MMS. In terms of overall agreement in interpretation of MICs by the two systems, an agreement of greater than 84% was seen for both gram-positive and gram-negative organisms when ampicillin and cephalothin (68 and 78% agreement for gram-positive cocci, respectively) were excluded. These disagreements in MIC interpretations may result in part from the small number of organisms tested per well (4,000 CFU) in the MMS, as compared with 10,000 CFU per test in the agar dilution method.


Tryptophan phosphorescence studies of the D-galactose/D-glucose-binding protein from Escherichia coli provide a molecular portrait with structural and dynamics features of the protein.
The D-galactose/D-glucose-binding protein (GGBP) from E. coli serves as an initial component for both chemotaxis toward glucose and high-affinity active transport of the sugar. In this work, we have used phosphorescence spectroscopy to investigate the effects of glucose and calcium on the dynamics and stability of GGBP. We found that GGBP exhibits a phosphorescence spectrum composed of two energetically distinct 0,0-vibrational bands centered at 404.43 and 409.61 nm; the large energy separation between them indicates two classes of chromophores making distinct dipolar interactions with their surrounding. Interestingly, the high-energy spectral component (404.43 nm) is one of the bluest spectra reported to date in proteins. Considering the ground state dipole direction, low-energy configurations for the indole side chain in proteins leading to blue-shifted spectra can arise from negative charges in proximity to the imidazole-ring nitrogen and/or positive charges near C4-C5 of the benzene ring. Among the five tryptophan residues of GGBP, Trp-284, located at the N-terminal domain of the protein, and Trp-183, located in the protein hinge region, make strong attractive charge interactions with surrounding side chains. Regarding Trp-284, the indole ring nitrogen is in contact with the negative charge of the Asp-267, whereas Trp-183 is next to the Glu-149 residue. In the latter, the ground state energy is further lowered by the proximity of the Arg-158 to the negative end (near C6) of the indole dipole. Regarding the red spectral component (409.61 nm), it is more intense than the blue component, presumably because more residues contribute to it. lambda 0,0 is typical of environments that are weakly polar or characterized by charges positioned near 90 degrees from the ground state dipole direction (the case of W195 and W127). The binding of glucose modifies the phosphorescence lifetime values as well as the spectrum of GGBP, shifting the blue band 0.54 nm to the blue and the red band 1 nm to the red. Finally, the removal of the calcium from GGBP structure causes variations in lifetime values and spectral shifts similar to those induced by glucose binding to the native protein. Aided by a detailed inspection of the three-dimensional structure of GGBP, these results contribute to a better understanding of the structure/function relationship of this protein.


Growth and toxin production of phomopsin A and ochratoxin A forming fungi under different storage conditions in a pea (Pisum sativum) model system.
Phomopsins are mycotoxins mainly infesting lupines, with phomopsin A (PHOA) being the main mycotoxin. PHOA is produced by Diaporthe toxica, formerly assigned as toxigenic Phomopsis leptostromiformis, causing infections in lupine plants and harvested seeds. However, Diaporthe species may also grow on other grain legumes, similar to Aspergillus westerdijkiae as an especially potent ochratoxin A (OTA) producer. Formation of PHOA and OTA was investigated on whole field peas as model system to assess fungal growth and toxin production at adverse storage conditions. Field pea samples were inoculated with the two fungal strains at two water activity (aw) values of 0.94 and 0.98 and three different levels of 30, 50, and 80% relative air humidity.After 14 days at an aw value of 0.98, the fungi produced 4.49 to 34.3 mg/kg PHOA and 1.44 to 3.35 g/kg OTA, respectively. Strains of D. toxica also tested showed higher PHOA concentrations of 28.3 to 32.4 mg/kg.D. toxica strains did not grow or produce PHOA at an aw values of 0.94, while A. westerdijkiae still showed growth and OTA production.Elevated water activity has a major impact both on OTA and, even more pronouncedly, on PHOA formation and thus, proper drying and storage of lupins as well as other grain legumes is crucial for product safety.


Complete sequence, subunit structure, and complexes with pancreatic alpha-amylase of an alpha-amylase inhibitor from Phaseolus vulgaris white kidney beans.
The complete amino acid sequence of a white kidney bean (Phaseolus vulgaris) alpha-amylase inhibitor (PHA-I), which is composed of two kinds of glycopolypeptide subunits, alpha and beta, was established by conventional methods. The polypeptide molecular weight of PHA-I determined by the light-scattering technique, considered together with the sequence molecular weights revealed for the subunits, indicated that PHA-I has the subunit stoichiometry of (alpha beta)2 complex. Inhibition test of PHA-I with increasing amounts of porcine pancreatic alpha-amylase (PPA) suggested that an inactive 2:1 complex is formed between PPA and PHA-I. In fact, two complexes differing from each other in the molar ratio of PPA to PHA-I were separated by gel filtration, and molecular weight estimation by the light-scattering technique confirmed that they are complexes of PHA-I with one or two PPA molecules. The binding of PPA to PHA-I appeared to follow simple binomial statistics, suggesting that two binding sites on PHA-I are independent and of high affinity for PPA.


Management issues in herpes zoster.
Herpes zoster is a common clinical illness, with acute and chronic pain a major feature. Antiviral agents started within 72 hours of the onset of the rash are effective in the acute illness and in post herpetic neuralgia or zoster associated pain. Newer agents offer advantages in efficacy and dosage convenience.


Identifying Explainable Machine Learning Models and a Novel SFRP2+ Fibroblast Signature as Predictors for Precision Medicine in Ovarian Cancer.
Ovarian cancer (OC) is a type of malignant tumor with a consistently high mortality rate. The diagnosis of early-stage OC and identification of functional subsets in the tumor microenvironment are essential to the development of patient management strategies. However, the development of robust models remains unsatisfactory. We aimed to utilize artificial intelligence and single-cell analysis to address this issue. Two independent datasets were screened from the Gene Expression Omnibus (GEO) database and processed to obtain overlapping differentially expressed genes (DEGs) in stage II-IV vs. stage I diseases. Three explainable machine learning algorithms were integrated to construct models that could determine the tumor stage and extract important characteristic genes as diagnostic biomarkers. Correlations between cancer-associated fibroblast (CAF) infiltration and characteristic gene expression were analyzed using TIMER2.0 and their relationship with survival rates was comprehensively explored via the Kaplan-Meier plotter (KM-plotter) online database. The specific expression of characteristic genes in fibroblast subsets was investigated through single-cell analysis. A novel fibroblast subset signature was explored to predict immune checkpoint inhibitor (ICI) response and oncogene mutation through Tumor Immune Dysfunction and Exclusion (TIDE) and artificial neural network algorithms, respectively. We found that Support Vector Machine-Shapley Additive Explanations (SVM-SHAP), Extreme Gradient Boosting (XGBoost), and Random Forest (RF) successfully diagnosed early-stage OC (stage I). The area under the receiver operating characteristic curves (AUCs) of these models exceeded 0.990. Their overlapping characteristic gene, secreted frizzled-related protein 2 (SFRP2), was a risk factor that affected the overall survival of OC patients with stage II-IV disease (log-rank test: p < 0.01) and was specifically expressed in a fibroblast subset. Finally, the SFRP2+ fibroblast signature served as a novel predictor in evaluating ICI response and exploring pan-cancer tumor protein P53 (TP53) mutation (AUC = 0.853, 95% confidence interval [CI]: 0.829-0.877). In conclusion, the models based on SVM-SHAP, XGBoost, and RF enabled the early detection of OC for clinical decision making, and SFRP2+ fibroblast signature used in diagnostic models can inform OC treatment selection and offer pan-cancer TP53 mutation detection.


Synaptic plasticity after chemical deafening and electrical stimulation of the auditory nerve in cats.
The effects of deafness on brain structure and function have been studied using animal models of congenital deafness that include surgical ablation of the organ of Corti, acoustic trauma, ototoxic drugs, and hereditary deafness. This report describes the morphologic plasticity of auditory nerve synapses in response to ototoxic deafening and chronic electrical stimulation of the auditory nerve. Normal kittens were deafened by neonatal administration of neomycin that eliminated auditory receptor cells. Some of these cats were raised deaf, whereas others were chronically implanted with cochlear electrodes at 2 months of age and electrically stimulated for up to 12 months. The large endings of the auditory nerve, endbulbs of Held, were studied because they hold a key position in the timing pathway for sound localization, are readily identifiable, and exhibit deafness-associated abnormalities. Compared with those of normal hearing cats, synapses of ototoxically deafened cats displayed expanded postsynaptic densities, a 35.4% decrease in synaptic vesicle (SV) density, and a reduction in the somatic size of spherical bushy cells (SBCs). In comparison with normal hearing cats, ototoxically deafened cats that received cochlear stimulation had endbulbs that expressed postsynaptic densities (PSDs) that were statistically identical in size, showed a 48.1% reduction in SV density, and whose target SBCs had a 25.5% reduction in soma area. These results demonstrate that electrical stimulation via a cochlear implant in chemically deafened cats preserves PSD size but not other aspects of synapse morphology. This determination further suggests that the effects of ototoxic deafness are not identical to those of hereditary deafness.


Protein denaturation and protein:drugs interactions from intrinsic protein fluorescence measurements at the nanolitre scale.
Protein stability and ligand-binding affinity measurements are widely required for the formulation of biopharmaceutical proteins, protein engineering and drug screening within life science research. Current techniques either consume too much of often precious biological or compound materials, in large sample volumes, or alternatively require chemical labeling with fluorescent tags to achieve measurements at submicrolitre volumes with less sample. Here we present a quantitative and accurate method for the determination of protein stability and the affinity for small molecules, at only 1.5-20 nL optical sample volumes without the need for fluorescent labeling, and that takes advantage of the intrinsic tryptophan fluorescence of most proteins. Coupled to appropriate microfluidic sample preparation methods, the sample requirements could thus be reduced 85,000-fold to just 10(8) molecules. The stability of wild-type FKBP-12 and a destabilizing binding-pocket mutant are studied in the presence and absence of rapamycin, to demonstrate the potential of the technique to both drug screening and protein engineering. The results show that 75% of the interaction energy between FKBP-12 and rapamycin originates from residue Phe99 in the binding site.


Trait Self-Compassion Reflects Emotional Flexibility Through an Association with High Vagally Mediated Heart Rate Variability.
Converging evidence shows a positive effect of self-compassion on self-reported well-being and mental health. However, few studies have examined the relation between self-compassion and psychophysiological measures. In the present study, we therefore examined the relation between trait self-compassion and vagally mediated heart rate variability (vmHRV) in 53 students (39 female, mean age = 23.63). Trait self-compassion was assessed using the Self-Compassion Scale, and resting vmHRV was measured during a 5-min ECG baseline period. We hypothesized that higher levels of trait self-compassion would predict higher levels of resting vmHRV. Controlling for potential covariates (including age, gender, and BMI), the results confirmed our hypotheses, showing that higher levels of trait self-compassion predicted higher vmHRV. These results were validated with a 24-h measure of vmHRV, acquired from a subsample of the participants (n = 26, 16 female, mean age = 23.85), confirming the positive correlation between high trait self-compassion and higher vmHRV. The relation between trait self-compassion, vmHRV, self-reported trait anxiety (the trait scale of the State-Trait Anxiety Inventory; STAI) and self-reported rumination (the Rumination subscale of the Rumination-Reflection Questionnaire; RRQ-Rum) was also investigated. Higher levels of trait anxiety and rumination were highly correlated with low levels of trait self-compassion. Trait anxiety, but not rumination, correlated marginally significantly with the level of vmHRV. The findings of the present study indicate that trait self-compassion predicts a better ability to physiologically and psychologically adapt emotional responses. Possible implications and limitations of the study are discussed.


Optimizing Discharge Summaries: A Multispecialty, Multicenter Survey of Primary Care Clinicians.
Patient care in the United States has become increasingly more fragmented, and the discharge summary serves as a critical tool for transmitting information on a patient's hospital admission to the primary care clinician. Some guidelines regarding how to write discharge summaries exist, but few are focused on prioritizing content that is most important to optimize a patient's transition of care. We conducted a national survey across various medical primary care specialties, including trainees and advanced practice providers, to understand the priorities of primary care clinicians. We distributed the survey to 2184 clinicians affiliated with 8 large academic institutions. Our response rate was 21%. Hospital course, discharge diagnoses, medication reconciliation, and follow-up sections were ranked as the most important categories with a 95.5% concordance rate among surveyed institutions. The least important sections were contact numbers for inpatient clinicians, ancillary services, weight-bearing status, and wound care. Similar themes were also identified via consensus review of the free-texted comments, adding that discharge summary style was also important. Other identified barriers to high-quality transition of care are both the limited time primary care clinicians can spend reviewing discharge summaries and lack of adequate communication between hospitalists and the outpatient clinician. High-yield content should be presented at the beginning of the discharge summary and conveyed in a brief, succinct manner to ensure maximal utility of the document as a transition of care tool.


A phase II study of the combination of gemcitabine and imatinib mesylate in pemetrexed-pretreated patients with malignant pleural mesothelioma.
Second-line chemotherapy is not a standard of care in patients with malignant pleural mesothelioma (MPM) that progresses after first-line treatment with cisplatin and pemetrexed. In pre-clinical models, the combination of gemcitabine (GEM) and imatinib mesylate (IM), compared with GEM alone, led to a further tumor growth inhibition and improved survival. This phase II study evaluates the antitumor activity of a combination of IM and GEM in platinum-pemetrexed-pretreated MPM patients expressing PDGFR-β and/or cKIT by immunohistochemistry (IHC). GEM (1000 mg/m2) was given on days 3 and 10; IM (400 mg) was taken orally on days 1-5 and 8-12 of a 21-day cycle. The primary endpoint was the 3-month progression-free survival (PFS) rate. The study follows the optimal two-stage design of Simon. A 3-month PFS target of 75 % was required. With a probability error α = 10 % and a power of 80 %, the calculated sample size was 22 patients. In particular, in the first step, six out of nine patients and globally 14/22 patients free from progressive disease at 3 months were required. Secondary endpoints included response rate, duration of response, toxicity and overall survival (OS). In total, 23 patients were enrolled (ECOG PS 0-1/2: 9/13; one previous line/≥two previous lines: 10/13). Partial response was achieved in four patients (17.4 %) and stable disease in 11 (47.8 %) with a disease control rate of 65.3 %. After a median follow-up of 34.5 months, median PFS and OS were 2.8 and 5.7 months, respectively. The 3-month PFS rate was 39.1 % (9/23 patients). All-grade drug-related adverse events occurred in 17 (73.9 %) patients. Grade 3 treatment-related adverse events were observed in four (17 %) patients. The combination of IM and GEM is well tolerated in platinum-pemetrexed-pretreated MPM patients expressing PDGFR-β and/or cKIT by IHC, but it does not show a significant PFS benefit.


Carbon fluxes of xylose-consuming Saccharomyces cerevisiae strains are affected differently by NADH and NADPH usage in HMF reduction.
Industrial Saccharomyces cerevisiae strains able to utilize xylose have been constructed by overexpression of XYL1 and XYL2 genes encoding the NADPH-preferring xylose reductase (XR) and the NAD(+)-dependent xylitol dehydrogenase (XDH), respectively, from Pichia stipitis. However, the use of different co-factors by XR and XDH leads to NAD(+) deficiency followed by xylitol excretion and reduced product yield. The furaldehydes 5-hydroxymethyl-furfural (HMF) and furfural inhibit yeast metabolism, prolong the lag phase, and reduce the ethanol productivity. Recently, genes encoding furaldehyde reductases were identified and their overexpression was shown to improve S. cerevisiae growth and fermentation rate in HMF containing media and in lignocellulosic hydrolysate. In the current study, we constructed a xylose-consuming S. cerevisiae strain using the XR/XDH pathway from P. stipitis. Then, the genes encoding the NADH- and the NADPH-dependent HMF reductases, ADH1-S110P-Y295C and ADH6, respectively, were individually overexpressed in this background. The performance of these strains, which differed in their co-factor usage for HMF reduction, was evaluated under anaerobic conditions in batch fermentation in absence or in presence of HMF. In anaerobic continuous culture, carbon fluxes were obtained for simultaneous xylose consumption and HMF reduction. Our results show that the co-factor used for HMF reduction primarily influenced formation of products other than ethanol, and that NADH-dependent HMF reduction influenced product formation more than NADPH-dependent HMF reduction. In particular, NADH-dependent HMF reduction contributed to carbon conservation so that biomass was produced at the expense of xylitol and glycerol formation.


High-resolution melting amplicon analysis as a method to detect c-kit and platelet-derived growth factor receptor alpha activating mutations in gastrointestinal stromal tumors.
High-resolution melting amplicon analysis (HRMAA) was used to detect c-kit and platelet-derived growth factor receptor alpha (PDGFRA) activating mutations in 96 gastrointestinal stromal tumors (GISTs). HRMAA detected mutations in 87 GISTs (91%). Of the 87 cases, 69 (79%) contained c-kit mutations and 18 (21%), PDGFRA mutations. One c-kit mutation-positive case contained an exon 9 mutation, ins FY at codon 503, that has not been previously described. One PDGFRA mutation-positive case contained mutation D842V del 843, also not previously described. Of 18 PDGFRA mutation-positive cases, 3 (17%) were strongly positive for kit expression as measured by CD117 immunohistochemical analysis. Of 69 c-kit mutation-positive cases, 66 (96%) showed strong kit immunohistochemical expression, but 3 (4%) showed negative to weak CD117 expression. Of 96 cases, 9 (9%) were wild type for c-kit and PDGFRA. Of the wild-type cases, 8 still showed strong immunohistochemical kit expression, whereas 1 showed weak kit expression. GISTs with PDGFRA mutations were found in the stomach, omentum, and peritoneum but not the small intestine. GISTs with c-kit exon 9 mutations were found primarily in the small intestine. HRMAA is a sensitive technique that can be used to rapidly identify c-kit and PDGFRA activating mutations in GISTs.


Identification of canine coronavirus strains from feces by S gene nested PCR and molecular characterization of a new Australian isolate.
A nested PCR (nPCR) assay for the detection of canine coronavirus (CCV) in fecal samples is described. The target sequence for the assay was a 514-bp fragment within the spike (S) glycoprotein gene. The sensitivity of the assay is extremely high, detecting as little as 25 50% tissue culture infective doses per g of unprocessed feces. A clinical trial using dogs challenged orally with CCV SA4 and CCV NVSL was used to compare viral isolation and the nPCR assay as detection techniques over a 2-week period of infection. Virus isolation detected CCV shedding from day 4 to 9 postchallenge, while the nPCR assay detected CCV shedding from day 4 to 13 postchallenge. Cloning and sequencing of the nPCR assay product enabled investigation of the evolutionary relationships between strains within the S gene. The simple and rapid procedure described here makes this assay an ideal alternative technique to electron microscopy and viral isolation in cell culture for detection of CCV shedding in feces. The described assay also provides a method of identifying new strains of CCV without the complicated and time-consuming practice of raising antibodies to individual strains. This is illustrated by the identification, for the first time, of an Australian isolate of CCV (UWSMN-1).


Differential effects of nuclear receptor corepressor (N-CoR) expression levels on retinoic acid receptor-mediated repression support the existence of dynamically regulated corepressor complexes.
Thyroid hormone and retinoic acid receptors are members of the nuclear receptor superfamily of ligand-dependent transcription factors that stimulate the transcription of target genes in the presence of activating ligands and repress transcription in their absence. Transcriptional repression by the thyroid hormone and retinoic acid receptors has been proposed to be mediated by the nuclear receptor corepressor, N-CoR, or the related factor, SMRT (silencing mediator of retinoic acid and thyroid hormone receptors). Recent studies have suggested that transcriptional repression by N-CoR involves a corepressor complex that also contains mSin3A/B and the histone deacetylase, RPD3. In this manuscript, we demonstrate that transcriptional repression by the retinoic acid receptor can be either positively or negatively regulated by changes in the levels of N-CoR expression, suggesting a relatively strict stoichiometric relationship between N-CoR and other components of the corepressor complex. Consistent with this interpretation, overexpression of several functionally defined domains of N-CoR also relieve repression by nuclear receptors. N-CoR is distributed throughout the nucleus in a nonuniform pattern, and a subpopulation becomes concentrated into several discrete dot structures when highly expressed. RPD3 is also widely distributed throughout the nucleus in a nonuniform pattern. Simultaneous imaging of RPD3 and N-CoR suggest that a subset of each of these proteins colocalize, consistent with the existence of coactivator complexes containing both proteins. In addition, a substantial fraction of both N-CoR and mSin3 A/B appear to be independently distributed. These observations suggest that interactions between RPD3 and Sin3/N-CoR complexes may be dynamically regulated.


Chordal transfer for repair of anterior leaflet prolapse.
Chordal transfer is a reliable technique for correction of anterior leaflet prolapse. In most cases, normal chordae and a strip of leaflet tissue are transferred from the posterior leaflet to the free edge of unsupported anterior leaflet; the posterior leaflet is repaired as after a quadrangular resection with either a sliding repair or a standard repair. Occasionally, secondary anterior leaflet chordae may be transferred from the undersurface of the anterior leaflet to its unsupported free edge, effecting a rapid and effective repair. After chordal transfer, annuloplasty completes the mitral valve repair.


Infant and Young Child Feeding Index and its association with nutritional status: A cross-sectional study of urban slums of Ahmedabad.
Infant and young child feeding (IYCF) practices are multidimensional and change rapidly in short intervals in the 1st year of life, asking for simultaneous assessment of various feeding dimensions in children of 6 months and older. Infant and Child Feeding Index (ICFI) is a composite index which measures complete feeding practices for infants and young children. The present study was conducted to assess IYCF practices for children aged 6-36 months in terms of ICFI and some sociodemographic factors and find out the association of ICFI with nutritional status. A cross-sectional study was conducted from July 2015 to October 2015 in Girdharnagar ward of Ahmedabad. Two hundred and ten mother-child pairs were selected by two-staged cluster sampling and were interviewed using a schedule adapted from Knowledge, Practices, and Coverage 2000+ model questionnaire. Appropriate anthropometric measurements were taken and nutritional indicators were calculated. Chi-square test, t- test, and regression analysis were applied wherever required. Epi info version 7.0 and MS Excel 2007 were used for statistical analysis. Nearly 65.2% of the children were stunted, 43.3% were underweight, and 11.9% were wasted. Only 38.3% of the children were initiated on breastfeeding within 1 h of birth. Only 19.1% of the children were breastfed for 2 years and beyond. Meal frequency was adequate in 64.3% and dietary diversity of >4 food groups was given to only 15.7% of the children. Significant higher proportions of children with low ICFI scores had illiterate mothers, were older, and belonged to lower socioeconomic strata. There was statistically significant association of ICFI with all the three nutritional status indicators. Higher proportion of children with lower ICFI scores had lower weight-for-height Z-scores, weight-for-age Z-scores, and height-for-age Z-scores. The present study revealed that ICFI can be used to measure IYCF practices in a single composite index, which in turn can reflect the nutritional status of the children.


[Differential evaluation of excitatory conduction rats along different segments of motor nerves and functional tests in the diagnosis of diseases of the human peripheral neuromotor apparatus].
In 61 patients with neurono- and neuropathies and affections of the synaptic apparatus and the muscular matter of various etiology, the state of the peripheral nerves was examined. It was found that various forms of the pathology are characterized by specific changes in the speed of the stimulus transmission along various parts of motor nerves. The changes of the functional state can be seen especially clearly on comparing the speed of the stimulus transmission along the proximal and the distal parts of a peripheral nerve (the difference being named "transmission speed gradient"). In neuronopathies the transmission speed changes are prevailing in the proximal, in neuropathies in the distal, and in synaptic structure abnormalities in the extreme distal parts of the nerve. A 15-minute artificial ischemia of the nerve (a functional test) led to characteristic changes of the transmission speed, these changes correlating with the process stage. In cases of mild changes of the transmission speed the nerve was more sensitive, while in cases of pronounced changes more resistant to the ischemia. It was shown that the transmission speed changes in ischemia manifested themselves along the whole nerve, but not only in the site of its affection. In cases of the carpal canal syndrome the ischemia-induced lowering of the transmission speed in the nerve distal part was extremely sharp.