The learning curve for EMR with circumferential mucosal incision in treating intramucosal gastric neoplasm.
EMR with circumferential mucosal incision facilitates en bloc resection, which is the most important predictor for complete resection in the treatment of intramucosal gastric neoplasm. The objective of the study was to evaluate the efficacy and the safety of EMR with circumferential mucosal incision in relation to the endoscopist's experience. In this retrospective study, we analyzed the outcome of 80 EMR procedures performed by a single endoscopist at the National Cancer Center, Goyang, Korea. The EMR procedure with circumferential mucosal incision was performed with a conventional needle knife. We compared the en bloc resection rate, the complete resection rate, the duration of the procedure time, and the associated complications by quartiles. From the first to the last quartile, en bloc resection rates were 55%, 45%, 85%, and 85% (p = 0.006), and complete resection rates were 65%, 60%, 90%, and 85% (p = 0.039). The increase in the mean en bloc and complete resection rates between the first two quartiles and the second two quartiles was significant (p = 0.002 and p = 0.019, respectively). Three perforations (15%) were reported in the first quartile and only one (1.7%) in the remaining 3 quartiles (p = 0.046). The procedure time also decreased after the first 20 cases (p = 0.004). For an experienced endoscopist, EMR with circumferential mucosal incision could be performed effectively and safely after the experience of 40 cases.


The BTNL2 A allele variant is frequent in Danish patients with sarcoidosis.
The butyrophilin-like 2 (BTNL2) gene is located on chromosome 6p21.3 close to the HLA-class II genes. An association has been reported between sarcoidosis and a single nucleotide polymorphism in BTNL2, rs2076530, also termed the A allele. To evaluate whether patients with sarcoidosis carry the A allele more frequently than healthy subjects. The series comprised 87 ethnic Danes with sarcoidosis and 113 healthy control subjects. Analysis of rs2076530 was performed by Taqman assay, polymerase chain reaction and sequencing of genomic DNA. Sarcoidosis patients had a higher frequency of the A allele than controls (73.9% vs 55.8%) (P < 0.025). The frequency of GG, GA and AA genotype was 5.7%, 40.2% and 54.0% in patients vs 16.0%, 56.6% and 27.4% in controls (P < 0.001). The AA genotype was associated with increased risk of sarcoidosis in both a dominant [odds ratio (OR) 3.1; 95% confidence interval (CI) 1.1-8.7; P < 0.03] and a recessive model (OR 3.1; 95% CI 1.72-5.61; P < 0.001). Population attributable fraction for disease was 50% in a dominant model and 25% in a recessive model. The BTNL2 A allele variant occurs with a high frequency in Danish patients with sarcoidosis and the AA genotype is associated with a ∼threefold higher risk of sarcoidosis than the GG genotype. Our results should encourage future studies on the interrelationship between the BTNL2 protein and granuloma formation in sarcoidosis.


Synthesis, Characterization, and Reactivity of Tris(imido)chromium(VI) Complexes.
Multiple tris(imido)chromium(VI) complexes, including neutral and ionic compounds, have been synthesized and characterized. (tBuN)2Cr(NHtBu)Cl can be deprotonated by KN(SiMe3)2, yielding K[(tBuN)3CrCl]. This tris(imido) anion undergoes nucleophilic substitution by PPh3 and tBuNH2 to form (tBuN)3Cr(PPh3) and (tBuN)2Cr(NHtBu)2, respectively. (tBuN)2Cr(NHtBu)2 loses one amido proton to form K[(tBuN)3Cr(NHtBu)] upon reaction with KN(SiMe3)2. The imido ligands of K[(tBuN)3CrCl] and (tBuN)3Cr(PPh3) are attacked by the electrophile MeI to produce (tBuN)2Cr(NMetBu)Cl and (tBuN)2Cr(NMetBu)I, respectively. An alternate way to make tris(imido) anions is deprotonation of (tBuN)2Cr(NHtBu)Cl by an alkyl lithium reagent, e.g., Me3SiCH2Li. The resulting Li[(tBuN)3CrCl] was alkylated by a second equivalent of Me3SiCH2Li to form Li[(tBuN)3Cr(CH2SiMe3)]. Reactivity studies of tris(imido) complexes show cycloaddition with PhNCO or CO2 to form metallacycles.


Changes of lipoprotein(a) levels with endogenous steroid hormones.
Lipoprotein(a) [Lp(a)] is an LDL-like molecule that is likely causal for cardiovascular events and Lp(a) variability has been shown to be mostly of genetic origin. Exogenous hormones (hormone replacement therapy) seem to influence Lp(a) levels, but the impact of endogenous hormone levels on Lp(a) is still unknown. The aim of the study was to assess the effect of endogenous steroid hormone metabolites on Lp(a). Lipoprotein(a) levels were measured in 1,021 participants from the Swiss Kidney Project on Genes in Hypertension, a family-based, multicentre, population-based prospective cohort study. Endogenous levels of 28 steroid hormone precursors were measured in 24-h urine collections from 883 individuals. Of the participants with Lp(a) data, 1,011 participants had also genotypes available. The participants had an average age of 51 years and 53% were female. Median Lp(a) levels were 62 mg/L, and the 90th percentile was 616 mg/L. The prevalence of a Lp(a) elevation ≥700 mg/L was 3.2%. Forty-three per cent of Lp(a) variability was explained respectively by: age (2%, p < .001), LDL-C (1%, p = .001), and two SNPs (39%, p value<2⋅10-16 ). Of the 28 endogenous steroid hormones assessed, androstenetriol, androsterone, 16α-OH-DHEA and estriol were nominatively associated with serum Lp(a) levels in univariable analyses and explained 0.4%-1% of Lp(a) variability, but none of them reached significance in multivariable models. In this contemporary population-based study, the prevalence of a Lp(a) elevation ≥700 mg/L was 3.2%. The effect of endogenous steroid hormone levels of Lp(a) variability was small at best, suggesting a negligible impact on the wide range of Lp(a) variability.


[Multidisciplinarity, interdisciplinarity, transdisciplinarity].
Working in a team, concepts in transdisciplinarity, multidisciplinarity, and interdisciplinarity. Integrated research in a society immersed in care, where the aim is none other than to increase the quality of life of those people who need it, the participation of all as links in the same chain where the failure or lack of commitment by one of the parties can endanger any project of the procedure to relate scientific knowledge, extra-scientific experience and the practice of solving problems.


Perceptions and motivations of an Australian cohort of women with or without congenital heart disease proceeding to pregnancy.
To assess the perceptions of women with congenital heart disease (CHD) regarding the severity of their cardiac abnormality and its implications in pregnancy, and whether their motivations to conceive were similar to those of women without CHD. A retrospective descriptive study of a cohort of women with (n = 20) or without (n = 20) CHD, aged over 18 years, who had had one or more successful pregnancies. The women were surveyed using a semi-structured questionnaire and their medical records were reviewed. Data were analyzed using thematic analysis. Women with CHD were concerned about the health risks associated with their CHD; however, they were not deterred from conceiving because 10 out of 20 women had a tendency to downplay the seriousness of their CHD. The women's motivations to conceive were similar in both groups. Although women with CHD were clinically identified as an at-risk group, with possible health complications for themselves and their infant, it was ultimately the women's inaccurate perception of the severity of their CHD and of the associated potential health risks that influenced their decision to pursue motherhood. This emphasizes the need for clinicians to have detailed knowledge of the cardiac abnormality and its implications in pregnancy.


Resolution of the clinical features of tyrosinemia following orthotopic liver transplantation for hepatoma.
The clinical history before transplantation and subsequent clinical and biochemical course of 3 children and one adult with hereditary tyrosinemia treated by orthotopic hepatic transplantation is described. All four patients are now free of their previous dietary restrictions and appear to be cured of both their metabolic disease and their hepatic neoplasm.


Establishment of functional epithelial organoids from human lacrimal glands.
Tear deficiency due to lacrimal gland (LG) dysfunction is one of the major causes of dry eye disease (DED). Therefore, LG stem cell-based therapies have been extensively reported to regenerate injured lacrimal tissue; however, the number of stem cells in the LG tissue is low, and 2D long-term cultivation reduces the differentiation capacity of stem cells. Nevertheless, 3D LG organoids could be an alternative for a DED therapy because it is capable of prolonged growth while maintaining the characteristics of the LG tissue. Here, we report the development of LG organoids and their application as cell therapeutics. Digested cells from human LG tissue were mixed with Matrigel and cultured in five different media modified from human prostate/salivary organoid culture media. After organoid formation, the growth, specific marker expression, and histological characteristics were analyzed to authenticate the formation of LG organoids. The secretory function of LG organoids was confirmed through calcium influx or proteomics analysis after pilocarpine treatment. To explore the curability of the developed organoids, mouse-derived LG organoids were fabricated and transplanted into the lacrimal tissue of a mouse model of DED. The histological features and specific marker expression of LG organoids were similar to those of normal LG tissue. In the pilocarpine-treated LG organoid, levels of internal Ca2+ ions and β-hexosaminidase, a lysosomal protein in tear fluid, were increased. In addition, the secreted proteins from pilocarpine-treated lacrimal organoids were identified through proteomics. More than 70% of the identified proteins were proven to exosome through gene ontology analysis. These results indicate that our developed organoid was pilocarpine reactive, demonstrating the function of LG. Additionally, we developed LG organoids from patients with Sjogren's syndrome patients (SS) and confirmed that their histological features were similar to those of SS-derived LG tissue. Finally, we confirmed that the mouse LG organoids were well engrafted in the lacrimal tissue two weeks after transplantation. This study demonstrates that the established LG organoids resemble the characteristics of normal LG tissue and may be used as a therapy for patients with DED.


FMRFamide is endogenous to the Aplysia heart.
The presence of the molluscan neuropeptide FMRFamide was investigated in the heart of the sea hare, Aplysia californica. Immunohistochemical localization and high performance liquid chromatography (HPLC) coupled with radioimmunoassays of HPLC fractions were used to demonstrate the presence of FMRFamide and FLRFamide in the heart. FMRFamide-immunoreactive (FMRFamide-IR) nerve fibers, varicosities, and neuronal somata were observed in whole-mounts of the hearts. The atrium and atrioventricular (AV) valve regions contained significantly higher densities (P < 0.05, ANOVA) of immunoreactive varicosities compared to the ventricle. The high density of FMRFamide-IR varicosities in the atrium and the lack of sensitivity of this region to FMRFamide suggest that the atrium may be a neurohemal organ for the release of FMRFamide. The presence of FMRFamide-IR somata in the Aplysia heart suggests that peripheral neurons may play a role in modifying heart activity, independent of the central nervous system.


Fyn is induced by Ras/PI3K/Akt signaling and is required for enhanced invasion/migration.
Src family kinases (SFKs) are frequently over-expressed and/or activated in human cancers, and play key roles in cancer cell invasion, metastasis, proliferation, survival, and angiogenesis. Allosteric activation of SFKs occurs through well-defined post-translational mechanisms, however the SFK member Fyn is over-expressed in multiple human cancers (prostate, melanoma, pancreatic, glioma, chronic myelogenous leukemia) and the mechanism of increased Fyn expression is unclear. Since activation of Ras oncogenes is a common oncogenic event leading to the activation of multiple effector pathways, we explored if Ras could induce Fyn expression. Retroviral transduction of the human keratinocyte cell line HaCaT with oncogenic H-Ras dramatically up-regulated Fyn mRNA (>100-fold, P < 0.001), protein, and kinase activity without affecting Src levels or activity. Activation of Akt, but not MAPK or EGFR, was necessary and sufficient for induction of Fyn by H-Ras. Expression of active Fyn was sufficient to increase HaCaT cell migration and invasion, and the enhanced migration and invasion induced by H-Ras could be significantly blocked (70% reduction, P < 0.01) by knockdown of Fyn with a specific siRNA or inhibition of SFKs with PP2. In addition, expression of Fyn in MDA-MB-231 breast cancer cells was dependent on PI3K activity and was involved in their invasive phenotype. Thus, the Ras/PI3K/Akt pathway can account for Fyn over-expression in cancers, and Fyn is a critical mediator of the Ras-stimulated invasive cell phenotype. These results support the development of therapeutic strategies targeting Akt/Fyn pathway to block migration and invasion of tumor cells.


Study on fluorination-toxicity relationships. Syntheses of 1-N-[(2R,3R)- and (2R,3S)-4-amino-3-fluoro-2-hydroxybutanoyl] derivatives of kanamycins.
(2R,3R)- And (2R,3S)-4-azido-3-fluoro-2-hydroxybutanoic acids (11 and 22) have been prepared from 3-deoxy-3-fluoro-1,2-O-isopropylidene-alpha-D-glucofuranose (1) and 3,5-di-O-benzyl-1,2-O-isopropylidene-alpha-D-xylofuranose (12), respectively. They were then coupled to the N2N-1 group of suitably protected kanamycin A or kanamycin B analogs to give, 1-N-[(2R,3R)- and (2R,3S)-4-amino-3-fluoro-2-hydroxybutanoyl]kanamycins (32-35). This group of compounds (32-34) exhibited similar antibacterial activity and toxicity level as those of the corresponding 1-N-[(S)-4-amino-2-hydroxybutanoyl] (AHB) derivatives of kanamycins. The base strength of the H2N-4"' group of 32 and 34, as determined by 13C NMR spectroscopy (in D2O) at varying pD values, was found to be lower when compared to the basicity for the corresponding AHB analogs. The relationship between observed toxicity and base strength of the H2N-4"' group is discussed.


Maternal serum Schwangerschafts protein-1 (SP1) and fetal chromosomal abnormalities at 10-13 weeks' gestation.
Maternal serum SP1 concentration was measured at 10-13 weeks' gestation in samples from 87 pregnancies with fetal chromosomal abnormalities (trisomy 21 n = 45; trisomy 18 n = 19; trisomy 13 n = 8; Turner syndrome n = 7; 47,XXX or 47,XXY n = 4; triploidy n = 4), and in samples from 348 matched controls. In the control group, SP1 increased significantly with fetal crown-rump length (r = 0.20, P < 0.0001) and there was no significant association with fetal nuchal translucency thickness (r = 0.03). Similarly, in the group with fetal chromosomal abnormalities, SP1 increased significantly with crown-rump length (r = 0.31, P < 0.01) and there was no significant association with nuchal translucency thickness (r = -0.08). In the groups with fetal trisomy 18 and trisomy 13, the median SP1 (0.76 MoM and 0.57 MoM, respectively) was significantly lower than in the controls (z = 2.64 and z = 3.27, respectively); in 21% and 25% of the cases, values were below the 5th centile. In the group with trisomy 21 and other chromosomal abnormalities the median SP1 (0.96 MoM and 0.93 MoM, respectively) was not significantly different from controls (z = 1.17 and z = 0.67, respectively). Measurement of SP1 concentration at 10-13 weeks' gestation is not likely to be useful in the prediction of fetal chromosomal abnormalities.


Subcellular distribution of cardiac 5'-nucleotidase: alteration of microsomal pool in hypertrophied pig heart.
The subcellular distribution of the 5'-nucleotidase activity was investigated in normal and hypertrophied pig hearts; normal rat hearts were used for comparison. The left ventricular hypertrophy was induced in pigs by banding the supravalvular aorta for 4, 8 and 12 weeks. By employing different procedures for the isolation of cardiac membranes, a major catalytic site for 5'-nucleotidase was found to be located at sarcolemma in rat heart and microsomes (sarcoplasmic reticulum) in pig heart. A progressive decrease in the homogenate and microsomal 5'-nucleotidase activity occurred upon the development of myocardial hypertrophy in pigs. This reduction in microsomal 5'-nucleotidase activity was characterized by a depression in both apparent Vmax and Km values. These results indicate that a primary 5'-nucleotidase pool is present in the intracellular compartment of the pig heart and is altered during the development of hypertrophy.


Extended clomiphene citrate (CC) and prednisone for the treatment of chronic anovulation resistant to CC alone.
To evaluate effectiveness and safety of a regimen of extended clomiphene citrate (CC) and prednisone for patients who fail treatment with CC alone. Retrospective observational analysis. University-based tertiary infertility center. Twenty-four anovulatory patients who failed to ovulate after CC 150 mg administered for 5 days. Treatment consisted of CC given on cycle days 3 through 9 (extended) at a starting dose of 100 to 150 mg/d. Additionally, patients were given prednisone 5 mg orally each night throughout the cycle. Ovulation was confirmed by luteal serum P. Pregnancy was confirmed by rising hCG levels and transvaginal ultrasound. A total of 60 cycles were available for review. Forty-four of these cycles were ovulatory (73%) and 11 patients (46%) conceived on this therapy. Logistic (two-parameter) pregnancy occurrence over time (cycles) revealed a maximum pregnancy probability of 0.66 and a cycle fecundity of 0.36. No complications of therapy were noted. Clomiphene citrate-resistant anovulatory patients have high rates of ovulation and pregnancy after treatment with extended CC and prednisone. This therapy offers a potential reduction in cost and risk and should be considered in this group of patients before gonadotropin stimulation or surgery.


Comparison of cystometric methods in female rats.
Rat cystometry is a common model used to investigate urinary storage and voiding function. The effect of cystometric instrumentation in rat studies might be a source of deviation from normal physiologic responses. We hypothesized that transurethral catheterization would produce obstruction-related changes, and that suprapubic catheterization would limit volume-related functions as well as disrupt normal urothelial sensory function. We investigated the influence of transurethral and suprapubic catheterization on storage and voiding in the rat model. Three groups of female SD rats 250-300 g under urethane anesthesia were studied. Cystometric and pseudoaffective responses to physiologic voiding with and without suprapubic catheter placement, and cystometry via suprapubic and transurethral catheterization were studied. In free-voiding animals, per-void volume was 1.8 +/- 0.2 ml with an average flow rate of 0.18 ml/sec, and intercontraction interval (ICI) 60 min. Suprapubic catheterization decreased the ICI and per-void volume consistent with capacity reduction. Suprapubic cystometry did not significantly alter parameters compared to voiding except for a shortened ICI. Bladder pressures and somatic responses were increased, and urine flow impaired by transurethral cystometry. Terazosin did not significantly improve voiding parameters. Other than volume-related parameter changes probably related to surgical compromise of bladder capacity, suprapubic catheterization does not alter the cystometric and physiologic responses to voiding when compared to normal, uninstrumented voiding. Transurethral cystometry appears to be obstructive and may activate nociceptive reflexes. For this reason, whenever possible, urodynamic testing using the rat model should employ suprapubic catheterization.


An inducible proline transport system in Candida albicans.
1. When Candida albicans cells were preincubated with proline or grown in the presence of proline as the sole nitrogen source they exhibited a rapid increase in the influx of proline (the inducible transport system). 2. The induction appeared to be specific for proline and also demonstrated in other Candida species. 3. Both the inducible and constitutive proline uptake systems exhibited similar characteristic features. 4. The nature of the inducer for proline uptake in C. albicans appeared to be free proline. 5. The development of the inducible proline transport system was dependent on concomitant synthesis of RNA and protein and the induction was not affected by glucose or any other carbon sources used.


Integration of multiple lineage measurements from the same cell reconstructs parallel tumor evolution.
Organoid evolution models complemented with integrated single-cell sequencing technology provide a powerful platform to characterize intra-tumor heterogeneity (ITH) and tumor evolution. Here, we conduct a parallel evolution experiment to mimic the tumor evolution process by evolving a colon cancer organoid model over 100 generations, spanning 6 months in time. We use single-cell whole-genome sequencing (WGS) in combination with viral lineage tracing at 12 time points to simultaneously monitor clone size, CNV states, SNV states, and viral lineage barcodes for 1,641 single cells. We integrate these measurements to construct clonal evolution trees with high resolution. We characterize the order of events in which chromosomal aberrations occur and identify aberrations that recur multiple times within the same tumor sub-population. We observe recurrent sequential loss of chromosome 4 after loss of chromosome 18 in four unique tumor clones. SNVs and CNVs identified in our organoid experiments are also frequently reported in colorectal carcinoma samples, and out of 334 patients with chromosome 18 loss in a Memorial Sloan Kettering colorectal cancer cohort, 99 (29.6%) also harbor chromosome 4 loss. Our study reconstructs tumor evolution in a colon cancer organoid model at high resolution, demonstrating an approach to identify potentially clinically relevant genomic aberrations in tumor evolution.


Species distribution of coagulase-negative staphylococci isolated from clinical sources.
The distribution of coagulase-negative staphylococcal species from clinical material was investigated in order to detect possible tropisms. 398 strains of staphylococci (336 coagulase-negative and 62 coagulase-positive strains) and 3 micrococcus strains were isolated. Differentiation between the two genera was achieved by testing sensitivity to lysostaphin and anaerobic growth in thioglycolate. Four S. hominis strains reacted like micrococci in these tests. Staphylococci were classified according to the Kloos and Schleifer biotyping scheme in a modified way on microtitre plates. Additionally coagulation and reduction of methylene blue milk were determined and the test proved to be useful. Identified coagulase-negative species included S. epidermidis (205 strains), S. hominis (64), S. haemolyticus (35), S. warneri (15), S. capitis (5), S. simulans (6), S. saprophyticus (5), and S. species (1). A high predominance of S. epidermidis was found among isolates from nasopharyngeal (91%) and tracheobronchial (80%) tracts and most other localizations. A different distribution of species was observed in urine and vaginal swabs where non-S. epidermidis species accounted for over 50% of identified strains. The majority of S. hominis and S. haemolyticus strains was isolated from these sources. S. simulans, like S. saprophyticus was found in urine specimens only and occurred predominantly in women of the sexually active age.


Stereochemistry of the dTDP-glucose oxidoreductase reaction.
The stereochemical course of the dTDP-glucose oxidoreductase (EC 188.8.131.52) reaction was studied using enzyme partially purified from Escherichia coli and dTDP-(6R)- and (6S)-[4-2H, 6-3H]glucose as substrate. The latter was prepared enzymatically by reduction of (3R)- and (3S)-3-P-[3-3H]glycerate to the 1-deuterated 3-P-glyceraldehyde with (4S)-[4-2H]NADH, followed first by conversion to glucose-1-P with the glycolytic enzymes, and then by transformation into the dTDP derivative. The stereospecifically labeled dTDP-glucose samples were mixed with nonlabeled carrier material and converted to dTDP-4-keto-6-deoxyglucose, which contained a chiral methyl group as shown by chirality analysis of the acetic acid resulting from Kuhn-Roth oxidation of the sugar nucleotide. These results confirm that the hydrogen transfer from C4 to C6 is intramolecular and show that the migrating hydrogen replaces the 6-hydroxyl group with inversion of configuration. Assuming that the hydrogen transfer, since it is intramolecular, must be suprafacial, it follows that the elimination of water from C5 and C6 is formally syn, whereas the reduction of the resulting delta5,6-double bond formally involves an anti addition of H+ and H-.


Urinary albumin and cardiovascular profile in the middle-aged population.
The moderate increase in urinary albumin excretion defined as microalbuminuria is not rare and is associated with cardiovascular risk factors. Microalbuminuria prevalence is low in the absence of cardiovascular risk factors and progressively increases with the number cardiovascular risk factors. The main correlate of microalbuminuria is blood pressure, either systolic or diastolic pressure. The relation between blood pressure and microalbuminuria is continuous and graded because the microalbuminuria prevalence increases with the severity of hypertension. Among hypertensive patients on drug treatment, blood pressure control is associated with a low prevalence of microalbuminuria. Thus, blood pressure appears as a determinant of microalbuminuria rather than a mere correlate. For hypercholesterolemia, smoking, and diabetes, data are less strong but point to an independent positive association with microalbuminuria. Altogether, data indicate that microalbuminuria in the population reflects the presence of cardiovascular risk factors. Data on microalbuminuria and coronary heart disease support this idea. There is a continuous and graded relation between urinary albumin excretion and coronary heart disease prevalence. High urinary albumin excretion is likely a sign of vascular damage existing both at the renal and cardiac levels and induced by 1 or more uncontrolled cardiovascular risk factors.


CH-Bin: A convex hull based approach for binning metagenomic contigs.
Metagenomics has enabled culture-independent analysis of micro-organisms present in environmental samples. Metagenomics binning, which involves the grouping of contigs into bins that represent different taxonomic groups, is an important step of a typical metagenomic workflow followed after assembly. The majority of the metagenomic binning tools represent the composition and coverage information of contigs as feature vectors consisting of a large number of dimensions. However, these tools use traditional Euclidean distance or Manhattan distance metrics which become unreliable in the high dimensional space. We propose CH-Bin, a binning approach that leverages the benefits of using convex hull distance for binning contigs represented by high dimensional feature vectors. We demonstrate using experimental evidence on simulated and real datasets that the use of high dimensional feature vectors to represent contigs can preserve additional information, and result in improved binning results. We further demonstrate that the convex hull distance based binning approach can be effectively utilized in binning such high dimensional data. To the best of our knowledge, this is the first time that composition information from oligonucleotides of multiple sizes has been used in representing the composition information of contigs and a convex hull distance based binning algorithm has been used to bin metagenomic contigs. The source code of CH-Bin is available at https://github.com/kdsuneraavinash/CH-Bin.


Teacher Awareness and Attitudes Regarding Adolescent Risk Behaviours: a Sample of Finnish Middle and High School Teachers.
Empirical evidence has shown that youth gamble on both regulated and unregulated games, despite legislative prohibitions. This study assesses middle and high school teachers' awareness and attitudes regarding adolescent gambling and other potentially high-risk behaviours in Finland. A convenience sample of teachers (N = 157) from 13 provinces participated in the survey. The results suggest that teachers in Finland were more knowledgeable of the age limits of other adolescent high-risk behaviours than the legal age for gambling. Teachers were somewhat familiar with the behaviours and consequences associated with adolescent gambling. All other risk behaviours were perceived as being more important than gambling. Teachers' awareness about gambling prevention material in Finnish schools was limited. Results suggest that initiatives are required to enhance teachers' knowledge of adolescent problem gambling and its harmful short- and long-term consequences. School policies and guidelines including gambling behavior should be implemented in middle and high schools globally.


Human striatal glia differentially contribute to AD- and PD-specific neurodegeneration.
The commonalities and differences in cell-type-specific pathways that lead to Alzheimer disease (AD) and Parkinson disease (PD) remain unknown. Here, we performed a single-nucleus transcriptome comparison of control, AD and PD striata. We describe three astrocyte subpopulations shared across different brain regions and evolutionarily conserved between humans and mice. We reveal common features between AD and PD astrocytes and regional differences that contribute toward amyloid pathology and neurodegeneration. In contrast, we found that transcriptomic changes in microglia are largely unique to each disorder. Our analysis identified a population of activated microglia that shared molecular signatures with murine disease-associated microglia (DAM) as well as disease-associated and regional differences in microglia transcriptomic changes linking microglia to disease-specific amyloid pathology, tauopathy and neuronal death. Finally, we delineate undescribed subpopulations of medium spiny neurons (MSNs) in the striatum and provide neuronal transcriptomic profiles suggesting disease-specific changes and selective neuronal vulnerability.


The forms of vitamin B12 in foods.
1. The forms of vitamin B12 were determined in foods, most of which had been prepared for consumption. 2. Five forms were detected: adenosylcobalamin, hydroxocobalamin, methylcobalamin, cyanocobalamin and sulphitocobalamin. Adenoxylcobalamin and hydroxocobalamin were the predominant forms. 3. The intestinal absorption of [57Co]sulphitocobalamin was estimated and found to be lower than that of [58Co]cyanocobalamin.


Risk factors for pre-adolescent onset suicidal behavior in a high-risk sample of youth.
To identify risk factors for preadolescent onset suicidal behavior compared with adolescent/young adult onset suicidal behavior in a longitudinal sample of youth with parental history of mood disorders. The sample includes 545 youth who were age 21 years or less at the time of their baseline assessment. Participants underwent baseline and yearly study assessments. Observations were censored at the time point closest to the first episode of suicidal behavior for youth with suicidal behavior and at the time of last observation for youth without suicidal behavior. Youth were categorized into 3 groups: first onset of suicidal behavior before the age of 13 (n = 32), first onset of suicidal behavior between the ages of 13-21 (n = 51) and those without suicide related behaviors (n = 462). ANOVA, Chi-square, Fisher's exact test and multinomial regression were used to test the hypotheses. Significant predictors of preadolescent onset suicidal behavior were diagnosis of depressive disorder (RRR = 11.41, p<.001) and diagnosis of ADHD (RRR = 2.86, p = .02). Adolescent onset was predicted by diagnosis of depressive disorder (RRR = 4.12, p = .008), female sex (RRR = 2.68, p = .02) and self-reported suicidal ideation (RRR = 1.48, p = .004). These results are most applicable to offspring of parents with significant mood disorders. The strongest predictor of suicidal behavior in both groups was a diagnosis of depressive disorder, and the risk was nearly 3 times higher in preadolescents. ADHD was a significant predictor only for preadolescents, while female sex and self-reported suicidal ideation predicted suicidal behavior in adolescents.


The stomach-brain axis.
The stomach has distinct functions in relation to the ingestion and handling of solids and liquids. These functions include storage of the food before it is gradually emptied into the duodenum, mechanical crushing of larger food particles to increase the surface area, secretion of an acidic enzyme rich gastric juice and mixing the ingested food with the gastric juice. In addition, the stomach 'senses' the composition of the gastric content and this information is passed via the vagal nerve to the lateral hypothalamus and the limbic system, most likely as palatability signals that influence eating behaviour. Other sensory qualities related to the stimulation of gastric tension receptors are satiety and fullness. Receptors that respond to macronutrient content or gastric wall tension influence appetite and meal related hormone responses. The ingestion of food - in contrast to an infusion of nutrients into the stomach - has distinct effects on the activation of specific brain regions. Brain areas such as thalamus, amygdala, putamen and praecuneus are activated by the ingestion of food. Gastric nutrient infusion evokes greater activation in the hippocampus and anterior cingulate. The brain integrates these interrelated neural and hormonal signals arising from the stomach as well as visual, olfactory and anticipatory stimuli that ultimately influence eating and other behavioural patterns. Furthermore, there is now good evidence from experimental studies that gastric afferents influence mood, and animal studies point towards the possibility that gastric dysfunction may be a risk factor for mood disorders such as anxiety and depression. The stomach is also not only colonised by Helicobacter pylori but a large array of bacteria. While there is sufficient evidence to suggest that H. pylori may alter caloric intake and mood, the role of other gastric microbiome for the brain function is unknown. To address this appropriate targeted gastric microbiome studies would be required instead of widely utilised opportunistic stool microbiome studies. In summary, it is now well established that there are important links between the brain and the stomach that have significant effects on gastric function. However, the stomach also influences the brain. Disturbances in the crosstalk between the stomach and the brain may manifest as functional GI disorders while disturbances in the stomach-brain communication may also result in an altered regulation of satiety and as a consequence may affect eating behaviour and mood. These observations may enable the identification of novel therapies targeted at the gastroduodenum that positively alter brain function and treat or prevent conditions such as obesity or functional gastrointestinal disorders.


Robotic versus laparoscopic right colectomy for colon cancer: a systematic review and meta-analysis.
The aim of the study was to compare the short-term surgical outcomes of robotic right colectomy (RRC) with laparoscopic right colectomy (LRC) for colon cancer, to evaluate the safety and feasibility of the robotic surgery system. A systematic literature review was conducted using the PubMed, Web of Science, Embase, and Cochrane Library databases regarding the comparison of RRC vs. LRC for colon cancer in the last 5 years. Studies were included as per the PICOS criteria, and relevant event data were extracted. Fifteen studies (RRC: 1116 patients; LRC: 4036 patients) were evaluated. RRC demonstrated lower conversion to laparotomy (p = 0.03) and shorter length of hospital stay (p = 0.01), compared with LRC. However, operation times were longer in RRC than in LRC (p < 0.001). The estimated blood loss, retrieved lymph nodes, and overall postoperative complications were similar between RRC and LRC (p > 0.05). RRC can be regarded as a feasible and safe technique for colon cancer.


Protocol to purify the histone deacetylase SIRT6 and assess its activity in vitro.
The histone deacetylase known as sirtuin 6 (SIRT6) deacetylates both histone and non-histone proteins but has low deacetylase activity in vitro. Here, we present a protocol to monitor SIRT6-mediated deacetylation of long-chain acyl-CoA synthase 5 in the presence of palmitic acid. We describe the purification of His-SIRT6 and a Flag-tagged substrate. We then detail a deacetylation assay protocol that can be widely applied to study other SIRT6-mediated deacetylation events and the effect of SIRT6 mutations on its activity. For complete details on the use and execution of this protocol, please refer to Hou et al. (2022).1.


Chemically defined and xeno-free culture condition for human extended pluripotent stem cells.
Extended pluripotent stem (EPS) cells have shown great applicative potentials in generating synthetic embryos, directed differentiation and disease modeling. However, the lack of a xeno-free culture condition has significantly limited their applications. Here, we report a chemically defined and xeno-free culture system for culturing and deriving human EPS cells in vitro. Xeno-free human EPS cells can be long-term and genetically stably maintained in vitro, as well as preserve their embryonic and extraembryonic developmental potentials. Furthermore, the xeno-free culturing system also permits efficient derivation of human EPS cells from human fibroblast through reprogramming. Our study could have broad utility in future applications of human EPS cells in biomedicine.


One-compartmental biometric blood loss calculation after cesarean section.
Blood loss during cesarean section was calculated based on post-operative decrement of hemoglobin (Hb) and hematocrit (Hct) level. The model used for pregnant women was previously validated for non-pregnant women who underwent gynecological surgery. 1,068 pregnant women who underwent cesarean section and 517 women who underwent gynecological surgery were included in the study. Regression lines between collected (x) and calculated (y) blood loss in pregnant women (y = 0.164 + 0.602x) and non-pregnant (y = 0.255 + 0.750x) women were statistically parallel. This study confirmed the applicability of one-compartmental model based on the drop in Hb levels for blood loss calculations for both gynecological surgery and cesarean section. Improved methods for calculating blood loss, V after cesarean section as V = [0.0115 × body weight (kg) × ln (preoperative Hb/postoperative Hb)] + [0.1905 × height³ (m) × ln (preoperative Hb/postoperative Hb)] + 0.3158 and after gynecological surgery V = [0.004 × body weight (kg) × ln (preoperative Hb/postoperative Hb)] + [0.4624 × height ³(m) × ln (preoperative Hb/postoperative Hb)] + 0.0472 were suggested.


Is there a relationship between proximity to industry and the occurrence of otitis media with effusion in school entrant children?
In a study of the relationship between the prevalence of persistent otitis media with effusion (OME) in school entrant children in Redcar and Cleveland and the distance of the homes of these children from known industrial emission points, 1156 school entrant children were screened for the presence of persistent OME. The social disadvantage scores and map references were determined from the postcode area of each of the study entrants and map references were also obtained of known industrial emission points in the locality. Analyses were conducted on the association between the presence of OME and distance from emission sources and between the presence of OME and disadvantage score. A significantly greater proportion of study entrants with OME lived within 1000 meters of an industrial emission point than further away. However, there was no trend established between the proportion of study entrants with OME and increasing distance from an emissions source, nor was there any significant relationship established between the social disadvantage score of the areas of residence of the study entrants and the presence of OME. Further research is required to establish the effect of confounding variables on this relationship.


Global transcriptome analysis of allopolyploidization reveals large-scale repression of the D-subgenome in synthetic hexaploid wheat.
Synthetic hexaploid wheat (SHW) lines are created as pre-breeding germplasm to diversify the D subgenome of hexaploid wheat and capitalize upon the untapped genetic diversity of the Aegilops tauschii gene pool. However, the phenotypes observed in the Ae. tauschii parents are not always recovered in the SHW lines, possibly due to inter-subgenome interactions. To elucidate this post-polyploidization genome reprogramming phenomenon, we performed RNA-seq of four SHW lines and their corresponding tetraploid and diploid parents, across ten tissues and three biological replicates. Homoeologue expression bias (HEB) analysis using more than 18,000 triads suggests massive suppression of homoeoalleles of the D subgenome in SHWs. Comparative transcriptome analysis of the whole-genome gene set further corroborated this finding. Alternative splicing analysis of the high-confidence genes indicates an additional layer of complexity where all five splice events are identified, and retained intron is predominant. Homoeologue expression upon resynthesis of hexaploid wheat has implications to the usage and handling of this germplasm in breeding as it relates to capturing the effects of epistatic interaction across subgenomes upon polyploidization. Special considerations must be given to this germplasm in pre-breeding activities to consider the extent of the inter-subgenome interactions on gene expression and their impact on traits for crop improvement.


Status of IL-4 and IL-10 driven markers in experimental models of Visceral Leishmaniasis.
Leishmania donovani, the causative agent for visceral leishmaniasis (VL), modulates host monocytes/macrophages to ensure its survival. However, knowledge regarding the host-parasite interactions underpinning the disease remains limited. As disease progression is associated with polarization of monocytes/macrophages towards M2, which is regulated by cytokines IL-4/IL-13 and IL-10, this study evaluated the status of key IL-4- and IL-10 driven markers in experimental models of VL, as also evaluated their correlation, if any, with parasite load. In liver and splenic tissues from L donovani-infected hamsters and BALB/c mice, the parasite burden was determined along with mRNA expression of IL-4-driven markers, that is CD206, Arginase-I, CCL17, CCL22, PPAR-γ, STAT6, KLF4, FIZZ1 and YM1 along with IL-10-driven markers, CXCL13, IL-10, TGF-β, VDR, CCR2 and CYP27A1. The mRNA expression of IL-4- and IL-10-driven markers was enhanced in both models, but only in the hamster model, the splenic tissues demonstrated a positive correlation between all the IL-10-driven markers and parasite load. Contrary to human VL, both models demonstrated an increased expression of IL-4- and IL-10-driven markers.


Succinylation Modified Ovalbumin: Structural, Interfacial, and Functional Properties.
In this study, ovalbumin (OVA) was succinylated with the addition of different levels of succinic anhydride, and the structural and functional properties of succinylated OVA (SOVA) were investigated. SDS-PAGE and FTIR spectrum confirmed the covalent attachment of the succinyl group to OVA. Thermal stability and the absolute value of zeta potential (pH 6.0) of SOVA were enhanced by 14.90% and 76.77% higher than that of the native OVA (NOVA), respectively. Circular dichroism (CD) spectra demonstrated that the succinylation decreased the α-helix and increased β-sheet content to 21.31% and 43.28%, respectively. The content of free sulfhydryl groups increased and intrinsic fluorescence spectra suggested the SOVA became more unfolded and flexible as the degree of succinylation enhanced. Furthermore, succinylation effectively enhanced the solubility and decreased the interface tension (oil-water and air-water interface) of OVA. Compared to NOVA, the emulsifying activity and stability of SOVA were increased by 1.6 times and 1.2 times, respectively, and foaming capacity and stability were enhanced by 2.7 times and 1.5 times, respectively.


Informing disease modelling with brain-relevant functional genomic annotations.
The past decade has seen a surge in the number of disease/trait-associated variants, largely because of the union of studies to share genetic data and the availability of electronic health records from large cohorts for research use. Variant discovery for neurological and neuropsychiatric genome-wide association studies, including schizophrenia, Parkinson's disease and Alzheimer's disease, has greatly benefitted; however, the translation of these genetic association results to interpretable biological mechanisms and models is lagging. Interpreting disease-associated variants requires knowledge of gene regulatory mechanisms and computational tools that permit integration of this knowledge with genome-wide association study results. Here, we summarize key conceptual advances in the generation of brain-relevant functional genomic annotations and amongst tools that allow integration of these annotations with association summary statistics, which together provide a new and exciting opportunity to identify disease-relevant genes, pathways and cell types in silico. We discuss the opportunities and challenges associated with these developments and conclude with our perspective on future advances in annotation generation, tool development and the union of the two.


Dorsal raphe nucleus and locus coeruleus neural networks and the elaboration of the sweet-substance-induced antinociception.
In order to investigate the effects of monoaminergic neurons of the dorsal raphe nucleus (DRN) and locus coeruleus (LC) on the elaboration and control of sweet-substance-induced antinociception, male albino Wistar rats weighing 180-200 g received sucrose solution (250 g/L) for 7-14 days as their only source of liquid. After the chronic consumption of sucrose solution, each animal was pretreated with unilateral microinjection of ibotenic acid (1.0 microg/0.2 microL) in the DRN or in the LC. The tail withdrawal latencies of the rats in the tail-flick test were measured immediately before and 7 days after this treatment. The neurochemical lesion of locus coeruleus, but not of DRN neural networks with ibotenic acid, after the chronic intake of sweetened solution, decreased the sweet-substance-induced antinociception. These results indicate the involvement of noradrenaline-containing neurons of the LC in the sucrose-induced antinociception. We also consider the possibility of DRN serotonergic neurons exerting some inhibitory effect on the LC neural networks involved with the elaboration of the sweet-substance-induced antinociception.


Fragmentation landscape of cell-free DNA revealed by deconvolutional analysis of end motifs.
Cell-free DNA (cfDNA) fragmentation is nonrandom, at least partially mediated by various DNA nucleases, forming characteristic cfDNA end motifs. However, there is a paucity of tools for deciphering the relative contributions of cfDNA cleavage patterns related to underlying fragmentation factors. In this study, through non-negative matrix factorization algorithm, we used 256 5' 4-mer end motifs to identify distinct types of cfDNA cleavage patterns, referred to as "founder" end-motif profiles (F-profiles). F-profiles were associated with different DNA nucleases based on whether such patterns were disrupted in nuclease-knockout mouse models. Contributions of individual F-profiles in a cfDNA sample could be determined by deconvolutional analysis. We analyzed 93 murine cfDNA samples of different nuclease-deficient mice and identified six types of F-profiles. F-profiles I, II, and III were linked to deoxyribonuclease 1 like 3 (DNASE1L3), deoxyribonuclease 1 (DNASE1), and DNA fragmentation factor subunit beta (DFFB), respectively. We revealed that 42.9% of plasma cfDNA molecules were attributed to DNASE1L3-mediated fragmentation, whereas 43.4% of urinary cfDNA molecules involved DNASE1-mediated fragmentation. We further demonstrated that the relative contributions of F-profiles were useful to inform pathological states, such as autoimmune disorders and cancer. Among the six F-profiles, the use of F-profile I could inform the human patients with systemic lupus erythematosus. F-profile VI could be used to detect individuals with hepatocellular carcinoma, with an area under the receiver operating characteristic curve of 0.97. F-profile VI was more prominent in patients with nasopharyngeal carcinoma undergoing chemoradiotherapy. We proposed that this profile might be related to oxidative stress.


Effects of long-term finasteride treatment on prostate cancer morphology and clinical outcome.
To examine the morphologic alterations of finasteride therapy on prostate cancer compared with no treatment or treatment with a luteinizing hormone-releasing hormone (LHRH) agonist and the clinical outcomes of patients treated with finasteride. The data of 56 patients with prostate cancer who had taken finasteride for at least 6 months were reviewed. A central pathology review was performed in a blinded manner comparing these patients with 56 matched controls and 44 patients who received a 3-month course of LHRH agonist before radical prostatectomy. The histologic hormonal treatment effects included apoptosis, vacuolated cytoplasm, pyknotic nuclei, and small irregular glands. An overall consensus was scored using a three-tiered system: no apparent effect (score = 0), suspicious for hormonal effect (score = 1), and highly suggestive of hormonal treatment (score = 2). A Gleason score was assigned to all prostatectomy specimens. The mean hormonal treatment score for the prostatectomy specimens was 0.4, 0.5, and 1.6 for the control, finasteride, and 3-month LHRH groups, respectively. Surprisingly, 20% of control patients had suspicious or highly suggestive hormonal effects and 26% of the finasteride-treated patients did so. For the LHRH-treated group, only 23% did not demonstrate classic features of hormonal treatment. Similar clinical outcomes were observed between the finasteride and control groups. No consistent hormonal therapy effects with finasteride treatment were observed compared with LHRH agonists. The hormonal effect was observed in the control group. Therefore, although other aspects of the Prostate Cancer Prevention Trial design might account for the greater percentage of Gleason grade tumors in the study arm, morphologic changes due to long-term finasteride treatment were not a likely cause.


Bandwidth of quantized surface plasmons: competition between radiative and nonradiative damping effects.
We investigate the damping effects of coherent electron oscillations on the bandwidth of a quantized nanoparticle plasmon resonance. The nanoparticle (NP) is treated as a two-level quantum system, and the total relaxation time involves both the population relaxation time associated with radiative processes and the collisional relaxation time associated with nonradiative processes that result in dephasing/decoherence of electron oscillations. We describe the optical response of NPs to an external electromagnetic field by the optical Bloch equations employing the density matrix formalism to capture the quantum description nature of dipolar plasmon resonance and suggest a generalized criterion for the validity of dipole approximation. Then we explore the competition between the radiative and nonradiative damping in determining the plasmon bandwidth of two typical NP models; metallic nanospheres and dielectric core-metal shell NPs (nanoshells). We show that the frequency of plasmon resonance, in addition to the NP size, plays an important role in the competition between the damping mechanisms. Consequently, the damping processes are significantly influenced by the factors that determine the resonance frequency, such as the core size, the dielectric constant of the medium, and the shell thickness (for nanoshells). For both models of NPs, we identify the optimum parameters that achieve a narrower plasmon bandwidth (minimal damping), which is a prerequisite for advanced sensing and medical applications. We demonstrate excellent agreement of the simulated spectral features of the plasmon resonance with previously reported experimental results for a single NP where the inhomogeneous broadening of the plasmon line is excluded. For NP ensembles where inhomogeneous broadenings and interface chemical effects are significant, our theoretical approach successfully predicts the overall trend of size-dependent damping rates.


Postpolymerization of a 3D-printed denture base polymer: Impact of post-curing methods on surface characteristics, flexural strength, and cytotoxicity.
This study investigated the influence of postpolymerization of a three-dimensional (3D) printed denture base polymer. The effect of post-curing methods on surface characteristics, flexural strength, and cytotoxicity was evaluated. A total of 172 specimens were additively manufactured using one denture base material (V-Print dentbase, VOCO) and further post-cured by different light-curing devices, including Otoflash G171 (OF), Labolight DUO (LL), PCU LED (PCU), and LC-3DPrintbox (PB), respectively. Polymethyl methacrylate resin (PalaExpress Ultra) was used as a reference (REF). Afterward, surface topography was observed using scanning electron microscopy, and surface roughness was measured (n = 6). Furthermore, flexural strength was tested (n = 20). Cytotoxicity was evaluated by the extract and direct contact tests. The data were analyzed using the Kolmogorov-Smirnov test and one-way ANOVA followed by Tukey's multiple comparisons and Kruskal-Wallis tests (p < 0.05). The different post-curing methods applied did not significantly influence surface topography and roughness (Ra). Meanwhile, specimens post-cured by PCU (162.3 ± 44.16 MPa) and PB (171.2 ± 34.41 MPa) showed significantly higher flexural strength than those post-cured by OF (131.3 ± 32.87 MPa) and REF (131.2 ± 19.19 MPa), respectively. Additionally, various post-curing methods effectively decreased the cytotoxic effects of 3D-printed denture base polymer. Different post-curing methods did not significantly alter the Ra values of the 3D-printed denture base material. However, flexural strength was significantly affected by the postpolymerization methods, which might be attributed to the different wavelengths of post-curing devices. In addition, various postpolymerization methods reduced the cytotoxic effects of the 3D-printed denture base polymer. Flexural strength of additively manufactured denture bases depends on the postpolymerization strategy. Therefore, an appropriate post-curing method is required to optimize the flexural strength of 3D-printed denture materials.


Reproducibility of the Wisconsin cataract grading system in the Blue Mountains Eye Study.
To measure the inter- and intra-grader reliability of the Wisconsin cataract grading system for grading of nuclear, cortical, and posterior subcapsular cataract (PSC) in the Blue Mountains Eye Study. Random samples of lens photographs from study subjects were regraded by each grader according to the Wisconsin cataract grading system. The results were compared using quadratic weighted kappa statistics to derive measures of inter- and intra-grader reliability for the grading of nuclear (5 grades), PSC (3 grades), and cortical cataract (3 grades). Inter-grader reliability (quadratic weighted kappa) for grading of nuclear cataract varied between 0.82 and 0.79; kappa was 0.78 for cortical cataract and 0.57 for PSC. Intra-grader reliability was slightly higher than inter-grader reliability for all three cataract types. In the Blue Mountains Eye Study, the Wisconsin cataract grading system provided good reproducibility, comparable to that reported from the Beaver Dam Eye Study, endorsing its usefulness in cataract epidemiology.


Tight hormonal phenotypic integration ensures honesty of the electric signal of male and female Brachyhypopomus gauderio.
Hormones mediate sexually selected traits including advertisement signals. Hormonal co-regulation links the signal to other hormonally-mediated traits such that the tighter the integration, the more reliable the signal is as a predictor of those other traits. Androgen administration increases the duration of the communication signal pulse in both sexes of the electric fish Brachyhypopomus gauderio. To determine whether the duration of the signal pulse could function as an honest indicator of androgen levels and other androgen-mediated traits, we measured the variation in sex steroids, signal pulse duration, and sexual development throughout the breeding season of B. gauderio in marshes in Uruguay. Although the sexes had different hormone titres and signal characteristics, in both sexes circulating levels of the androgens testosterone (T) and 11-ketotestosterone (11-KT) were strongly related to signal pulse duration. Consequently, signal pulse duration can serve as an honest indicator of circulating androgens in males and females alike. Additionally, through phenotypic integration, signal pulse duration also predicts other sexual traits directly related to androgen production: gonad size in males and estradiol (E2) levels in females. Our findings show that tight hormonal phenotypic integration between advertisement signal and other sex steroid-mediated traits renders the advertisement signal an honest indicator of a suite of reproductive traits.


Retrocecal hernia: A case report.
Retroperitoneal Retrocecal hernias are a rare variety of internal hernias and represent an unusual cause of bowel obstruction. Early diagnosis is based on CT scan and requires knowledge of the pathology in order to avoid small bowel resection. We report a case of retrocecal hernia treated surgically and review the characteristics and treatment of retrocecal hernias in the literature. Our work is a retrospective case report with a descriptive aim concerning a patient operated for retrocecal hernia within the department of general surgery of CHU Ibn Rochd Casablanca. A 72-year-old man presented to the emergency department with abdominal pain and vomiting that have been evolving for 9 days complicated by an occlusive syndrome 36 hours before the admission. The patient was apyretic, and the abdominal examination noted abdominal meteorism predominantly in the right iliac fossa, absence of abdominal scarring, and free hernial orifices. The abdominal X-ray showed air-fluid levels and the abdominopelvic CT scan found clumping of the dilated small intestines posteriorly and below the cecum. The diagnosis of retrocecal hernia was suspected and the patient was taken to the operating room. The operation was performed by laparotomy through a midline incision. On exploration, the cecum and ascending colon were pushed forward and viable bowel loops were incarcerated in a fossa located posteriorly and below the cecum. The procedure consisted of a collapse of the retrocecal ligaments by right coloparietal collapse. A bowel obstruction in an apyretic patient without abdominal scarring or parietal hernia should suggest the diagnosis of internal hernia, which must be investigated.


RASAL2 promotes lung cancer metastasis through epithelial-mesenchymal transition.
It was reported that genetically-engineered RASAL2 knockout mice are prone to development of several sporadic tumor, including lung adenocarcinoma. However, a causative relationship between RASAL2 deficiency and lung adenocarcinoma development still remains unknown. In the present study, RASAL2 level was determined in patients with lung adenocarcinoma and control subjects in an attempt to explore its potential clinical diagnostic and prognostic value. Low RASAL2 expression levels were found in 71% (37 of 52) of lung adenocarcinoma, which were correlated with lymph node metastasis in lung adenocarcinoma. Moreover, Low RASAL2 expression levels were correlated with reduced overall survival (OS) in lung adenocarcinoma. We find that inactivation of RASAL2 promotes lung cancer cell migration through the induction of epithelial mesenchymal transition (EMT) and promoted lung metastasis in nude mice. Our results suggest that the down-regulation of RASAL2 promotes metastatic progression of lung adenocarcinoma, hence it could serve as a potential target for the development of lung cancer therapies.


Evaluation of qPCR reference genes in GH-overexpressing transgenic zebrafish (Danio rerio).
Reference genes (RGs) must have a stable expression in tissues in all experimental conditions to normalize real-time quantitative reverse transcription PCR (qRT-PCR) data. F0104 is a highly studied lineage of zebrafish developed to overexpress the growth hormone (GH). It is assumed that the transgenic process may influence the expression levels of commonly used RGs. The objective of the present study was to make a comprehensive analysis of stability of canditade RGs actb1, actb2, b2m, eif2s2, eef1a1, gapdh, rplp2, rpl7, rpl13α, tuba1, and rps18, in gh-transgenic and non-transgenic zebrafish. Liver, brain, intestine and muscle samples from both groups had qRT-PCR results analyzed by dCt, geNorm, NormFinder, BestKeeper, and RefFinder softwares. Consensus analyses among software concluded that rpl13α, rpl7, and eef1a1 are the most stable genes for zebrafish, considering the studied groups and tissues. Gapdh, rps18, and tuba1 suffered variations in stability among different tissues of both groups, and so, they were listed as the genes with lowest stability. Results from an average pairwise variations test indicated that the use of two RGs would generate reliable results for gene expression analysis in the studied tissues. We conclude that genes that are commonly used in mammals for qRT-PCR assays have low stability in both non-transgenic and gh-transgenic zebrafish reinforcing the importance of using species-specific RGs.


Genetic polymorphisms of estrogen receptors in patients with premature coronary artery disease.
Estrogen protects against atherosclerosis through its genomic/nongenomic effects. Estrogen receptors (ESR) alpha (1) and beta (2) mediate much estrogen action. Both receptors exist in the arterial wall, but the extent of their distribution in arterial layers is unknown. Allelic variants of the gene encoding ESR1 and ESR2 may alter their expression and function, resulting in genetic variability. In the present age-adjusted, case-control study, the prevalence of four mutations in estrogen receptors was analyzed in patients with premature CAD and controls. Mutation in the ESR1 (PvuII) was more prevalent in the controls (18 vs. 11%; p=0.062) than in CAD patients, and the mutation identified by the XbaI enzyme in the same receptor was associated with reduced apolipoprotein B levels and low body mass index. Mutation of the ESR2 (AluI) was more prevalent in CAD patients (0.6 vs. 18%; p=0.008). Homozygosis for this mutation involved increased body mass index, elevated serum triglycerides and apolipoprotein B, and reduced HDL-cholesterol. Multivariate logistic regression analysis showed dyslipidemia, low serum HDL levels, and ESR2 polymorphism (AluI) [OR=1.89 (95% CI: 1.08-3.34); p=0.034] to be independent risk factors for CAD. Our data suggest that mutation of the ESR2 is an independent risk marker for premature CAD.


The relationship between the fish consumption and blood total/methyl-mercury concentration of costal area in Korea.
The purpose of this study was to evaluate the relationship between fish consumption and blood THg/MeHg concentration in Korean adults by measuring MeHg concentration in blood directly. The study subjects consisted of 400 adults aged 20 or older from 30 subareas in Busan, Ulsan and Gyeongsangnam-do province in Korea from August to October, 2010. We tried to recruit the same number of male and female participants in different age groups (20s, 30s, 40s, 50s and 60s) and allocated 13-16 subjects by district to represent Hg concentration in the research areas. The geometric means of THg and MeHg concentration in blood were 5.27 μg/L (5.00-5.57) and 4.05 μg/L (3.81-4.32), respectively. The proportion of MeHg/THg concentration was 78.53% (77.09-79.97). MeHg concentration was higher in coastal areas (4.26 μg/L) than in inland areas (3.52 μg/L) and was higher in men (4.68 μg/L) than in women (3.52 μg/L). In male participants, blood MeHg concentration increased with increasing annual fish consumption, and the proportion of MeHg/THg concentration showed an upward trend as THg concentration increased. However, none of the measures of the proportion of MeHg/THg showed significant differences. This is the first report in Korea about the relationship between blood MeHg concentration and related factors. Our findings suggest that MeHg concentration is affected by fish consumption as well as by gender difference and drinking status. Since the pathological mechanism has not been clarified, additional studies are needed for explaining the biological and lifestyle differences in the risk of adverse health effects by Hg exposure.


Assessing scrub typhus and rickettsioses transmission risks in the Chiang Rai province of northern Thailand.
Scrub typhus is an important disease in the Asia-Pacific countries with increasing relevance for public health worldwide. Entomological risk assessment for scrub typhus and rickettsial disease in Phu Chi Fah village-Chiang Rai was performed to determine areas at greatest risk for disease transmission in order to increase awareness of diseases to travelers and healthcare workers. From 2016 to 2018, rodents and chiggers were collected from 7 sites covering residential, grassland, and forest areas to determine the prevalence of pathogen of interest. The correlation between land type and vector-host-pathogen interaction system was investigated. High prevalence of O. tsutsugamushi-infected and Rickettsia-infected chiggers was observed especially in areas with grassland and forest ecotones. Chigger and rodent species composition were negatively associated with the level of human disturbance. Increased density of rodents was responsible for a higher abundance of chigger population and increased prevalence of O. tsutsugmaushi infection in chigger in the following year. Communities in the study areas have an increased exposure risk to scrub typhus and potentially spotted fever group rickettsiosis. Travellers to this endemic area should pay more attention pathogen risks so as to avoid vector and disease exposure. Seasonal rodenticidal activity may help migitate the risk of pathogen transmission.


Unconventional protein secretion (UPS) pathways in plants.
As in yeast and mammalian cells, novel unconventional protein secretion (UPS) or unconventional membrane trafficking pathways are now known to operate in plants. UPS in plants is generally associated with stress conditions such as pathogen attack, but little is known about its underlying mechanism and function. Here, we present an update on the current knowledge of UPS in the plants in terms of its transport pathways, possible functions and its relationship to autophagy.


The value of transvaginal ultrasonography in the endometrial evaluation of breast cancer patients using tamoxifen.
To estimate the value of transvaginal ultrasonography (TVU) in evaluating the endometrium in breast cancer patients on tamoxifen, and to investigate the relationship between TVU and histologic endometrial findings in these patients. 107 breast cancer patients taking tamoxifen were included in this study. TVU was performed twice for each patient: prior to starting tamoxifen therapy and 1 year after taking tamoxifen. An endometrial thickness of >or=5 mm was considered abnormal. Endometrial biopsy was performed at the same time the 2nd TVU was done. The patients were divided into 2 groups: symptomatic (group A) and asymptomatic (group B), based on whether or not there was abnormal vaginal bleeding. 31 patients (29%) were symptomatic, while the remaining 76 (71%) were asymptomatic. The endometrial thickness increased after 1 year of taking tamoxifen from 4.84+/-0.4 mm to 6.34+/-2.1 mm in group A and from 4.73+/-0.3 mm to 5.67+/-1.95 mm in group B (p>0.05). Endometrial biopsy revealed 77 (71.96%) atrophic endometria and 21 (19.62%) polyps. A comparison between the 2 groups showed a significant difference in patients with endometrial atrophy and atypical hyperplasia. Patients who had an endometrial thickness of >5 mm had a significantly higher prevalence of atypical hyperplasia (p=0.003) and polyps (p=0.041). The sensitivity, specificity, positive predictive and negative predictive values of TVU were 63.3, 28.57, 25.67 and 66.66%, respectively. Our study showed a discrepancy between TVU and endometrial biopsy findings, due to the specific histology of the endometrium in breast cancer patients using tamoxifen. Due to this discrepancy, TVU alone is not an effective screening test for endometrial pathology and its application alone might lead to an undesirably high frequency of invasive diagnostic procedures.


Recruitment of general practitioners in China: a scoping review of strategies and challenges.
China is rapidly expanding its general practitioner (GP) workforce as part of recent healthcare reform, with an extra 400,000 GPs by 2030. This scoping review identifies the published strategies for GP recruitment that are being implemented and the challenges encountered. We searched six English and three Chinese databases from 2015 to April 2022, following Arksey and O'Malley's framework and the PRISMA ScR reporting guidelines. A total of 40 Chinese-language and 5 English-language records were included. We identified multiple policies, pathways and programmes focused on expanding GP recruitment. Twenty-two evaluations of these initiatives show varying degrees of effectiveness. Selecting general practice as a career is affected by many factors, including individual's background, remuneration and benefits, career prospects, working environment, self-fulfilment, and current national developments and reorganisations of primary care. The challenge most frequently reported was the adequate provision of qualified GP in rural regions. The targeting of students from rural areas and provision of free education in return for an obligatory six-years' working in their hometown upon graduation appears to be effective. Extracted records mostly studied views of trainees in a defined locality, and we identified a paucity of studies which explored the perspectives of organisations and institutions, similarly there were areas of China not contributing to the literature and there were no records taking a national perspective. Long-term monitoring is required to assess policy changes and to systematically evaluate the effectiveness of the interventions nationally. The monitoring of the challenges influencing GP recruitment can be used to inform the design of future initiatives. Development of a minimum agreed standardised set of outcomes used to measure and report evaluations will help assess the relative contributions and cost effectiveness of different approaches being used to boost GP numbers. We provide suggestions for improving the benefits and rewards for GPs and how to promote recruitment to the more rural or less attractive areas.


Surgical repair of a tibialis anterior tendon rupture : a case report.
A 61-year-old man who stretched in the morning presented a rupture of the tibialis anterior tendon treated by extensor hallucis longus transfer. Rupture of the tibialis anterior tendon is rare. Surgical treatment seems to be more efficient in improving the function.


Incidence and genetic variability of Listeria monocytogenes isolated from vegetables in Poland.
The aim of the present study is to investigate the prevalence and genetic diversity of Listeria monocytogenes in various fresh and frozen vegetable products available in Poland. The samples were collected at retail market within the framework of national official control and monitoring program. In the years 2016-2019 a total of 49 samples out of 8712 collected vegetable samples were positive for L. monocytogenes. Our findings demonstrated that the occurrence of L. monocytogenes in various vegetable products was generally low, on average only 0.56% in the studied years. All isolates were susceptible to 11 antimicrobial agents: penicillin, ampicillin, meropenem, erythromycin, sulfamethoxazole-trimethoprim, amoxicillin-clavulanic acid, ciprofloxacin, chloramphenicol, gentamicin, vancomycin, and tetracycline. All of them harbored virulence-associated genes (inlA, inlC, and lmo2672), 82% harbored inlJ gene and few of them (22%) also possessed the llsX gene. The majority of collected isolates (65%) belonged to molecular serogroup 1/2a-3a, followed by 4ab-4b-4d-4e (33%), and only one to serogroup 1/2b-3b-7 (2%). Isolates yielded 18 different restriction profiles, revealing a large cluster of contamination linked to frozen corn (21 strains) and distributed in 3 pulsotypes. MLST analysis classified selected isolates into nine clonal complexes (CCs). The obtained results contribute to characterizing the diversity of L. monocytogenes isolated from various vegetable products in Poland and their impact on food safety and public health.


Splenic artery aneurysms in pregnancy--a systematic review.
Visceral artery aneurysm is an uncommon pathology, with a potential for rupture. Splenic artery aneurysms (SAA) are most commonly (60%) associated with a high mortality rate of 25% in case of aneurysm rupture. This increases disproportionately to 75% among pregnant women with fetal mortality of 95%. Although this is a rare event, because of the associated catastrophic consequences, prompt management of splenic artery aneurysms (SAA) is of prime importance. This systematic review provides up-to-date information about the management of splenic artery aneurysms in pregnancy.


Sex differences in CSF biomarkers for neurodegeneration and blood-brain barrier integrity.
As cerebrospinal fluid (CSF) neurofilament light protein (NfL) and the CSF/serum albumin ratio (QAlb) are used in the clinical routine, the impact of demographic factors on these biomarkers is important to understand. Participants were derived from two Swedish samples: the population-based H70 Study (n = 308, age 70) and a clinical routine cohort (CSF NfL, n = 8995, QAlb, n = 39252, age 0 to 95). In the population-based study, QAlb and NfL were examined in relation to sex, cardiovascular risk factors, and cerebral white matter lesions (WMLs). In the clinical cohort, QAlb and NfL sex differences were tested in relation to age. Men had higher QAlb and NfL concentrations and had higher QAlb and NfL concentrations from adolescence throughout life. NfL was not related to WML, but QAlb correlated positively with WMLs. The CSF NfL sex difference could not be explained by vascular pathology. Future studies should consider using different reference limits for men and women.


Roles of exosomes as drug delivery systems in cancer immunotherapy: a mini-review.
Exosomes can be released by a variety of cells and participate in intercellular communication in many physiological processes in the body. They can be used as carriers of cancer therapeutic drugs and have natural delivery capabilities. Some biologically active substances on exosomes, such as major histocompatibility complex (MHC), have been shown to be involved in exosome-mediated anticancer immune responses and have important regulatory effects on the immune system. Exosome-based drug delivery systems hold great promise in future cancer immunotherapy. However, there are still substantial challenges to be overcome in the clinical application of exosomes as drug carriers. This article reviews the biological characteristics of exosome drug delivery systems and their potential applications and challenges in cancer immunotherapy.


Stimulation of vascular prostacyclin synthesis by extracellular ADP and ATP.
Extracellular ADP and ATP stimulated the synthesis of prostacyclin - as reflected by the release of 6-keto-PGF1 alpha - in the rabbit aorta, the rabbit pulmonary artery and the rat aorta. A doubling of 6-keto-PGF1 alpha output was produced by 3 microM ADP. Adenosine had no effect and the stimulation by ADP was blocked by quinidine, but not by theophylline. This stimulation was abolished by indomethacin and lost after mechanical removal of the endothelium. Stimulation of vascular prostacyclin synthesis by ADP released from aggregating platelets could help localize thrombus formation to areas of vascular damage.


Ultrasonics as a tool for development of pine-needle extract loaded bee wax edible packaging for value addition of Himalayan cheese.
In the present study, Himalayan cheese, kradi was coated with beeswax loaded with pine needle extract (PNE) to increase its shelf life and nutraceutical potential. PNE was extracted via ultrasonication and incorporated into beeswax at concentrations, 2:1, 1:1, and 2:3 (grams of beeswax to mL of PNE). The dispersion of PNE in the coatings was carried out using an ultrasonic probe at a frequency of 20 kHz for 15 min and at power rating of 500 W. The coatings were characterised using scanning electron microscopy, light microscopy, dynamic light scattering (DLS), fourier transmission infrared spectroscopy. DLS revealed a hydrodynamic diameter and zeta potential of 12.11 ± 0.41 µm and -19.32 ± 0.61 mV for coating loaded with highest concentration of PNE. The bioactivities of the coating including antioxidant, antidiabetic and antibacterial assays revealed significantly higher values with the increase in PNE concentration. Shelf life and sensory evaluation study including microbiological and sensory analysis revealed inhibition of mould growth and good score of texture and appearance with the increase in concentration of PNE. The study provides a future perspective for application of beeswax loaded PNE coatings in cheese industry.


Cold atmospheric plasma coupled with air abrasion in liquid medium for the treatment of peri-implantitis model grown with a complex human biofilm: an in vitro study.
Treatment of implants with peri-implantitis is often unsuccessful due to residual microbial biofilm hindering re-osseointegration. The aim of this study was to treat biofilm-grown titanium (Ti) implants with different modalities involving air abrasion (AA) and cold atmospheric plasma (CAP) to compare the effectiveness in surface decontamination and the alteration/preservation of surface topography. Saliva collected from a peri-implantitis patient was used to in vitro develop human biofilm over 35 implants with moderately rough surface. The implants were then mounted onto standardized acrylic blocks simulating peri-implantitis defects and treated with AA (erythritol powder), CAP in a liquid medium, or a combination (COM) of both modalities. The remaining biofilm was measured by crystal violet (CV). Surface features and roughness before and after treatment were assessed by scanning electron microscope (SEM). The data were statistically analyzed using Kruskal-Wallis followed by Tukey's multiple comparison test. In the present peri-implantitis model, the human complex biofilm growth was successful as indicated by the statistical significance between the negative and positive controls. All the treatment groups resulted in a remarkable implant surface decontamination, with values very close to the negative control for AA and COM. Indeed, statistically significant differences in the comparison between the positive control vs. all the treatment groups were found. SEM analysis showed no post-treatment alterations on the implant surface in all the groups. Decontamination with AA delivering erythritol with or without CAP in liquid medium demonstrated compelling efficacy in the removal of biofilm from implants. All the tested treatments did not cause qualitative alterations to the Ti surface features. No specific effects of the CAP were observed, although further studies are necessary to assess its potential as monotherapy with different settings or in combination with other decontamination procedures. CAP is a promising option in the treatment of peri-implantitis because it has potential to improve the elimination of bacterial plaque from implant surfaces, in inaccessible pockets or during open-flap debridement, and should stimulate the process of the re-osseointegration of affected dental implants by not altering surface features and roughness.


A quantitative study of the effect of topical indomethacin on cutaneous erythema induced by UVB and UVC radiation.
The effect of indomethacin gel on UVB and UVC erythema was assessed objectively using a reflectance instrument. When indomethacin was applied immediately after irradiation, UVR (ultraviolet radiation) dose-dependent suppression of erythema was demonstrated for both wavelengths until 36 h after irradiation when both indomethacin and control gel base-treated sites were equally erythematous. Suppression of erythema also occurred when application of indomethacin was delayed until 24 h after irradiation, showing that for both wavelengths prostaglandin synthesis remains increased throughout this period. The degree of suppression at any time, however, was no greater than that achieved by a single application immediately after irradiation, indicating that the eventual equal erythema of indomethacin and gel base-treated sites was not due to tachyphylaxis or inadequate dosage. Construction of dose-response curves for the indomethacin-responsive and indomethacin-unresponsive components of erythema shows that in human skin the difference in erythemal response to UVB and UVC radiation is not due to the formation of different mediators at these wavelengths.


Evaluation of the cutaneous wound healing potential of tamanu oil in wounds induced in rats.
Tamanu is a plant oil derived from the fruit and seeds of the Calophyllum inophyllum tree. Although scientific data on tamanu oil are limited, it is recommended worldwide for the treatment of abrasions, burns, diabetic wounds and scars. This study aimed to compare the wound healing efficacy of the topical use of tamanu oil with a reference drug in rats. Uniform wounds were induced on the dorsum of 21 rats, randomly divided into three groups. The control group received normal saline; the tamanu group received tamanu oil; and the centella group was treated with Centella asiatica. Wound healing was clinically evaluated using wound healing scoring and wound contraction. A biopsy was taken from the wound sites of each rat on days 7, 14 and 21 for histopathological evaluation. Wound contraction was significantly lower in the tamanu group compared with the other groups. On day 7, the intensity of macrophage infiltration and mature granulation tissues were significantly higher in the centella and tamanu groups than in the control group. Fibrosis and collagen density were higher in the tamanu group than the other groups on day 7. In wound healing in rats, tamanu oil accelerated the formation of macrophage-granulation tissues-fibrosis and resulted in less wound contraction.


Radix Salviae miltiorrhizae improves bone microstructure and strength through Wnt/β-catenin and osteoprotegerin/receptor activator for nuclear factor-κB ligand/cathepsin K signaling in ovariectomized rats.
Although radix Salviae miltiorrhizae (RSM) is reported to exhibit the antiosteoporotic effect in preclinical study, the underlying mechanism is unclear. To this end, ovariectomized (OVX) rats were employed with administration of RSM (5 g/kg) for 14 weeks. The disturbed serum levels of alkaline phosphatase (ALP), osteoprotegerin (OPG), tartrate-resistant acid phosphatase, and receptor activator of nuclear factor-κB ligand (RANKL) in OVX rats were improved by RSM treatment. Furthermore, supplement of RSM to OVX rats resulted in an increase in femoral bone mineral density and bone strength as well as an improvement in bone microstructures. Moreover, the decreased expression of phosphor (p)-LRP6, insulin-like growth factor-1(IGF-1), ALP, and OPG, as well as increased expression of RANKL and cathepsin K in the tibias and femurs of OVX rats were shifted by RSM treatment. Additionally, RSM reversed the decreased ratio of p-glycogen synthase kinase 3β (GSK3β) to GSK3β and increased ratio of p-β-catenin to β-catenin in OVX rats. Altogether, it is suggestive that RSM improves bone quantity and quality by favoring Wnt/β-catenin and OPG/RANKL/cathepsin K signaling pathways in OVX rats thereby suggesting the potential of this herb to be a novel source of antiosteoporosis drugs.


Neural network based 3D tracking with a graphene transparent focal stack imaging system.
Recent years have seen the rapid growth of new approaches to optical imaging, with an emphasis on extracting three-dimensional (3D) information from what is normally a two-dimensional (2D) image capture. Perhaps most importantly, the rise of computational imaging enables both new physical layouts of optical components and new algorithms to be implemented. This paper concerns the convergence of two advances: the development of a transparent focal stack imaging system using graphene photodetector arrays, and the rapid expansion of the capabilities of machine learning including the development of powerful neural networks. This paper demonstrates 3D tracking of point-like objects with multilayer feedforward neural networks and the extension to tracking positions of multi-point objects. Computer simulations further demonstrate how this optical system can track extended objects in 3D, highlighting the promise of combining nanophotonic devices, new optical system designs, and machine learning for new frontiers in 3D imaging.


α-Chaconine Affects the Apoptosis, Mechanical Barrier Function, and Antioxidant Ability of Mouse Small Intestinal Epithelial Cells.
α-Chaconine is the most abundant glycoalkaloid in potato and toxic to the animal digestive system, but the mechanisms underlying the toxicity are unclear. In this study, mouse small intestinal epithelial cells were incubated with α-chaconine at 0, 0.4, and 0.8 μg/mL for 24, 48, and 72 h to examine apoptosis, mechanical barrier function, and antioxidant ability of the cells using a cell metabolic activity assay, flow cytometry, Western blot, immunofluorescence, and fluorescence quantitative PCR. The results showed that α-chaconine significantly decreased cell proliferation rate, increased apoptosis rate, decreased transepithelial electrical resistance (TEER) value, and increased alkaline phosphatase (AKP) and lactate dehydrogenase (LDH) activities, and there were interactions between α-chaconine concentration and incubation time. α-Chaconine significantly reduced the relative and mRNA expressions of genes coding tight junction proteins zonula occludens-1 (ZO-1) and occludin, increased malondialdehyde (MDA) content, decreased total glutathione (T-GSH) content, reduced the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), and γ-glutamylcysteine synthetase (γ-GCS) and the mRNA expressions of SOD, CAT, GSH-Px, and γ-GCS genes. In conclusion, α-chaconine disrupts the cell cycle, destroys the mechanical barrier and permeability of mucosal epithelium, inhibits cell proliferation, and accelerates cell apoptosis.


Parallel recovery of chromatin accessibility and gene expression dynamics from frozen human regulatory T cells.
Epigenetic features such as DNA accessibility dictate transcriptional regulation in a cell type- and cell state- specific manner, and mapping this in health vs. disease in clinically relevant material is opening the door to new mechanistic insights and new targets for therapy. Assay for Transposase Accessible Chromatin Sequencing (ATAC-seq) allows chromatin accessibility profiling from low cell input, making it tractable on rare cell populations, such as regulatory T (Treg) cells. However, little is known about the compatibility of the assay with cryopreserved rare cell populations. Here we demonstrate the robustness of an ATAC-seq protocol comparing primary Treg cells recovered from fresh or cryopreserved PBMC samples, in the steady state and in response to stimulation. We extend this method to explore the feasibility of conducting simultaneous quantitation of chromatin accessibility and transcriptome from a single aliquot of 50,000 cryopreserved Treg cells. Profiling of chromatin accessibility and gene expression in parallel within the same pool of cells controls for cellular heterogeneity and is particularly beneficial when constrained by limited input material. Overall, we observed a high correlation of accessibility patterns and transcription factor dynamics between fresh and cryopreserved samples. Furthermore, highly similar transcriptomic profiles were obtained from whole cells and from the supernatants recovered from ATAC-seq reactions. We highlight the feasibility of applying these techniques to profile the epigenomic landscape of cells recovered from cryopreservation biorepositories.


Platelet gel in the treatment of severe scleroderma skin ulcers.
Systemic sclerosis (SSc) is characterized by microvascular damage and fibrosis of the skin and internal organs. Non-healing skin ulcers, mainly non-venous leg ulcers, represent one of the most challenging complications. Platelet gel (PG) is a hemocomponent containing numerous growth factors, potentially useful for tissue reparation. This pilot open study aimed to evaluate the effect of PG in 12 SSc patients with skin ulcers resistant to conventional therapies from at least 6 months. PG was applied in the wound bed twice weekly for 2 weeks, then once a week for 12 weeks; in all cases, the ongoing treatments remained unchanged at the time of PG applications. Skin ulcers were evaluated at 0, 12 and 24 weeks; the patient's quality of life was also evaluated using the visual analogical scale (VAS) and the health assessment questionnaire (HAQ). During the 6-month follow-up, the skin ulcers consistently improved in 10/12 patients, with complete healing in 4. At the last evaluation, wound size reduced from 23.4 ± 14.9 SD to 2.3 ± 2.2 SD cm(2) (p < 0.0001). Patient's quality of life markedly improved: VAS decreased from 87.08 ± 13.5 to 57.9 ± 12.6; p < 0.0001 and HAQ from 0.73 ± 0.43 to 0.57 ± 0.22; p < 0.0001. PG may represent a novel therapeutic option for SSc skin ulcers refractory to conventional treatments.


Family and Caregiver Characteristics Contribute to Caregiver Change in Use of Strategies and Growth in Child Spoken Language in a Parent-Implemented Language Intervention in Fragile X Syndrome.
This study examined relationships among family characteristics, caregiver change in use of strategies, and child growth in spoken language over the course of a parent-implemented language intervention (PILI) that was developed to address some of the challenges associated with the fragile X syndrome (FXS) phenotype. Participants were 43 parent-child dyads from two different PILI studies, both of which taught parents various language facilitation strategies to support child language. Before starting the intervention, parents reported on their mental health, parenting stress, and parenting competence. This study focused on potential barriers to treatment gains by examining correlations between the measures of parent well-being and (a) parent change in use of intervention strategies taught in the PILI and (b) changes in child language outcomes from preto post-intervention. Parents in this study had elevated mental health symptoms across several domains and increased rates of parenting stress. Furthermore, although PILI resulted in treatment gains for both parents and children, a variety of parent mental health symptoms were found to be significantly and negatively associated with change in use of strategies and growth in child language over the course of the intervention. Some inconsistent findings also emerged regarding the relationships between parenting stress and competence and change in parent strategy use and growth in child language. This study provides preliminary evidence that parents who are experiencing significant mental health challenges may have a more difficult time participating fully in PILIs and that there may be subsequent effects on child outcomes. Future PILIs could benefit from addressing parent well-being as a substantial part of the intervention program.


Artificial Intelligence against COVID-19 Pandemic: A Comprehensive Insight.
COVID-19 is a pandemic initially identified in Wuhan, China, which is caused by a novel coronavirus, also recognized as the Severe Acute Respiratory Syndrome (SARS-nCoV-2). Unlike other coronaviruses, this novel pathogen may cause unusual contagious pain, which results in viral pneumonia, serious heart problems, and even death. Researchers worldwide are continuously striving to develop a cure for this highly infectious disease, yet there are no well-defined absolute treatments available at present. Several vaccination drives using emergency use authorisation vaccines have been held across many countries; however, their long-term efficacy and side-effects studies are yet to be studied. Various analytical and statistical models have been developed, however, their outcome rate is prolonged. Thus, modern science stresses the application of state-of-the-art methods to combat COVID-19. This paper aims to provide a deep insight into the comprehensive literature about AI and AI-driven tools in the battle against the COVID-19 pandemic. The high efficacy of these AI systems can be observed in terms of highly accurate results, i.e., > 95%, as reported in various studies. The extensive literature reviewed in this paper is divided into five sections, each describing the application of AI against COVID-19 viz. COVID-19 prevention, diagnostic, infection spread trend prediction, therapeutic and drug repurposing. The application of Artificial Intelligence (AI) and AI-driven tools are proving to be useful in managing and fighting against the COVID-19 pandemic, especially by analysing the X-Ray and CT-Scan imaging data of infected subjects, infection trend predictions, etc.


Human cytochrome P450 expression in bacteria: Whole-cell high-throughput activity assay for CYP1A2, 2A6 and 3A4.
Cytochrome P450s (CYPs) are key enzymes involved in drug and xenobiotic metabolism. A wide array of in vitro methodologies, including recombinant sources, are currently been used to assess CYP catalysis, to identify the metabolic profile of compounds, potential drug-drug interactions, protein-protein interactions in the CYP enzyme complex and the role of polymorphic enzymes. We report here on a bacterial whole-cells high-throughput method for the activity evaluation of human CYP1A2, 2A6, and 3A4, when sustained by NADPH cytochrome P450 oxidoreductase (CPR), in the absence or presence of cytochrome b5 (CYB5). This new assay consists of a microplate real-time fluorometric method, with direct measurement of metabolite formation, in a suspension of Escherichia coli BTC-CYP bacteria, a human CYP competent tester strain when incubated with specific fluorogenic substrates. Overall, the maximum turnover (kcat) velocities of the three human CYPs resulting from the whole-BTC cells assays were similar to those obtained when applying the corresponding standard reference membrane fractions assays. CYP activity screening with co-expression of CYB5 suggests an enhancing effect of CYB5 on the kcat of specific isoforms, when using the whole-BTC cells assay. Our results demonstrate that this new approach can offer an efficient high-throughput method for screening of CYP1A2, 2A6 and 3A4 activity and can be potentially applicable for other human CYPs. This can be of particular use for timely and efficient screening of chemical libraries or mutant libraries of CYP enzyme complex proteins, without the necessity for labor intensive isolation of subcellular fractions.


Clinical, dosimetric, and radiographic correlation of radiation injury involving the brainstem and the medial temporal lobes following stereotactic radiotherapy for neoplasms of central skull base.
Stereotactic Radiotherapy (SRT) is more commonly used for skull base tumors in conjunction with the technical development of radiation intensity modulation. Purpose of this study is to correlate clinical and radiographic characteristics of delayed radiation injury (RI) occurring around central skull base following SRT with SRT dosimetric data. Total of six patients were identified to have developed RI in the vicinity of SRT target volume out of 141 patients who received SRT in he center or near-center of the skull base. The images and medical records were retrospectively reviewed. The analysis was performed for RI location, time of development, imaging and clinical characteristics and evolution of RI and correlated with SRT dosimetric analysis using image fusion with follow-up MRI scans. Mean follow-up time was 24 +/- 9 months. During the follow-up period, twelve sites of RI were found in 6 patients. They were clinically symptomatic in 4/6 patients (66.6%) at median 12.5 months after SRT. Mean time interval between SRT and detection of RI was 9 +/- 3, 18.5 +/- 5, and 13.5 months for brainstem, temporal lobe, and cerebellum/labyrinth lesions, respectively. All RI lesions were included in the region of high SRT doses. After steroid and symptomatic treatment, 50% of RI lesions showed complete response, and 40% showed partial response. RI can occur around the skull base because of irregular shape of target tumor, its close proximity to normal brain parenchyma, and inhomogeneity of dose distribution. Brainstem lesions occurred earlier than temporal lobe RI. The majority of the RI lesions, not mixed with the tumor in this study, showed radiographic and clinical improvement with steroid and symptomatic treatments.