Large Cell Neuroendocrine Carcinoma of the Bladder with Adenocarcinomatous Component.
Large cell neuroendocrine carcinoma (LCNC) is one of the rarest types of bladder cancer occurring in <1%. Either pure or mixed with another component, it remains one of the most aggressive types of bladder cancer. We report a case of LCNC of the bladder with an adenocarcinomatous component. The patient was a 64-year-old smoker male, who presented for the first time with dysuria and hematuria. A bladder tumor invading the anterior and right lateral bladder walls was discovered, without any secondary localizations. Tumor biopsy showed an LCNC with adenocarcinomatous components. The patient was treated by recurrent tumor resections, chemotherapy, and radiotherapy. No improvement was noted despite close follow-up and adequate treatment. Neuroendocrine bladder tumor is known to have an aggressive, rapid, and disadvantageous evolution. Multiple case reports were published so far, and a recent review was conducted in March 2020 by Sanguedolce et al. (2020). More cases are needed to establish the best management plan for this type of tumor.


The transcriptional programme of human heart valves reveals the natural history of infective endocarditis.
Infective endocarditis (IE) is an infectious disease that is mainly caused by Staphylococcus aureus and Streptococcus sp. It usually leads to valvular destruction and vegetation formation. Its pathophysiology is badly understood and likely involves immune and coagulation systems with close interactions with the microorganism. Our objective was to evaluate host response by comparing transcriptional profiles of cardiac valves from IE patients with controls. Hierarchical clustering revealed a signature of IE consisting of 146 genes. Among the 89 up-regulated genes, we identified two genes strongly associated with IE: metalloproteinase 12 (MMP-12) and aquaporin-9, a member of the aquaglyceroporin membrane channel family. The up-regulation of MMP-12 gene is strengthened by the down-modulation of the gene encoding its inhibitor TIMP3. In addition, MMP-12 was expressed in macrophages infiltrating EI valves. We also found that aquaporin-9 was expressed in endothelial cells lining neo-vessel lumen, suggesting that aquaporin-9 might be associated with neovascularization of infected valves leading to tissue oedema secondary to the inflammatory process. The Gene Ontology annotation and the resulting functional classification showed that most up-regulated genes account for recruitment of inflammatory cells in vegetations, angiogenesis and remodelling of endocardium tissue. A network analysis confirmed the involvement of molecules related to the remodelling of endocardium tissue and angiogenesis in IE. It also evidenced the role of caspases, especially that of caspase-9 and intrinsic apoptotic pathway in IE. Based on this study we propose a scenario for the natural history of IE in humans. Some parameters identified in this work could become tools for measuring the disease activity and should be tested as biomarkers for diagnosis or prognosis assessment in future studies.


Cloning and expression of the chicken type 2 iodothyronine 5'-deiodinase.
The type 2 iodothyronine deiodinase (D2) is critical for the intracellular production of 3,5,3'-triiodothyronine from thyroxine. The D2 mRNA of higher vertebrates is over 6 kilobases (kb), and no complete cDNA clones have been reported. Using 5'- and 3'-rapid amplification of cDNA ends and two cDNA libraries, we have cloned the 6094-base pair full-length chicken D2 cDNA. The deduced protein is approximately 31 kDa and contains two in-frame UGA codons presumably encoding selenocysteine. One of these is in the highly conserved active catalytic center; the other is near the carboxyl terminus. Unusual features of the cDNA include a selenocysteine insertion sequence element approximately 4.8 kb 3' to the UGA codon in the active center and three short open reading frames in the 5'-untranslated region. The Km of D2 is approximately 1.0 nM for thyroxine, and the reaction is insensitive to inhibition by 6-n-propylthiouracil. Chicken D2 is expressed as a single transcript of approximately 6 kb in different brain regions and in the thyroid and lung. Hypothyroidism increases D2 mRNA in the telencephalon. Unlike in mammals, D2 mRNA and activity are expressed in the liver of the chicken, suggesting a role for D2 in the generation of plasma 3,5,3'-triiodothyronine in this species.


Overview of facial aging.
Facial aging is a multidimensional, multifactorial process. The aging face has traditionally been treated by each specialty in a different manner. However, by understanding the process from the perspective of different specialties, each physician may better treat the spectrum of facial aging. Whether or not the facial plastic surgeon injects products to restore volume, uses lasers to resurface the epidermis and dermis, incorporates cosmeceuticals to enhance and maintain improvements in the skin integrity and appearance, or relaxes muscles with botulinum toxins, he or she can best advise patients and address facial aging by having a functional understanding of these various modalities. With this knowledge, the facial plastic surgeon can parse the component of facial aging that enables him or her to correct each with the appropriate treatment.


[Biochemical and pharmacological mechanisms of different types of hypoxic preconditioning in cerebral ischemia in mice].
Different types of hypoxic preconditioning (hypoxic, circulatory, hemic and tissue hypoxia) increase the tolerance to complete global cerebral ischemia at early terms (hours). Biochemico-pharmacological analysis with the use of selective agonists and antagonists showed the importance of adenosine A1-receptors and K+(ATP)-channels in the mechanisms of the neuroprotective effect and natural tolerance. The general scheme of the investigated mechanisms of different types of hypoxic preconditioning has been proposed.


Parental Socioeconomic Status and Autism Spectrum Disorder in Offspring: A Population-Based Cohort Study in Taiwan.
Studies from the United States have shown increasing incidence of autism spectrum disorder (ASD) with increasing socioeconomic status (SES), whereas in Scandinavian countries, no such relation was identified. We investigated how ASD risk in offspring varied according to parental SES in Taiwan, where there is universal health care. Through linking birth reporting data and data from Taiwan's national health insurance program, we studied 706,111 singleton births from 2004 to 2007 and followed them until 2015. Parental SES was determined by monthly salary at the time of childbirth, and child neuropsychiatric outcomes were defined using International Classification of Diseases codes. We identified 7,323 ASD cases and 7,438 intellectual disability (ID) cases; 17% of ASD cases had co-occurring ID. In multivariable Cox regression analysis, higher SES was independently associated with higher risk of ASD after we took into account urbanization levels, child sex, parental age, and other covariates. By contrast, higher SES was independently associated with lower risk of ID. Besides the SES disparity in ASD case ascertainment and in the access to health care, findings from Taiwan suggest that other social, environmental, biological, and immunological factors linked with parental SES levels may contribute to the positive relation of SES and ASD risk.


Impact of formulary restrictions on the cost-effectiveness of antidepressant treatment.
Newer antidepressants are associated with higher costs of treatment of anxiety and depression. Managed care organizations are challenged to control treatment costs by implementing restricted formularies based on price and perceived medical value. Despite unfavorable side effects of efficacious tricyclic antidepressants, the low acquisition cost rationalizes the inclusion of this older class of agents on a formulary. On the other hand, cost-containment approaches have been taken toward more expensive drug classes (e.g., selective serotonin reuptake inhibitors) despite a superior safety profile of these drug classes over tricyclics. There is compelling evidence that dual reuptake inhibitors (e.g., venlafaxine extended-release), which have acquisition costs similar to serotonin reuptake inhibitors, have a broad spectrum of efficacy and thus added value, contributing to the cost-effectiveness of including this agent in the managed care formulary. Assessment of overall cost-effectiveness should not be limited by acquisition costs but should take total healthcare costs into consideration.


Synthesis, anticancer, structural, and computational docking studies of 3-benzylchroman-4-one derivatives.
A series of 3-Benzylchroman-4-ones were synthesized and screened for anticancer activity by MTT assay. The compounds were evaluated against two cancerous cell lines BT549 (human breast carcinoma), HeLa (human cervical carcinoma), and one noncancerous cell line vero (normal kidney epithelial cells). 3b was found to be the most active molecule against BT549 cells (IC50 = 20.1 µM) and 3h against HeLa cells (IC50 = 20.45 µM). 3b also exhibited moderate activity against HeLa cells (IC50 = 42.8 µM). The molecular structures of 3h and 3i were solved by single crystal X-ray crystallographic technique. Additionally, the molecular docking studies between the tumour suppressor protein p53 with the lead compound 3h, which exhibited better anticancer activity against HeLa cells was examined.


Outer membrane permeability of Acinetobacter calcoaceticus and its implication in antibiotic resistance.
In order to understand high and broad antibiotic resistance of Acinetobacter calcoaceticus, the outer membrane permeability was studied. The permeability coefficients of zwitterionic cephalosporins in the intact cell outer membrane were 0.14-1.12 x 10(-4) cm3/min/mg protein. These values were two to seven times lower than the permeability coefficients of the same beta-lactams in the outer membrane of Pseudomonas aeruginosa. The diffusion rates of carbapenems and zwitterionic cephalosporins into liposomes containing purified outer membrane appeared to be about 1-3%, that of the Escherichia coli B outer membrane. These results indicate that the outer membrane of A. calcoaceticus acts as a substantial barrier against the penetration of these antibiotics. About 80 times of the A. calcoaceticus outer membrane was needed in the liposome compared with E. coli B to show the same extent of saccharide permeability. Two minor outer membrane proteins (less than 5% of total outer membrane protein) were identified to be the porins. These results suggest that one of the causes for the high antibiotic resistance of A. calcoaceticus is attributable to the presence of a small number of small-sized porins.


Urolithiasis and pregnancy.
The diagnosis of urolithiasis during pregnancy is common, even though no additional measures are required in asymptomatic cases. Renal colic or complications of urinary lithiasis occur more frequently during the last months of pregnancy, and there are several particularities for the diagnosis and treatment of this subset of women. The present manuscript aim to review the current knowledge concerning this subject and present authors personal experience.


Antenatal and postnatal corticosteroid and resuscitation induced lung injury in preterm sheep.
Initiation of ventilation using high tidal volumes in preterm lambs causes lung injury and inflammation. Antenatal corticosteroids mature the lungs of preterm infants and postnatal corticosteroids are used to treat bronchopulmonary dysplasia. To test if antenatal or postnatal corticosteroids would decrease resuscitation induced lung injury. 129 d gestational age lambs (n = 5-8/gp; term = 150 d) were operatively delivered and ventilated after exposure to either 1) no medication, 2) antenatal maternal IM Betamethasone 0.5 mg/kg 24 h prior to delivery, 3) 0.5 mg/kg Dexamethasone IV at delivery or 4) Cortisol 2 mg/kg IV at delivery. Lambs then were ventilated with no PEEP and escalating tidal volumes (VT) to 15 mL/kg for 15 min and then given surfactant. The lambs were ventilated with VT 8 mL/kg and PEEP 5 cmH20 for 2 h 45 min. High VT ventilation caused a deterioration of lung physiology, lung inflammation and injury. Antenatal betamethasone improved ventilation, decreased inflammatory cytokine mRNA expression and alveolar protein leak, but did not prevent neutrophil influx. Postnatal dexamethasone decreased pro-inflammatory cytokine expression, but had no beneficial effect on ventilation, and postnatal cortisol had no effect. Ventilation increased liver serum amyloid mRNA expression, which was unaffected by corticosteroids. Antenatal betamethasone decreased lung injury without decreasing lung inflammatory cells or systemic acute phase responses. Postnatal dexamethasone or cortisol, at the doses tested, did not have important effects on lung function or injury, suggesting that corticosteroids given at birth will not decrease resuscitation mediated injury.


Diauxic shift-induced stress resistance against hydroperoxides in Saccharomyces cerevisiae is not an adaptive stress response and does not depend on functional mitochondria.
Respiring Saccharomyces cerevisiae cells grown on a non-fermentable carbon source are intrinsically more resistant to several stresses, including oxidative stress. The mechanisms leading to increased stress resistance are not yet well understood. Active mitochondria are the major source of intracellular reactive oxygen species (ROS), which could cause the up-regulation of the antioxidant defense systems. We investigated the role of mitochondria in the intrinsic stress resistance against the hydroperoxides H2O2 and tert-butylhydroperoxide 4 h after a shift in carbon source. We found that, independently of functional mitochondria, the yeast acquired the intrinsic resistance of respiring cells against hydroperoxides solely as a response to a change of carbon source in the growth medium. Furthermore, utilizing reporter gene fusion constructs, we monitored the expression of the gamma-glutamylcysteinyl synthetase (encoded by GSH1) and the two superoxide dismutases (encoded by SOD1 and SOD2) during the metabolic transition from fermentation to respiration; and we detected an up-regulation of all three genes during the diauxic shift. Overall available data allowed us to propose that the antioxidant system of S. cerevisiae could be considered as a class of genes under glucose/carbon catabolite regulation. This control system is different from the well-known adaptive response to oxidative stress.


Anomalous effect of permeant ion concentration on peak open probability of cardiac Na+ channels.
Human heart Na+ channels were expressed transiently in both mammalian cells and Xenopus oocytes, and Na+ currents measured using 150 mM intracellular Na+. Decreasing extracellular permeant ion concentration decreases outward Na+ current at positive voltages while increasing the driving force for the current. This anomalous effect of permeant ion concentration, especially obvious in a mutant (F1485Q) in which fast inactivation is partially abolished, is due to an alteration of open probability. The effect is only observed when a highly permeant cation (Na+, Li+, or hydrazinium) is substituted for a relatively impermeant cation (K+, Rb+, Cs+, N-methylglucamine, Tris, choline, or tetramethylammonium). With high concentrations of extracellular permeant cations, the peak open probability of Na+ channels increases with depolarization and then saturates at positive voltages. By contrast, with low concentrations of permeant ions, the open probability reaches a maximum at approximately 0 mV and then decreases with further depolarization. There is little effect of permeant ion concentration on activation kinetics at depolarized voltages. Furthermore, the lowered open probability caused by a brief depolarization to +60 mV recovers within 5 ms upon repolarization to -140 mV, indicative of a gating process with rapid kinetics. Tail currents at reduced temperatures reveal the rapid onset of this gating process during a large depolarization. A large depolarization may drive a permeant cation out of a site within the extracellular mouth of the pore, reducing the efficiency with which the channel opens.


Origins of peripheral B cells in IL-7 receptor-deficient mice.
The interleukin 7 (IL-7) signaling pathway is critical for early lymphoid differentiation. We found dramatic perturbations in fetal liver B cell development and confirmed a complete absence of developing B cells in the adult bone marrow in mice lacking the IL-7 receptor alpha (IL-7Ralpha) gene. We show that peripheral B-2 and B-1 cell populations are deficient in IL-7Ralpha-/- mice. B-2 follicular cell and peritoneal B-1 cell percentages are reduced, while B-2 marginal zone cell percentages are increased. A comparison of bone marrow and splenic populations at different ages revealed that the splenic B cell populations seen in adult IL-7Ralpha-/- mice first appear during neonatal development. We have measured N-nucleotide addition at the joints of V(D)J rearrangements in splenic B cells and have used it as a somatic marker to define and separate bone marrow-derived B cells from fetal liver-derived B cells. B cells isolated from the bone marrow and spleen of adult and neonatal IL-7Ralpha-deficient mice harbor high levels of N-nucleotide additions similar to those found in equivalent wild-type B cell populations. We conclude that the majority of splenic B cells in IL-7Ralpha-deficient mice originate from the bone marrow and not the fetal liver.


Human papillomavirus-16 E5 protein: oncogenic role and therapeutic value.
Human papillomavirus (HPV) is a non-enveloped, double-stranded DNA virus. HPV infection occurs through sexual route, and the virus infects mucosal and cutaneous epithelial cells. Inside the cell, the viral DNA replicates extrachromosomally. HPV is the major cause of cervical cancer worldwide. HPVs infecting mucosal epithelial cells are sub-grouped into low-risk or high-risk by virtue of them causing benign warts or cancer, respectively. The early oncoproteins of HPV, namely E4, E5, E6 and E7, are known to contribute to tumorigenesis. The roles and functions of HPV E6 and E7 have been thoroughly studied over the years. However, limited studies have been done on E5 regarding its intracellular functions. This review attempts to discuss the positive role of HPV16 E5 in the form of therapeutic target for cervical cancer, as well as its role in modulation of several intracellular signalling pathways leading to transformed phenotype of the host cell.


Towards the dietary prevention of cancer: contributions of epidemiology.
There may be a considerable potential for the prevention of cancer through dietary modifications or chemo-prevention strategies. The epidemiological methods which have been used in the study of the relation between diet and cancer are briefly reviewed and the scientific basis for major hypotheses linking diet to cancer are summarized. Clinical trials, now being considered to assess the potential for the prevention of cancer by dietary supplementation or modification, do not have many of the uncertainities of other methods, yet there are many potential difficulties in both the conduct and interpretation of such trials. These limitations are discussed as well as the problem in making general dietary recommendations for the prevention of cancer based on current knowledge. A more complete understanding of the effect of diet on cancer risk will likely emerge only through a more integrated and interdisciplinary approach in future studies.


Polyomavirus infection in pediatric renal transplant recipients: evaluation using a quantitative real-time PCR technique.
Polyomavirus infection and related nephropathy is being increasingly recognized as an important cause of allograft dysfunction in adult renal transplant recipients. We prospectively monitored pediatric renal transplant recipients for the presence of BK and JC polyomavirus in urine and blood using a quantitative PCR assay to evaluate the prevalence and clinical relevance of polyomavirus infection in the pediatric renal transplant population. Of 46 pediatric renal recipients who were evaluated, nine (20%) demonstrated isolated BKV viruria, while five (11%) had concomitant BKV viremia and viruria. JCV viruria was found in eight (17%) patients. BKV viremia was associated with a significantly higher urinary BKV viral load: median urinary viral load 1.9 x 10(9) copies/mL (range 6.7 x 10(2)-1.8 x 10(11)) for the group with concomitant viremia and viruria vs. 1.8 x 10(3) copies/mL (range 2.5 x 10(2)-4.5 x 10(6)) for the group with isolated viruria (p < 0.0001). In children that were followed prospectively since their transplantation, the BKV urinary viral load increased markedly before viremia became detectable a few weeks later. None of the patients with JCV viruria or isolated BKV viruria had renal dysfunction. Among the five patients with BKV in both urine and blood, two developed biopsy-proven BKV nephropathy associated with deterioration of the renal function. Management of the BKV nephropathy consisted of reduction of immunosuppression alone or in combination with antiviral treatment with cidofovir. This study shows that polyomavirus infection and related interstitial nephritis is a relevant clinical issue in the pediatric renal transplant population. Monitoring the polyomaviral load in the urine and the blood of the patients using a quantitative PCR technique is a useful tool in the diagnosis and subsequent management of this infection. Even before viremia is present, an important rise in the urinary viral load should draw the attention of the transplant clinician and raise the issue of adapting the immunosuppression.


Nitric oxide donors for patients undergoing IVF. A prospective, double-blind, randomized, placebo-controlled trial.
Nitric oxide (NO) is involved in local control of the uterine cycle and in preparation of the uterus for pregnancy. NO donors, acting as vasodilating agents, may therefore have possible therapeutic uses, for example they may be of benefit to patients with a history of implantation failure. In a prospective, comparative, randomized, placebo-controlled study, we assessed the efficacy of nitroglycerin (NTG) administered to 138 IVF patients with a history of implantation failure. Controlled ovarian stimulation was performed with long agonist protocol combined with recombinant FSH. Embryos were transferred on day 2 or 3 after oocyte retrieval. Eligible patients were those who had at least two 'good quality' embryos. The NTG patch was administered the day before embryo transfer and continued until either the results of the pregnancy test were known or until menstruation occurred. Ovarian response, implantation rate and pregnancy rate were comparable between both groups. No difference was observed in uterine Doppler findings, particularly the mean pulsatility index. NTG treatment the day before embryo transfer was no more effective than placebo in improving the implantation or pregnancy rates in a population of IVF patients with a previous history of implantation failures.


[Maternal cannabis use and intra-uterine growth restriction].
Marijuana is the most commonly used illegal drug, especially among young women in Western societies. The effects of cannabis use during pregnancy have been studied for many years. The vast majority of studies have shown a link between maternal consumption of cannabis and foetal development. Foetal growth restriction seems to be the major complication of cannabis exposure. Nevertheless, all these studies have suffered from several methodological biases. The maternal marijuana use should be first and foremost detected in pregnant women for a specific addiction management and pregnancy follow-up.


Organizational Best Practices Supporting Mental Health in the Workplace.
To provide a narrative review of best and promising practices for achieving exemplary mental health in the workplace as the foundation for the inaugural Carolyn C. Mattingly Award for Mental Health in the Workplace. Research was drawn from peer-reviewed articles using the search terms associated with workplace mental health. Eight categories of best practices were identified: (1) culture, (2) robust mental health benefits, (3) mental health resources, (4) workplace policies and practices, (5) healthy work environment, (6) leadership support, (7) outcomes measurement, and (8) innovation. The review provided the scientific backing to support criteria developed for the Carolyn C. Mattingly Award for Mental Health in the Workplace. By recognizing organizations that apply evidence-based practices in their health and well-being programs, the Mattingly Award may inspire employers to adopt best practices.


Phosphorylation of nuclear protein is an early event in TGF beta 1 action.
Transforming growth factor beta (TGF beta) is a family of polypeptides that modulate growth and differentiation. TGF beta exerts its effects on target cells through interaction with specific cell surface receptors, but the signal transduction pathways are as yet largely unresolved. In this study we report that the growth inhibitory action of TGF beta on mink lung CCl 64 cells is associated with a rapid and transient phosphorylation of a number of nuclear proteins. In parallel, a transient expression of the immediate early gene jun B is observed. The expression of jun B can be inhibited by the protein kinase inhibitor H7 and can be augmented by the phosphatase inhibitor okadaic acid. Thus, protein phosphorylation can be a possible mechanism through which TGF beta 1 initiates early genomic responses.


[Segmental infarction of testicle presenting as right acute scrotum: a case report].
We report a case of segmental infarction of a testicle seen in a 28-year-old man with right lower abdominal pain who visited our hospital. On physical examination, the right testicle was slightly elevated and Prehn's sign was positive. As torsion of the right spermatic cord was suspected, ultrasonography and color Doppler sonography were performed, revealing sufficient blood supply in the right spermatic cord and in the most part of the right testicle. However, a low echogenic area without blood flow was noted in the upper pole of testicle. Since CT and MRI findings couldn't rule out a testicular tumor, right high orchiectomy was performed. The specimen revealed a hemorrhagic area, histologically proved to be segmental hemorrhagic infarction of the testicle. Segmental infarction of a testicle is rare and our case is the 31st case in the world literature (the 7th case in Japan).


Many growth factors may not be growth factors.
Both organ growth and tumor growth are dependent upon a biased ratio of cell births to cell deaths; this ratio is independent of the frequency of mitosis. Thus, so-called growth factors that affect only the frequency of mitosis are not really growth factors. I shall advance two hypotheses: that the normal adult ratio of cell births to cell deaths is maintained by the activity of a factor or factors produced by the stem cells; and that the differentiating cells, as well as the stem cells, produce a different factor that limits the frequency of mitosis.


Receptor signal output mediated by the ETR1 N terminus is primarily subfamily I receptor dependent.
etr1-1 is a dominant ethylene receptor gene in Arabidopsis (Arabidopsis thaliana) and confers ethylene insensitivity. The truncated etr1-1(1-349) protein is capable of repressing ethylene responses, whereas etr1(1-349) is not, lending support to a hypothesis that the dominant etr1-1(1-349) could convert wild-type receptors to an ethylene-insensitive state. Assuming that etr1-1(1-349) and etr1(1-349) would share the same signaling mechanism, we hypothesize that the etr1(1-349) protein is capable of repressing ethylene responses when not bound with ethylene. In this study, we show that both etr1(1-349) and etr1-1(1-349) are capable of receptor signal output, which is primarily dependent on subfamily I receptors. The etr1(1-349) and etr1-1(1-349) clones were individually transformed to mutants and the resulting phenotypes were scored. Each of those transgenes restored the rosette growth and flower fertility of etr1-7 ers1-2 to a similar extent. In contrast, neither etr1(1-349) nor etr1-1(1-349) was capable of signal output in etr1-7 ers1-3. The ERS1 transcript was detectable in ers1-2 but not in ers1-3, implying that ETR1 N-terminal signaling is subfamily I dependent. Loss of the subfamily II receptor genes did not perturb etr1-1(1-349)-mediated ethylene insensitivity. Possible roles of subfamily I receptors and disulfide linkages in ETR1 receptor signal output mediated through the N terminus are discussed.


Pemphigus Vulgaris and Squamous Cell Carcinoma of the Larynx.
A 40 year old male patient who developed pemphigus two years after developing squamous cell carcinoma of larynx is presented. The tumour was active at the time of development of pemphigus vulgaris Pemphigus lesions were controlled with corticosteroids even though the tumour continued to show activity.


Synergism of nitric oxide and iron in killing the transformed murine oligodendrocyte cell line N20.1.
Nitric oxide (NO) produced in inflammatory lesions may play a major role in the destruction of oligodendrocytes in multiple sclerosis and experimental allergic encephalomyelitis. The transformed murine oligodendroglial line N20.1 is much more resistant than primary oligodendrocytes to killing by the NO generator S-nitroso-N-acetyl-DL-penicillamine (SNAP). This observation prompted investigation of the mechanisms leading to cell death in the N20.1 cells and comparison of SNAP with another NO donor, sodium nitroprusside (SNP). We observed that N20.1 cells were 30 times more sensitive to SNP than to SNAP. The specific NO scavenger 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (PTIO) protected against SNP only, not against SNAP. However, dithiothreitol protected against both SNAP and SNP, indicating that S-nitrosylation of cysteines plays a major role in the cytotoxicity of both NO donors. We did not observe any formation of peroxynitrite or increase of Ca2+ concentration with either SNAP or SNP, thus excluding their involvement in the mechanisms leading to N20.1 cell death. Based on two observations, (a) potentiation of the cytotoxic effect of SNP when coincubated with ferricyanide or ferrocyanide, but not sodium cyanide, and (b) protection by deferoxamine, an iron cyanide chelator, we conclude that the greater sensitivity of N20.1 cells to SNP compared with SNAP is due to synergism between NO released and the iron cyanide portion of SNP, with the cyanide accounting for very little of the cytotoxicity. Finally, SNP but not SNAP induces some apoptosis, as shown by DNA laddering and protection by a caspase-3 inhibitor. These results suggest that low levels of NO in combination with increased iron content lead to apoptotic cell death rather than the necrotic cell death seen with higher levels of NO generated by SNAP.


Application of WST-8 based colorimetric NAD(P)H detection for quantitative dehydrogenase assays.
The reduction of tetrazolium salts by NAD(P)H to formazan product has been widely used to determine the metabolic activity of cells, and as an indicator of cell viability. However, the application of a WST-8 based assay for the quantitative measurement of dehydrogenase enzyme activity has not been described before. In this study, we reported the application of an assay based on the tetrazolium salt WST-8 for the quantitative measurement of dehydrogenase activity. The assay is performed in a microplate format, where a single endpoint is measured at 450 nm. The optimized dehydrogenase-WST-8 assay conditions, the limit of detection (LOD), accuracy, and precision for measuring NAD(P)H, were demonstrated. The sensitivity of the WST-8 assay for detecting NAD(P)H was 5-fold greater than the spectrophotometric measurement of NAD(P)H absorption at 340 nm (LOD of 0.3 nmole vs 1.7 nmole, respectively). In the dehydrogenase assay, the colorimetric WST-8 method exhibits excellent assay reproducibility with a Z' factor of 0.9. The WST-8 assay was also used to determine dehydrogenase activity in biological samples, and for screening the substrate of uncharacterized short-chain dehydrogenase/oxidoreductase from Burkholderia pseudomallei. The results suggest that the WST-8 assay is a sensitive and rapid method for determining NAD(P)H concentration and dehydrogenase enzyme activity, which can be further applied for the high-throughput screening of dehydrogenases.


Gene expression in rat lungs during early response to paraquat-induced oxidative stress.
Paraquat (PQ) is a well-known pneumotoxicant and provides an established model of oxidative stress. This study focused on the transcriptional response to PQ-driven oxidative stress in rat lungs during an early phase post-injection. Rats were sacrificed at 3 h and 24 h after PQ injection (i.p., 20 mg/kg b.w.), and at 3 h after a second injection (i.p., 20 mg/kg b.w.). The left lungs were rapidly excised and used immediately for RNA preparation. The lung tissues did not show any pathological damage microscopically. Differential expression of RNAs in the lung at 3 h was investigated using a DNA array system. Fifteen genes showed a >1.7-fold change in expression level, which was confirmed by real-time PCR. Five genes related to oxidative stress, TRX, HO-1, GST-Yc, NQO-1, and RL/IF-1, and one gene, CLK3, whose function is unknown, showed a significant increase in their expression due to PQ injection. Two genes, HO-1 and NQO-1, that showed 3- and 2-fold increases at 3-h post-injection, were localized by immunohistochemistry. HO-1 was expressed in the bronchial epithelial cells, some type II cells and macrophages of control lungs, and the cells, especially the bronchial epithelial cells, were strongly stained 3 h following PQ treatment. Immunohistochemical analysis of NQO-1 also showed an increase in positive staining in the bronchial epithelial cells of PQ-treated lung sections. The expression of CYP2C6, 2C7, and 2C12, which are specific to or dominant in female liver, decreased markedly, while the male-specific CYP2C13 and 2C11 showed an increase or no effect. Further investigation is needed to clarify the role of these CYP2C family genes on the early phase of PQ toxicity. These results indicate that the acclimation to oxidative stress is already a highly complex process at the onset of PQ-induced damage and that the genes described herein may prove to be major contributors to the subsequent pulmonary fibrosis.


Will cytokines alter the treatment of myelodysplastic syndrome?
Myelodysplastic syndrome is a frustrating disorder, which until recently lacked effective treatment. Patients usually succumb to infection, bleeding complications, or progression to acute leukemia. Recombinant cytokines such as granulocyte macrophage-colony stimulating factor, granulocyte-colony stimulating factor, interleukin-3, and erythropoietin have been used to ameliorate the cytopenias associated with this disease. Small clinical trials in myelodysplastic syndrome patients, using cytokines with myeloid activity (G-CSF, GMCSF, IL-3), have shown consistent elevations in the white blood cell counts with little success in elevating hemoglobin or platelets. Erythropoietin is able to increase the hemoglobin in a small group of myelodysplastic syndrome patients. Future trials using combinations of these cytokines may lead to multilineage effects.


[Geriatrics and gerontology in Senegal].
Senegal is dealing positively with its demographic transition. On September 1st 2006, the Senegalese government introduced the "Plan Sesame", a national free health care program for elderly people aged 60 years and over. The University of Dakar academic authorities support the Sesame plan through an innovative training program in geriatrics and gerontology. Such programs aim to address the challenge of ageing in a developing country.


The Impact of COVID-19 on Medical Examinations.
Background and aim The coronavirus-19 (COVID-19) pandemic has had a profound impact on many aspects of our lives, including medical education. The suspension of clinical placements and cancellation of objective structured clinical examinations (OSCE) is likely to have an influence on students' performance. Methods Using a questionnaire, a retrospective observational study was conducted. Our primary focus was 3rd year medical students following their examinations in May 2020. Results Out of 46 responses obtained, the results demonstrated over 2/3 felt they perform better in clinical OSCE compared to written examinations and the majority performed worse this year during the pandemic. On a Likert scale, a mean result of 3/5 was obtained for confidence upon returning to placement and to address this, most stated they would benefit from extra optional teaching. Additionally, a further average of 3.82 was derived to represent how greatly students believed the absence of third year clinical OSCE would negatively impact their performance in the OSCE for the following academic year. Conclusion The results of our study highlight that the cancellation of clinical placements and OSCE due to coronavirus has negatively impacted on medical education and if we are to be faced with future pandemics, we must be better prepared to train future doctors.


Improving T-cell assays for the diagnosis of latent TB infection: potential of a diagnostic test based on IP-10.
There is a need for simple tools such as the M.tuberculosis specific IFN-gamma release assays (IGRA) to improve diagnosis of M.tuberculosis-infection in children. The aim of the study was to evaluate the performance of an IP-10 and IL-2 based tests for the diagnosis of M.tuberculosis-infection in recently exposed children from Nigeria. Samples were obtained from 59 children at high risk of infection with M.tuberculosis (contacts of adults with smear and culture-positive tuberculosis) and 61 at low risk (contacts of smear-negative/culture-positive tuberculosis or community controls). IP-10 and IL-2 was measured in plasma after stimulation of whole-blood with M.tuberculosis specific antigens and mitogen. Previously developed criteria for positive IP-10 and IL-2 tests were used and the diagnostic performances of the IP-10 and IL-2 tests were compared with the Quantiferon In-Tube (QFT-IT) and the Tuberculin Skin Tests (TST). In response to M.tuberculosis specific antigens, the high-risk children expressed significantly higher levels of IP-10 (1358 pg/ml[IQR 278-2535 pg/ml]) and IL-2 (164 pg/ml[11-590 pg/ml]) than low risk groups 149 pg/ml(25-497 pg/ml), and 0 pg/ml(0-3 pg/ml), respectively. There was excellent agreement (>89%,k>0.80) between IP-10, IL-2 tests and QFT-IT, better than with TST (>74%,k>0.49). The IP-10 and IL-2 responses were strongly associated with M.tuberculosis exposure and with grade of infectiousness of the index cases (p<0.0001). IP-10, IL-2, and TST but not QFT-IT was associated with age of the child in the low risk groups (p<0.02). IP-10 is expressed in high levels and results of the IP-10 test were comparable to the QFT-IT. IL-2 was released in low amounts in response to the antigens and not in response to the mitogen therefore IL-2 seems a less useful marker. We have demonstrated that IP-10 and possibly IL-2 could be alternative or adjunct markers to IFN-gamma in the diagnosis infection with M.tuberculosis.


The Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) Pandemic: Are Africa's Prevalence and Mortality Rates Relatively Low?
Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), the cause of coronavirus disease 19 (COVID-19), has been rapidly spreading since December 2019, and within a few months, it turned out to be a global pandemic. The disease affects primarily the lungs, but its pathogenesis spreads to other organs as well. However, its mortality rates vary, and in the majority of infected people, there are no serious consequences. Many factors including advanced age, preexisting health conditions, and genetic predispositions are believed to exacerbate outcomes of COVID-19. The virus contains several structural proteins including the spike (S) protein with subunits for binding, fusion, and internalization into host cells following interaction with host cell receptors and proteases (ACE2 and TMPRSS2, respectively) to cause the subsequent pathology. Although the pandemic has spread into all countries, most of Africa is thought of as having relatively less prevalence and mortality. Several hypotheses have been forwarded as reasons for this and include warmer weather conditions, vaccination with BCG (i.e., trained immunity), and previous malaria infection. From genetics or metabolic points of view, it has been proposed that most African populations could be protected to some degree because they lack some genetic susceptibility risk factors or have low-level expression of allelic variants, such as ACE2 and TMPRSS2 that are thought to be involved in increased infection risk or disease severity. The frequency of occurrence of α-1 antitrypsin (an inhibitor of a tissue-degrading protease, thereby protecting target host tissues including the lung) deficiency is also reported to be low in most African populations. More recently, infections in Africa appear to be on the rise. In general, there are few studies on the epidemiology and pathogenesis of the disease in African contexts, and the overall costs and human life losses due to the pandemic in Africa will be determined by all factors and conditions interacting in complex ways.


The complete mitochondrial genome of pronghorn spiny lobster Panulirus penicillatus (Olivier, 1791).
In this paper, we determined and characterized the complete mitochondrial genome of Pronghorn spiny lobster Panulirus penicillatus for the first time from South China Sea. The P. penicillatus mitogenome is 15,671 bp long, and consists of 22 tRNA genes, 2 rRNA genes, 13 protein-coding genes (PCGs), and 1 control region. The nucleotide composition of P. penicillatus mitogenome is significantly biased (A, G, T, and C was 33.62, 13.32, 32.31, and 20.75%, respectively) with A + T contents of 65.93%. Almost PCGs used a standard initiation codon or stop codon, except COX2, ND3, ND4 and ND1 were terminated with an incomplete stop codon T and ND5 ended with TA. One microsatellite (C)12 was identified in the control region of P. penicillatus mitogenome sequences. Phylogenetic tree showed that P. penicillatus was first clustered with P. polyphagus and P. versicolor.


SNP- and Haplotype-Based GWAS of Flowering-Related Traits in Brassica napus.
Traits related to flowering time are the most promising agronomic traits that directly impact the seed yield and oil quality of rapeseed (Brassica napus L.). Developing early flowering and maturity rapeseed varieties is an important breeding objective in B. napus. Many studies have reported on days to flowering, but few have reported on budding, bolting, and the interval between bolting and DTF. Therefore, elucidating the genetic architecture of QTLs and genes regulating flowering time, we presented an integrated investigation on SNP and haplotype-based genome-wide association study of 373 diverse B. napus germplasm, which were genotyped by the 60K SNP array and were phenotyped in the four environments. The results showed that a total of 15 and 37 QTLs were detected from SNP and haplotype-based GWAS, respectively. Among them, seven QTL clusters were identified by haplotype-based GWAS. Moreover, three and eight environmentally stable QTLs were detected by SNP-GWAS and haplotype-based GWAS, respectively. By integrating the above two approaches and by co-localizing the four traits, ten (10) genomic regions were under selection on chromosomes A03, A07, A08, A10, C06, C07, and C08. Interestingly, the genomic regions FT.A07.1, FT.A08, FT.C06, and FT.C07 were identified as novel. In these ten regions, a total of 197 genes controlling FT were detected, of which 14 highly expressed DEGs were orthologous to 13 Arabidopsis thaliana genes after integration with transcriptome results. In a nutshell, the above results uncovered the genetic architecture of important agronomic traits related to flowering time and provided a basis for multiple molecular marker-trait associations in B. napus.


Congenital sinus of the upper lip with idiopathic precocious puberty.
An extremely rare case of congenital sinus of the upper lip is presented in a 22-year-old woman with idiopathic precocious puberty. The sinus has a pin-like orifice located in the midline of the upper lip philtum just below the base of the columella. Sexual precocity was found out when she was 7 years old. A series of exact examinations had been performed at that time. Neither neurogenic lesions of the central nervous system nor other systemic abnormalities were observed. Oral Diseases


Employing Molecular Phylodynamic Methods to Identify and Forecast HIV Transmission Clusters in Public Health Settings: A Qualitative Study.
Molecular HIV surveillance is a promising public health strategy for curbing the HIV epidemic. Clustering technologies used by health departments to date are limited in their ability to infer/forecast cluster growth trajectories. Resolution of the spatiotemporal dynamics of clusters, through phylodynamic and phylogeographic modelling, is one potential strategy to develop a forecasting tool; however, the projected utility of this approach needs assessment. Prior to incorporating novel phylodynamic-based molecular surveillance tools, we sought to identify possible issues related to their feasibility, acceptability, interpretation, and utility. Qualitative data were collected via focus groups among field experts (n = 17, 52.9% female) using semi-structured, open-ended questions. Data were coded using an iterative process, first through the development of provisional themes and subthemes, followed by independent line-by-line coding by two coders. Most participants routinely used molecular methods for HIV surveillance. All agreed that linking molecular sequences to epidemiological data is important for improving HIV surveillance. We found that, in addition to methodological challenges, a variety of implementation barriers are expected in relation to the uptake of phylodynamic methods for HIV surveillance. The participants identified several opportunities to enhance current methods, as well as increase the usability and utility of promising works-in-progress.


Mixed Convective Peristaltic Flow of Water Based Nanofluids with Joule Heating and Convective Boundary Conditions.
Main objective of present study is to analyze the mixed convective peristaltic transport of water based nanofluids using five different nanoparticles i.e. (Al2O3, CuO, Cu, Ag and TiO2). Two thermal conductivity models namely the Maxwell's and Hamilton-Crosser's are used in this study. Hall and Joule heating effects are also given consideration. Convection boundary conditions are employed. Furthermore, viscous dissipation and heat generation/absorption are used to model the energy equation. Problem is simplified by employing lubrication approach. System of equations are solved numerically. Influence of pertinent parameters on the velocity and temperature are discussed. Also the heat transfer rate at the wall is observed for considered five nanofluids using the two phase models via graphs.


Successful engraftment after hematopoietic stem cell transplantation with infusion of donor stem cells through the extracorporeal membrane oxygenation circuit.
Wiskott-Aldrich syndrome (WAS) is a rare X-linked primary immunodeficiency due to mutations in the WAS gene expressed in hematopoietic cells. Hematopoietic stem cell transplantation (HSCT) is the treatment of choice when an appropriate human leukocyte antigen-matched donor is available. The use of the extracorporeal membrane oxygenation (ECMO) circuit to infuse donor cells for HSCT has not been previously published in the literature. We describe a case of a child who had successful engraftment after HSCT with infusion of the donor stem cells through the ECMO circuit.


Nitrile versus Latex for Glove Juice Sampling.
The objective of this study was to explore the utility of nitrile gloves as a replacement for latex surgical gloves in recovering bacteria from the hands. Two types of nitrile gloves were compared to latex gloves using the parallel streak method. Streaks of Klebsiella pneumoniae and Staphylococcus aureus were made on tryptic soy agar plates, and the zones of inhibition were measured around pieces of glove material placed on the plates. Latex gloves produced a mean zone of inhibition of 0.28 mm, compared to 0.002 mm for nitrile gloves (p<.001). While the parallel streak method is not intended as a quantitative estimate of antimicrobial properties, these results suggest that nitrile may be a viable alternative to latex in glove juice sampling methods, since nitrile avoids the risk of latex exposure.


The tet(Q) gene in bacteria isolated from patients with refractory periodontal disease.
Twenty-two tetracycline-resistant (tetr) anaerobic and facultative anaerobic bacteria isolated from periodontal pockets of 12 patients with refractory periodontitis were examined for the presence of the Tet Q determinant by DNA-DNA hybridization. Dot blots of bacterial DNA were tested with an intragenic digoxigenin-labelled tet(Q) probe consisting of a 1.45 kb EcoRI/PvuII fragment from plasmid pNFD13-2. Southern blots of chromosomal DNA digested with the restriction enzyme EcoRI were also examined. The tet(Q) probe hybridized with DNA from 8 of the 22 tetr strains, including 2 Prevotella intermedia strains and one strain each of Prevotella nigrescens, Prevotella loescheii, Prevotella veroralis and Prevotella melaninogenica. The tetr strains of Mitsuokella dentalis and Capnocytophaga ochracea also hybridized with the probe. The lack of discernible plasmid DNA in all the probe-positive isolates suggests that these tetracycline-resistance genes were chromosomally encoded. The probe hybridized with a different size fragment in all the isolates. This study extends the number of species that carry the tet(Q) gene to include several outside the genera Prevotella and Bacteroides.


The Korean Medication Algorithm Project for Depressive Disorder 2021: Comparisons with Other Treatment Guidelines.
The Korean Medication Algorithm Project for Depressive Disorder (KMAP-DD) first was published in 2002, and has been revised four times, in 2006, 2012, 2017, and 2021. In this review, we compared recommendations from the recently revised KMAP-DD 2021 to four global clinical practice guidelines (CPGs) for depression published after 2010. The recommendations from the KMAP-DD 2021 were similar to those from other CPGs, although there were some differences. The KMAP-DD 2021 reflected social culture and the healthcare system in Korea and recent evidence about pharmacotherapy for depression, as did other recently published evidence-based guidelines. Despite some intrinsic limitations as an expert consensus-based guideline, the KMAP-DD 2021 can be helpful for Korean psychiatrists making decisions in clinical settings by complementing previously published evidence-based guidelines, especially for some clinical situations lacking evidence from rigorously designed clinical trials.


Piezoresponse force microscopy at sub-room temperatures.
Piezoresponse force microscopy is demonstrated at temperatures between -80 °C and +120 °C using a commercial room temperature atomic force microscope upgraded with a home-built cooling/heating-stage. We applied temperature-ramp-synchronized piezoresponse force microscope (PFM) for tracing the temperature dependence of the formation of ferroelectric domains. The potential of our sub-room temperature PFM is demonstrated by investigating the formation and evolution of ferroelectric domains in RbHSO4 as a function of temperature and time, respectively.


Birmingham Hip Resurfacing at 20 years.
The Birmingham Hip Resurfacing (BHR) arthroplasty has been used as a surgical treatment of coxarthrosis since 1997. We present 20-year results of 234 consecutive BHRs performed in our unit. Between 1999 and 2001, there were 217 patients: 142 males (65.4%), mean age 52 years (18 to 68) who had 234 implants (17 bilateral). They had patient-reported outcome measures collected, imaging (radiograph and ultrasound), and serum metal ion assessment. Survivorship analysis was performed using Kaplan-Meier estimates. Revision for any cause was considered as an endpoint for the analysis. Mean follow-up was 20.9 years (19.3 to 22.4). Registry data revealed that 19 hips (8.1%) had been revised and 26 patients (12%) had died from causes unrelated to the BHR. Among the remaining 189 hips, 61% were available for clinical follow-up at 20 years (n = 115) and 70% of patients had biochemical follow-up (n = 132). The cumulative implant survival rate at 20 years for male patients was 96.5% (95% confidence interval (CI) 93.5 to 99.6), and for female patients 87% (95% CI 79.7 to 94.9). The difference was statistically significant (p = 0.029). The mean Oxford Hip Score, Hip disability and Osteoarthritis Outcome Score, and Forgotten Joint Score were 45 (29 to 48), 89 (43 to 100), and 84 (19 to 100), respectively. The mean scores for each of the five domains of the EuroQol five-dimension three-level questionnaire were 1.2, 1.0, 1.2, 1.3, and 1.1, and mean overall score 82.6 (50 to 100). Ultrasound showed no pseudotumour. Mean cobalt and chromium levels were 32.1 nmol/l (1 to 374) and 45.5 nmol/l (9 to 408), respectively. This study shows that BHRs provide excellent survivorship and functional outcomes in young male patients. At 20 years, soft-tissue imaging and serum metal ion studies suggest that a metal-on-metal resurfacing implant can be well tolerated in a group of young patients.


[Teratogenic effects of arsenic on rats].
To further clarify the effects of arsenic on pregnant rats, fetuses, fetal cells and to verify the experimental results in vitro, and to study the developmental toxicity and its mechanisms of arsenic in depth. Teratogenic methods in vivo combined with scanning electron microscopy and histochemical techniques were used in rats. No apparently toxic symptoms or deaths were observed in rats exposed to various doses of arsenic trioxide of 0, 1, 4 and 10 mg/kg. Fetal death rates in rats were 10.2%, 17.4%, 24.8% and 61.1%, proportions of skeletal deformity were 1.89%, 3.12%, 17.28% and 78.57%, and embryo deformity rates were 3.16%, 5.32%, 13.79% and 40.48% 48 hours after exposure, respectively for those exposed to various doses of arsenic. Damage in fetal epidermal cells and cavities of different size between cell surface and cell junction in multiple sites could be seen under scanning microscope. Dysrhaphia, body anomalies and small limb bud, etc. were main manifestation of embryo deformity. There was a close relationship between nitric oxide and anomalies in fetal development, with a dose-response pattern, showed by inducible nitric oxide synthetase (iNOS) histochemical examinations. In vivo staining showed that excessive apoptosis could be induced by arsenic. And, teratogenic mechanisms of arsenic was analyzed in various ways. Arsenic has certain teratogenic effects on rats in vivo.


Tumor-infiltrating B lymphocytes as a potential source of identifying tumor antigen in human lung cancer.
Functional studies using freshly isolated tumor-infiltrating B lymphocytes(TIB) are difficult to perform and interpret. Here we document a novel function of TIB using fresh human lung cancer tissues engrafted in SCID mice; they are at activated state and produce tumor-specific antibodies in tumor microenvironment: (a) TIB engrafted in SCID mice produced human IgG; (b) IgG derived from TIB highly bound intracellular and membrane-bound antigens of autologous cancer cells; and (c) less recognition of autoantigens expressed on normal lymphocytes by IgG derived from TIB compared with IgG from the serum of the patient. On the basis of the novel findings presented in this study, we modified the original serological analysis of antigens by recombinant cDNA expression cloning design in a patient with lung cancer who expressed unusually favorable clinical evolution and analyzed humoral immunity against identified mutated p53 antigen. This study provides the first demonstration that antibodies derived from TIB recognize tumor antigens by serological analysis of antigens by recombinant cDNA expression cloning methodology and circulating anti-p53 antibodies in sera derived from TIB in tumor microenvironment. Our approach using TIB may allow the identification of key antigens in the humoral cancer-related immune system.


Increased vasopressin and adrenocorticotropin responses to stress in the midluteal phase of the menstrual cycle.
Accumulating evidence indicates that gonadal steroids modulate functioning of the hypothalamic-pituitary-adrenal (HPA) axis, which has been closely linked to the pathophysiology of anxiety and depression. However, the effect of the natural menstrual cycle on HPA axis responsivity to stress has not been clearly described. In nine healthy women, metabolic and hormonal responses to treadmill exercise stress during the early follicular phase of the menstrual cycle, when gonadal steroid levels are low, were compared with responses in the midluteal phase of the cycle, when both progesterone and estrogen levels are relatively high. Exercise intensity was gradually increased over 20 min to reach 90% of each subject's maximal oxygen consumption during the final 5 min of exercise. Basal plasma lactate, glucose, ACTH, vasopressin, oxytocin, and cortisol levels were similar in the two cycle phases. However, in response to exercise stress, women in the midluteal phase had enhanced ACTH (P < 0.0001), vasopressin (P < 0.01), and glucose (P < 0.001) secretion. These findings suggest that relatively low levels of gonadal steroids during the early follicular phase of the menstrual cycle provide protection from the impact of stress on the HPA axis.


Surface Molecularly Imprinted Polymer Film with Poly(p-aminothiophenol) Outer Layer Coated on Gold Nanoparticles Inner Layer for Highly Sensitive and Selective Sensing Paraoxon.
This paper presents the fabrication of a molecularly imprinted, polymer-based disposable electrochemical sensor for paraoxon (PO) determination. The sensor was based on a screen-printed carbon electrode (SPCE) modified with a surface molecularly imprinted poly (p-aminothiophenol) (PATP)/gold nanoparticles (AuNPs) composite film, which consisted of a PATP outer layer and an AuNPs inner layer. We report a novel strategy, combining surface molecularly imprinting and self-assembly directed electro-polymerization with high densely imprinting PO molecules in the PATP/AuNPs film. Firstly, AuNPs were in situ electrodeposited at the electrode surface, and then assembled with electropolmerizable functional monomer p-aminothiophenol (ATP). Subsequently, PO molecules were assembled onto the ATP monolayer-modified AuNPs, forming a basis of surface molecular imprinting. After that, replenished PO molecules were embedded in the PATP/AuNPs film by PO and the ATP molecular self-assembly directed electro-polymerization in the polymerization precursor mixture. The resulting imprinted PATP/AuNPs/SPCE possesses high sensitivity, affinity, and selectivity toward PO, with a low detection limit of 1 × 10-9 M. The proposed sensor was successfully applied for the determination of PO in fruit and vegetables, giving satisfactory recoveries. The strategy reported herein can be further expected to fabricate various molecular imprinted sensors for the determination of other pesticide residuals.


T-cell-mediated immune response in murine malaria: differential effects of antigen-specific Lyt T-cell subsets in recovery from Plasmodium yoelii infection in normal and T-cell-deficient mice.
For analysis of the role of immune T cells in protective immunity against murine malaria, Plasmodium yoelii-immune Lyt T-cell subsets were functionally characterized in vitro and in vivo. Selected Lyt2- and Lyt2+ T cells from P. yoelii-immune C57BL/10 mice differed in their capability to proliferate in response to P. yoelii antigen in vitro. Only the Lyt2- T-cell population produced T-cell growth factor upon restimulation, and none of the selected T-cell subsets produced detectable amounts of macrophage activating factor. Lyt2- but not Lyt2+ lymphocytes were capable of transferring protection to normal C57BL/10 mice. When transferred into T-cell-deficient C57BL/6-nu/nu mice, adoptive resistance to P. yoelii by Lyt2- lymphocytes was only demonstrable after prior reconstitution of recipients with normal T cells. These results suggest an interaction between P. yoelii-immune Lyt2- T cells and normal T lymphocytes via T-cell growth factor in the development of protective immunity to malaria.


Design of fMet-tRNA and calculation of its bonding properties by quantum mechanics.
It is generally accepted that initiation of protein synthesis in Escherichia coli starts with formyl-methionine, directed by the codons of AUG or GUG. In one case, reinitiating on the mRNA of amber mutants of UUG is used as the initiation codon. Early studies indeed showed that the triplets AUG, GUG, and UUG are the most effective in stimulating fMet-tRNA binding to ribosomes in vitro. We study the bonding properties of fMet-tRNA. The structure was optimized at the Hartree-Fock (HF) level of theory. We performed nonempirical quantum mechanical calculations at the HF and BLYP and B3LYP/3-21G, 6-31G, and 6-31G* levels of theory in the gas phase and water solvent at temperature of 310 K. Finally, we employed the density functional theory (DFT) and HF to calculate nuclear magnetic resonance spectra and infrared spectra.


Integration host factor is required for positive regulation of the tdc operon of Escherichia coli.
A 14-bp segment in the promoter region of the tdcABC operon of Escherichia coli shows sequence identity with the consensus binding site for the E. coli integration host factor (IHF). In an himA (IHF-deficient) strain, expression of beta-galactosidase from a tdcB'-'lacZ protein fusion plasmid was about 10% of that seen with an isogenic himA+ strain. Threonine dehydratase activity from the chromosomal tdcB gene in the himA mutant was also about 10% of the wild-type enzyme level. Two different mutations introduced into the putative IHF-binding site in the fusion plasmid greatly reduced the plasmid-coded beta-galactosidase activity in cells containing IHF. In vitro gel retardation and DNase I footprinting analyses showed binding of purified IHF to the wild-type but not to the mutant promoter. IHF protected a 31-bp region between -118 and -88 encompassing the conserved IHF consensus sequence. These results suggest that efficient expression of the tdc operon in vivo requires a functional IHF and an IHF-binding site in the tdc promoter.


A safe percutaneous method to recanalize sharp venous occlusions that resist the passage of guidewires.
It is occasionally necessary to cross a sharp venous occlusion (a thin web of tissue without a lumen) and sometimes it is not possible to cross the stricture with a wire, so puncturing it with a sheathed needle is then required to enable stenting. Other methods have been reviewed; however, we think this balloon technique makes it a safer procedure.


Determination of melatonin and monoamines in rat pineal using reversed-phase ion-interaction chromatography with fluorescence detection.
A method is reported for the ion-interaction, reversed-phase separation of 24 compounds (chiefly monoamines) arising from the metabolism of tyrosine and tryptophan. These compounds were separated as two groups. The first group comprised 3,4-dihydroxyphenylethylene glycol, tyrosine, 3-methoxy-4-hydroxyphenyl glycol, 5-hydroxytryptophan, norepinephrine, 3,4-dihydroxyphenylacetic acid, epinephrine, 5-hydroxyindole-3-acetic acid, homovanillic acid, 5-hydroxytryptophol, dopamine, tryptophan. N-acetylserotonin, N-acetyltryptophan, 5-methoxytryptophan and serotonin. The mobile phase consisted of a 6.8:93.2 (v/v) mixture of acetonitrile and an aqueous solution containing 0.16 M ammonium phosphate, 0.06 M citric acid, 0.15 mM disodium EDTA, 10 mM dibutylamine and 6 mM sodium 1-octanesulphonate at pH 4.50. The second group of compounds comprised 6-hydroxymelatonin, 5-methoxyindole-3-acetic acid, indole-3-acetic acid, 5-methoxytryptamine, tryptamine, 5-methoxytryptophol, melatonin and tryptophol. The mobile phase consisted of a 16:84 (v/v) mixture of acetonitrile and an aqueous solution containing 0.05 M ammonium phosphate, 0.05 M citric acid, 0.15 mM disodium EDTA, 25 mM dibutylamine and 5 mM sodium 1-octanesulphonate at pH 5.30. Detection was by fluorescence measurement (lambda ex = 280 nm, lambda em = 340 nm). The proposed method exhibited linear calibration over the biochemically significant concentration range, with detection limits in the 10-200 pg range. Excellent precision for peak areas and retention times was observed, even over a period of 24 h. The applicability of amperometric detection (at 0.72V) is also demonstrated. The method is applied to the determination of monoamines in individual rat pineals. Low nanogram levels of tyrosine, norepinephrine, 5-hydroxyindole-3-acetic acid, tryptophan, serotonin and 6-hydroxymelatonin, and picogram levels of 5-hydroxytryptophan, 5-hydroxytryptophol, 5-methoxyindole-3-acetic acid, indole 3-acetic acid, 5-methoxytryptophol and melatonin were indicated in most of the samples.


Computerized nuclear morphometry for the prediction of inguinal lymph nodes metastases in squamous cell carcinoma of the vulva.
This study evaluated the sensitivity and specificity of computerized morphometry in predicting lymph nodes metastases (LNM) in patients with squamous cell carcinoma (SCC) of the vulva. Histologic samples obtained from 20 consecutive cases of SCC of the vulva with positive inguinal LNM were morphometrically assessed and compared with samples from 20 consecutive cases of vulvar SCC negative for LNM. Computerized morphometry was performed on tumor cells and on adjacent nonneoplastic epithelial cells located 2-4 mm from the tumor margins. Computerized morphometric variables of tumor cell nuclei in patients with negative LNM significantly differed from those in patients with positive LNM. Morphometric differences in nuclear size and contour regularity were detected when comparing the nonneoplastic nuclei adjacent to the tumor of both groups. Multivariate analysis showed that the only independent predictors of LNM were the depth of the invasion (P= 0.005) and the mean nuclear roundness of the nonneoplastic nuclei adjacent to the tumors (P= 0.008). Using these variables, a discriminant score revealed a sensitivity of 90% and a specificity of 86.4% for predicting LNM in SCC of the vulva. Our data suggest that cells from the primary tumors with LNM differ morphometrically from primary tumors with no LNM. In addition, normal epithelial cells adjacent to the tumor express morphometric changes between the two groups. The results of our study justify the need for a prospective study of a larger number of patients to evaluate the reproducibility and the clinical use of the data.


Signaling modulation of bile salt-induced necrosis in isolated rat hepatocytes.
Hydrophobic bile salts induce either necrosis or apoptosis depending on the severity of the injury caused by them. Since bile salt-induced apoptosis is influenced by Ca2+- and protein kinase-signaling pathways, and both necrosis and apoptosis share common initiating mechanisms, we analyzed whether these signaling cascades also influence bile salt-induced necrosis in isolated rat hepatocytes. Taurochenodeoxycholate (TCDC, 0.25-1.50 mM, 2 h) reduced, in a dose-dependent manner, the percentage of viable hepatocytes, and increased the release of the cytosolic enzyme, lactate dehydrogenase (LDH) and alanine aminotransferase (ALAT), and that of the plasma membrane enzyme, alkaline phosphatase (AP). The PKC inhibitors, H7 (100 microM) and chelerythrine (2.5 microM), both prevented significantly TCDC-induced necrosis. On the contrary, the PKA activator, dibutyryl-cAMP, exacerbated TCDC-induced cell damage in a dose-dependent manner; this effect was more likely due to cAMP-mediated PKA activation, as the PKA inhibitor, KT5720 (1 microM), counteracted this effect. Instead, the intracellular Ca2+ chelator, BAPTA/AM (20 microM), was without effect. TCDC (1 mM) increased lipid peroxidation from 0.7 +/- 0.2 to 7.5 +/- 0.9 nmol of malondialdehyde per mg of protein, p < 0.001; the addition of the free radical scavenger, diphenyl-p-phenylendiamine, completely blocked this increase and prevented significantly TCDC-induced necrosis. PKC inhibition induced only a slight attenuation of TCDC-induced lipid peroxidation. Possible mechanisms accounting for the modulatory effect of signal transduction pathways on TCDC-induced necrosis, including signaling influence on TCDC transport events and TCDC-induced oxidative stress, are discussed.


ROCK1 and 2 differentially regulate actomyosin organization to drive cell and synaptic polarity.
RhoGTPases organize the actin cytoskeleton to generate diverse polarities, from front-back polarity in migrating cells to dendritic spine morphology in neurons. For example, RhoA through its effector kinase, RhoA kinase (ROCK), activates myosin II to form actomyosin filament bundles and large adhesions that locally inhibit and thereby polarize Rac1-driven actin polymerization to the protrusions of migratory fibroblasts and the head of dendritic spines. We have found that the two ROCK isoforms, ROCK1 and ROCK2, differentially regulate distinct molecular pathways downstream of RhoA, and their coordinated activities drive polarity in both cell migration and synapse formation. In particular, ROCK1 forms the stable actomyosin filament bundles that initiate front-back and dendritic spine polarity. In contrast, ROCK2 regulates contractile force and Rac1 activity at the leading edge of migratory cells and the spine head of neurons; it also specifically regulates cofilin-mediated actin remodeling that underlies the maturation of adhesions and the postsynaptic density of dendritic spines.


Disparities in Awareness of Myocardial Infarction and Stroke Symptoms and Response Among United States- and Foreign-Born Adults in the National Health Interview Survey.
Background Atherosclerotic cardiovascular disease, defined as nonfatal myocardial infarction (MI), coronary heart disease death, or fatal or nonfatal stroke, is the leading cause of death in the United States. MI and stroke symptom awareness and response reduce delays in hospitalization and mortality. Methods and Results We analyzed cross-sectional data from the 2014 and 2017 National Health Interview Surveys on US- and foreign-born adults from 9 regions of birth (Europe, South America, Mexico/Central America/Caribbean, Russia, Africa, Middle East, Indian subcontinent, Asia, and Southeast Asia). The outcomes were recommended MI and stroke knowledge, defined as knowing all 5 symptoms of MI or stroke, respectively, and choosing "call 9-1-1" as the best response. We included 63 059 participants, with a mean age 49.4 years; 54.1% were women, and 38.5% had a high school education or less. Recommended MI and stroke knowledge were highest in US-born people. In both 2014 and 2017, MI knowledge was lowest in individuals born in Asia (23.9%±2.5% and 32.1%±3.3%, respectively), and stroke knowledge lowest for the Indian subcontinent (44.4%±2.4% and 46.0%±3.2%, respectively). Among foreign-born adults, people from Russia and Europe had the highest prevalence of recommended MI knowledge in 2014 (37.4%±5.4%) and 2017 (43.5%±2.5%), respectively, and recommended stroke knowledge was highest in people from Europe (61.0%±2.6% and 67.2%±2.5%). Improvement in knowledge was not significant in all groups between 2014 and 2017. Conclusions These findings suggest a disparity in MI and stroke symptom awareness and response among immigrants in the United States. Culturally tailored public health education and health literacy initiatives are needed to help reduce these disparities in awareness.


Anticonvulsants lamotrigine and riluzole disrupt maternal behavior in postpartum female rats.
Maternal behavior is a highly motivated and well-organized social behavior. Previous studies have reported that anticonvulsants are frequently used in postpartum bipolar disorder. However, the maternal disruptive effect of the anticonvulsants has not been explored. The purpose of the present study was to examine the effect of anticonvulsants lamotrigine and riluzole on maternal behavior in postpartum female rats. On postpartum Day 3, Sprague-Dawley mother rats were given a single intraperitoneal injection of vehicle, lamotrigine (15, 25, 35 mg/kg), or riluzole (2, 4, 8 mg/kg). Maternal behavior was tested 30 min before and after injection. Animals treated with lamotrigine or riluzole had a longer pup retrieval latency, retrieved fewer pups into the nest, spent less time on nursing pups, as well as on building the disturbed nest, and animals treated with riluzole spent less time on pup licking. Whereas, the drugs in the tested doses did not shorten the total duration of behavior unrelated to maternal behavior. Overall, these data indicate that lamotrigine and riluzole disrupt major components of maternal behavior in postpartum female rats, but do not inhibit the behaviors unrelated to maternal behavior, which indicates that the maternal disruptive effect is not due to nonspecific sedative effect.


Reinduction of hyponatremia improves survival in rats with myelinolysis-related neurologic symptoms.
Brain myelinolysis occurs after excessive correction (delta SNa > 20 mEq/1/24 hours) of chronic hyponatremia. However, we showed recently that the mechanisms leading to brain myelinolysis remain reversible. Indeed, reinduction of the hyponatremia by water administration despite 12 hours of sustained excessive correction could prevent the development of demyelination in rats still asymptomatic at that time. Whether this therapeutic maneuver could be also beneficial to rats with preexisting myelinolysis-related neurologic symptoms is unknown. Therefore we evaluated here the effect of reinduction of the hyponatremia on the survival and on brain damage in rats presenting obvious neurologic symptoms after excessive correction of hyponatremia. After 3 days of severe hyponatremia induced by 2.5 D-glucose in water and continuous infusion of AVP, rats were submitted to a large correction (delta SNa approximately 30 mEq/l) by 2 i.p. injections of hypertonic saline given over 24 hours. In group I (n = 15) the rats developing neurologic symptoms during the first 24 hours of correction received one i.p. injection of distilled water which rapidly decreased the natremia to a final correction gradient <20 mEq/l/24 hour. In group II (n = 13, controls) the symptomatic rats were left permanently overcorrected. In group I, after water administration, the neurological manifestations were generally attenuated or disappeared. Seven of the 15 rats (47%) in this group survived up to day 10 with a mean survival time of 7.5 +/- 2 days, an outcome clearly improved as compared to group II (controls): only 1 of the 13 rats (7%, p < 0.03) was still alive on day 10 and the mean survival time was 3.3 +/- 2 days (p < 0.001) in this group II. The duration of the symptoms also influences the prognosis. In group I, in 9 rats the water administration was performed 4 hours after symptoms onset. These rats had a better outcome than the 6 rats with more sustained (8-10 hours) neurologic symptoms before water loading. Brain analysis in the 7 surviving rats of group I demonstrated demyelinating lesions in only 2 of them, suggesting the reversibility of the process even when neurologic manifestation developed. In conclusion, after exposure to an excessive correction of chronic hyponatremia, even when rats have developed myelinolysis-related neurologic symptoms, hypotonic fluids administration could improve survival and could prevent the subsequent development of brain myelinolysis.


Solvation and rotation dynamics in the trihexyl(tetradecyl)phosphonium chloride ionic liquid/methanol cosolvent system.
The interactions and solvent structure in trihexyl(tetradecyl)phosphonium chloride ionic liquid ([P(14,6,6,6)(+)][Cl(-)], "PIL-Cl")/methanol (MeOH) solutions across the entire range of mole fraction PIL-Cl (x(IL) = 0-1) are discussed. Viscosity and conductivity measurements are used to characterize the bulk solvent properties. At x(IL) < 0.1, the log(η) data show a nonlinear dependence on mole fraction in contrast to the data for x(IL) > 0.1 where the data vary linearly with mole fraction. Conductivity data show a maximum at x(IL) = 0.03 in good agreement with conductivity measurements in imidazolium ILs. Steady-state and time-resolved fluorescence spectroscopies were used to measure the equilibrium, lifetime, and rotational response of coumarin 153 (C153) in neat and MeOH cosolvent modified PIL-Cl. The collective set of data depicts the formation of an increasingly aggregated solvent structure that changes in proportion to the amount of PIL-Cl present in MeOH. Average solvation and rotation times are found to scale with solution viscosity. At x(IL) values of 0.02-0.2, the rotation times are at or near the hydrodynamic stick limit, whereas for x(IL) > 0.2 rotation times drop to between 40 and 70% of the stick limit, consistent with the IL literature. In this cosolvent system, the most dramatic changes in solution behavior occur between 0 and 10% PIL-Cl.


Dilated cardiomyopathy of Becker-type muscular dystrophy with exon 4 deletion--a case report.
The authors report a 47-year-old man with Becker-type muscular dystrophy presenting with dilated cardiomyopathy. Left ventriculography showed diffuse severe hypokinesia: left ventricular end-diastolic volume index 193 mL/m2, left ventricular end-systolic volume index 143 mL/m2, and left ventricular ejection fraction 26%. Skeletal muscle biopsy demonstrated a dystrophic process. Genetic analysis revealed a deletion of exon 4. There was a difference in immunostaining pattern between skeletal muscles and cardiac muscles. Severe cardiac dysfunction in this case may be associated with the damage in dystrophin-deficient fibers.


Identification of a small-molecule inhibitor of the interaction between Survivin and Smac/DIABLO.
The protein Survivin is selectively overexpressed in a variety of cancers, but not in normal tissues. It has been reported to be involved in cell survival and cell division. However, the molecular mechanisms involved in its function are not clear, although several binding partner proteins have been proposed to date. Here, we report the identification of a novel small molecule Survivin antagonist, which disrupts the Survivin-Smac/DIABLO interaction in cells. In order to identify Survivin-directed antagonists, we developed a high-throughput screening system based on AlphaScreen technology, which allows the identification of small molecules with the ability to inhibit the interaction of Survivin with Smac/DIABLO or INCENP in vitro. We screened chemical libraries, generated in-house, using this system and identified a 5-deazaflavin analog (compound 1) as a hit compound that selectively inhibited the interaction of Survivin with Smac/DIABLO but not INCENP. In cultured cells, compound 1 abrogated the formation of the complex between Survivin and Smac/DIABLO. In addition, this compound was able to sensitize cultured cells to doxorubicin-mediated DNA damage stress and synergistically enhance apoptotic cell death. Thus, the small-molecule inhibitor described here may serve as a proof-of-principle agent for discriminating between the multiple functions of Survivin.


Borderline Metal Centers on Nonporous Metal-Organic Framework Nanowire Boost Fast Li-Ion Interfacial Transport of Composite Polymer Electrolyte.
Metal-organic frameworks (MOFs) fillers are emerging for composite polymer electrolytes (CPEs). Enhancing Lewis acid-base interaction (LABI) among MOFs, polymer and Li-salt is expected to promote Li+ -transport. However, it is unclear how to customize a strong LABI interface. The large surface-area of classical MOFs also interferes with clarifying the LABI influence on Li+ -transport. Herein, Bi3+ as metal centers to design colloidal-dispersed nonporous MOFs (Bi/HMT-MOFs) nanowire with a surface-area of only 17.13 m2 g-1 to prepare polyethylene oxide (PEO)-based CPEs (BMCPE) is chosen. The nonporous feature can exclude the surface-area effect on Li+ -transport. More interestingly, Bi3+ is a typical borderline acid, which can interact with both hard-basic PEO and soft-basic Li-salt anion. Accordingly, Bi/HMT-MOFs are uniformly dispersed in the BMCPE to form a strong LABI interface with PEO and Li-salt, promoting Li-salt dissociation and providing rapid Li+ -transport channels. Despite the ultralow surface-area of Bi/HMT-MOFs, BMCPE exhibits significantly enhanced ion-conductivity and Li+ transference number, which completely rival traditional MOFs-filled CPEs. BMCPE also enables symmetric and full cells with excellent high-rate performance and long-term cycling stability. In contrast, when Bi3+ sites are obscured, electrochemical performances are obviously decreased. Therefore, employing borderline metal centers will be an effective strategy to construct a LABI interface for high-performance MOFs-filled CPEs.


Expression of small-conductance calcium-activated potassium channels (SK3) in skeletal muscle: regulation by muscle activity.
The type 3 small conductance calcium-activated potassium channel (SK3) is expressed in embryonic and adult denervated skeletal muscles where it contributes to hyperexcitability. This study aimed at determining the role of muscle activity in regulating SK3 channels. Soleus muscles of adult rats were denervated by cutting the sciatic nerve. In reinnervation studies, the soleus nerve was crushed: in one group, muscles were reinnervated with electrically silent axons, by chronic sciatic nerve perfusion with tetrodotoxin. Several groups of denervated muscles were subjected to chronic direct electrical stimulation, using either fast (100 Hz) or slower patterns (20 or 30 Hz). The SK3 mRNA and protein levels in soleus muscle were determined by reverse transcriptional-PCR, Western blot and immunofluorescence. Both denervated and reinnervated-paralysed soleus muscles displayed similar up-regulation of SK3 mRNA and protein. Reinnervation with electrically active axons instead inhibited SK3 up-regulation. Chronic muscle direct stimulation in vivo, irrespective of the pattern used, reversed the denervation-induced up-regulation of SK3 expression or prevented it when initiated at the time of denervation. Chronic electrical stimulation of denervated muscles also completely prevented the development of the after-hyperpolarization (AHP) following the action potential, normally induced in the muscle fibres by denervation. We conclude that action potential activity evoked by motor neurones in muscle fibres is both necessary and sufficient to account for the physiological down-regulation of SK3 channels in the non-junctional membrane of skeletal muscle.


Transinfection of Wolbachia in planthoppers: nymphal injection of cultured Wolbachia and infection dynamics.
Wolbachia species are intracellular symbionts that cause reproductive alterations in arthropods. Transinfection experiments have been performed in many arthropod species to elucidate the interaction between Wolbachia and a new host. To ease transinfection of this bacterium to new arthropod hosts, we introduced two techniques: nymphal injection instead of embryonic injection and the use of a cultured source of Wolbachia instead of direct transfer from donors to recipients. Wolbachia in the small brown planthopper Laodelphax striatellus was cultivated in a cell line and injected into the nymphal body cavity of the brown planthopper Nilaparvata lugens together with the cells. By using these techniques, two transinfected planthopper lines were obtained. In one line, Wolbachia disappeared after several generations; in the other line, Wolbachia was retained for >7 yr. Infection rates in this latter transinfected line were approximately 80% in early generations after transinjection but decreased to <10% through 40-60 generations. Subsequent selection for Wolbachia-infected females in this line did not increase the infection rate as a temporary effect. Thus, this transinfected line of N. lugens showed cytoplasmic incompatibility, although the incompatibility level was lower than in L. striatellus, the original host. The method of transinfection presented herein is useful for transmitting intracellular symbionts between small arthropod hosts.


Structure characterization and antioxidant activity of carboxymethylated polysaccharide from Pholiota nameko.
The objective of this paper was to prepare the carboxymethyl polysaccharide of Pholiota nameko (CPPN) and further evaluate its structural characteristics, water solubility, and antioxidant activities. The optimal carboxymethylation conditions were as follows: 20% NaOH concentration, 1.5 hr alkalization, 4 hr etherification time, and 60°C with 2 g chloroacetic acid. Under these conditions, a substitution degree of 0.72 and a yield of 88.3% were achieved. The results of NMR and FT-IR spectra showed that carboxymethylation modification was successful. The protein, total sugar, and total phenol content decreased, while the molecular weight increased, and the monosaccharide composition content and surface structure changed upon carboxymethylation. Compared with unmodified polysaccharides, CPPN had significant antioxidant activity and water solubility. Thus, this study could provide a potential antioxidant for functional foods and pharmaceuticals. PRACTICAL APPLICATIONS: Pholiota nameko is an edible and medicinal mushroom, rich in carbohydrates, cellulose, and minerals. Pharmacological studies have shown that P. nameko polysaccharides have many medicinal effects on the human body. In this study, CPPN was synthesized by carboxymethylation of P. nameko polysaccharides. Through structural characterization, antioxidant activity, and solubility studies, carboxymethylated polysaccharides shown significant antioxidant properties and increased solubility. Therefore, this research lays the foundation for the high-value application of P. nameko polysaccharides in functional food and pharmaceutical industry.


Differential induction of midazolam metabolism in the small intestine and liver by oral and intravenous dexamethasone pretreatment in rat.
1. Midazolam is metabolized in the rat by CYP3A enzymes to 4-OH-midazolam (4-OH-MDZ) and 1'-OH-midazolam (1'-OH-MDZ). The induction of midazolam metabolism was studied in male Wistar rats treated with dexamethasone (50 mg kg(-1) day(-1)) during 4 days via the oral or intravenous routes. Microsomes were prepared from the liver and the proximal small intestine and in vitro metabolism of midazolam was determined. In addition, CYP3A1- and CYP3A2-like protein levels were measured by gel electrophoresis and immunoblotting. 2. The V(max)'s (mean SEM) for 4-OH-MDZ and 1'-OH-MDZ formation were much lower in intestinal (0.078 +/- 0.002 and 0.074 +/- 0.002 microM min(-1) mg(-1) protein, respectively) compared with hepatic microsomes prepared from the uninduced rat (0.870 +/- 0.007 and 0.310 +/- 0.020 microM min(-1) mg(-1) protein, respectively). Induction by oral or intravenous dexamethasone pretreatment led to significant increases in V(max) for 4-OH-MDZ and 1'-OH-MDZ by both intestinal and hepatic microsomes. Oral dexamethasone pretreatment via the oral route resulted in a more pronounced increase in V(max) compared with intravenous administration of the inducer. 3. CYP3A1 and CYP3A2 protein levels in liver microsomes were significantly increased following oral (3.7- and 3.2-fold, respectively) or intravenous (2.6- and 2.1-fold, respectively) pretreatment with dexamethasone. On the contrary, only oral dexamethasone pretreatment resulted in a significant change in intestinal CYP3A2-like protein (7.3-fold). A slight difference in the migration distance of the immunoreactive band for CYP3A2 was also observed for intestinal microsomes. 4. These results suggest that intestinal CYP3A enzymes in the rat differ from hepatic CYP3A1 and CYP3A2. They also demonstrate that systemic dexamethasone administration can induce intestinal microsome activity.


[The diagnostic value of radial multiplanar reformatting (MPR) in the CT-diagnosis of pulmonary embolism].
To evaluate the diagnostic value of radial reformatting of axial multislice CT (MS-CT) data sets in the presumptive diagnosis of pulmonary embolism. In 126 cases with the presumptive diagnosis of an acute pulmonary embolism, 4- and 8-slice axial MS-CT data sets were radially reformatted. The pulmonary vessels were evaluated by five experienced radiologists who determined the number of thrombi at the level of the segmental and subsegmental arteries. A pulmonary artery was considered as thrombosed if it showed at least one unambiguous filling defect on two consecutive sections. It was determined whether the artery was uniformly opacified without filling defects, contained thrombotic material or was completely occluded by a blood clot. Pulmonary vessels that could not be evaluated because of moving artifacts or inadequate opacification were classified as negative. Finally, all experts gave a subjective estimate concerning the simplicity of the diagnosis for both reconstructions. The diagnosis of thromboembolism at a subsegmental level was significantly different for axial sections and radial reconstruction: at 4-slice CT, 77 subsegmental thromboembolisms were found in axial sections vs. 98.6 in radial reconstructions; at 8-slice CT, 23.6 in axial sections vs. 31.2 in radial reconstructions; and using the combined evaluation, 103.2 in axial sections vs. 130.6 in radial reconstructions, with a significance of p = 0.043. The five judges did not find any significant differences in the number of the blood clots in each pulmonary segment. Concerning the simplicity of the diagnosis, the subjective judgment classified the radial reconstructions as better in 49.7 % and the axial sections as better in 22 %. The radial reconstructions of the 4-slice CT were classified as better in 45.7 % vs. 25 % and the 8-slice CT as better in 66 % vs. 16 %. Using radial multiplanar reformatting of the MS-CT data sets improves significantly the recognition of subsegmental pulmonary embolism.


Flukes without snails: advances in the in vitro cultivation of intramolluscan stages of trematodes.
In vitro cultivation of parasitic helminths, including the digenetic trematodes, has long been a valuable tool in medical and veterinary parasitology, permitting and/or facilitating the development of diagnostic reagents, chemotherapeutic agents, and vaccines and providing insights into naturally complex host-parasite interactions. In vitro cultivation of the intramolluscan stages of trematodes has been particularly challenging, given the ontogenic complexities involved in the production of multiple larval generations from germinal tissues through an asexual "budding" process. Recently, however, advanced larval development has been achieved by incorporating the Biomphalaria glabrata embryonic (Bge) cell line into cocultivation systems. Most notably, the entire intramolluscan cycle (from miracidium to cercaria) has been completed for the human blood fluke Schistosoma mansoni, while significant primary sporocyst development has been attained for several other digeneans including S. japonicum and Fascioloides magna. Here we review recent advances in the cultivation of several larval trematode species and discuss the potential use of this culture system for addressing fundamental questions of host-parasite compatibility.


Use of abasic site-containing DNA strands for nucleobase recognition in water.
Nucleobase recognition in water is successfully achieved by the use of an abasic site (AP site) as the molecular recognition field. We intentionally construct the AP site in DNA duplex so as to orient the AP site toward a target nucleobase and examine the complexation of 2-amino-7-methylnaphthyridine (AMND) with nucleobases at the AP site. AMND is found to selectively bind to cytosine (C) base with a 1:1 binding constant of >106 M-1, accompanied by remarkable quenching of its fluorescence. In addition to hydrogen bonding, a stacking interaction with nucleobases flanking the AP site seems responsible for the binding properties of AMND at the AP site. Possible use of AMND is also presented for selective and visible detection of a single-base alternation related to the cytosine base.


Direct X-ray and electron-beam lithography of halogenated zeolitic imidazolate frameworks.
Metal-organic frameworks (MOFs) offer disruptive potential in micro- and optoelectronics because of the unique properties of these microporous materials. Nanoscale patterning is a fundamental step in the implementation of MOFs in miniaturized solid-state devices. Conventional MOF patterning methods suffer from low resolution and poorly defined pattern edges. Here, we demonstrate the resist-free, direct X-ray and electron-beam lithography of MOFs. This process avoids etching damage and contamination and leaves the porosity and crystallinity of the patterned MOFs intact. The resulting high-quality patterns have excellent sub-50-nm resolution, and approach the mesopore regime. The compatibility of X-ray and electron-beam lithography with existing micro- and nanofabrication processes will facilitate the integration of MOFs in miniaturized devices.


Examining the link between information processing speed and executive functioning in multiple sclerosis.
Slowed information processing speed (IPS) is frequently reported in those with multiple sclerosis (MS), and at least 20% are compromised on some aspect of executive functioning also. However, any relationship between these two processes has not been examined. The Sternberg Memory Scanning Test, Processing Speed Index (WAIS-III), Delis Kaplan Executive Function System (D.KEFS), and Working Memory Index (WMS-III) were administered to 90 participants with MS. Their performance on the PSI was significantly below the normative scores but no deficits in memory scanning speed were evident. The initial response speed of the Sternberg and the PSI were more closely related to D.KEFS performance, particularly in timed tasks with a high cognitive demand (switching tasks). In contrast, memory scanning speed was related to working memory. This study reinforces the link between IPS and working memory in MS, and supports the suggestion that IPS is not a unitary construct.


Predictors of recurrence after surgical treatment of idiopathic progressive subglottic stenosis.
Idiopathic progressive subglottic stenosis is a rare cause of tracheal narrowing. Partial cricotracheal resection and anastomosis can cure idiopathic stenosis, even if some patients may require multiple interventions and experience voice and swallowing deterioration. We investigated risk factors for retreatment and assessed the impact of crico-tracheal resection on functional parameters. We conducted a retrospective multicentric study on 44 female patients (mean age 52.6 ± 13.1 years) affected by idiopathic stenosis and treated by crico-tracheal resection between 2002 and 2016. Functional outcomes after crico-tracheal resection were assessed by the airway-dyspnoea-voice-swallowing score (range 1-5, with "1" expressing normal and "5" completely altered function). Previous treatments, grade of stenosis, site, airway comorbidities, age and resection length were tested as predictors of postoperative complications and number of additional treatments, using bivariate and multivariate analysis. The overall decannulation rate was 97.3%. The dyspnoea score improved (mean variation 1.4 ± 1.0; p < 0.001), while voice and swallowing were negatively affected (mean variation 1.6 ± 0.9 and 0.5 ± 0.7, respectively; p < 0.001). Airway comorbidities were associated with a higher rate of complications (p < 0.05). Retreatments were more frequent in patients with postoperative complications (p < 0.05). The length of resection correlated with the number of subsequent treatments (R = 0.52; p < 0.01). At multivariate analysis, post-operative complications were predicted by comorbidities and disease stage (p < 0.05); number of retreatments was linked to the length of resection (p < 0.05) as well as with the application of mitomycin C (p < 0.001). Crico-tracheal resection for idiopathic progressive subglottic stenosis offers good functional results in terms of airway patency. These data suggest that a higher complication rate can be expected in patients affected by comorbidities. Moreover, more extensive surgical resection seems to be associated with the occurrence and number of subsequent retreatments. On the contrary, the local application of an anti-proliferative drug does not seem to be of use in preventing recurrences.


Symptomatic severe acute respiratory syndrome coronavirus 2 reinfection in a lupus patient treated with hydroxychloroquine: a case report.
Hydroxychloroquine and chloroquine have been used for hospitalized coronavirus disease 2019 patients because of their antiviral and anti-inflammatory function. However, little research has been published on the impact of the immunomodulatory effect of (hydroxy)chloroquine on humoral immunity. We report a case of symptomatic severe acute respiratory syndrome coronavirus 2 reinfection, diagnosed 141 days after the first episode, in a 56-year-old man of Black African origin treated with hydroxychloroquine for lupus erythematosus. No anti-severe acute respiratory syndrome coronavirus 2 IgG antibodies could be detected 127 days after the initial episode of coronavirus disease 2019. The treatment with hydroxychloroquine probably explains the decreased immune response with negative serology and subsequent reinfection in our patient. As humoral immunity is crucial to fight a severe acute respiratory syndrome coronavirus 2 infection, the use of (hydroxy)chloroquine is likely to have a detrimental effect on the spread of the virus. This case emphasizes that more needs to be learned about the role of antibodies in protecting against severe acute respiratory syndrome coronavirus 2 (re)infection and the role of (hydroxy)chloroquine on humoral immunity.