The neurobiology of antiepileptic drugs for the treatment of nonepileptic conditions.
Antiepileptic drugs (AEDs) are commonly prescribed for nonepileptic conditions, including migraine headache, chronic neuropathic pain, mood disorders, schizophrenia and various neuromuscular syndromes. In many of these conditions, as in epilepsy, the drugs act by modifying the excitability of nerve (or muscle) through effects on voltage-gated sodium and calcium channels or by promoting inhibition mediated by gamma-aminobutyric acid (GABA) A receptors. In neuropathic pain, chronic nerve injury is associated with the redistribution and altered subunit compositions of sodium and calcium channels that predispose neurons in sensory pathways to fire spontaneously or at inappropriately high frequencies, often from ectopic sites. AEDs may counteract this abnormal activity by selectively affecting pain-specific firing; for example, many AEDs suppress high-frequency action potentials by blocking voltage-activated sodium channels in a use-dependent fashion. Alternatively, AEDs may specifically target pathological channels; for example, gabapentin is a ligand of alpha2delta voltage-activated calcium channel subunits that are overexpressed in sensory neurons after nerve injury. Emerging evidence suggests that effects on signaling pathways that regulate neuronal plasticity and survival may be a factor in the delayed clinical efficacy of AEDs in some neuropsychiatric conditions, including bipolar affective disorder.


Serum and tissue expression of activin a in postmenopausal women with breast cancer.
Activins are growth factors involved in the control of cell proliferation and differentiation. Human breast tissues express immunoreactive activin subunits, and activin A is able to inhibit the replication of mammary cells in vitro. The aim of the present study was to evaluate 1) whether breast cancer expresses activin betaA subunit mRNA, 2) whether serum activin A levels are altered in postmenopausal women with breast cancer, and 3) how circulating activin A levels change after tumor removal. Four groups of women (n = 158) were enrolled for the present prospective study: two groups were composed of postmenopausal women with breast cancer (n = 74) or benign lesions (n = 15); the third was a control group composed of healthy postmenopausal women (n = 62); and the fourth group included healthy fertile women (n = 7) undergoing plastic surgery with removal of non-neoplastic mammary tissue. RT-PCR showed that betaA subunit mRNA was expressed in breast carcinoma, fibroadenoma, and normal mammary tissue, and the level of expression was higher in carcinoma than in normal tissue (P < 0.05). Dimeric activin A was detectable in homogenates of breast cancer tissue at concentrations twice as high as in non-neoplastic tissue (P < 0.01). In women with breast cancer, median serum activin A levels were significantly higher than in controls (P < 0.001). The high serum activin A levels in patients with breast cancer were not correlated with the presence of lymph node metastasis, tumor grade, or tumor diameter. After tumor excision, a significant decrease of activin A in the first and second postoperative days was observed (P < 0.01; Friedman's ANOVA). Conversely, activin A levels remained unchanged after plastic surgery in healthy women. The present results suggest that activin A is expressed and secreted in postmenopausal women with breast cancer. The pathophysiological and possible clinical implications of this finding remain to be investigated.


Use of cultured epithelia to study transport and its regulation.
Epithelial cells from a variety of species and organs form polarized epithelia in culture. When epithelia are grown on a porous surface, such as a millipore filter, transport can be studied using adaptations of standard techniques. In the few years in which cultured epithelia have been studied by transport physiologists, most work has been focused on identification and description of the differentiated transport exhibited by cultured epithelia. Epithelia formed by a continuous line of cells derived from pig kidney (LLC-PK1) exhibit sodium-coupled glucose transport similar to that of the proximal tubule and have vasopressin-sensitive adenylate cyclase that has been studied in great detail. Also of interest are epithelia formed by continuous lines of cells derived from amphibian kidney (A6) and from amphibian urinary bladder (TBM). Each line forms epithelia that have high electrical resistance and amiloride-sensitive sodium transport. Transport is stimulated by aldosterone and by cAMP or hormones that raise cell cAMP levels. In LLC-PK1 and in A6 epithelia, transport and the response to hormones can be manipulated by manipulating the culture conditions. Cultured epithelia have also been used to explore the cell biology of epithelia. Most interesting in this regard are studies of the development and maintenance of epithelial cell polarity. This approach should be especially valuable.


Overexpression of a novel imprinted gene, PEG10, in human hepatocellular carcinoma and in regenerating mouse livers.
Many of the promising applications of the microarray technology are pertinent to identifying abnormalities in gene expression that contribute to malignant progression. We developed a bioinformatics tool to identify differentially expressed genes in human hepatocellular carcinoma (HCC). This involved the construction of a liver EST database (http://lestdb.nhri.org.tw) and in silico verification of differentially expressed genes with a human hepatoma microarray database. The stringency of the search was reinforced with a statistical analysis. A novel imprinted gene, paternally expressed 10(PEG10) was identified as having an elevated level of expression in the majority of the HCC samples and was also induced to express during G2/M phase of regenerating mouse liver. Ectopic expression of PEG10 in 293T cells affects cell cycle progression. PEG10 is distributed in the cytosol and associates with the nuclear membrane. This is the first time that an imprinted gene has been found to reexpress in both human HCC and in the regenerating mouse liver. This result indicates that the induction of the paternally imprinted gene may play an important role during liver regeneration or carcinogenesis of the human hepatocyte. Understanding the molecular basis of the abnormal imprinting of PEG10 will shed new light on the process that leads to liver disease.


Autophagy is involved in the initiation and progression of Graves' orbitopathy.
Differentiation of orbital fibroblasts into mature adipocytes and subsequent accumulation of adipose tissue has been shown in the progression of Graves' orbitopathy (GO). Autophagy is involved in adipogenesis, but little is known about the role of autophagy in the initiation and progression of GO. The aim of this study is to investigate the role of autophagy in the pathogenesis of GO. Orbital adipose/connective tissue explants from patients with GO and from normal subjects, as well as isolated orbital fibroblasts, were analyzed. Adipogenesis was induced using differentiating medium with or without hydrogen peroxide, and autophagy was manipulated using bafilomycin A1 and Atg5-targeted short hairpin RNA (shRNA). Autophagosomes were identified by electron microscopy. Expression of autophagy-related genes and adipogenesis-related transcription factors were analyzed by real time reverse transcription-polymerase chain reaction and/or Western blot analysis. Lipid droplet accumulation was examined by Oil Red O staining. Autophagic vacuoles were more abundant in GO cells than in non-GO cells (p<0.05). Expression of autophagy-related genes was significantly higher in GO tissues and cells than in their non-GO counterparts, respectively. Interleukin-1β increased LC3-II, p62, and Atg7 protein in GO cells. Autophagosome accumulation was shown at day 4 of adipogenesis and decreased by day 10, along with lipid droplet formation. Expression of LC3 and p62 proteins increased within 48 hours of differentiation and diminished gradually from day 4 to 10. Bafilomycin A1 treatment and Atg5 knockdown by shRNA inhibited lipid droplet accumulation and suppressed expression of adipogenic markers. Autophagy was increased in GO tissue and cells compared to non-GO tissue and cells, suggesting that autophagy plays a role in GO pathogenesis. Autophagy manipulation may be a therapeutic target for GO.


[Dynamics of blood fibronectin level in myocardial infarction].
The fibronectin level in the blood of patients with myocardial infarction was measured at varying times from the onset of an angina pectoris attack in order to elucidate the diagnostic importance of blood fibronectin. At the same time these patients were examined over time for the blood content of myoglobin, MB creatine kinase protein and C-reactive protein playing a well-known role in the diagnosis. The blood concentrations of these substances reached the maximal values at different times of myocardial infarction. The mean concentrations of fibronectin in the blood of patients with myocardial infarction ranged within normal starting from the first till the 28th day since the onset of an angina pectoris attack. Moreover, the mean blood fibronectin level in myocardial infarction patients did not differ within the first-third days since the disease onset from that in patients with a clinical picture of unstable angina pectoris which was not accompanied by the development of myocardial infarction. Based on the data obtained it is concluded that measurement of blood fibronectin level does not play any diagnostic role in myocardial infarction. On the other hand, progressive increase in blood fibronectin level throughout 4 weeks starting from the 3d day of the disease and a significantly higher fibronectin content on the 28th day as compared with that on the 3d day is likely to mirror the activity of repair processes occurring in the myocardium.


Tympanometric measurements in conscious sheep - a diagnostic tool for pre-clinical middle ear implant studies.
To investigate and establish the use of tympanometry in conscious sheep to provide a means of objective assessment of tympanic membrane integrity, middle ear ventilation and functioning of the Eustachian tube (ET). After conditioning the sheep for four weeks, tympanometric measurements at 226 Hz were carried out weekly for 13 weeks. Before measurements, the external ear canal had been cleaned. Resultant curves were classified according to human reference values. Tests were performed on 12 female blackface sheep. After cleaning of the external ear canal under otoscopic control, tympanic membranes were intact with no evidence of acute or chronic middle ear inflammation, middle ear effusion or retraction. Cleaning ensured valid, objective and reproducible measurements. As the majority of normal tympanograms were notched without the appearance of any malformation, an additional tympanogram type (AN) was introduced. The notched appearance can most likely be explained by the anatomy of the middle ear of the sheep and the test frequency that was used. The current study demonstrated how tympanometry can be used to evaluate treatment modalities for middle ear and ET function in conscious sheep. This provided a large animal model for further human research in otology.


[Meningoencephalitis in the acute phase of measles. Report of 6 cases].
We present the clinical and laboratory manifestations of encephalitis following measles in six patients which were diagnosed during the epidemics that occurred in the city of São Paulo, Brazil, in 1997. We performed retrospective case analysis of the six patients diagnosed as having encephalitis due to measles. Encephalitis was diagnosed based on clinical grounds and on the cerebrospinal fluid (CSF) alterations. All the cases were serologically confirmed. Of 467 patients with measles who presented themselves for medical care at the Instituto de Infectologia Emílio Ribas six were diagnosed with encephalitis. Patient's age was 2 months to 28 years old. The most frequent symptoms were drowsiness and nuchal rigidity. CSF showed an increased of white cell count in all cases. Four patients were admitted to the intensive care unit. Two of them required mechanical ventilation. In only two patients did the computerized tomography show abnormalities. All showed good recovery without sequelae.


Whole-Genome Differentially Hydroxymethylated DNA Regions among Twins Discordant for Cardiovascular Death.
Epigenetics is a mechanism underlying cardiovascular disease. It is unknown whether DNA hydroxymethylation is prospectively associated with the risk for cardiovascular death independent of germline and common environment. Male twin pairs middle-aged in 1969-1973 and discordant for cardiovascular death through December 31, 2014, were included. Hydroxymethylation was quantified in buffy coat DNA collected in 1986-1987. The 1893 differentially hydroxymethylated regions (DhMRs) were identified after controlling for blood leukocyte subtypes and age among 12 monozygotic (MZ) pairs (Benjamini-Hochberg False Discovery Rate < 0.01), of which the 102 DhMRs were confirmed with directionally consistent log2-fold changes and p < 0.01 among additional 7 MZ pairs. These signature 102 DhMRs, independent of the germline, were located on all chromosomes except for chromosome 21 and the Y chromosome, mainly within/overlapped with intergenic regions and introns, and predominantly hyper-hydroxymethylated. A binary linear classifier predicting cardiovascular death among 19 dizygotic pairs was identified and equivalent to that generated from MZ via the 2D transformation. Computational bioinformatics discovered pathways, phenotypes, and DNA motifs for these DhMRs or their subtypes, suggesting that hydroxymethylation was a pathophysiological mechanism underlying cardiovascular death that might be influenced by genetic factors and warranted further investigations of mechanisms of these signature regions in vivo and in vitro.


The simian-virus-40 large-tumor antigen in replicating viral chromatin. A salt-resistant protein-DNA interaction.
The large tumor antigen (T antigen) is a genome regulation protein, coded by simian virus 40, that binds with high affinity to specific binding sites on viral DNA. The specifically bound T antigen is released from these sites in 0.2-0.3 M NaCl. Immunoprecipitation techniques were used to show that T antigen also dissociates in 0.2-0.3 M NaCl from mature viral chromatin but not from replicating viral chromatin. In fact, a considerable fraction of T antigen remains associated with replicating chromatin at NaCl concentrations as high as 1.2 M NaCl when most chromatin proteins, including histones, dissociate. However, T antigen binding to both replicating DNA and mature DNA is sensitive to intercalating drugs such as caffeine and ethidium bromide. We consider the possibility that the unexpectedly tight binding of T antigen to replicating DNA is related to the function that T antigen performs during viral DNA replication.


Carbon monoxide from neighbouring restaurants: the need for an integrated multi-agency response.
Carbon monoxide (CO) is a colourless, odourless toxic gas produced during incomplete combustion of carbon-based fuels. Most CO incidents reported to the UK Health Protection Agency (HPA) are due to faulty gas appliances, and legislation exists to ensure gas appliances are properly installed. We present three CO poisoning incidents of unusual origin reported to the HPA. In each, residents living above restaurants were poisoned after workers left charcoal smouldering overnight in specialist or traditional ovens whilst ventilation systems were turned off. This led to production of CO, which travelled through floorboards and built up to dangerous concentrations in the flats. Working with local authorities, these incidents were investigated and resolved, and work was conducted to prevent further occurrences. The novel nature of these CO incidents led to delays in recognition and subsequent remedial action. Although previously undescribed, it is likely that due to the number of residences built above restaurants and the rising popularity of traditional cooking methods, similar incidents may be occurring and could increase in frequency. Multi-agency response and reporting mechanisms could be strengthened. Awareness raising in professional groups and the public on the importance of correct ventilation of such appliances is vital.


Addition of azathioprine to corticosteroids does not benefit patients with IgA nephropathy.
The optimal treatment for IgA nephropathy (IgAN) remains unknown. Some patients respond to corticosteroids, suggesting that more aggressive treatment may provide additional benefit. We performed a randomized, multicenter, controlled trial to determine whether adding azathioprine to steroids improves renal outcome. We randomly assigned 207 IgAN patients with creatinine ≤2.0 mg/dl and proteinuria ≥1.0 g/d to either (1) a 3-day pulse of methylprednisolone in months 1, 3, and 5 in addition to both oral prednisone 0.5 mg/kg every other day and azathioprine 1.5 mg/kg per day for 6 months (n = 101, group 1) or (2) steroids alone on the same schedule (n = 106, group 2). The primary outcome was renal survival (time to 50% increase in plasma creatinine from baseline); secondary outcomes were changes in proteinuria over time and safety. After a median follow-up of 4.9 years, the primary endpoint occurred in 13 patients in group 1 (12.9%, 95% CI 7.5 to 20.9%) and 12 patients in group 2 (11.3%, CI 6.5 to 18.9%) (P = 0.83). Five-year cumulative renal survival was similar between groups (88 versus 89%; P = 0.83). Multivariate Cox regression analysis revealed that female gender, systolic BP, number of antihypertensive drugs, ACE inhibitor use, and proteinuria during follow-up predicted the risk of reaching the primary endpoint. Treatment significantly decreased proteinuria from 2.00 to 1.07 g/d during follow-up (P < 0.001) on average, with no difference between groups. Treatment-related adverse events were more frequent among those receiving azathioprine. In summary, adding low-dose azathioprine to corticosteroids for 6 months does not provide additional benefit to patients with IgAN and may increase the risk for adverse events.


Sinus of Valsalva thrombosis causing peripheral embolism.
A 44-year-old woman was admitted with the diagnosis of peripheral arterial emboli. Peripheral angiography demonstrated total occlusion of the popliteal artery. The obstruction was successfully resolved with a Fogarty arterial embolectomy catheter. Both transthoracic and transesophageal echocardiography showed a mobile, round thrombus in the noncoronary sinus of Valsalva. The patient did not accept surgery, and anticoagulation with warfarin was initiated. One month after treatment, transthoracic echocardiography demonstrated disappearance of the thrombus in the noncoronary sinus of Valsalva. The patient did not experience any recurrent episode of systemic embolization. This is a rare case of peripheral embolism caused by a thrombus in the noncoronary sinus of Valsalva without aneurysm.


The T cell factor/beta-catenin antagonist PKF115-584 inhibits proliferation of adrenocortical carcinoma cells.
Mutations of the beta-catenin (CTNNB1) gene are frequently found in adrenocortical tumors. This has important consequences to deregulate the expression of transcriptional targets of the Wnt pathway, which may contribute to tumorigenesis. The objective of the study was to investigate the effect of the small-molecule inhibitor of the T cell factor (Tcf)/beta-catenin complex PKF115-584 on beta-catenin-dependent transcription and proliferation of H295R adrenocortical tumor cells, which harbor mutations in CTNNB1 as well as the TP53 tumor suppressor gene. Immunofluorescence, transient transfection, proliferation assays, and flow cytometric analyses were used. Nuclear localization of beta-catenin and constitutive activation of beta-catenin-dependent transcription was observed in H295R cells. PKF115-584 dose-dependently inhibited beta-catenin-dependent transcription and H295R proliferation, even in the presence of increased steroidogenic factor-1 levels, which augment proliferation in this cell line. The drug had no effect on HeLa cells, a cell line in which the beta-catenin pathway is not activated. PKF115-584 decreased the percentage of H295R cells in S-phase and increased the percentage of apoptotic cells. Inhibitors of the Tcf/beta-catenin complex may prove useful in the treatment of adrenocortical tumors in which multiple genetic alterations have accumulated.


Urinary tract infection after vaginal surgery. Effect of prophylactic treatment with methenamine hippurate.
One hundred and nine consecutive patients undergoing surgery for uterovaginal prolapse followed by indwelling urinary catheter for 3 days were randomized for prophylactic treatment with methenamine hippurate (MH) or no MH prophylaxis. Significantly less bacteriuria occurred in the MH-treated patient group. In particular, the opportunistic hospital flora appeared to be suppressed by MH treatment. It is suggested that MH prophylaxis, 1 g three times daily, be used in gynecological surgery followed by short-term urinary catheterization.


Serum insulin-like growth factor I reference values for an automated chemiluminescence immunoassay system: results from a multicenter study.
Analysis of insulin-like growth factor I in serum (S-IGF-I) is an integral component in the diagnosis of GH-related disorders and is going to be of interest in the diagnosis and follow-up of many disorders. The objective of the present study was to develop cross-sectional reference values for S-IGF-I measured by an automated chemiluminescence immunoassay (Nichols Advantage). The study included samples from 3,961 healthy subjects (2,201 males, 1,760 females) aged 1 month to 88 years. Six laboratories were involved in this study and the samples were analyzed by one of seven automated immunoassay systems run in these laboratories. For data analysis, polynomial age and sex-specific models were fitted after transformation of S-IGF-I values. The results show the well-known age dependency of S-IGF-I levels. At ages <20, higher S-IGF-I levels were seen in girls with an estimated mean peak of 410 microg/l at age 14 and an estimated mean peak of 382 microg/l at age 16 in boys. Thereafter, a rapid decrease was seen to approximately 25 years of age, followed by a slow age-dependent decrease. In adulthood, S-IGF-I in males were slightly, but significantly higher than in females. It could be shown that the mean values of some reference sample subgroups differed significantly from the total mean. However, the multicenter approach used in this study reduces the impact of systematic population, sample handling and laboratory differences on the calculated reference mean. The present study establishes age- and sex-specific reference values for a fully automated immunoassay system based on a large population of healthy subjects. The established reference values may be used for this immunoassay system in different laboratories provided that the systematic difference between systems is low.


[Perinatal outcome of pregnancies in advanced maternal age].
Traditionally an advanced maternal age have been associated with a higher risk of unfavorable perinatal results. to know the perinatal outcomes in women giving birth at age 40 or older during a year in a concentration hospital located at Monterrey, Nuevo León. In a retrospective, cross-sectional, study. 163 women and 170 newborn were included. A newborn death (5.8 x 1000 RN) and no one maternal death was observed. The most frequent maternal associated disease was gestational diabetes (16%) followed by the hypertensive disorders (10%). The cesarean rate was 71.1% and the most frequent indication was previous cesarean. The most frequent hospitalization cause was early premature labour (4.2%). Premature membrane was observed in 11% of patients. The mean delivery time was at 38 weeks with a mean body weight of 3032 g. There were 5 newborn with congenital malformations (2.9%). 21 newborns were hospitalized (12%) and the most frequent indication was prematurity. Women giving birth at age 40 or older observed a higher gestational diabetes incidence followed by the pregnancy-induced hypertension. The cesarean delivery was the most frequent mode of delivery. We consider that the perinatal outcomes in this study were favorable for both mother and child.


Distribution of fucosylated N-acetyl lactosamine carbohydrate determinants during embryogenesis of the kidney in man.
The monoclonal antibodies AGF4.48 and AGF4.36 raised against the promyeloid cell line HL60 recognise a fucosylated N-acetyl lactosamine determinant. This oligosaccharide sequence has been shown to be present on a variety of tissues at different developmental stages. Using the immunoperoxidase technique and the AGF4.48 and AGF4.36 antibodies on formalin-fixed, paraffin-embedded tissues, the distribution of the corresponding antigenic determinants during human renal embryogenesis has been studied. Both antibodies bind to the surface of the cells of the ampullae of the ingrowing ureteric bud branches, but not to the remainder of the ureteric bud. Reactivity at this site persists until after fusion of the ureteric bud with the S-shaped tubule, but is then lost. The determinants are also found on different segments of the proximal convoluted tubule in the foetal and adult kidney. The determinants are thus found on the cells responsible for induction of renal tubulogenesis, and separately at specific stages and functionally distinct sites on the developing tubule.


Contribution of the CHEK2 1100delC variant to risk of multiple colorectal adenoma and carcinoma.
Aneuploidy is a characteristic of a subset of colorectal tumours. CHEK2 (also known as CHK2) is one of the cell cycle checkpoint genes coding for a family of proteins that sense damage in eukaryotic cells. Germline variation in CHEK2 has recently been shown to confer cancer susceptibility. Heterozygous mutations have been identified in patients with TP53-negative Li-Fraumeni syndrome. Furthermore, the CHEK2 1100delC variant carried by 1% of the population has been shown to act as a low penetrance allele for both breast and prostate cancers. To further our knowledge about the contribution of CHEK2 1100delC to cancer incidence we have analysed a series of 149 patients with multiple colorectal adenomas some of whom developed colorectal cancer. The CHEK2 1100delC allele was not over-represented in cases suggesting that this variant is not associated with an increased risk of colorectal disease.


Quantitative comparison of inflammatory infiltrate and linear contraction in human skin treated with 90-microsecond pulsed and 900-microsecond dwell time carbon dioxide lasers.
Skin resurfacing with 90-microsecond pulse duration carbon dioxide (CO2) resurfacing lasers has been reported to have shorter duration of erythema compared with skin resurfacing with 900-microsecond dwell time lasers. The presence of inflammatory infiltrate following resurfacing may correlate with the persistence of this erythema. Furthermore, skin treated with the 90-microsecond pulse duration laser and the 900-microsecond dwell time lasers both result in equivalent improvement of rhytids in the treated skin. To quantitative the inflammatory cell infiltrate and linear contraction of skin treated with the 90-microsecond pulsed and 900-microsecond dwell time CO2 lasers at intervals of 2 and 4 weeks after treatment. Volunteers were recruited from patients who were planning to undergo full face laser resurfacing under general anesthesia. Informed consent was obtained from all volunteers. In the posterior auricular areas of all volunteers, four separate rectangular areas were marked using a skin marking pen and a template. Two rectangular areas behind the right ear were treated with 6 passes of the 90-microsecond laser and two rectangular areas behind the left ear were treated with the 900-microsecond dwell time laser. The resurfaced areas were wiped with a moist cotton swab and then patted dry with dry gauze between passes. Contraction measurements of the resurfaced areas were taken before and immediately after laser treatment and again at 2 and 4 weeks following treatment. Punch biopsies were also performed at 2 and 4 weeks after treatment in an area of skin different from where contraction measurements were taken. The number of inflammatory cells present in the skin at 2 and 4 weeks after laser resurfacing are greater for skin resurfaced with a 900-microsecond dwell time laser than a 90-microsecond pulse time laser. Linear contraction of skin immediately after treatment was 18% greater with the 900-microsecond dwell time laser than with the 90-microsecond pulsed laser. The difference in the amount of contraction produced by the lasers tended to decrease over time. At 4 weeks there was a 10% difference in mean linear contraction between the two laser types. Increased numbers of inflammatory cells in skin resurfaced with the 900-microsecond dwell time laser may explain the observed persistence of erythema associated with the 900-microsecond dwell time laser. Measurable linear contraction produced by the 900-microsecond dwell time laser was initially 18% greater than the 90-microsecond pulse laser. This difference tends to decrease over time.


Auditory-induced bouncing is a perceptual (rather than a cognitive) phenomenon: Evidence from illusory crescents.
A central task for vision is to identify objects as the same persisting individuals over time and motion. The need for such processing is made especially clear in ambiguous situations such as the bouncing/streaming display: two discs move toward each other, superimpose, and then continue along their trajectories. Did the discs stream past each other, or bounce off each other? When people are likely to perceive streaming, playing a brief tone at the moment of overlap can readily cause them to see bouncing instead. Recent research has attributed this effect to decisional (rather than perceptual) processes by showing that auditory tones alter response biases but not the underlying sensitivity for detecting objective bounces. Here we explore the nature of this phenomenon using 'illusory causal crescents': when people perceive bouncing (or causal 'launching'), they also perceive the second disc to begin moving before being fully overlapped with the first disc (i.e. leaving an uncovered crescent). Here we demonstrate that merely playing a sound coincident to the moment of overlap can also reliably induce the perception of such illusory crescents. Moreover, this effect is due to the coincidence of the tone, per se, since the effect disappears when the tone is embedded in a larger regular tone sequence. Because observers never have to explicitly categorize their percept (e.g. as streaming)-and because the effect involves a subtle quantitative influence on another clearly visual property (i.e. the crescent's width)-we conclude that this audiovisual influence on the perception of identity over time reflects perceptual processing rather than higher-level decisions.


Associations Between Postpartum Depression, Breastfeeding, and Oxytocin Levels in Latina Mothers.
Postpartum depression (PPD), often comorbid with anxiety, is the leading medical complication among new mothers. Latinas have elevated risk of PPD, which has been associated with early breastfeeding cessation. Lower plasma oxytocin (OT) levels have also been associated with PPD in non-Latinas. This pilot study explores associations between PPD, anxiety, breastfeeding, and OT in Latinas. Thirty-four Latinas were enrolled during their third trimester of pregnancy and followed through 8 weeks postpartum. Demographic data were collected at enrollment. Depression was assessed using the Edinburgh Postnatal Depression Scale (EPDS) at each time point (third trimester of pregnancy, 4 and 8 weeks postpartum). The Spielberger State-Trait Anxiety Inventory (STAI) was administered postpartum and EPDS anxiety subscale was used to assess anxiety at each time point. Breastfeeding status was assessed at 4 and 8 weeks postpartum. At 8 weeks, OT was collected before, during, and after a 10-minute breast/bottle feeding session from 28 women who completed the procedures. Descriptive statistics are provided and comparisons by mood and breastfeeding status were conducted. Analyses of variance were used to explore associations between PPD, anxiety, breastfeeding status, and OT. Just under one-third of women were depressed at enrollment. Prenatal depression, PPD, and anxiety were significantly associated with early breastfeeding cessation (i.e., stopped breastfeeding before 2 months) (p < 0.05). There was a significant interaction between early breastfeeding cessation and depression status on OT at 8 weeks postpartum (p < 0.05). Lower levels of OT were observed in women who had PPD at 8 weeks and who had stopped breastfeeding their infant by 8 weeks postpartum. Future studies should investigate the short- and long-term effects of lower OT levels and early breastfeeding cessation on maternal and child well-being.


SLAMF1 Promotes Methotrexate Resistance via Activating Autophagy in Choriocarcinoma Cells.
The acquisition of chemoresistance to methotrexate (MTX) still remains one of the major challenges for choriocarcinoma treatment. Herein, we aimed to evaluate the potential role of Signaling Lymphocytic Activation Molecule Family Member 1 (SLAMF1) as a possible regulator of chemoresistance to MTX in choriocarcinoma. MTX-resistant JEG3 and JAR sublines (JEG3/MTX, JAR/MTX) were used to study SLAMF1 function. CCK8 assay and soft agar assay were conducted to measure the cell viability and clonogenesis of choriocarcinoma cells, respectively; MDC incorporation assay was conducted for the quantification of intracellular autophagy; BrdU labeling was used to assess the proliferative potential of choriocarcinoma cells; SLAMF1 protein expression was analyzed by Western blotting. Upregulation of SLAMF1 expression was observed in MTX-resistant JEG3/MTX and JAR/MTX sublines compared to their parental JEG3 and JAR cell lines, respectively. Knockdown of SLAMF1 markedly attenuated cell viability and soft agar clonogenesis after incubation with MTX in JEG3/MTX and JAR/MTX cells. In contrast, constitutive expression of SLAMF1 rescued cell survival soft agar clonogenesis in JEG3 and JAR cells treated with MTX. Moreover, autophagy is apparently activated in MTX-resistant JEG3/MTX and JAR/MTX sublines compared to their parental cell lines. Autophagy inhibitor 3-methyladenine and bafilomycin A1 enhanced MTX-induced cytotoxicity in MTX-resistant JEG3 and JAR sublines. Further, SLAMF1 might activate autophagy-related mechanism to promote resistance to MTX in choriocarcinoma cells. Depletion of SLAMF1 suppressed autophagy and induced apoptosis in MTX-treated JEG3/MTX and JAR/MTX cells. SLAMF1 might promote MTX resistance via activating protective autophagy in choriocarcinoma cell lines. Targeting SLAMF1 might be a useful therapeutic strategy to sensitize choriocarcinoma cells to MTX-based regimens.


Pregnancy during residency: a literature review.
It is estimated that by 2010 30% of U.S. physicians will be women. Pregnancy during residency can and does happen in all programs, and continues to provide problems for many. The author reviews the issues surrounding pregnancy during residency by evaluating published commentaries and research reports. A literature search was conducted using Medline (January 1984-October 2001). Published articles were categorized as research or commentary. Research reports were sorted by content and summarized under three headings: mother and infant health, sources of stress and support for the pregnant resident, and reactions of colleagues to the pregnant resident. A total of 27 research reports were located; two additional reports published before 1984 were added because they complemented included studies. The majority of the studies in this review used retrospective self-report questionnaires, mostly completed by female residents and physicians. All reports suggested an increased risk of complications, especially adverse late-pregnancy events, for pregnant physicians. Pregnant residents found the physical demands of residency and lack of support from fellow residents and their departments most stressful. Anger and resentment toward the pregnant resident were common among not-pregnant residents, feelings particularly associated with expectations of increased workload. Individual maternity/parental leave policies were inconsistent. Policy development is discussed. The studies in this review supported planning for residents' pregnancies, and the author advocates clear maternity/parental leave policies. The author comments on the use of existing data to make common sense changes and on the need for further studies to help clarify the issues and evaluate program changes.


Orai1 (CRACM1) is the platelet SOC channel and essential for pathological thrombus formation.
Platelet activation and aggregation at sites of vascular injury are essential for primary hemostasis, but are also major pathomechanisms underlying myocardial infarction and stroke. Changes in [Ca(2+)](i) are a central step in platelet activation. In nonexcitable cells, receptor-mediated depletion of intracellular Ca(2+) stores triggers Ca(2+) entry through store-operated calcium (SOC) channels. STIM1 has been identified as an endoplasmic reticulum (ER)-resident Ca(2+) sensor that regulates store-operated calcium entry (SOCE) in immune cells and platelets, but the identity of the platelet SOC channel has remained elusive. Orai1 (CRACM1) is the recently discovered SOC (CRAC) channel in T cells and mast cells but its role in mammalian physiology is unknown. Here we report that Orai1 is strongly expressed in human and mouse platelets. To test its role in blood clotting, we generated Orai1-deficient mice and found that their platelets display severely defective SOCE, agonist-induced Ca(2+) responses, and impaired activation and thrombus formation under flow in vitro. As a direct consequence, Orai1 deficiency in mice results in resistance to pulmonary thromboembolism, arterial thrombosis, and ischemic brain infarction, but only mild bleeding time prolongation. These results establish Orai1 as the long-sought platelet SOC channel and a crucial mediator of ischemic cardiovascular and cerebrovascular events.


Intravitreal bevacizumab (Avastin) in combination with verteporfin photodynamic therapy for choroidal neovascularization associated with age-related macular degeneration (IBeVe Study).
A novel alternative for combined treatment using verteporfin photodynamic therapy (PDT) has emerged as preliminary safety and efficacy data of the intravitreal use of the anti-angiogenic bevacizumab became available. In the current study we investigate the feasibility of intravitreal bevacizumab combined with verteporfin PDT for the treatment of choroidal neovascularization (CNV) secondary to age-related macular degeneration (AMD). A single-centre, prospective, open-label study of 11 patients with documented CNV progression after PDT treatment who underwent combined PDT and intravitreal injection of 1.5 mg of bevacizumab was undertaken. Standardized ophthalmic evaluation was performed at baseline and at weeks 1, 2, 12 and 24. Clinical evidence of complications and changes in logarithm of the minimum angle of resolution (logMAR) best-corrected visual acuity (BCVA) using Early Treatment Diabetic Retinopathy Study (ETDRS) charts and in fluorescein leakage from CNV were evaluated. The mean (+/-SD) age of the 11 patients was 74 (+/-5) years. Seven eyes had been treated with one previous PDT session and four eyes had two previous PDT sessions. The mean baseline logMAR ETDRS BCVA was 1.031 (Snellen equivalent, 20/200(-2)). At follow-up weeks 1, 2, 12 and 24, the mean logMAR ETDRS BCVA (Snellen equivalent) was 0.944 (20/160(-2)), 0.924 (20/160(-1)), 0.882 (20/160(+1)), and 0.933 (20/160(-2)), respectively. The change in BCVA from baseline was significant at each study follow-up interval (P < or = 0.001); at 12 and 24 weeks, the mean change in BCVA from baseline was an improvement of 1.49 and of 0.98 ETDRS line, respectively. Fluorescein leakage from CNV was absent in all eyes at week 12. One additional treatment session was required in seven (63.6%) eyes at week 24 due to recurrent fluorescein leakage from CNV ("minimum" [<50% of the leaking area noted at baseline], n = 4; and "moderate" [>50% of the leaking area noted at baseline], n = 3). No progression of the neovascular lesion was observed at week 24. No safety issues were identified throughout the period of the study. The overall changes in vision and fluorescein leakage from CNV throughout the study suggest that a possible synergistic effect may arise from the combination of intravitreal bevacizumab with verteporfin PDT for the treatment of neovascular AMD.


Microstructure and chemical analysis of four calcium silicate-based cements in different environmental conditions.
The objective of this study was to analyze the microstructure and crystalline structures of ProRoot MTA, Biodentine, CEM Cement, and Retro MTA when exposed to phosphate-buffered saline, butyric acid, and blood. Mixed samples of ProRoot MTA, Biodentine, CEM Cement, and Retro MTA were exposed to either phosphate-buffered saline, butyric acid, or blood. Scanning electron microscope (SEM) and energy-dispersive X-ray spectroscopic (EDX) evaluations were conducted of specimens. X-ray diffraction (XRD) analysis was also performed for both hydrated and powder forms of evaluated calcium silicate cements. The peak of tricalcium silicate and dicalcium silicate detected in all hydrated cements was smaller than that seen in their unhydrated powders. The peak of calcium hydroxide (Ca(OH)2) in blood- and acid-exposed ProRoot MTA, CEM Cement, and Retro MTA specimens were smaller than that of specimens exposed to PBS. The peak of Ca(OH)2 seen in Biodentine™ specimens exposed to blood was similar to that of PBS-exposed specimens. On the other hand, those exposed to acid exhibited smaller peaks of Ca(OH)2. Exposure to blood or acidic pH decreased Ca(OH)2 crystalline formation in ProRoot MTA, CEM Cement and Retro MTA. However, a decrease in Ca(OH)2 was only seen when Biodentine™ exposed to acid. The formation of Ca(OH)2 which influences the biological properties of calcium silicate cements was impaired by blood and acid exposures in ProRoot MTA, CEM Cement, and Retro MTA; however, in the case of Biodentine, only exposure to acid had this detrimental effect.


Gene expression, oxidative stress and apoptotic changes in rabbit ileum experimentally infected with Eimeria intestinalis.
Coccidiosis is a parasitic disease caused by protists (apicomplexans) of the genus Eimeria Schneider, 1875 and is considered to be the most important disease faced by rabbit breeders due to its high morbidity. In the present study, the antioxidant status and changes in apoptosis and in the expression of some genes were quantified in rabbits' ilea following infection with Eimeria intestinalis Cheissin, 1948. Rabbits, orally infected with 1 × 105 sporulated oocysts of E. intestinalis, started to shed oocysts in their faeces on 8 days post infection (dpi) and reached maximum excretion on 10 dpi, with approximately 5 million oocysts. This was accompanied by a significant decrease in the live body weight of infected rabbits. Also, malondialdehyde and nitric oxide were significantly increased while catalase and glutathione were significantly decreased in the ileum tissues of the infected rabbits. In addition, a significant increase was observed in the percentages of apoptotic cells in the ilea of the infected rabbits. Furthermore, interleukin-1β and interleukin-2 mRNA levels were significantly down-regulated and mRNA levels of interleukin-6, interferon gamma and inducible nitric oxide synthase were significantly up-regulated, while those of C-reactive protein remained unchanged. We conclude that infection with E. intestinalis induces oxidative stress, a significant increase in the percentage of apoptotic cells and a diverse and robust Th1 and Th1-related cytokine response in the ileum tissues.


The 2.1A crystal structure of the far-red fluorescent protein HcRed: inherent conformational flexibility of the chromophore.
We have determined the crystal structure of HcRed, a far-red fluorescent protein isolated from Heteractis crispa, to 2.1A resolution. HcRed was observed to form a dimer, in contrast to the monomeric form of green fluorescent protein (GFP) or the tetrameric forms of the GFP-like proteins (eqFP611, Rtms5 and DsRed). Unlike the well-defined chromophore conformation observed in GFP and the GFP-like proteins, the HcRed chromophore was observed to be considerably mobile. Within the HcRed structure, the cyclic tripeptide chromophore, Glu(64)-Tyr(65)-Gly(66), was observed to adopt both a cis coplanar and a trans non-coplanar conformation. As a result of these two conformations, the hydroxyphenyl moiety of the chromophore makes distinct interactions within the interior of the beta-can. These data together with a quantum chemical model of the chromophore, suggest the cis coplanar conformation to be consistent with the fluorescent properties of HcRed, and the trans non-coplanar conformation to be consistent with non-fluorescent properties of hcCP, the chromoprotein parent of HcRed. Moreover, within the GFP-like family, it appears that where conformational freedom is permissible then flexibility in the chromophore conformation is possible.


Aragonite-Polylysine: Neuro-Regenerative Scaffolds with Diverse Effects on Astrogliosis.
Biomaterials, especially when coated with adhesive polymers, are a key tool for restorative medicine, being biocompatible and supportive for cell adherence, growth, and function. Aragonite skeletons of corals are biomaterials that support survival and growth of a range of cell types, including neurons and glia. However, it is not known if this scaffold affects neural cell migration or elongation of neuronal and astrocytic processes, prerequisites for initiating repair of damage in the nervous system. To address this, hippocampal cells were aggregated into neurospheres and cultivated on aragonite skeleton of the coral Trachyphyllia geoffroyi (Coral Skeleton (CS)), on naturally occurring aragonite (Geological Aragonite (GA)), and on glass, all pre-coated with the oligomer poly-D-lysine (PDL). The two aragonite matrices promoted equally strong cell migration (4.8 and 4.3-fold above glass-PDL, respectively) and axonal sprouting (1.96 and 1.95-fold above glass-PDL, respectively). However, CS-PDL had a stronger effect than GA-PDL on the promotion of astrocytic processes elongation (1.7 vs. 1.2-fold above glass-PDL, respectively) and expression of the glial fibrillary acidic protein (3.8 vs. and 1.8-fold above glass-PDL, respectively). These differences are likely to emerge from a reaction of astrocytes to the degree of roughness of the surface of the scaffold, which is higher on CS than on GA. Hence, CS-PDL and GA-PDL are scaffolds of strong capacity to derive neural cell movements and growth required for regeneration, while controlling the extent of astrocytic involvement. As such, implants of PDL-aragonites have significant potential as tools for damage repair and the reduction of scar formation in the brain following trauma or disease.


A perfect smoother.
The well-known and popular Savitzky-Golay filter has several disadvantages. A very attractive alternative is a smoother based on penalized least squares, extending ideas presented by Whittaker 80 years ago. This smoother is extremely fast, gives continuous control over smoothness, interpolates automatically, and allows fast leave-one-out cross-validation. It can be programmed in a few lines of Matlab code. Theory, implementation, and applications are presented.


Effectiveness of single posterior decompressive laminectomy in symptomatic lumbar spinal stenosis: a retrospective study.
The best method for surgical intervention in symptomatic lumbar stenosis is not clear. The present study aims to assess first year outcomes and complication rates of patients treated with single posterior decompressive laminectomy. Patients requiring surgery for severe, symptomatic, lumbar spinal stenosis were evaluated retrospectively. Oswestry disability index scores as well as the complications attributable to surgery were recorded before, at the sixth month and at the twelfth month of the surgery. Eighty patients were enrolled to the study. The mean age of the population was 63,14 ± 11,57. Neurogenic claudication was the most common finding (65%). Of the patients, 67.5% had severe spinal stenosis. The mean ODI score at the baseline was relatively high than in the literature and was measured as 74.30 ± 5.38. At the end of the 6 months follow-up period, all patients' ODI scores significantly improved. Moreover, this improvement continued till the end of the 12 month. The mean change in ODI at the end of the first year was 41.80% ± 12.73. In selected cases of symptomatic lumbar spinal stenosis, single posterior decompression using laminectomy is safe and effective.


Cecocentral scotomas in gliomatosis cerebri.
A previously healthy 69-year-old white male presented with slowly progressive, painless loss of vision in both eyes. Bilateral cecocentral scotomas were found, and this was his only chief complaint for 9 months. Signs of progressive neurologic dysfunction ensued, and the patient was followed through a complete neuropathologic study at postmortem examination. To our knowledge, this is the only reported instance of pathologically confirmed gliomatosis cerebri with an initial presentation of bilateral chronic cecocentral scotomas.


Polymeric Nanoparticles Active against Dual-Species Bacterial Biofilms.
Biofilm infections are a global public health threat, necessitating new treatment strategies. Biofilm formation also contributes to the development and spread of multidrug-resistant (MDR) bacterial strains. Biofilm-associated chronic infections typically involve colonization by more than one bacterial species. The co-existence of multiple species of bacteria in biofilms exacerbates therapeutic challenges and can render traditional antibiotics ineffective. Polymeric nanoparticles offer alternative antimicrobial approaches to antibiotics, owing to their tunable physico-chemical properties. Here, we report the efficacy of poly(oxanorborneneimide) (PONI)-based antimicrobial polymeric nanoparticles (PNPs) against multi-species bacterial biofilms. PNPs showed good dual-species biofilm penetration profiles as confirmed by confocal laser scanning microscopy. Broad-spectrum antimicrobial activity was observed, with reduction in both bacterial viability and overall biofilm mass. Further, PNPs displayed minimal fibroblast toxicity and high antimicrobial activity in an in vitro co-culture model comprising fibroblast cells and dual-species biofilms of Escherichia coli and Pseudomonas aeruginosa. This study highlights a potential clinical application of the presented polymeric platform.


Review article: the modern management of portal hypertension--primary and secondary prophylaxis of variceal bleeding in cirrhotic patients.
Variceal bleeding is a life-threatening complication of liver cirrhosis with a high probability of recurrence. Treatment to prevent first bleeding or rebleeding is mandatory. To provide an overview of the current knowledge on the best evidence-based therapeutic options to prevent first or recurrent bleeding from oesophageal varices in patients with cirrhosis. For the preparation of this narrative review, we sought to analyse randomized controlled trials that examined the efficacy and side effects of pharmacological or endoscopic therapy for the primary and secondary prophylaxis of oesophageal variceal bleeding. Endoscopic band ligation (EBL) and nonselective beta-blockers are both effective in preventing first bleeding. Until more long-term data are available, nonselective beta-blockers should be the first treatment option because of less severe side effects. EBL is an alternative when beta-blockers are contraindicated or not tolerated. Patient preference may also be considered. For prevention of rebleeding, nonselective beta-blockers (preferably in association with isosorbide-5-mononitrate) or EBL are both effective and good alternative treatments. A combination of both treatments may be the best alternative. A great improvement in the prevention of variceal bleeding has emerged over the last years. However, further therapeutic options that combine higher efficacy, better tolerance and fewer side effects are needed.


Pediatric-onset mastocytosis: a long term clinical follow-up and correlation with bone marrow histopathology.
Pediatric onset mastocytosis usually presents as urticaria pigmentosa; and less often as diffuse cutaneous mastocytosis. While the literature indicates that disease often resolves, there has been a move to more aggressive therapy for mastocytosis early in life. We addressed the long term prognosis of pediatric-onset disease by examining 17 children with mastocytosis which we had reported on in 1989 [1]. We successfully contacted 15 of these patients and data was collected regarding their clinical status. Original bone marrow specimens were re-stained, re-examined, and correlated with disease outcome using consensus criteria. Three of five patients with persistent disease underwent repeat bone marrow biopsies. There was complete regression of disease as defined by cutaneous findings and symptoms (clinical disease severity) in 10 of 15 patients (67%). Three patients had major (20%) and two had partial regression of disease (13%). Repeat marrow examinations on three patients with persistent disease documented systemic mastocytosis based on marrow findings in one patient who had partial regression of disease and was the only patient with initial morphologic evidence of systemic disease. Of the remaining two patients, one demonstrated partial regression and the other major regression of disease; and neither had evidence of systemic mastocytosis. This study demonstrates that initial bone marrow biopsies were prognostic in that those without evidence of systemic disease experienced disease regression; and that the long term prognosis for children managed symptomatically with mastocytosis is highly encouraging.


Myopic regression after phakic intraocular lens implantation and LASIK.
In myopia, biometry including the axial length is important, along with the refractive data. We compared the rates of myopic regression 3 years after phakic intraocular lens (pIOL) implantation and laser in situ keratomileusis (LASIK) after matching the preoperative axial length in highly myopic eyes of Japanese patients. This retrospective nonrandomized study included 133 eyes of 84 patients with myopia exceeding -6.00 diopters (D) who underwent implantation of two iris-fixated pIOLs (pIOL group, 66 eyes/46 patients) or myopic LASIK (LASIK group, 67 eyes/38 patients) who were followed for more than 3 years postoperatively. The patient age, preoperative refraction, and preoperative axial length were matched between the study groups. There were no significant differences preoperatively between the groups in age, intraocular pressure, refraction, keratometry, or axial length. The mean regression values after 3 years compared with the 1-month postoperative refractions were -0.12 ± 0.47 (SD) D in the pIOL group and -0.82 ± 0.69 D in the LASIK group (p < 0.001). The differences in the mean regression rates between 1 and 12 months, 12 and 24 months, and 24 and 36 months of follow-up were, respectively, 0.09 ± 0.38 D, -0.11 ± 0.35 D, and -0.11 ± 0.30 D in the pIOL group and -0.57 ± 0.84 D, -0.24 ± 0.47 D, and 0.00 ± 0.53 D in the LASIK group (p < 0.001, p = 0.07, p = 0.15, respectively). There was a significant difference in myopic regression 3 years postoperatively between the groups matched for preoperative axial length in Japanese patients. This result has the potential to elucidate myopia in the future.


Impact of community pharmacists in COPD management: Inhalation technique and medication adherence.
Inhalation technique and medication adherence are highly important for the management of chronic obstructive pulmonary disease (COPD) since they are essential pre-requisites for achieving full therapeutic effect in patients. Community pharmacists are in the best position to deliver services in these two areas. This is a ten-year period review of studies looking into the impact of community pharmacists in the management of COPD in relation to: inhalation technique and medication adherence in the period from 2005 to 2015. Ten studies are included in the review. The studies show that community pharmacists' interventions had a positive impact on improving patients' inhalation technique and adherence to inhaled medications. This was shown in some studies to be cost-effective in terms of reducing hospitalisation and severe exacerbation rate. Scarcity of studies in this domain is noted through this review. This review showed that community pharmacists can have a positive impact in the management of COPD especially on inhaler technique education and medication adherence. Nevertheless, their role is still not fully recognised in this area, thus there is a need for more research. There is also a need for more research to identify the optimal frequency for inhaler technique re-checking and education as a pre-emptive measure against technique deterioration in patients. The results highlight the need for healthcare systems to recognise more the role of community pharmacists in COPD management in two critical areas that are still challenging in real practice.


Inhibition Effect of Triglyceride Accumulation by Large Yellow Croaker Roe DHA-PC in HepG2 Cells.
The phospholipids (PLs) of large yellow croaker (Pseudosciaena crocea, P. crocea) roe contain a high level of polyunsaturated fatty acids, especially docosahexaenoic acid (DHA), which can lower blood lipid levels. In previous research, PLs of P. crocea roe were found able to regulate the accumulation of triglycerides. However, none of these involve the function of DHA-containing phosphatidylcholine (DHA-PC), which is the main component of PLs derived from P. crocea roe. The function by which DHA-PC from P. crocea roe exerts its effects has not yet been clarified. Herein, we used purified DHA-PC and oleic acid (OA) induced HepG2 cells to establish a high-fat model, and the cell activity and intracellular lipid levels were then measured. The mRNA and protein expression of Fatty Acid Synthase (FAS), Carnitine Palmitoyl Transferase 1A (CPT1A) and Peroxisome Proliferator-Activated Receptor α (PPARα) in HepG2 cells were detected via RT-qPCR and western blot as well. It was found that DHA-PC can significantly regulate triglyceride accumulation in HepG2 cells, the effect of which was related to the activation of PPARα receptor activity, upregulation of CPT1A, and downregulation of FAS expression. These results can improve the understanding of the biofunction of hyperlipidemia mediated by DHA-PC from P. crocea roe, as well as provide a theoretical basis for the utilization of DHA-PC from P. crocea roe as a functional food additive.


Changes in turtle brain neurotransmitters and related substances during anoxia.
Freshwater turtles (Pseudemys scripta elegans) were exposed to 0.5-13 h of anoxia at 25 degrees C, whereupon the brain concentrations of 14 amino acid and monoamine neurotransmitters and related substances were measured. Monoamines are of particular interest, because their synthesis and (in part) degradation require molecular oxygen. During anoxia, the level of the inhibitory transmitter gamma-aminobutyric acid (GABA) increased (2.3-fold after 13 h) and the level of the excitatory transmitter Glu fell. Furthermore, anoxia caused increases in the levels of Ala (14 times after 13 h), Tau, Gly, and Ser, whereas the Gln level fell. The increase in Ala is likely to inhibit pyruvate kinase, thereby mediating the decreased rate of glycolysis seen after prolonged anoxia. The increased level of Tau might protect the turtle brain against Ca2(+)-mediated anoxic damage. The monoamine metabolites almost vanished within a few hours of anoxia, indicating a halt in monoamine synthesis and breakdown, and the dopamine level fell. Nevertheless, serotonin, norepinephrine, and epinephrine levels were maintained during 13 h of anoxia, at levels extremely high compared with mammals, suggesting adaptive mechanisms such as stockpiling. It is hypothesized that the pattern of change in levels of amino acids (notably GABA and Glu) and monoamines is of functional significance, because it promotes the decrease in brain activity and energy consumption seen in anoxic turtles.


The Nuclear Matrix Protein Megator Regulates Stem Cell Asymmetric Division through the Mitotic Checkpoint Complex in Drosophila Testes.
In adult Drosophila testis, asymmetric division of germline stem cells (GSCs) is specified by an oriented spindle and cortically localized adenomatous coli tumor suppressor homolog 2 (Apc2). However, the molecular mechanism underlying these events remains unclear. Here we identified Megator (Mtor), a nuclear matrix protein, which regulates GSC maintenance and asymmetric division through the spindle assembly checkpoint (SAC) complex. Loss of Mtor function results in Apc2 mis-localization, incorrect centrosome orientation, defective mitotic spindle formation, and abnormal chromosome segregation that lead to the eventual GSC loss. Expression of mitotic arrest-deficient-2 (Mad2) and monopolar spindle 1 (Mps1) of the SAC complex effectively rescued the GSC loss phenotype associated with loss of Mtor function. Collectively our results define a new role of the nuclear matrix-SAC axis in regulating stem cell maintenance and asymmetric division.


Dopamine depresses excitatory synaptic transmission onto rat subicular neurons via presynaptic D1-like dopamine receptors.
Schizophrenia is considered to be associated with an abnormal functioning of the hippocampal output. The high clinical potency of antipsychotics that act as antagonists at dopamine (DA) receptors indicate a hyperfunction of the dopaminergic system. The subiculum obtains information from area CA1 and the entorhinal cortex and represents the major output region of the hippocampal complex. To clarify whether an enhanced dopaminergic activity alters the hippocampal output, the effect of DA on alveus- and perforant path-evoked excitatory postsynaptic currents (EPSCs) in subicular neurons was examined using conventional intracellular and whole cell voltage-clamp recordings. Dopamine (100 microM) depressed alveus-elicited (S)-alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor-mediated EPSCs to 56 +/- 8% of control while perforant path-evoked EPSCs were attenuated to only 76 +/- 7% of control. Dopamine had no effect on the EPSC kinetics. Dopamine reduced the frequency of spontaneous miniature EPSCs without affecting their amplitudes. The sensitivity of subicular neurons to the glutamate receptor agonist (S)-alpha-amino-3-hydoxy-5-methyl-4-isoxazolepropionic acid was unchanged by DA pretreatment, excluding a postsynaptic mechanism for the observed reduction of excitatory synaptic transmission. The effect of DA on evoked EPSCs was mimicked by the D1 receptor agonist SFK 38393 and partially antagonized by the D1 receptor antagonist SCH 23390. While the D2 receptor agonist quinelorane failed to reduce the EPSCs, the D2 receptor antagonist sulpiride did not block the action of DA. The results indicate that DA strongly depresses the hippocampal and the entorhinal excitatory input onto subicular neurons by decreasing the glutamate release following activation of presynaptic D1-like DA receptors.


The procyanidin-induced pseudo laminar shear stress response: a new concept for the reversal of endothelial dysfunction.
Reduced endothelium-dependent vasodilator responses with increased synthesis of ET-1 (endothelin-1) are characteristics of endothelial dysfunction in heart failure and are predictive of mortality. Identification of treatments that correct these abnormalities may have particular benefit for patients who become refractory to current regimens. Hawthorn preparations have a long history in the treatment of heart failure. Therefore we tested their inhibitory effects on ET-1 synthesis by cultured endothelial cells. These actions were compared with that of GSE (grape seed extract), as the vasoactive components of both these herbal remedies are mainly oligomeric flavan-3-ols called procyanidins. This showed extracts of hawthorn and grape seed were equipotent as inhibitors of ET-1 synthesis. GSE also produced a potent endothelium-dependent vasodilator response on preparations of isolated aorta. Suppression of ET-1 synthesis at the same time as induction of endothelium-dependent vasodilation is a similar response to that triggered by laminar shear stress. Based on these results and previous findings, we hypothesize that through their pharmacological properties procyanidins stimulate a pseudo laminar shear stress response in endothelial cells, which helps restore endothelial function and underlies the benefit from treatment with hawthorn extract in heart failure.


Interactive effects of food quality, temperature and rearing time on condition of juvenile black bream Acanthopagrus butcheri.
A laboratory experiment was conducted to determine the interactive effects of temperature and diet on condition indices of juvenile black bream Acanthopagrus butcheri, reared for time periods ranging from 2 to 42 days. After fish were reared for varying periods, growth, morphometric (Fulton's K) and biochemical [RNA:DNA (R:D) ratios] indices were measured. Fulton's K responded primarily to temperature, with progressive decrease in condition over time for fish reared at high temperatures. In contrast, R:D ratios were primarily affected by diet composition, with the highest values observed for fish reared on fish-based diets as opposed to vegetable-based diets. Significant effects of rearing time were also observed for Fulton's K and R:D ratios, as were some interactive treatment effects. In addition, Fulton's K and R:D ratios were not significantly correlated, perhaps due to the different periods of time integrated by each index or their relative sensitivity to lipid and protein deposition. These results highlight the complex responses of these condition indices to environmental variables and nutritional status.


Sources of intrapopulation variability in causative agents of nosocomial infection.
The mechanisms of heterogeneity of S. aureus and P. aeruginosa populations of pathological foci in patients with wound, traumatic, and burns infections were studied. It has been established that populations of the mentioned species in the sources of infection (patients and carriers) are pronouncedly heterogeneous and objects in the external environment serving as factors of transmission are contaminated with a whole complex of diverse variants. The conclusion can therefore be drawn that the first source of intrapopulation variability of wound populations of microbes is the presence of varying strains and variants in the infecting dose of the causative agent. The second source consists in repeated superinfection (immigration) of pathological processes by new, usually nosocomial, strains and variants of the same species with their subsequent selection during the patients' stay in the health-service establishment. Other sources of population variability, such as mutation and recombination, are of minor importance. With respect to the obtained results it is necessary to increase the volume of the representative sample of cultures in bacteriological diagnosis of open processes, to protect them reliably from superinfection, to follow up dynamically changes in the composition of causative agents and to provide for suitable correction of chemotherapy.


Developmental changes in the response of cardiac Purkinje fibers to SC-72-14.
We used standard microelectrode techniques to study the effects of SC-72-14, a monoclonal antibody to eel electroplax, and tetrodotoxin (TTX) on the electrophysiological properties of neonatal and adult canine Purkinje fibers. SC-72-14, 7 ng/ml, depressed maximum upstroke potential (Vmax) and membrane responsiveness of adult but not neonatal fibers. In contrast, we previously had shown TTX to have a greater effect on neonatal than adult fibers (29). Even in the presence of SC-72-14, TTX continued to have a greater effect on the neonatal fibers. The effects of TTX and SC-72-14 were cumulative over the concentration range studied, and our results are consistent with their acting at different sites.


Fatty acids in the portal vein of the rat regulate hepatic insulin clearance.
The effects of FFA on hepatic insulin clearance were studied in the in situ perfused rat liver. Clearance decreased with increasing body weight (age) of the rats. When FFA were added to the perfusate a 40% reduction of hepatic removal of insulin was found over the normal, physiological range (less than 1,000 mumol/liter), less pronounced in heavier rats. When perfusion was started with high concentrations of FFA, inhibition was rapidly reversible, a phenomenon again blunted in heavier rats. In contrast to FFA, different glucose concentrations in the perfusate did not affect the hepatic insulin uptake in the presence of FFA within physiological concentrations. Thus, hepatic clearance of insulin is proportional to rat weight (age) and portal FFA concentrations. Other studies have recently shown that fatty acids inhibit insulin binding, degradation, and function in isolated rat hepatocytes, and that hepatic clearance is inversely dependent on hepatic triglyceride concentrations, both inhibitions reversible by prevention of fatty acid oxidation. It is suggested that the diminished hepatic clearance of insulin in heavier (older) rats is at least partly due to their relative obesity and increased hepatic triglyceride contents. This effect as well as that of portal FFA is probably mediated via fatty acid oxidation in the liver. This mechanism may have implications for the regulation of hepatic metabolism, and peripheral insulin concentrations.


Calpain 4 is not necessary for LFA-1-mediated function in CD4+ T cells.
T cell activation and immune synapse formation require the appropriate activation and clustering of the integrin, LFA-1. Previous work has reported that the calpain family of calcium-dependent proteases are important regulators of integrin activation and modulate T cell adhesion and migration. However, these studies have been limited by the use of calpain inhibitors, which have known off-target effects. Here, we used a LoxP/CRE system to specifically deplete calpain 4, a small regulatory calpain subunit required for expression and activity of ubiquitously expressed calpains 1 and 2, in CD4+ T cells. CD4+ and CD8+ T cells developed normally in Capn4(F/F):CD4-CRE mice and had severely diminished expression of Calpain 1 and 2, diminished talin proteolysis and impaired casein degradation. Calpain 4-deficient T cells showed no difference in adhesion or migration on the LFA-1 ligand ICAM-1 compared to control T cells. Moreover, there was no impairment in conjugation between Capn4(F/F):CD4-CRE T cells and antigen presenting cells, and the conjugates were still capable of polarizing LFA-1, PKC-theta and actin to the immune synapse. Furthermore, T cells from Capn4(F/F):CD4-CRE mice showed normal proliferation in response to either anti-CD3/CD28 coated beads or cognate antigen-loaded splenocytes. Finally, there were no differences in the rates of apoptosis following extrinsic and intrinsic apoptotic stimuli. Our findings demonstrate that calpain 4 is not necessary for LFA-1-mediated adhesion, conjugation or migration. These results challenge previous reports that implicate a central role for calpains in the regulation of T cell LFA-1 function.


Role of acidosis in regulating hepatic nitrogen metabolism during fasting in conscious dog.
This study was designed to investigate the role that acidosis plays in the metabolic responses to fasting. Eighteen conscious dogs with surgically implanted catheters in the femoral artery and in the hepatic, portal, and renal veins were studied. Six were fasted for 24 h and 12 were fasted for 4 days (96 h). On the day of the study, six 4-day fasted dogs were infused intravenously with NaHCO3 (10 mumol X kg-1 X min-1) for 3 h, while the rest received saline and acted as controls. Splanchnic balances of glutamine, alanine, blood urea nitrogen, ammonia, lactate, beta-hydroxybutyrate, and acetoacetate were estimated using the Fick principle. Blood flow to the splanchnic and renal beds were estimated using indocyanine green and p-aminohippurate extraction methods, respectively. The infusion of NaHCO3 nearly abolished the base deficit associated with fasting and normalized arterial bicarbonate levels but did not alter blood pH. It suppressed but did not abolish hepatic glutamine output by 60%. This was associated with a shift in cytoplasmic and mitochondrial redox potentials of the hepatocyte as evident by a decrease in hepatic production of beta-hydroxybutyrate and an increase in hepatic production of acetoacetate and a decrease in hepatic lactate utilization. Concomitantly, renal glutamine uptake decreased. Glutamine release of skeletal muscle was unchanged. The data suggest that hepatic glutamine synthesis and release seen with 4-day fasting has two components: a bicarbonate-dependent component that is influenced by the redox potential of the hepatocyte and a bicarbonate-independent component, the nature of which is not yet clear.(ABSTRACT TRUNCATED AT 250 WORDS)


Use of computed tomography in the management of colorectal cancer.
Computed tomography (CT) plays an important role in the management of colorectal cancer (CRC). The use of CT (colonography) as a screening tool for CRC has been validated and is expected to rise over time. The results of prior studies suggest that CT is suboptimal for assessment of local T stage and moderate for N stage disease. Recent advances in CT technology are expected to lead to some improvement in staging accuracy. At present, the main role of CT in pre-treatment imaging assessment lies in its use for the detection of distant metastases, especially in the liver. In a select group of patients, routine post-treatment surveillance with CT confers survival benefits. The role of CT for post-treatment assessment has been radically altered and improved with the advent of fusion positron emission tomography/CT. Perfusion CT shows promise as another functional imaging modality but further experience with this technique is necessary before it can be applied to routine clinical practice.


Rate-dependent changes in excitability of depressed cardiac Purkinje fibers as a mechanism of intermittent bundle branch block.
When the heart rate is accelerated, rate-dependent intraventricular block may occur. This block has been attributed to abnormal action potential prolongation in a diseased conducting pathway. Less often, intraventricular block develops during slowing of the heart rate and has been explained in terms of phase 4 depolarization in potentially automatic cells within the diseased fascicle. We tested these hypotheses in isolated bundles of Purkinje fibers placed in a three-chambered tissue bath. In one group of experiments, conditions of localized injury and depressed excitability were mimicked by superfusing the central segment with sucrose solution. Action potentials were initiated in the proximal segment while the slope of phase 4 of cells in the distal end was controlled by intracellular ramps of current of either polarity. In these preparations, phase 4 depolarization facilitated rather than retarded propagation across the depressed segment, even at takeoff potentials as low as -45 mV. In a second group, depressed excitability was induced by exposing the three fiber segments to Tyrode's solution that contained high concentrations of KCl and CaCl2 or isoproterenol (0.1 microgram/ml). Under these conditions, Purkinje fibers did not undergo phase 4 depolarization and did not generate abnormally prolonged action potentials. These preparations showed a biphasic time dependence of conduction during premature stimulation or in response to changes in the basic cycle length. Conduction impairment and block were manifest at either side of an optimal interval or cycle length. Our results suggest that phase 4 depolarization and abnormally prolonged action potentials are not necessary conditions for intermittent block. Both tachycardia and bradycardia-dependent intraventricular conduction abnormalities may be associated with time-dependent variations in the excitability of depolarized conducting fibers as well as in the amplitude of the slow responses generated by these fibers. These alterations can be explained in terms of regulation of slow inward current by the intracellular calcium concentration.


The changing spectrum of surgically treated cystic neoplasms of the pancreas.
Introduction. While the incidence of pancreatic cystic lesions has steadily increased, we sought to evaluate the changes in their surgical management. Methods. Patients with pancreatic cystic lesions who underwent surgical resection from 2003 to 2013 were identified. Clinicopathologic factors were analyzed and compared to a similar cohort from 1992 to 2002. Results. There were 134 patients with pancreatic cystic lesions who underwent surgical resection from 2003 to 2013, compared to 73 from 1992 to 2002. The most common preoperative imaging was a CT scan, although 66% underwent EUS and 63% underwent biopsy. Pathology included 18 serous, 47 mucinous, 11 pseudopapillary, and 58 intraductal papillary mucinous neoplasms (IPMN). In comparing cohorts, there were significantly fewer serous lesions and more IPMN. Postoperative complication rates were similar, and perioperative mortality rates were comparable. Conclusion. There has been a dramatic change in surgically treated pancreatic cystic tumors over the past two decades. Our data suggests that the incorporation of new imaging and diagnostic tests has led to greater detection of cystic tumors and a decreased rate of potentially unnecessary resections. Therefore, all patients with cystic pancreatic lesions should undergo a focused CT-pancreas, and an EUS biopsy should be considered, in order to best select those that would benefit from surgical resection.


Impact of postoperative chemoradiotherapy and chemoradiotherapy alone for esophageal cancer in North-West Iran.
To investigate the role of surgical treatment for locally advanced esophageal cancer, we compared the outcomes of chemoradiotherapy alone (CRT) to postoperative chemoradiotherapy (S/CRT), using, Regional Radiotherapy Center, database. This retrospective study was conducted in North-West of Iran, included of 255 consecutive patients with esophageal cancer. Eligible operable and non-operable, were treated with S/CRT and CRT respectively. Radiotherapy (RT) was delivered at 1.8-2 Gy/day for five consecutive days in a given week. Chemotherapy (CT) consisted of cisplatin and 5-fluorouracil. From March 2006 to March 2011 255 patients: male/female 129/96, median age 68 (35-90), squamous/adeno 213/12, received CRT /S+CRT 166/59, median radiation dose 45 ± 13.6 Gy, Median survival 13.5 (11-15), overall survival (OS) One/ Two/Three 57/21/16%, Died/alive 158/97, Univariate analysis prognostic factors: age/stag/differentiation/dose of RT/fraction/treatment, Multivariate analysis predictor factor: dose of RT/fraction. Although this treatment offers some possibility for improvement of patients with esophageal cancer, there remains a significant need for development of new drug and new therapeutic approaches that can substantially impact survival.


Allosteric Effect of Nanobody Binding on Ligand-Specific Active States of the β2 Adrenergic Receptor.
Nanobody binding stabilizes G-protein-coupled receptors (GPCR) in a fully active state and modulates their affinity for bound ligands. However, the atomic-level basis for this allosteric regulation remains elusive. Here, we investigate the conformational changes induced by the binding of a nanobody (Nb80) on the active-like β2 adrenergic receptor (β2AR) via enhanced sampling molecular dynamics simulations. Dimensionality reduction analysis shows that Nb80 stabilizes structural features of the β2AR with an ∼14 Å outward movement of transmembrane helix 6 and a close proximity of transmembrane (TM) helices 5 and 7, and favors the fully active-like conformation of the receptor, independent of ligand binding, in contrast to the conditions under which no intracellular binding partner is bound, in which case the receptor is only stabilized in an intermediate-active state. This activation is supported by the residues located at hotspots located on TMs 5, 6, and 7, as shown by supervised machine learning methods. Besides, ligand-specific subtle differences in the conformations assumed by intracellular loop 2 and extracellular loop 2 are captured from the trajectories of various ligand-bound receptors in the presence of Nb80. Dynamic network analysis further reveals that Nb80 binding triggers tighter and stronger local communication networks between the Nb80 and the ligand-binding sites, primarily involving residues around ICL2 and the intracellular end of TM3, TM5, TM6, as well as ECL2, ECL3, and the extracellular ends of TM6 and TM7. In particular, we identify unique allosteric signal transmission mechanisms between the Nb80-binding site and the extracellular domains in conformations modulated by a full agonist, BI167107, and a G-protein-biased partial agonist, salmeterol, involving mainly TM1 and TM2, and TM5, respectively. Altogether, our results provide insights into the effect of intracellular binding partners on the GPCR activation mechanism, which should be taken into account in structure-based drug discovery.


Cadmium exposure and cancer mortality in the Third National Health and Nutrition Examination Survey cohort.
This study examined prospective data from the Third National Health and Nutrition Examination Survey (NHANES III) cohort to investigate the relationship between cadmium exposure and cancer mortality, and the specific cancers associated with cadmium exposure, in the general population. Vital status and cause of death through 31 December 2006 were obtained by the National Center for Health Statistics for NHANES III participants. The cadmium concentration of spot urine samples was measured and corrected for urine creatinine (uCd). Weighted Cox proportional hazards regression with age as the time metric was applied to estimate sex-specific adjusted HRs (aHRs) of mortality associated with uCd for all cancers and the cancers responsible for the most deaths in the USA. Estimates were stratified by smoking history and adjusted for education, body mass index and race. uCd was associated with cancer mortality (aHR per twofold higher uCd (95% CI), men: 1.26 (1.07 to 1.48); women: 1.21 (1.04 to 1.42)). In men, mortality from lung cancer, pancreatic cancer and non-Hodgkin lymphoma was associated with uCd; an association with leukaemia mortality was suggested. In women, associations were suggested with mortality due to lung cancer, leukaemia, ovarian and uterine cancer, but evidence was weaker than in men. Cadmium appears to be associated with overall cancer mortality in men and women, but the specific cancers associated differ between men and women, suggesting avenues for future research. Limitations of the study include the possibility of uncontrolled confounding by cigarette smoking or other factors, and the limited number of deaths due to some cancers.


Estimates of the Nutritional Impact of Non-Participation in the National School Lunch Program during COVID-19 School Closures.
The COVID-19 pandemic resulted in widespread school closures, reducing access to school meals for millions of students previously participating in the US Department of Agriculture (USDA) National School Lunch Program (NSLP). School-prepared meals are, on average, more nutritious than home-prepared meals. In the absence of recent data measuring changes in children's diets during the pandemic, this article aims to provide conservative, back-of-the-envelope estimates of the nutritional impacts of the pandemic for school-aged children in the United States. We used administrative data from the USDA on the number of NSLP lunches served in 2019 and 2020 and nationally representative data from the USDA School Nutrition and Meal Cost Study on the quality of school-prepared and home-prepared lunches. We estimate changes in lunchtime calories and nutrients consumed by NSLP participants from March to November 2020, compared to the same months in 2019. We estimate that an NSLP participant receiving no school meals would increase their caloric consumption by 640 calories per week and reduce their consumption of nutrients such as calcium and vitamin D. Because 27 to 78 million fewer lunches were served per week in March-November 2020 compared to the previous year, nationally, students may have consumed 3 to 10 billion additional calories per week. As students return to school, it is vital to increase school meal participation and update nutrition policies to address potentially widening nutrition disparities.


Multidrug-resistant genes of aminoglycoside-modifying enzymes and 16S rRNA methylases in Acinetobacter baumannii strains.
We examined the distribution of genes of aminoglycoside-modifying enzymes and 16S rRNA methylases in multidrug-resistant Acinetobacter baumannii to explore the association of these genes with drug resistance. Strains isolated from clinical specimens were screened using an automatic microbial identification system, and 9 aminoglycoside-modifying enzyme and 6 16S rRNA methylase genes were analyzed using polymerase chain reaction and verified by DNA sequencing. Next, sequence alignment was carried out using the Chromas software and a susceptibility test was performed using the Kirby-Bauer disk diffusion method. Genes encoding aminoglycoside-modifying enzymes were detected in all 20 strains of multidrug-resistant A. baumannii. The positive rates of aac(3')-I, aac(6')-Ib, ant(3'')-I, and aph(3')-I were 90.0, 90.0, 85.0, and 35.0%, respectively. However, genes encoding 16S rRNA methylases were not positively detected in the 20 strains of multidrug-resistant A. baumannii. The resistance of multidrug-resistant A. baumannii may be associated with aminoglycoside-modifying enzyme genes but not with 16S rRNA methylase genes.


Cav3.2 overexpression in L4 dorsal root ganglion neurons after L5 spinal nerve cutting involves Egr-1, USP5 and HMGB1 in rats: An emerging signaling pathway for neuropathic pain.
Overexpression of Cav3.2 T-type Ca2+ channels in L4 dorsal root ganglion (DRG) participates in neuropathic pain after L5 spinal nerve cutting (L5SNC) in rats. The L5SNC-induced neuropathic pain also involves high mobility group box 1 (HMGB1), a damage-associated molecular pattern protein, and its target, the receptor for advanced glycation end-products (RAGE). We thus studied the molecular mechanisms for the L5SNC-induced Cav3.2 overexpression as well as neuropathic pain in rats by focusing on; 1) possible involvement of early growth response 1 (Egr-1), known to regulate transcriptional expression of Cav3.2, and ubiquitin-specific protease 5 (USP5) that protects Cav3.2 from proteasomal degradation, and 2) possible role of HMGB1/RAGE as an upstream signal. Protein levels of Cav3.2 as well as Egr-1 in L4 DRG significantly increased in the early (day 6) and persistent (day 14) phases of neuropathy after L5SNC, while USP5 protein in L4 DRG did not increase on day 6, but day 14. An anti-HMGB1-neutralizing antibody or a low molecular weight heparin, a RAGE antagonist, prevented the development of neuropathic pain and upregulation of Egr-1 and Cav3.2 in L4 DRG after L5SNC. L5SNC increased macrophages accumulating in the sciatic nerves, and the cytoplasm/nuclear ratio of immunoreactive HMGB1 in those macrophages. Our findings suggest that L5SNC-induced Cav3.2 overexpression in L4 DRG and neuropathic pain involves Egr-1 upregulation downstream of the macrophage-derived HMGB1/RAGE pathway, and that the delayed upregulation of USP5 might contribute to the persistent Cav3.2 overexpression and neuropathy.


Evaluation of the food grade expression systems NICE and pSIP for the production of 2,5-diketo-D-gluconic acid reductase from Corynebacterium glutamicum.
2,5-diketo-D-gluconic acid reductase (2,5-DKG reductase) catalyses the reduction of 2,5-diketo-D-gluconic acid (2,5-DKG) to 2-keto-L-gulonic acid (2-KLG), a direct precursor (lactone) of L-ascorbic acid (vitamin C). This reaction is an essential step in the biocatalytic production of the food supplement vitamin C from D-glucose or D-gluconic acid. As 2,5-DKG reductase is usually produced recombinantly, it is of interest to establish an efficient process for 2,5-DKG reductase production that also satisfies food safety requirements. In the present study, three recently described food grade variants of the Lactobacillales based expression systems pSIP (Lactobacillus plantarum) and NICE (Lactococcus lactis) were evaluated with regard to their effictiveness to produce 2,5-DKG reductase from Corynebacterium glutamicum. Our results indicate that both systems are suitable for 2,5-DKG reductase expression. Maximum production yields were obtained with Lb. plantarum/pSIP609 by pH control at 6.5. With 262 U per litre of broth, this represents the highest heterologous expression level so far reported for 2,5-DKG reductase from C. glutamicum. Accordingly, Lb. plantarum/pSIP609 might be an interesting alternative to Escherichia coli expression systems for industrial 2,5-DKG reductase production.


Asymmetric All-Femtosecond Laser-Cut Corneal Allogenic Intrastromal Ring Segments.
To describe a novel technique for cutting asymmetric allogenic segments using the femtosecond laser for the management of cases of keratoconus with non-coinciding astigmatism and coma axes. Four eyes of 2 patients with irregular keratoconus and asymmetric allogenic segments were included. Visual, refractive, tomographic, and aberrometric outcomes, and optical coherence tomography (OCT) sections were measured preoperatively and 6 months postoperatively. Evaluations included slit-lamp examination, manifest refraction, uncorrected (UDVA) and corrected (CDVA) distance visual acuity, and simulated and maximum anterior keratometry (Kmax) using anterior segment OCT. Spherical and cylindrical refractive errors decreased from -2.38 ± 2.96 and -2.94 ± 2.16 to -1.81 ± 2.77 (P = .04) and -1.75 ± 2.07 (P = .01) diopters (D), respectively, 6 months postoperatively. There was an average gain of three lines of CDVA. Kmax decreased from 50.02 ± 1.99 to 47.89 ± 3.05 D (P= .03) and coma from 1.05 ± 0.21 to 0.21 ± 0.19 D (P = .01). Asymmetric femtosecond laser-cut allogenic segments allow a higher level of customization based on size, shape, and arc length, in contrast to the limited range of available synthetic asymmetrical segments. [J Refract Surg. 2023;39(12):856-862.].


Case Report: The First Case of COVID-19 in Bhutan.
The initial cases of novel coronavirus disease-19 (COVID-19) in a country are of utmost importance given their impact on healthcare providers, the country's preparedness response, and the initial molding of the public perception toward this pandemic. In Bhutan, the index case was a 76-year-old immunocompromised man who had traveled from the United States and entered Bhutan as a tourist. He presented initially with vague gastrointerestinal symptoms and later a cough. His atypical presentation led to a delay in diagnosis, but ultimately he was isolated and tested. On confirming the diagnosis of COVID-19, the patient was isolated in a separate hospital with a dedicated medical care team. All contacts were traced and quarantined. The patient's respiratory status deteriorated despite broad-spectrum antivirals, antibiotics, and intensive supportive care. He required intubation and was given a trial of intravenous immunoglobulin to modulate his likely aberrant immune response. Subsequently, the patient's clinical status improved, and after 8 days of hospitalization, he was transferred out of the country, where he recovered. This was a learning experience for the treating medical staff, the government, and the people of Bhutan.


Chemical Design of Palladium-Based Nanoarchitectures for Catalytic Applications.
Palladium (Pd) plays an important role in numerous catalytic reactions, such as methanol and ethanol oxidation, oxygen reduction, hydrogenation, coupling reactions, and carbon monoxide oxidation. Creating Pd-based nanoarchitectures with increased active surface sites, higher density of low-coordinated atoms, and maximized surface coverage for the reactants is important. To address the limitations of pure Pd, various Pd-based nanoarchitectures, including alloys, intermetallics, and supported Pd nanomaterials, have been fabricated by combining Pd with other elements with similar or higher catalytic activity for many catalytic reactions. Herein, recent advances in the preparation of Pd-based nanoarchitectures through solution-phase chemical reduction and electrochemical deposition methods are summarized. Finally, the trend and future outlook in the development of Pd nanocatalysts toward practical catalytic applications are discussed.


Phosphoryl choline introduces dual activity in biomimetic ionomers.
Dual activity of phosphoryl choline (PC) functional poly(trimethylene carbonate) (PTMC) was found which induces the zwitterionic biomimetic PC group to form physical cross-links with ionomers in the bulk, and at the same time enrich at the surface of cast films. The formation of zwitterionic domains from a bifunctional PC-PTMC-PC (ionomer) provided firm films with a low elastic modulus in contrast to the tacky PTMC starting material (Mn approximately 3900 g/mol) with poor mechanical performance. In addition, the ionomer possessed improved hemocompatible properties that was explained by the enrichment of PC at the surface, suggesting a way to tailor the mechanical performance of biodegradable PTMC-based ionomers while providing its bioactivity. Tailored elasticity while maintaining hemocompatibility of a biodegradable ionomer should be of particular interest for a variety of in vivo applications.


The transcriptional repressor even-skipped interacts directly with TATA-binding protein.
The Drosophila homeodomain protein Even-skipped (Eve) has previously been shown to function as a sequence-specific transcriptional repressor, and in vitro and in vivo experiments have shown that the protein can actively block basal transcription. However, the mechanism of repression is not known. Here, we present evidence establishing a direct interaction between Eve and the TATA-binding protein (TBP). Using cotransfection assays with minimal basal promoters whose activity can be enhanced by coexpression of TBP, we found that Eve could efficiently block, or squelch, this enhancement. Squelching did not require Eve DNA-binding sites on the reporter plasmids but was dependent on the presence of the Eve repression domain. Further support for an in vivo interaction between the Eve repression domain and TBP was derived from a two-hybrid-type assay with transfected cells. Evidence that Eve and TBP interact directly was provided by in vitro binding assays, which revealed a specific protein-protein interaction that required an intact Eve repression domain and the conserved C terminus of TBP. The Eve homeodomain was also required for these associations, suggesting that it may function in protein-protein interactions. We also show that a previously characterized artificial repression region behaves in a manner similar to that of the Eve repression domain, including its ability to squelch TBP-enhanced expression in vivo and to bind TBP specifically in vitro. Our results suggest a model for transcriptional repression that involves an interaction between Eve and TBP.


Stem cells in amyotrophic lateral sclerosis: motor neuron protection or replacement?
Given the lack of effective drug treatments for amyotrophic lateral sclerosis (ALS), compelling preclinical data on stem cell research has targeted this disease as a candidate for stem cell treatment. Stem cell transplantation has been effective in several animal models, but the underlying biological pathways of restorative processes are still unresolved. Several mechanisms such as cell fusion, neurotrophic factor release, endogenous stem cell proliferation, and transdifferentiation may explain positive therapeutic results in preclinical animal models, in addition to replacement of lost motor neurons. The clinical target in ALS has shifted from being neuroncentered to focus on the interaction between motor neurons and non-neuronal cells (mainly astroglial or microglial). In fact, one of the fundamental unanswered questions in ALS is whether and how much motor neuron death depends on neighboring cells, and how wildtype non-neuronal cells may protect motor neurons expressing an ALS-causing mutation. Lately, motor neuron replacement has been successfully achieved in animal models with reinnervation of the muscle target. Even if many biological issues need to be solved in preclinical models, preliminary stem cell transplantation trials have been performed in ALS patients with conflicting results. The review discusses relevant topics regarding the application of stem cell research to ALS focusing on their therapeutic relevance and mechanisms of action.