Further evidence of the etiology of prune belly syndrome provided by a transient massive intraabdominal cyst in a female.
We present a female neonate born with prune belly syndrome (PBS) in whom a large intraabdominal cyst was diagnosed at 12weeks of gestation. Rapid and exponential growth of the cyst caused pressure effects on the intraabdominal organs and stretching of the anterior abdominal wall by 19weeks of gestation. This led to drainage of the massive cyst at 20weeks of gestation to prevent fetal demise. This case provides further clues to the likely etiology of PBS: transient stretching and attenuation of the fetal abdominal wall secondary to gross fetal abdominal distension - from any cause.


Mutant phenotypes support a trans-splicing mechanism for the expression of the tripartite psaA gene in the C. reinhardtii chloroplast.
The chloroplast psaA gene of the green unicellular alga Chlamydomonas reinhardtii consists of three exons that are transcribed from different strands. Analysis of numerous nuclear and chloroplast mutants that are deficient in photosystem I activity reveals that roughly one-quarter of them are specifically affected in psaA mRNA maturation. These mutants can be grouped into three phenotypic classes, based on their inability to perform either one or both splicing reactions. The data indicate that the three exons are transcribed independently as precursors which are normally assembled in trans and that the splicing reactions can occur in either order. While some chloroplast mutations could act in cis, the nuclear mutations that fall into several complementation groups probably affect factors specifically required for assembling psaA mRNA.


An Introduction to the Science of Information Transfer, Uncertainty, and a Personal View of Organizational Intelligence.
An introduction to the concepts of entropy, information transfer, and the uncertainty in transfer of information is given. The specific target is information transfer in voice and speech. The entropies of a square wave, a sinusoid, and a sawtooth are calculated because these waveshapes approximate information carriers in vocalization. This is followed by a less-known concept that the increase of organizational intelligence gleaned from physical systems is proportional to the gradient of entropy. That leads to a multi-dimensional interpretation of diversity. Finally, a personal meta-physical extension is made to a universal reservoir of intelligence from which species can draw to advance civilizations.


Androgen receptors and testosterone in men--effects of protein ingestion, resistance exercise and fiber type.
The purpose of this study was to examine the impact of protein ingestion on circulating testosterone and muscle androgen receptor (AR) as well as on insulin-like growth factor-I (MGF and IGF-IEa) responses to a resistance exercise (RE) bout in (57-72 year) men. Protein (15 g whey) (n=9) or placebo (n=9) was consumed before and after a RE bout (5 sets of 10 repetition maximums), and vastus lateralis muscle biopsies were taken pre, 1 and 48 h post-RE. The protein ingestion blunted the RE-induced increase in serum free and total testosterone while the RE bout significantly increased muscle AR mRNA levels in older men (P<0.05). However, protein ingestion did not significantly affect AR mRNA or protein expression, or MGF and IGF-IEa mRNA expression at 1 and 48 h post-RE. Immunohistochemical staining of muscle cross-sections was done with antibodies specific to AR and MyHC I and II and showed that there seems to be within or near the type-I muscle fibers a greater staining of ARs than within or near the type-II fibres. In conclusion, the protein ingestion hinders RE-induced increase in serum testosterone in older men but may not significantly affect muscle AR, MGF or IGF-IEa gene expression. Furthermore, the present study shows that even older men are able to increase muscle AR mRNA expression in response to a RE bout.


Fibrinogen synthesis by megakaryocyte rich human marrow cell concentrates.
A rapid method is described for the production of a human marrow cell suspension highly enriched in megakaryocytes. These concentrates were incubated with radiolabelled amino-acids, and cell lysates were then analysed for fibrinogen synthesis. Neosynthesized proteins were detected by immunoprecipitation, immuno-affinity chromatography and electrophoresis. Fluorography of the electrophoresis gels showed three radioactive bands corresponding to the three chains of cold fibrinogen. Immunoblotting and autoradiography of bidimensional, nonreduced-reduced electrophoresis gels showed that these three proteins were joined by disulfide bonds in the cell. These results suggest that megakaryocytes synthesize fibrinogen, and imply that platelet fibrinogen is of megakaryocytic origin.


Development and in vitro evaluation of donepezil hydrochloride-loaded thermo-responsive polymer grafted molybdenum disulfide nanosheets: Modeling using response surface methodology.
Nanocarriers are utilized to deliver bioactive substances in the treatment of neurodegenerative diseases such as Alzheimer's. In this work, we prepared donepezil hydrochloride-loaded molybdenum disulfide modified thermo-responsive polymer as the thermo-responsive nanocarrier. Then, glycine was grafted to the surface of the polymer to improve the targeting and sustained release. The morphology, crystallinity, chemical bonding, and thermal behavior of nanoadsorbent were fully characterized by field emission scanning electron microscopes, energy dispersive X-ray, X-ray diffraction, Fourier-transform infrared spectroscopy, and thermo-gravimetric measurement. Response surface methodology with the central composite design was applied to optimize the sorption key factors such as pH solution (A: 5-9), contact time (B: 10-30 min), and temperature (C: 30-50 °C). Non-linear isotherm modeling confirmed that the sorption of the drug follows the Ferundlich model based on higher correlation coefficient values (R2 =0.9923) and lower errors values (root means square errors: 0.16 and Chi-square: 0.10), suggesting a heterogeneous multilayer surface sorption. The non-linear sorption kinetic modeling revealed that the pseudo-second-order kinetic model well-fitted the sorption data of the drug on the nanoadsorbent surface based on higher R2 values (R2 =0.9876) and lower errors values (root means square errors: 0.05 and Chi-square: 0.02). The in vitro drug release experiment of donepezil hydrochloride shown that about 99.74 % of drug release was found to be occurred at pH= 7.4 (T = 45 °C) within 6 h, whereas about 66.32 % of drug release occurred at pH= 7.4 (T = 37 °C). The release of donepezil hydrochloride from as prepared drug delivery system has shown a sustained release profile, which was fitted to Korsmeyer-Peppas kinetics.


A genetic study of the role of the Wnt/beta-catenin signalling in Paneth cell differentiation.
Wnt/beta-catenin signalling plays a key role in the homeostasis of the intestinal epithelium. Whereas its role in the maintenance of the stem cell compartment has been clearly demonstrated, its role in the Paneth cell fate remains unclear. We performed genetic studies to elucidate the functions of the Wnt/beta-catenin pathway in Paneth cell differentiation. We analysed mice with inducible gain-of-function mutations in the Wnt/beta-catenin pathway and mice with a hypomorphic beta-catenin allele that have not been previously described. We demonstrated that acute activation of Wnt/beta-catenin signalling induces de novo specification of Paneth cells in both the small intestine and colon and that colon cancers resulting from Apc mutations expressed many genes involved in Paneth cell differentiation. This suggests a key role for the Wnt/beta-catenin pathway in Paneth cell differentiation. We also showed that a slight decrease in beta-catenin gene dosage induced a major defect in Paneth cell differentiation, but only a modest effect on crypt morphogenesis. Overall, our findings show that a high level of beta-catenin activation is required to determine Paneth cell fate and that fine tuning of beta-catenin signalling is critical for correct Paneth cell lineage.


Pneumoperitoneum in the presence of acute and chronic kidney injury: an experimental model in rats.
There is evidence that pneumoperitoneum induces transient changes in renal function in healthy individuals. Its effect on malfunctioning kidneys is not well known. We investigated effects caused by pneumoperitoneum in animals with impaired renal function. Male Wistar rats underwent subtotal (5/6) nephrectomy in 2 sequential steps to induce renal failure. Two and 10 weeks postoperatively rats were classified with acute and chronic kidney injury, respectively. At those time points all rats were exposed to 0, 5 and 8 mm Hg pneumoperitoneum for 60 minutes. Changes in creatinine, blood urea nitrogen and creatinine clearance were measured. Histopathological changes and apoptosis were also evaluated in the subgroups. A total of 18 rats with acute and 18 with chronic kidney injury completed the study. Creatinine and blood urea nitrogen did not change after applying pneumoperitoneum in the different pressure subgroups but creatinine clearance significantly decreased in the 5 and 8 mm Hg subgroups in rats with acute and chronic kidney injury. Histopathological findings in the acute kidney injury subgroups that underwent 5 and 8 mm Hg pressure revealed ischemic changes while compensatory hypertrophy was noticed in the chronic injury pressurized subgroups. The apoptotic count was significantly higher in the chronic injury subgroups compared to their acute injury pressurized counterparts. Pneumoperitoneum seems feasible in rats with impaired baseline renal function. Particularly chronic cases should not be considered a contraindication to pneumoperitoneum while in acute cases pneumoperitoneum might be detrimental.


The spatiotemporal spread of cervical spinal cord contusion injury pathology revealed by 3D in-line phase contrast synchrotron X-ray microtomography.
Extensive structural changes occur within the spinal cord following traumatic injury. Acute tissue debris and necrotic tissue are broken down, proliferating local glia and infiltrating leukocytes remodel tissue biochemical and biophysical properties, and a chronic cavity surrounded by a scar forms at the injury epicentre. Serial-section 2D histology has traditionally assessed these features in experimental models of spinal cord injury (SCI) to measure the extent of tissue pathology and evaluate efficacy of novel therapies. However, this 2D snapshot approach overlooks slice intervening features, with accurate representation of tissue compromised by mechanical processing artefacts. 3D imaging avoids these caveats and allows full exploration of the injured tissue volume to characterise whole tissue pathology. Amongst 3D imaging modalities, Synchrotron Radiation X-ray microtomography (SRμCT) is advantageous for its speed, ability to cover large tissue volumes at high resolution, and need for minimal sample processing. Here we demonstrate how extended lengths of formalin-fixed, paraffin-embedded (FFPE) rat spinal cord can be completely imaged by SRμCT with micron resolution. Label-free contrast derived from X-ray phase interactions with low-density soft tissues, reveals spinal cord white matter, gray matter, tissue damage and vasculature, with tissue still viable for targeted 2D-histology after 3D imaging. We used SRμCT to quantify tissue pathology after a midline, cervical level (C6), 225 kDyne contusion injury over acute-to-chronic (24 h to 5 weeks) post injury time points. Quantification revealed acute tissue swelling prior to chronic atrophy across the whole imaged region (spanning 2 spinal segments above and below injury), along with rostro-caudal asymmetries in white and gray matter volume loss. 3D volumes revealed satellite damage in tissue far removed from the epicentre, and extensive rostro-caudal spread of damage through the base of the dorsal columns at 24 h post injury. This damage overlapped regions of vasogenic oedema, confirmed with subsequent histology. Tissue damage at later time points in border regions was most prominent in the dorsal columns, where it overlapped sites of damaged venous vasculature. Elaborating rostro-caudal and spatiotemporal asymmetries in reduced traumatic injury models centred on these regions may inform future treatments that seek to limit the spread of tissue pathology to these 'at-risk' regions.


Variola minor in Bragança Paulista County, 1956. Distribution of onset of cases in the two schools including most school children with variola.
Class attendance during illness was confirmed for numerous pupils of two schools. Successive generations whose median cases were separated by an interval consistent with the "serial interval" of variola minor were clearly found in the epidemic curve for II of the 29 classes with cases from both schools. Twenty two other classes had no case at all.


Amelioration of murine dextran sulfate sodium-induced colitis by nuclear factor-kappaB decoy oligonucleotides.
Activation of nuclear factor (NF)-kappaB has been shown to play a critical role in the pathogenesis of ulcerative colitis (UC). The purpose of the current study was to investigate the effects of NF-kappaB decoy oligonucleotides (ODNs) on an experimental model of UC. NF-kappaB decoy ODNs were administered in experimental colitis induced by dextran sulfate sodium (DSS). The disease activity index (DAI) and histological score were observed. NF-kappaB DNA binding activity was assessed by electrophoretic mobility shift assay (EMSA). The expression of tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) were measured by reverse transcription polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA). A significant improvement was observed in DAI and histological score in mice with NF-kappaB decoy ODNs, and the increase in NF-kappaB DNA binding activity, myeloperoxidase (MPO) activity, IL-1beta, and TNF-a in mice with DSS-induced colitis was significantly reduced following administration of NF-kappaB decoy ODNs. The administration of NF-kappaB decoy ODNs leads to an amelioration of DSS-induced colitis, suggesting administration of NF-kappaB decoy ODNs may provide a therapeutic approach for UC.


Differentiation of amyloid fibril proteins in tissue sections. Two simple and reliable histological methods applied to fifty-one cases of systemic amyloidosis.
The potassium permanganate method and the unlabeled immunoperoxidase (PAP) method were applied for distinguishing different types of amyloid fibril proteins in conventionally fixed, paraffin-embedded tissue sections obtained from fifty-one autopsied cases of systemic amyloidosis and three control cases of well-analysed fibril proteins. All of the eighteen cases "sensitive" to permanganate treatment, whose amyloid deposits lost completely their affinity to Congo red and birefringence under polarized light, were shown to have AA antigenic determinants by the PAP method. Meanwhile, all of the remaining thirty-three "resistant" cases, where Congo red affinity and birefringence were retained at various degree even only in minimal areas, were negative for AA antigenicity. This indicated the feasibility of potassium permanganate method for the identification of AA protein based on this criterion of "sensitivity". Twenty-eight cases were classified as AA, A lambda, A kappa or AA+[A kappa] the remaining twenty-three cases were unclassified, and there were some discrepancies between the preliminary clinicopathological classification and the protein nature of the amyloid. It is important to differentiate the types of amyloid fibril protein of individual patients because the expedience of selective therapeutic approaches had been suggested. The two methods applied herein are handy and useful for this purpose.


Distinct roles of NF-kappaB p50 in the regulation of acetaminophen-induced inflammatory mediator production and hepatotoxicity.
Oxidative stress plays an important role in acetaminophen (APAP)-induced hepatotoxicity. In addition to inducing direct cellular damage, oxidants can activate transcription factors including NF-kappaB, which regulate the production of inflammatory mediators implicated in hepatotoxicity. Here, we investigated the role of APAP-induced oxidative stress and NF-kappaB in inflammatory mediator production. Treatment of mice with APAP (300 mg/kg, i.p.) resulted in centrilobular hepatic necrosis and increased serum aminotransferase levels. This was correlated with depletion of hepatic glutathione and CuZn superoxide dismutase (SOD). APAP administration also increased expression of the proinflammatory mediators, interleukin-1beta (IL-1beta), tumor necrosis factor-alpha (TNFalpha), macrophage chemotactic protein-1 (MCP-1), and KC/gro, and the anti-inflammatory cytokine, interleukin-10 (IL-10). Pretreatment of mice with the antioxidant, N-acetylcysteine (NAC) prevented APAP-induced depletion of glutathione and CuZnSOD, as well as hepatotoxicity. NAC also abrogated APAP-induced increases in TNFalpha, KC/gro, and IL-10, but augmented expression of the anti-inflammatory cytokines interleukin-4 (IL-4) and transforming growth factor-beta (TGFbeta). No effects were observed on IL-1beta or MCP-1 expression. To determine if NF-kappaB plays a role in regulating mediator production, we used transgenic mice with a targeted disruption of the gene for NF-kappaB p50. As observed with NAC pretreatment, the loss of NF-kappaB p50 was associated with decreased ability of APAP to upregulate TNFalpha, KC/gro, and IL-10 expression and increased expression of IL-4 and TGFbeta. However, in contrast to NAC pretreatment, the loss of p50 had no effect on APAP-induced hepatotoxicity. These data demonstrate that APAP-induced cytokine expression in the liver is influenced by oxidative stress and that this is dependent, in part, on NF-kappaB. However, NF-kappaB p50-dependent responses do not appear to play a major role in the pathogenesis of toxicity in this model.


APOE loss-of-function variants: Compatible with longevity and associated with resistance to Alzheimer's Disease pathology.
The ε4 allele of apolipoprotein E (APOE) is the strongest genetic risk factor for sporadic Alzheimer's Disease (AD). Knockdown of this allele may provide a therapeutic strategy for AD, but the effect of APOE loss-of-function (LoF) on AD pathogenesis is unknown. We searched for APOE LoF variants in a large cohort of older controls and patients with AD and identified six heterozygote carriers of APOE LoF variants. Five carriers were controls (ages 71-90) and one was an AD case with an unremarkable age-at-onset between 75-79. Two APOE ε3/ε4 controls (Subjects 1 and 2) carried a stop-gain affecting the ε4 allele. Subject 1 was cognitively normal at 90+ and had no neuritic plaques at autopsy. Subject 2 was cognitively healthy within the age range 75-79 and underwent lumbar puncture at between ages 75-79 with normal levels of amyloid. The results provide the strongest human genetics evidence yet available suggesting that ε4 drives AD risk through a gain of abnormal function and support knockdown of APOE ε4 or its protein product as a viable therapeutic option.


Equine chlamydiosis-An emerging infectious disease requiring a one health surveillance approach.
Psittacosis is a rare but potentially fatal zoonosis caused by Chlamydia psittaci, an organism that is typically associated with bird contact. However C. psittaci is capable of infecting other non-avian hosts, such as horses, sheep, cattle and goats. Stud staff and veterinarians have significant exposure to parturient animals and reproductive materials in their routine work. To investigate the zoonotic potential associated with the emergence of C. psittaci as an abortifacient agent in horses, we established a programme of joint human and animal surveillance in a sentinel horse-breeding region in Australia. This programme comprised cross-notification of equine cases to public health agencies, and active follow-up of known human contacts, including stud workers, foaling staff, veterinarians and laboratory staff. We identified no confirmed cases of acute psittacosis despite intensive surveillance and testing of heavily exposed contacts; however, further work in the area is needed.


Inhibition of the mechanistic target of rapamycin induces cell survival via MAPK in tuberous sclerosis complex.
Tuberous sclerosis complex (TSC) is a genetic disorder that cause tumors to form in many organs. These lesions may lead to epilepsy, autism, developmental delay, renal, and pulmonary failure. Loss of function mutations in TSC1 and TSC2 genes by aberrant activation of the mechanistic target of rapamycin (mTORC1) signaling pathway are the known causes of TSC. Therefore, targeting mTORC1 becomes a most available therapeutic strategy for TSC. Although mTORC1 inhibitor rapamycin and Rapalogs have demonstrated exciting results in the recent clinical trials, however, tumors rebound and upon the discontinuation of the mTORC1 inhibition. Thus, understanding the underlying molecular mechanisms responsible for rapamycin-induced cell survival becomes an urgent need. Identification of additional molecular targets and development more effective remission-inducing therapeutic strategies are necessary for TSC patients. We have discovered an Mitogen-activated protein kinase (MAPK)-evoked positive feedback loop that dampens the efficacy of mTORC1 inhibition. Mechanistically, mTORC1 inhibition increased MEK1-dependent activation of MAPK in TSC-deficient cells. Pharmacological inhibition of MAPK abrogated this feedback loop activation. Importantly, the combinatorial inhibition of mTORC1 and MAPK induces the death of TSC2-deficient cells. Our results provide a rationale for dual targeting of mTORC1 and MAPK pathways in TSC and other mTORC1 hyperactive neoplasm.


Clinical outcome of 36 male patients with primary urethral carcinoma: a single center experience.
Retrospective analysis of male urethral carcinoma to assess the best therapeutic approach to the management of this tumor. A review of 36 cases of male urethral carcinoma diagnosed and treated at our center was performed. Clinical features, treatment modality and outcomes were analysed. The overall median survival time was 55.16 months. The 5-year overall and disease-free survival rate for the cohort was 49% and 23%, respectively. The 5-year survival is 67% for low-stage versus 33% for high-stage tumors and is significantly different (P = 0.001). The survival was 72% for tumors of the distal urethra versus 36% for tumors of the proximal, with a P-value of 0.02. The tumor location and clinicopathological stage were the most important predictors of the disease-free and overall survival. Multimodal approach is necessary for achieving local control especially for proximal and higher stage tumors.


The effects of insulin-induced hypoglycaemia on cardiovascular function in normal man: studies using radionuclide ventriculography.
The cardiovascular effects of an intravenous injection of soluble insulin and of acute hypoglycaemia were examined in six normal male subjects using multiple-gated radionuclide ventriculography. The basal left ventricular ejection fraction rose significantly from 47 +/- 3% (mean +/- SEM) to 54 +/- 3% p less than 0.01, within 5 min of the intravenous injection of insulin, and before any significant changes occurred in the blood glucose concentration. The ejection fraction subsequently rose to a peak of 72 +/- 5% coinciding with the onset of the acute hypoglycaemic reaction. This corresponded to the nadir of blood glucose and was associated with rises in heart rate, stroke volume and cardiac output. The heart rate returned to the resting value within 30 min of the acute hypoglycaemic reaction, but the ejection fraction, stroke volume and cardiac output were still elevated 90 min later. The peak ejection fraction value immediately preceded the maximal increment of plasma catecholamines released in response to hypoglycaemia. Thus, administration of intravenous insulin had a small, immediate, discernible effect on the cardiovascular system. A subsequent rise in left ventricular ejection fraction of much greater magnitude was stimulated by the development of acute hypoglycaemia, and was associated temporally with sympatho-adrenal activation. The use of radionuclide ventriculography showed that the haemodynamic changes provoked by hypoglycaemia produced a sustained effect on cardiac contractility.


ZipN is an essential FtsZ membrane tether and contributes to the septal localization of SepJ in the filamentous cyanobacterium Anabaena.
The organismic unit of heterocyst-forming cyanobacteria is a filament of communicating cells connected by septal junctions, proteinaceous structures bridging the cytoplasms of contiguous cells. This distinct bacterial organization is preserved during cell division. In Anabaena, deletion of the zipN gene could not be segregated. We generated strain CSL109 that expresses zipN from a synthetic regulatable promoter. Under conditions of ZipN depletion, cells progressively enlarged, reflecting restricted cell division, and showed drastic morphological alterations including cell detachment from the filaments, to finish lysing. In contrast to the wild-type localization in midcell Z-rings, FtsZ was found in delocalized aggregates in strain CSL109. Consistently, the proportion of membrane-associated to soluble FtsZ in fractionated cell extracts was lower in CSL109. Bacterial two-hybrid analysis showed that ZipN interacts with FtsZ and other cell-division proteins including cytoplasmic Ftn6 and SepF, and polytopic FtsW, FtsX, FtsQ and FtsI. Additionally, ZipN interacted with the septal protein SepJ, and in CSL109 depletion of ZipN was concomitant with a progressive loss of septal specificity of SepJ. Thus, in Anabaena ZipN represents an essential FtsZ membrane tether and an organizer of the divisome, and it contributes to the conformation of septal structures for filament integrity and intercellular communication.


Detection of adenovirus nucleic acid sequences in human tonsils in the absence of infectious virus.
Human tonsillar and adenoid tissues from surgical specimens and cell cultures established therefrom were screened for adenovirus type 2 (Ad2) sequences by in situ hybridization. For labeling we have utilized biotinylated DNA probes. We report detection of adenoviral sequences after hybridization with adenovirus type 2 DNA probes in tissues as well as in cell cultures from specimens without any signs of infectious virus even after long-term cultivation. In the infected tonsils only some of the cells appear to carry viral sequences. In conclusion, truly latent adenovirus infections in man seem to occur.


Trends in infant mortality in Austria between 1984 and 2002.
Infant mortality rate is an important medical indicator and is often used for comparing countries with respect to welfare and public health. Among other factors, effective medical technology, better access to pre- and postnatal care for all socioeconomic groups and better nutrition have decreased infant mortality in Austria from about 200 deaths per 1000 live births at the beginning of the 20th century to about 5 deaths per 1000 live births at the end. In this study we present the trends in infant mortality, based on 1,654,519 individual birth records, in Austria since 1984. The infant mortality rate dropped rapidly from about 12 per 1000 live births in 1985 to 4.6 per 1000 live births during the last two years of our study (2001/02). Infant mortality rates stratified by cause of death show somewhat differing trends. In particular, the number of deaths due to peripartal problems decreased as the result of improvements in obstetrics and neonatology, but in 1995 a change in the definition of live birth led to a rise of about 20% in the stillbirth rate. At present, Austria has one of the lowest infant mortality rates of all European countries; however, between 1999 and 2002 the mortality rate has been fairly static. A further reduction in mortality clearly cannot be achieved by advances in medicine alone. It remains a challenge for health politicians, physicians and society at large to reduce the prevalence of well-known risk factors such as alcohol abuse, heavy overweight and smoking during pregnancy.


Human placental 3 beta-hydroxy-5-ene-steroid dehydrogenase and steroid 5----4-ene-isomerase: purification from mitochondria and kinetic profiles, biophysical characterization of the purified mitochondrial and microsomal enzymes.
In human placenta, 3 beta-hydroxy-5-ene-steroid dehydrogenase and steroid 5----4-ene-isomerase, an enzyme complex found in microsomes and mitochondria, synthesizes progesterone from pregnenolone and androstenedione from fetal dehydroepiandrosterone sulfate. The dehydrogenase and isomerase activities of the mitochondrial enzyme were copurified (733-fold) using sequential cholate solubilization, ion exchange chromatography (DEAE-Toyopearl 650S), and hydroxylapatite chromatography (Bio-Gel HT). Enzyme homogeneity was demonstrated by a single protein band in SDS-polyacrylamide gel electrophoresis (monomeric Mr = 41,000), gel filtration at constant specific enzyme activity (Mr = 77,000), and a single NH2-terminal sequence. Kinetic constants were determined for the oxidation of pregnenolone (Km = 1.6 microM, Vmax = 48.6 nmol/min/mg) and dehydroepiandrosterone (Km = 2.4 microM, Vmax = 48.5 nmol/min/mg) and for the isomerization of 5-pregnene-3,20-dione (Km = 9.3 microM, Vmax = 914.2 nmol/min/mg) and 5-androstene-3,17-dione (Km = 27.6 microM, Vmax = 888.4 nmol/min/mg. Mixed substrate studies showed that the dehydrogenase and isomerase activities utilize their respective pregnene and androstene substrates competitively. Dixon analysis demonstrated that the product steroids, progesterone and androstenedione, are competitive inhibitors of the C-21 and C-19 dehydrogenase activities. Enzyme purified from mitochondria and microsomes had similar kinetic profiles with respect to substrate utilization, product inhibition, and cofactor (NAD+) reduction (mean Km +/- SD using C-19 and C-21 dehydrogenase substrates = 26.4 +/- 0.8 microM, mean Vmax = 73.2 +/- 1.3 nmol/min/mg). Pure enzyme from both organelles exhibited identical biophysical properties in terms of molecular weight and subunit composition, pH optima (pH 9.8, dehydrogenase; pH 7.5, isomerase), temperature optimum (37 degrees C), stability in storage and solution, effects of divalent cations, and the single NH2-terminal sequence of 27 amino acids. These results suggest that the mitochondrial and microsomal enzymes are the same protein localized in different organelles.


Serum-specific antibodies for Chlamydia trachomatis in premature contractions.
A case-control study was undertaken to examine the possible morbidity associated with Chlamydia trachomatis in 28 pregnant women having idiopathic premature contractions, in 35 healthy preterm pregnant women, and in 43 healthy pregnant women at term. Serum C. trachomatis IgG- and IgA-specific antibodies were determined by the single serovar inclusion immunoperoxidase assay. There were no significant differences in the prevalence rate of elevated C. trachomatis IgG-specific antibodies (titer greater than or equal to 1:128) between pregnant women suffering from idiopathic premature contractions as compared with healthy preterm and term pregnant women (11%, 28%, and 26%, respectively). The known prevalence rate in a normal healthy population is 23%. The prevalence rate of elevated C. trachomatis IgA-specific antibodies (titer greater than or equal to 1:16) was significantly lower in the pregnant women with idiopathic premature contractions as compared with the healthy preterm and term pregnant women (0%, 20%, and 17%, respectively; p less than 0.002 and p less than 0.012). These findings do not support the assumption that C. trachomatis has a role in premature contractions.


Pulmonary glutamine production: effects of sepsis and pulmonary infiltrates.
To define the role of the lung in the production of glutamine in the critically ill and to determine the effects of the presence of pulmonary infiltrates and the presence and severity of sepsis. Prospective clinical study in a single center; interdisciplinary intensive care unit at a university hospital. Eleven critically ill patients were compared to ten patients prior to cardiac bypass surgery. Fluxes of glutamine and other amino acids were measured. Chest radiography was performed, and APACHE II and multiple-organ failure scores were calculated. Septic patients showed significantly higher glutamine efflux from the lungs than controls. At least one-half of this glutamine is estimated to result from protein breakdown. Severity of illness had no impact on glutamine fluxes. In the presence of pulmonary infiltrates on chest radiographs glutamine efflux did not differ from zero. The lungs produce significant amounts of glutamine in septic patients. Pulmonary infiltrates decrease the glutamine efflux from the lung in septic patients. We suggest that this is caused by uptake of glutamine by white cells in the lung exerting immunological functions.


WNT and NOTCH signaling in human trophoblast development and differentiation.
Correct development of the human placenta and its differentiated epithelial cells, syncytial trophoblasts (STBs) and extravillous trophoblasts (EVTs), is crucial for a successful pregnancy outcome. STBs develop by cell fusion of mononuclear cytotrophoblasts (CTBs) in placental floating villi, whereas migratory EVTs originate from specialized villi anchoring to the maternal decidua. Defects in trophoblast differentiation have been associated with severe pregnancy disorders such as early-onset preeclampsia and fetal growth restriction. However, the evolutionary pathways underlying normal and adverse placentation are poorly understood. Herein, we discuss Wingless (WNT) and NOTCH signaling, two pathways that play pivotal roles in human placenta and trophoblast development. Whereas WNT is necessary for expansion of trophoblast progenitors and stem cells, NOTCH1 is required for proliferation and survival of EVT precursors. Differentiation of the latter is orchestrated by a switch in NOTCH receptor expression as well as by changes in WNT ligands and their downstream effectors.


Chemokine receptors expression and migration potential of tumor-infiltrating and peripheral-expanded Vgamma9Vdelta2 T cells from renal cell carcinoma patients.
We previously showed that Vdelta2 T cells infiltrate renal tumors and can be expanded as potent cytotoxic effectors from peripheral blood mononuclear cells of most renal cell carcinoma (RCC) patients, using a structural analog of nonconventional T-cell receptor gamma9delta2 ligand, bromohydrin pyrophosphate, and interleukin-2 (IL-2). In this study, we have further investigated the differentiation status and the migration potential of circulating and tumor-infiltrating Vgamma9Vdelta2 T lymphocytes from RCC patients. The repertoire of tumor-infiltrating and peripheral Vgamma9Vdelta2 T cells from RCC patients was characterized by a dominant CD27- CD45RA- subset. These effector memory Vgamma9Vdelta2 T cells were efficiently expanded using bromohydrin pyrophosphate combined with IL-15, but not IL-2. In addition, peripheral Vgamma9Vdelta2 T cells from RCC patients present a modified chemotactic pattern compared with donors. After ex vivo activation, peripheral expanded Vgamma9Vdelta2 T cells acquire low-migration capacities toward renal cells. Tumor-infiltrating Vgamma9Vdelta2 T cells migrated with higher efficiency toward primary renal tumor cells. The traffic toward tumor cells required the CXCL12/CXCR4 interaction. Altogether, these results outline that those Vgamma9Vdelta2 effectors exhibit differential migration capacities according to their localization, their differentiation status, and the tumor microenvironment parameters that may influence their use in immunotherapy.


[Thrombelastometric detection of factor XIII deficiency].
While undergoing Whipple's operation (pancreaticoduodenectomy) a patient developed diffuse bleeding and an unexpectedly high blood loss. An intraoperatively performed thrombelastometry with ROTEM (Pentapharm, Munich, Germany) showed an aprotonin-resistant mild fibrinolysis and suggested the presence of an isolated deficiency of coagulation factor XIII. This was confirmed by a second thrombelastometry, where no lysis was seen after in vitro substitution of factor XIII. After administration of 1250 IU factor XIII concentrate the diffuse bleeding ceased and further substitution of coagulation factor concentrates or fresh frozen plasma was not necessary. A postoperatively performed analysis confirmed the factor XIII deficiency (52%).


Effects of gene tranfection with CH50 polypeptide on the invasion ability of bladder cancer cell line BIU-87.
The expression of CH50 polypoptide in bladder cancer cell line BIU-87 and the effects on the invasion ability of BIU-87 were investigated. The eukaryotic expressing vector pCH510 of polypeptide CH50 was introduced into BIU-87 cells by gene transfection in vitro. The expression of CH50 polypeptide was detected by using immunohistochemical S-P method. The expression of the transfected gene was identified by RT-PCR. Cell invasion assay kit was applied to detect the effect of CH50 polypeptide on the invasion ability of BIU-87. The results showed that the BIU-87 cells transfected with pCH510 could express the CH50 polypeptide, while in the control group, no CH50 polypeptide was detectable. In the transfection group, the invasion ability of BIU-87 in vitro was lower than in control group (P<0.05). It was concluded that CH50 polypeptide was successfully expressed in BIU-87 cells by gene transfection, by which the in vitro invasion ability of BIU-87 was inhibited.


Increasing prevalence of multiple sclerosis in Tuscany, Italy.
An increase of prevalence and incidence of multiple sclerosis (MS) has been reported in several countries, especially taking into account a long-term evaluation. This increasing trend often reflects improved case identification and ascertainment due to the refinement of diagnostic criteria. The aim of this study was to update the prevalence rate of MS in Tuscany (central Italy) as of 2017, and to assess if there has been an increasing trend of prevalence in this Region considering a short period of analysis, from 2014 to 2017. To capture prevalent cases, a case-finding algorithm based on administrative data, previously created and validated, was used. As data sources, we considered hospital discharge records, drug-dispensing records, disease-specific exemptions from copayment to health care, home and residential long-term care, and inhabitant registry. As of January 1, 2017, 7809 cases were identified, of which 69.4% were females and 30.6% were males. Considering temporal variation, an increasing trend was observed, with standardized rates rising from 189.2 in 2014 to 208.7 per 100,000 in 2017. Results confirm that prevalence increases every year, probably mainly due to the difference between incidence and mortality, resulting in an increasing trend. Moreover, administrative data may accurately identify MS patients in a routinary way and monitor this cohort along disease care pathways.


Interference of cardiac implantable electronic devices and computed tomography imaging in the current era with a phantom model.
Cardiac implantable electronic devices are used in patients with cardiac rhythm disorders. Computed tomography irradiation is not prohibited for patients with cardiac implantable electronic devices, despite adverse events being reported. Hence, appropriate preparation and knowledge are required before computed tomography irradiation can be carried out in these patients. Since there is limited knowledge or literature about the influence of computed tomography irradiation in cases with recent cardiac implantable electronic devices, we aimed to evaluate the adverse events and elucidate the necessary and sufficient safety measures associated with this therapy. We placed cardiac implantable electronic devices on an anthropomorphic phantom model and observed their electrical activity in electrograms, while various protocols of computed tomography irradiation were implemented and adverse events evaluated. Oversensing with pauses of up to 3.2 s was observed in standard computed tomography protocols, but ventricular tachyarrhythmia or other clinically significant events could not be confirmed. Oversensing with pauses of up to 8.0 s was observed and ventricular tachyarrhythmia was detected in the maximum-dose protocols. However, treatments such as antitachycardia pacing or shock therapy for ventricular tachyarrhythmia were not observed because of their absence. Computed tomography irradiation for patients using cardiac implantable electronic devices is highly unlikely to cause clinically significant adverse events with the device settings and computed tomography protocols currently being used. Changing or monitoring the device settings routinely before computed tomography irradiation is not necessarily required for most patients.


Correction to: Assessment of vascular leakage and its development with FFA among patients treated with intravitreal anti-VEGF due to aggressive posterior ROP.
In the original publication, the authors' affiliations were published incorrectly.


Physiological promoters reduce the genotoxic risk of integrating gene vectors.
The possible activation of cellular proto-oncogenes as a result of clonal transformation is a potential limitation in a therapeutic approach involving random integration of gene vectors. Given that enhancer promiscuity represents an important mechanism of insertional transformation, we assessed the enhancer activities of various cellular and retroviral promoters in transient transfection assays, and also in a novel experimental system designed to measure the activation of a minigene cassette contained in stably integrating retroviral vectors. Retroviral enhancer-promoters showed a significantly greater potential to activate neighboring promoters than did cellular promoters derived from human genes, elongation factor-1alpha (EF1alpha) and phosphoglycerate kinase (PGK). Self-inactivating (SIN) vector design reduced but did not abolish enhancer interactions. Using a recently established cell culture assay that detects insertional transformation by serial replating of primary hematopoietic cells, we found that SIN vectors containing the EF1alpha promoter greatly decrease the risk of insertional transformation. Despite integration of multiple copies per cell, activation of the crucial proto-oncogene Evi1 was not detectable when using SIN-EF1alpha vectors. On the basis of several quantitative indicators, the decrease in transforming activity was highly significant (more than tenfold, P < 0.01) when compared with similarly designed vectors containing a retroviral enhancer-promoter with or without a well-characterized genetic insulator core element. In this manner, the insertional biosafety of therapeutic gene vectors can be greatly enhanced and proactively evaluated in sensitive cell-based assays.


Elevated skeletal muscle irisin precursor FNDC5 mRNA in obese OLETF rats.
There is debate as to whether fibronectin type III domain containing 5 (FNDC5) and its protein product irisin are therapeutic targets for obesity-associated maladies. Thus, we sought to examine FNDC5 mRNA within skeletal muscle of obese/diabetic-prone Otsuka Long-Evans Tokushima Fatty (OLETF) rats versus lean/healthy Long Evans Tokushima Otsuka (LETO) rats. We hypothesized that FNDC5 expression would be greater in obese (OLETF) versus lean (LETO) animals. Triceps muscle of 30-32week old OLETF and LETO rats were assayed for FNDC5 and PGC1α mRNA levels. Body composition and circulating biomarkers of the OLETF and LETO rats were also correlated with skeletal muscle FNDC5 mRNA expression patterns in order to examine potential relationships that may exist. OLETF rats exhibited twice the amount of triceps FNDC5 mRNA compared to LETO rats (p<0.01). Significant positive correlations existed between triceps muscle FNDC5 mRNA expression patterns versus fat mass (r=0.70, p=0.008), as well as plasma leptin (r=0.82, p<0.001). PGC1α mRNA levels were also highly correlated with FNDC5 mRNA (r=0.85, p<0.001). In subsequent culture experiments, low and high physiological doses of leptin had no effect on PGC1α mRNA or FNDC5 mRNA levels in C2C12 myotubes. Paradoxically, circulating irisin concentrations tended to be higher in a second cohort of LETO versus OLETF rats (p=0.085). These results reveal a positive association between total body adiposity and skeletal muscle FNDC5 gene expression. Of interest, circulating irisin levels tended to be lower in OLETF rats. Further research is needed to examine whether other adipose tissue-derived factors up-regulate FNDC5 transcription and/or inhibit irisin biosynthesis from FNDC5.


Risk assessment visualization of rubidium compounds: comparison of renal and hepatic toxicities, in vivo.
Rubidium has been considered to be nontoxic. Its use includes thin film on glass deposition and as medical contrast medium. Recent technology innovations also involve the use of rubidium, but there is limited information about the biological effects of its various compounds. In the present risk assessment study, a series of rubidium compounds with different counter anions-acetate, bromide, carbonate, chloride, and fluoride-were orally administrated in a single dose to several groups of rats. Cumulative 24-h urine samples were obtained, and the levels of rubidium, fluoride, N-acetyl-β-D-glucosaminidase and creatinine were measured to evaluate possible acute renal effects. Daily samples of serum were also obtained to determine the levels of aspartate and alanine aminotransferases to assess possible acute hepatic effects. Urinary rubidium excretion recovery of 8.0-10.5% shows that urine can be a useful diagnostic tool for rubidium exposure. The present results reveal that rubidium shows different biological effects depending on the counter anion. A pattern of large significant NAG leakage and elevation of ALT observed in rats treated with anhydrous rubidium fluoride indicates renal and hepatic toxicities that can be attributed to fluoride. The techniques reported in this study will be of help to assess the potential risks of toxicity of rubidium compounds with a variety of anions.


Disrupted caregiving and maternal HIV disease: a proposed model for evaluating HIV-affected children's psychosocial adjustment.
A current concern among social science researchers is the impact of human immunodeficiency virus (HIV) disease on the family, especially on the parenting abilities of mothers with HIV. Beginning research suggests that the children of HIV-positive parents are vulnerable, pointing to the impact of HIV disease on children's social and emotional adjustment. The objective of this article is to propose a model of the HIV disease-related disruption of care on children's psychosocial adjustment. The schematic model can be used to evaluate the mechanisms by which maternal HIV disease may influence the psychosocial adjustment of elementary school-age children.


Self-Cleaning Properties of Electrospun PVA/TiO₂ and PVA/ZnO Nanofibers Composites.
In this report, polyvinyl alcohol/zinoxide (PVA/ZnO) & polyvinyl alcohol/titanium dioxide (PVA/TiO₂) nanofibers were manufactured in three different concentrations of ZnO and TiO₂ NPs for the application of self-cleaning properties because metallic oxides, specifically ZnO & TiO₂, have the properties to remove the contaminants by hydroxyl radical (OH-1), which degrades the contaminants into small molecules and finally into CO₂ and H₂O. Therefore, these composites were manufactured by electrospinning. The resultant nanofibers were characterized for morphology by scan electron microscopy (SEM) & transmission electron microscopy (TEM), chemical interactions by Fourier-transform infrared (FT-IR) spectra, crystalline structure by X-ray diffraction (XRD) spectra water absorbency was evaluated by water contact angle, self-cleaning by solar simulator, and thermal degradation was done by thermogravimetric analysis (TGA) for the sake of nanoparticles the content. On the base of the characterization results it was concluded that these PVA/ZnO & PVA/TiO₂ nanofibers have self cleaning properties, but PVA/ZnO nanofibers have higher self-cleaning properties than PVA/TiO₂ nanofibers because PVA/ZnO nanofibers have 95% self-cleaning properties, which is higher than PVA/TiO₂ nanofibers.


Digital technologies to facilitate minimally invasive rehabilitation of a severely worn dentition: A dental technique.
This technique report describes digital technologies to facilitate minimally invasive rehabilitation with computer-aided design and computer-aided manufacturing (CAD-CAM) veneers for a severely worn dentition. The lingual veneers without tooth preparation and occlusal veneers with minimal tooth preparation were designed from the digital scan of the interim restorations. To facilitate the cementation of multiunit lingual veneers on unprepared teeth, a custom 3D-printed guide was designed from the digital cast of the natural dentition with the lingual veneers. The guide reduced the technique sensitivity of freehand practice and allowed the verification of the complete seating of multiunit lingual veneers on unprepared teeth. The digital technique provided a valid and reliable approach to making and cementing multiple CAD-CAM veneers for the minimally invasive restoration of worn teeth.


Echocardiographic diagnosis of left anterior descending coronary artery disease.
To determine the usefulness of the standard echocardiogram in the diagnosis of left anterior descending coronary artery disease proximal or distal to the first septal branch, coronary arteriograms and echocardiograms were performed in 77 patients with a chest pain syndrome. Seventy-nine percent of patients with proximal disease (15 of 19) had an abnormal septal motion measured as a posterior wall/interventricular septal (PW/IVS) excursion ratio greater than or equal to 2.5 compared with 10% of patients with distal disease (2 of 20) who had abnormal septal motion. Only 5% of patients without obstructive disease of the left anterior descending coronary artery (2 of 38) had abnormal septal motion. Proximal disease was found in 79% with abnormal septal motion in the echocardiogram (15 of 19) but in only 7% of patients with normal septal motion (4 of 58). Therefore, abnormal septal motion as measured by the PW/IVS excursion ratio in the echocardiogram is a useful index for the diagnosis of disease of the left anterior descending coronary artery when that disease is proximal to the first septal branch.


Researching continuity of care: can quality of life outcomes be linked to nursing care?
Research that informs nursing interventions across the care continuum is vital, especially with shortened hospital stays. Measuring Quality of Life (QOL) and Health Related Quality of Life (HRQOL) helps identfy health status improvements, but fails to provide insight into the effectiveness of nursing interventions aimed at continuity of care. Four research examples illustrate the need for complementary, qualitative studies of what patients and their families think, feel, need and want. These indicate a need to reconceptualise the research agenda in terms of the complexity and settings of nursing practice, and the need for informational as well as statistical significance.


A preliminary study on the occurrence of Aspergillus spp. and aflatoxin B1 in imported wheat and barley in Penang, Malaysia.
Thirty samples consisting of wheat (15) and barley (15) were collected from different markets in Penang, Malaysia, originating from India and Thailand, respectively. All samples were analyzed for occurrence of Aspergillus spp. and aflatoxin B1 (AFB1). Aspergillus flavus was dominant in all samples followed by A. niger. AFB1 could be detected in three wheat samples ranging from 0.42 to 1.89 μg/kg and one barley sample had 0.58 μg/kg of AFB1. The AFB1 levels in all the samples were below the Malaysian regulatory limits (<35 μg/kg). The frequency and quantity of AFB1 levels in this study were very low in wheat and barley samples compared to other agricultural commodities reported in India and Thailand. This is the first report on determination of Aspergillus spp. and AFB1 in imported wheat and barley grains in Penang, Malaysia.


An assessment of quality of randomized controlled trials in shoulder instability surgery using a modification of the clear CLEAR-NPT score.
The present study aimed to evaluate the methodological quality and determine the quality of reporting of randomized controlled trials (RCTs) that assess surgical treatment for shoulder instability. A Cochrane, Pubmed, EMBASE and Trip database search was performed, including the relevant literature, regarding RCTs that report on shoulder instability published between January 1994 and January 2017. Methodological quality was assessed with a modification of the Checklist to Evaluate A Report of a Nonpharmacologic Trial (CLEAR-NPT). Points were assigned based on 18 items regarding patient characteristics, randomization, care provider characteristics, surgical details and blinding, with a total score ranging from 0 points to 18 points. Missing items were verified with the corresponding authors of the studies. Quality of reporting corresponds to the total scores including the items that were additionally provided by the authors. We included 22 studies. Of these, nine corresponding authors provided additional information. The average methodological quality was 16.9 points (11 studies) and the average quality of reporting was 9.5 points (22 studies). Items scoring worst included information regarding the surgeon's experience, the patients' level of activity, comorbidities, analyzing according to 'intention-to-treat' principles, and blinding of care providers, participants and assessors. RCTs reporting on shoulder instability surgery are well performed but poorly reported.


Reaching movements are attracted by stimuli that signal reward.
When presented with a set of possible reach targets, the movement trajectory can reveal aspects of the underlying competition for action selection. Current goals and physical salience can affect the trajectory of reaching movements to be attracted towards a distractor. Some studies demonstrated that stimuli associated with reward can also cause an attraction when reaching towards the reward stimulus was previously rewarded and the reward stimulus was physically salient. Here we demonstrate that a non-salient stimulus that signals the availability of reward attracts reaching movements even when moving towards it was never necessary nor rewarded. Moreover, the attraction by reward is particularly evident with short-latency movements. We conclude that neither physical salience nor reinforcing the movement towards a stimulus is necessary for reward to gain priority in the selection for action.


Frequency and Effects of Excess Dosing of Anticoagulants in Patients ≤55 Years With Acute Myocardial Infarction Who Underwent Percutaneous Coronary Intervention (from the VIRGO Study).
Excess dosing of anticoagulant agents has been linked to increased risk of bleeding after percutaneous coronary intervention (PCI) for women compared with men, but these studies have largely included older patients. We sought to determine the prevalence and gender-based differences of excess dosing of anticoagulants including glycoprotein IIb/IIIa inhibitors, bivalirudin, and unfractionated heparin in young patients with acute myocardial infarction who underwent PCI and to examine its association with bleeding. Of 2,076 patients enrolled in the Variation in Recovery: Role of Gender on Outcomes of Young Acute Myocardial Infarction Patients study who underwent PCI, we abstracted doses of unfractionated heparin, bivalirudin, and glycoprotein IIb/IIIa inhibitors administered during PCI from the medical records. At least 47.2% received at least 1 excess dose of an anticoagulant, which did not differ by gender. We used logistic regression to determine the predictors of excess dosing and the association of excess dosing with bleeding. In multivariable analysis, only lower body weight and younger age were significant predictors of excess dosing. Bleeding was higher in young women who received excess dosing versus those who did not (9.3% vs 6.0%, p = 0.03) but was comparable among men (5.2% vs 5.9%, p = 0.69) in univariate analysis. In multivariable analysis, there was a trend to an association between excess dosing and bleeding (odds ratio 1.33, 95% confidence interval 0.92 to 1.91) although not statistically significant. In conclusion, approximately half of the patients received excess dosing of anticoagulant drugs during PCI, which did not vary based on gender. There was a trend toward an association between excess dosing and increased bleeding, although not statistically significant.


Synergistic Optimization of Buried Interface by Multifunctional Organic-Inorganic Complexes for Highly Efficient Planar Perovskite Solar Cells.
For the further improvement of the power conversion efficiency (PCE) and stability of perovskite solar cells (PSCs), the buried interface between the perovskite and the electron transport layer is crucial. However, it is challenging to effectively optimize this interface as it is buried beneath the perovskite film. Herein, we have designed and synthesized a series of multifunctional organic-inorganic (OI) complexes as buried interfacial material to promote electron extraction, as well as the crystal growth of the perovskite. The OI complex with BF4- group not only eliminates oxygen vacancies on the SnO2 surface but also balances energy level alignment between SnO2 and perovskite, providing a favorable environment for charge carrier extraction. Moreover, OI complex with amine (- NH2) functional group can regulate the crystallization of the perovskite film via interaction with PbI2, resulting in highly crystallized perovskite film with large grains and low defect density. Consequently, with rational molecular design, the PSCs with optimal OI complex buried interface layer which contains both BF4- and -NH2 functional groups yield a champion device efficiency of 23.69%. More importantly, the resulting unencapsulated device performs excellent ambient stability, maintaining over 90% of its initial efficiency after 2000 h storage, and excellent light stability of 91.5% remaining PCE in the maximum power point tracking measurement (under continuous 100 mW cm-2 light illumination in N2 atmosphere) after 500 h.


Modelling vaccination capacity at mass vaccination hubs and general practice clinics: a simulation study.
COVID-19 mass vaccination programs place an additional burden on healthcare services. We aim to model the queueing process at vaccination sites to inform service delivery. We use stochastic queue network models to simulate queue dynamics in larger mass vaccination hubs and smaller general practice (GP) clinics. We estimate waiting times and daily capacity based on a range of assumptions about appointment schedules, service times and staffing and stress-test these models to assess the impact of increased demand and staff shortages. We also provide an interactive applet, allowing users to explore vaccine administration under their own assumptions. Based on our assumed service times, the daily throughput for an eight-hour clinic at a mass vaccination hub ranged from 500 doses for a small hub to 1400 doses for a large hub. For GP clinics, the estimated daily throughput ranged from about 100 doses for a small practice to almost 300 doses for a large practice. What-if scenario analysis showed that sites with higher staff numbers were more robust to system pressures and mass vaccination sites were more robust than GP clinics. With the requirement for ongoing COVID-19 booster shots, mass vaccination is likely to be a continuing feature of healthcare delivery. Different vaccine sites are useful for reaching different populations and maximising coverage. Stochastic queue networks offer a flexible and computationally efficient approach to simulate vaccination queues and estimate waiting times and daily throughput to inform service delivery.


Chronic treatment with choline or scopolamine indicates the presence of muscarinic cholinergic receptor plasticity in the frontal cortex of young but not of aged mice.
Chronic treatment (2 weeks) with either scopolamine (4 mg/kg, once daily p.o.) or choline (200 mg/kg, once daily p.o.) resulted in a pronounced muscarinic cholinergic receptor up- or down-regulation in the frontal cortex of young (4 weeks) but not of aged (18 months) female mice. It is speculated that a similar age-related decline of muscarinic receptor plasticity might contribute to the profound dysfunction of cholinergic neurotransmission in Alzheimer's disease.


Major histocompatibility complex class I-binding peptides are recycled to the cell surface after internalization.
Cytotoxic T lymphocytes (CTL) recognize target antigens as short, processed peptides bound to major histocompatibility complex class I (MHC-I) heavy and light chains (beta 2-microglobulin; beta 2-m). The heavy chain, which comprise the actual peptide binding alpha-1 and alpha-2 domains, can exist at the cell surface in different forms, either free, bound to beta 2-m or as a ternary complex with beta 2-m and peptides. MHC-I chains are also known to internalize, and recycle to the cell surface, and this has been suggested to be important in peptide presentation. Whether MHC-I-bound peptides also can recycle is not known. We have investigated this by using both peptide transporter mutant RMA-S cells and EL4 cells loaded with Db-binding peptides, by two different approaches. First, peptides were covalently linked with galabiose (Gal alpha 4Gal) at a position which did not interfere with Db binding or immunogenicity, and peptide recycling tested with Gal2-specific monoclonal antibodies. By flow cytometry, a return of Gal2 epitopes to the cell surface was found, after cellular internalization and cell surface clearance by pronase treatment. This peptide recycling could be discriminated from free fluid-phase uptake and was inhibited by methylamine, chloroquine and low temperature (18 degrees C) but not by leupeptin. Second, specific CTL were reacted with peptide-loaded target cells after complete removal of surface Db molecules by pronase, and after different times of incubation at 37 degrees C to allow reexpression. By this procedure, reappearance of target cell susceptibility was confirmed. The results are in agreement with a model for optimizing peptide presentation by recycling through an intracellular compartment similar to early endosomes in certain antigen-presenting cells.


Monomer surface modifications for rapid peptide analysis by capillary electrophoresis and capillary electrochromatography coupled to electrospray ionization-mass spectrometry.
In this study, positively charged alkylaminosilyl monomers were used to modify the inner surface of fused silica capillaries, which subsequently were employed in capillary electrophoresis (CE) and capillary electrochromatography (CEC). The obtained surfaces yield a reversed electroosmotic flow (EOF) and have varying carbon chain lengths, that interact with the analytes and give chromatographic retention. The coating procedure is very simple and fast. The performance of the modified capillaries was evaluated regarding pH influence on EOF and chromatographic interactions. The experiments were conducted with UV and mass spectrometry (MS) and applied to the separation of various neuropeptides. The derivatized surfaces showed a linear (R(2) approximately 0.99) pH dependence with isoelectric points (pI) at 8.6-8.8. Rapid separations of peptide standards and a protein digest with efficiencies as high as 5 x 10(5) plates/m were performed.


Huanglian Ganjiang decoction alleviates ulcerative colitis by restoring gut barrier via APOC1-JNK/P38 MAPK signal pathway based on proteomic analysis.
Ulcerative colitis (UC) is a kind of chronic intestinal inflammation accompanied with abdominal pain, diarrhea and hematochezia. Huanglian Ganjiang decoction (HGD) derived from "Beiji Qianjin Yao Fang" was used for UC patients clinically. However, the specific mechanism of HGD in treating UC remain unclear. Our study devoted to demonstrating the therapeutic effect of HGD for colitis and clarifying the underlying mechanism. UPLC-MS was carried out to identify the ingredients of HGD. UC mice were induced by giving 3% dextran sulfate sodium (DSS) solution for one week and treated by HGD for another week. Body weight fluctuation, disease activity index (DAI), colon length and pathological change of colon tissues were observed to evaluate therapeutical effect of HGD. ELISA and qPCR were carried out to estimate the inflammatory state. Western blot, qPCR and immunofluorescence were used to access the expression of tight junction proteins. Tandem mass tag (TMT)-Based proteomics and network pharmacology was launched to screen and predict the potential targets and pathway regulated by HGD. Based on the UPLC-MS/MS analysis, 100 components were identified in HGD. After 7-day treatment, HGD significantly alleviated colitis-associated symptoms including body weight loss, shorted colon, increase of DAI score, histopathologic lesions. HGD also reduced inflammatory cytokines IL-6 and IL-1β levels, increased the number of goblet cells and restored tight junction proteins Occludin, Claudin-1 in colon. Network pharmacology study predicted that tight junction and MAPK pathway might be affected by HGD in colitis mice. APOC1 was screened out as key target in HGD-treated mice using TMT-based proteomics study. Further Western blot results showed that HGD reduced expressions of APOC1, p-P38 and p-JNK. HGD improves general symptoms of colitis mice at medium and high doses, which may be associated with restoring tight junction and intestinal barrier integrity and function through suppression of APOC1-JNK/P38 MAPK signal pathway.


Increasing binding of a transcription factor immediately downstream of the cap site of a cytomegalovirus gene represses expression.
A closely related family of ubiquitous DNA binding proteins, called MDBP, binds with high affinity to two 14 base pair (bp) sites within the human cytomegalovirus immediate early gene 1 (CMV IE1) enhancer and with low affinity to one site beginning 5 bp downstream of the CMV IE1 transcription start point (+5 site). Unlike several cap position downstream MDBP sites in mammalian genes, these MDBP sites do not require cytosine methylation for optimal binding. Mutation of one of the enhancer MDBP sites to prevent MDBP recognition modestly increased the function of a neighboring CREB binding site in a transient transfection assay in the context of one promoter construct. A much larger effect on reporter gene expression (a 10-fold reduction) was seen when the low affinity MDBP recognition sequence at position +5 was converted to a high affinity site in a plasmid containing the CMV IE1 promoter upstream of the reporter gene. Evidence that the increased binding of MDBP at the mutant site is largely responsible for the observed results was provided by transfection experiments with this high affinity MDBP +5 site re-mutated to a non-binding site and by in vitro transcription assay.


Glucocorticoid regulation of glycosaminoglycan synthesis in cultured human skin fibroblasts: evidence for a receptor-mediated mechanism involving effects on specific de novo protein synthesis.
Human skin fibroblast cultures synthesize and accumulate glycosaminoglycan (GAG). This laboratory has reported recently that the synthetic rate of hyaluronate (HA), the most abundant GAG produced by these cells, is inhibited by glucocorticoids. The purpose of these studies was to characterize further that hormonal response. Effects of potent glucocorticoid agonists dexamethasone (Dex) and RU28362 on GAG synthesis were studied. Both steroids inhibited [3H]GAG synthesis when added 24 hours before labeling with [3H]acetate. RU28362 inhibited HA synthesis by 58% and Dex by 60%. The half-maximal effect for each was achieved at a concentration of 0.5 nmol/L and maximal effects at 10 nmol/L. The recently described glucocorticoid antagonist RU38486 failed to alter [3H]GAG synthesis at concentrations up to 1,000 nmol/L. However, this compound did block the inhibition of the agonists. At concentrations of RU28362 and Dex that maximally inhibited [3H]GAG synthesis (10 nmol/L), RU38486 attenuated the glucocorticoid effect in a dose-dependent manner, with half-maximal inhibition at 5 nmol/L and maximal inhibition at 100 nmol/L. The antagonist blocked as much as approximately 80% of the Dex inhibition and approximately 70% of the RU28362 inhibition. Cycloheximide (25 micrograms/mL) inhibited [3H]HA synthesis by 55% within four hours of its addition to the culture. When cultures were pretreated with cycloheximide without or with Dex (100 nmol/L) for three hours and then treated with actinomycin D (2 micrograms/mL) followed by [3H]acetate labeling, Dex pretreated cultures synthesized 33% less [3H]GAG, implying that Dex had decreased the abundance of a translatable mRNA.(ABSTRACT TRUNCATED AT 250 WORDS)


Identification of new DNA adducts in human bladder epithelia exposed to the proximate metabolite of 4-aminobiphenyl using 32P-postlabeling method.
The DNA adducts were analyzed by 32P-postlabeling method following exposure of human uroepithelial cells (HUC) to N-hydroxy-4-aminobiphenyl (N-OH-ABP), the proximate metabolite of the human bladder carcinogen 4-aminobiphenyl (ABP). TLC of the postlabeled products on the first dimension revealed several products, the majority of which stayed close to the origin and were earlier identified as the 3',5' -bisphospho derivatives of N-(deoxyguanosin-8-yl)-4-aminobiphenyl and N-(deoxyadenosin-8-yl)-4-aminobiphenyl (Carcinogenesis 13 (1993) 955; Carcinogenesis 16 (1995) 295). Here we report characterization of two additional adducts that amounted to less than 5% of the total adducts. Autoradiography of D1 chromatogram of the postlabeled products of calf thymus DNA chemically interacted with N-OH-ABP under acidic conditions revealed two adducts, #1 and #2, with R(f) values of about 0.2 and 0.3, respectively. Two adducts with D1 thin layer chromatographic properties similar to those of adducts #1 and #2 were obtained on postlabeling analyses of products generated by chemical interaction of N-acetoxy-4-aminobiphenyl (N-OAc-ABP) with deoxyguanosine-3' -monophosphate (dGp). Based on proton NMR and mass spectroscopic analyses of the synthetic products derived from N-OAc-ABP, the chemical structures of adducts #1 and #2 have been identified as 3-(deoxyguanosin-N(2)-yl)-4-aminobiphenyl, and N-(deoxyguanosin-N(2)-yl)-4-aminobiphenyl, respectively. Both of these adducts were insensitive to digestion with nuclease P1. 32P-Postlabeling analysis of the nuclease P1 enriched DNA hydrolysate of HUC cells treated with N-OH-ABP showed the presence of adduct #2 but not adduct #1. Adduct #2 was also detected in calf thymus DNA incubated with HUC cytosol and N-OH-ABP in the presence of acetyl CoA. These results suggest that in the target cells for ABP carcinogenesis in vivo, N-OH-ABP is bioactivated by acetyl CoA-dependent acyltransferases to reactive arylnitrenium ions that covalently interact at N(2)-position of deoxyguanosine in DNA.


31P MRS evaluation of fatigue in anterior tibial muscle in postpoliomyelitis patients and healthy volunteers.
Changes in concentration of high energy phosphates and pH were studied during rest, exercise and subsequent recovery in the anterior tibial muscle of 10 patients with late effects of poliomyelitis and 10 age- and sex-matched healthy volunteers using 31P MRS. The exercise was dynamic and isometric, and the force levels were individually adapted to each subject and stepwise increased. In general, there were no differences in metabolite changes between the groups, except for lower Pi and Pi/PCr for the volunteers during the recovery phase, also reflected by shorter recovery half-time for Pi. The interindividual variation was much higher for the patient group. Some of the patients showed deviating results probably because of differences in muscle fibre type.


Unrecognized myocardial infarction and risk of atrial fibrillation: the Rotterdam Study.
Persons with a clinically recognized myocardial infarction are at increased risk for atrial fibrillation. However a large proportion of all myocardial infarctions remain clinically unrecognized. Whether subjects with electrocardiographic signs of an unrecognized myocardial infarction are also at an increased risk of developing atrial fibrillation is unknown. The objective of this study was to investigate whether unrecognized myocardial infarction was associated with an increased risk of atrial fibrillation in a prospective population-based cohort study. The study is set within the prospective population-based Rotterdam Study. The study population comprised 2505 men and 3670 women without atrial fibrillation at baseline. Participants were classified based on electrocardiography, interview, and clinical data into those with recognized myocardial infarction, those with ECG based unrecognized myocardial infarction and those without myocardial infarction. Atrial fibrillation was ascertained from ECG assessments as well as medical records. During a mean follow-up of 11.7 years (SD 5.0), 329 men and 398 women developed atrial fibrillation. Unrecognized myocardial infarction was associated with a two-fold risk of developing atrial fibrillation in men (HR: 2.21, 95%CI:1.51 to 3.23) compared to men without a history of myocardial infarction, independent of age, and cardiovascular risk factors. In women, unrecognized myocardial infarction was not associated with atrial fibrillation (HR: 0.92, 95%CI:0.59 to 1.44). The presence of an unrecognized myocardial infarction is associated with a twofold increased risk of atrial fibrillation in men, independent of known cardiovascular risk factors.


Factor IX Kawachinagano: impaired function of the Gla-domain caused by attached propeptide region due to substitution of arginine by glutamine at position -4.
Factor IX Kawachinagano (KWC) is a mutant factor IX protein initially recognized in a patient with severe haemophilia B, who had 46% of normal factor IX antigen and no detectable clotting activity. Previous studies indicated that factor IX KWC was not activated by factor XIa in the presence of Ca ions. In the present study, we purified and analysed factor IX KWC at a structural level in an attempt to clarify the nature of the impaired reaction with factor XIa. Kinetic studies showed that activation of factor IX KWC by factor X activator from Russell's viper venom (RVV-X) was normal, whereas activation by factor XIa was defective. Amino acid sequence analysis of tryptic peptides and direct analysis of the NH2-terminal sequence of factor IX KWC demonstrated that this mutant factor IX retained the propeptide region of 18 amino acids due to a substitution of arginine-(-4) by glutamine. These data suggested that the Gla-domain of factor IX KWC was dysfunctional, although the total gamma-Gla content, measured by alkaline-hydrolysis, was normal. We assumed that this attached propeptide region of the molecule directly interferes with the adjacent NH2-terminus and prevents the metal-induced conformational changes which are essential for biological activity of normal factor IX.


[Risk factors for developing laryngeal cancer in adult population at the Hospital Español in Mexico City].
Laryngeal cancer is the most frequent head and neck cancer. Considerable geographic differences exist in its incidence by gender. In Spain, there is male predominance (>90%). Numerous authors have documented exposure to tobacco and/or alcohol as the main risk factors for laryngeal cancer. Gastroesophageal reflux, genetics, occupational factors and also human papilloma virus are also cited. The present study involved 43 patients diagnosed with laryngeal cancer and 130 healthy control subjects. The goal was to establish the frequency affecting both sexes and to identify the risk factors of those born in Spain but living in Mexico. Odds ratios (OR) for each risk factor were analyzed using univariate analysis. A considerable predominance of laryngeal cancer was found in males (90.6%). Tobacco (OR=6.56) and alcohol consumption (OR=3.04) are significant risk factors with a multiplier effect. Gastroesophageal reflux does not show any significant OR. Occupational exposure had a significant OR=37.28. The main advantage of this type of studies is the ability to design strategies to modify the risk factors. Male predominance and risk factors were no different from other findings reported in Spain, except for the considerable risk relating to occupational exposure.


Sleep apnea in the elderly.
Obstructive sleep apnea (OSA) is a common clinical condition associated with obesity. A high prevalence of sleep apnea exists in the elderly, presumably because of changes in oropharyngeal collapsibility. Elderly patients may be less likely to seek medical attention for this condition, because they are less likely to be symptomatic. Excessive daytime somnolence is a known consequence of untreated OSA, but adverse cardiovascular consequences, such as hypertension, arrhythmias, and congestive heart failure, are more serious in older patients. Continuous positive airway pressure therapy is the most effective treatment of OSA, although compliance remains an issue.


Childhood central nervous system vasculitis.
Childhood CNS vasculitis is a newly recognized disease entity, which can occur in isolation as a primary disorder - childhood primary angiitis of the CNS (cPACNS)-or secondary to systemic diseases. Since early initiation of treatment is associated with improved disease outcome, rapid evaluation and timely diagnosis are essential. The clinical presentation can be highly variable. Children present with newly acquired focal and/or diffuse neurological deficits and/or psychiatric symptoms. Inflammatory markers and CSF analysis are essential, but show limited sensitivity and specificity. MRI studies are highly sensitive for areas of focal inflammation, however lack specificity. Negative MRI studies do not exclude inflammatory brain disease. Angiography is the key diagnostic modality for large to medium vessel cPACNS. Characteristic angiographic findings include segmental cerebral vessel stenoses and corresponding contrast enhancement in the vessel wall. In children with normal angiography, confirmation of small vessel cPACNS requires brain biopsy. A thorough diagnostic evaluation for cPACNS should include a workup for causes of secondary CNS vasculitis, evaluation for CNS vasculitis mimics, and analysis of markers of nonvasculitic inflammatory brain diseases when appropriate. Early recognition with timely treatment of childhood CNS vasculitis can significantly alter the course of this potentially devastating disease.


Physical activity, dietary habits and Coronary Heart Disease risk factor knowledge amongst people with severe mental illness: a cross sectional comparative study in primary care.
Evidence regarding Coronary Heart Disease (CHD) related lifestyle in people with severe mental illnesses (SMI) such as schizophrenia is sparse. We aimed to quantify adverse CHD knowledge, diet and exercise in a representative primary care sample, and to determine whether socio-economic deprivation explained any findings. We compared CHD lifestyle and CHD knowledge in 74 people with SMI and 148 without from seven general practices. We measured CHD knowledge, dietary fibre, fats and exercise using validated instruments and adjusted for socio-economic status. Fewer people with SMI had higher CHD knowledge: OR 0.49 (95% CI: 0.27-0.88), higher total exercise scores 0.49 (0.27-0.86), higher fibre 0.46 (0.26-0.82) or lower saturated fat diets 0.53 (0.30-0.94). These results were stable irrespective of antipsychotic medication, socio-economic status or type of statistical analysis. High fat, low fibre diets, lack of exercise and smoking are the likely causes of the majority of CHD in this high-risk group, irrespective of medication and socio-economic deprivation. This lifestyle and particularly the lower CHD knowledge provides a theoretical focus for more comprehensive preventative CHD interventions in SMI.


Cervical Lymph Node Fine-Needle Aspiration and Needle-Wash Thyroglobulin Reflex Test for Papillary Thyroid Carcinoma.
Fine-needle aspiration (FNA) cytology coupled with needle-wash thyroglobulin (FNA-Tg) testing is recommended for cervical lymph node (LN) biopsies in patients with a history of papillary thyroid carcinoma (PTC). However, the procedure has not been standardized with the assay for FNA-Tg testing. A standard operating procedure (SOP) has been generated at our facility for cervical LN FNAs with Tg reflex testing on patients with a history of PTC. The procedure requires FNA cytology to be reviewed first, and all cases not positive for PTC are reflexed for FNA-Tg testing with the Beckman Access thyroglobulin assay. The thyroglobulin cutoff value is ≤ 1.0 ng/mL. From 2016 to 2017, 117 patients, including 71 women and 46 men, were identified as having a history of PTC. Patients' clinical characteristics were collected from medical records. A total of 143 LN biopsies were investigated for these patients. The results show that four out of 11 (36.4%) non-diagnostic LNs and five out of five (100%) atypical/suspicious LNs tested positive for FNA-Tg. Among these nine patients with positive thyroglobulin testing, LN metastases were proven histologically for all nine patients, and two patients were treated with LN ablation. Out of 68 LNs positive for PTC, three had FNA-Tg results. FNA-Tg testing was ordered for unknown reasons on two positive LNs (> 5000 ng/mL thyroglobulin) from one patient. The third LN was tested due to non-classic morphology, and the result was less than the cutoff value. Three patients with negative LN biopsies were tested to have elevated (> 1.0 ng/mL) thyroglobulin levels. One patient (FNA-Tg ng/mL) was proven to have multiple metastatic LNs through follow-up surgery. However, no positive LN was identified for the other two patients who had FNA-Tg level of 4.1 ng/mL and 37 ng/mL respectively. This is likely due to contamination, as these two patients had intact thyroids. In our practice, the FNA-Tg test is a very useful adjunct test to LN FNA specimens with a non-positive diagnosis in patients with a history of PTC. Furthermore, FNA-Tg testing increases diagnostic sensitivity among non-diagnostic and atypical/suspicious LNs. However, FNA-Tg testing should not substitute conventional cytology due to the following reasons: (1) false-negative thyroglobulin lab results; (2) PTC with loss of thyroglobulin expression; (3) LN metastasis from other origins; and (4) false-positive thyroglobulin testing due to blood contamination in patients who are not completely athyrotic.


Persistent cryoglobulinemia after antiviral treatment is associated with advanced fibrosis in chronic hepatitis C patients.
High dosage and longer duration of antiviral treatment has been suggested to treat cryoglobulinemia patients. We aimed to investigate the efficacy of antiviral treatment in cryoglobulinemia patients and analyze the associated factors of persistent cryoglobulinemia. Totally 148 patients after completion of anti-HCV treatment were enrolled in our study. Serum cryoglobulinemia precipitation was assessed and analyzed for the associated factors after antiviral therapy. Fifty-one (34.5%) out of 148 patients were positive for serum cryoglobulinemia after completion of antiviral therapy. In multivariate analysis, advanced fibrosis (Odds Ratio [OR]- 4.13, 95% Confidence Interval [95% CI]- 1.53-11.17, p = 0.005) and platelet counts (OR-0.98, 95% CI- 0.97-0.99, p = 0.010) were independently and significantly associated with persistent cryoglobulinemia. The factors associated with the persistent cryoglobulinemia in SVR patients were advanced fibrosis (OR-1.93, 95% CI- 1.02-3.65, p = 0.041) and platelet count (OR-0.98, 95% CI- 0.96-0.99, p = 0.041) by multivariate analysis. Multivariate logistic regression analysis showed persistent (OR-4.83, 95% CI- 1.75-13.36, p = 0.002) was significantly associated with advanced fibrosis in patients with cryoglobulinemia follow up after antiviral therapy. The prevalence of the persistent cryoglobulinemia is 34.5% after completing antiviral therapy and it is associated with advanced fibrosis, also HCV clearance.


The new NO donor Terpy induces similar relaxation in mesenteric resistance arteries of renal hypertensive and normotensive rats.
The present work aimed to investigate the cellular mechanisms involved on the vasorelaxation induced by the new nitric oxide donor [Ru(terpy)(bdq)NO](3+) (Terpy) in isolated mesenteric resistance artery and to compare the vascular responses in isolated vessels from 2K and 2K-1C hypertensive rats. We have used this artery because it is important to the control of vascular resistance and consequently to the blood pressure control. The NO donor Terpy induced relaxation in a concentration-dependent way in mesenteric resistance arteries. There were no differences between renal hypertensive (2K-1C) and normotensive (2K) in Terpy-induced relaxation neither in NO released. The relaxation induced by Terpy was inhibited by the soluble guanylyl-cyclase (sGC) inhibitor ODQ both in 2K and in 2K-1C with similar amplitude. In agreement with these data, the protein expression of the subunits α1 and β1 of the enzyme sGC was not different between 2K-1C and 2K mesenteric bed. The relaxation induced by Terpy was inhibited by the cGMP-dependent protein kinase (G kinase) inhibitor or by the non-selective K(+) channel blocker tetraethylamonium (TEA), but with no difference between 2K-1C and 2K arteries. The relaxation induced by Terpy was also inhibited by the SERCA inhibitor thapsigargin in both groups. Taken together, these results show that the vascular relaxation induced by the NO donor [Ru(terpy)(bdq)NO](3+) involves the activation of NO/sGC/cGMP/GK pathway, activation of K(+) channels sensitive to TEA and SERCA in normotensive and renal hypertensive rat mesenteric resistance arteries. Surprisingly, Terpy-induced vasorelaxation is similar in mesenteric resistance arteries of renal hypertensive and normotensive rats.


Transgenic expression of CTLA4-Ig by fetal pig neurons for xenotransplantation.
The transplantation of fetal porcine neurons is a potential therapeutic strategy for the treatment of human neurodegenerative disorders. A major obstacle to xenotransplantation, however, is the immune-mediated rejection that is resistant to conventional immunosuppression. To determine whether genetically modified donor pig neurons could be used to deliver immunosuppressive proteins locally in the brain, transgenic pigs were developed that express the human T cell inhibitory molecule hCTLA4-Ig under the control of the neuron-specific enolase promoter. Expression was found in various areas of the brain of transgenic pigs, including the mesencephalon, hippocampus and cortex. Neurons from 28-day old embryos secreted hCTLA4-Ig in vitro and this resulted in a 50% reduction of the proliferative response of human T lymphocytes in xenogenic proliferation assays. Transgenic embryonic neurons also secreted hCTLA4-Ig and had developed normally in vivo several weeks after transplantation into the striatum of immunosuppressed rats that were used here to study the engraftment in the absence of immunity. In conclusion, these data show that neurons from our transgenic pigs express hCTLA4-Ig in situ and support the use of this material in future pre-clinical trials in neuron xenotransplantation.


Acute recanalization treatments in postnatal paediatric ischaemic arterial stroke. Paediatric stroke code.
In children, arterial ischemic stroke is a much less understood disease compared to in adults due to its lower frequency and different aetiology. However, it is also a serious disease, with a high incidence of severe and permanent sequelae that exceeds 50% of total cases. The acute management of postnatal arterial ischaemic stroke (MNAIS) has changed drastically in recent years, chiefly on account of recanalization treatments (thrombolysis and endovascular therapies). These treatments, which used to not be recommended in childhood, are increasingly implemented in everyday clinical practice. Although the evidence from studies carried out in children is not of high quality due to their retrospective design and the small number of reported cases, they support the hypothesis that these treatments are as safe and effective as they are in adults as long as appropriate eligibility criteria are applied and they are used within a certain time from the onset of symptoms (therapeutic window). This article reviews the MNAIS based on the current scientific evidence. Since the efficacy of these treatments is highly dependent on their early initiation, a paediatric stroke code needs to be in place as an extension of the stroke code applied to adults. It has started to be introduced in Spain since 2019, although there are still large areas of the country where it has yet to be applied.


Phenotypic and phylogenetic analysis of lactic acid bacteria isolated from forage crops and grasses in the Tibetan Plateau.
A total of 140 lactic acid bacteria (LAB) strains were isolated from corn, alfalfa, clover, sainfoin, and Indian goosegrass in the Tibetan Plateau. According to phenotypic and chemotaxonomic characteristics, 16S rDNA sequence, and recA gene PCR amplification, these LAB isolates were identified as belonging to five genera and nine species. Corn contained more LAB species than other forage crops. Leuconostoc pseudomesenteroides, Lactococcus lactis subsp. lactis, Lactobacillus brevis, and Weissella paramesenteroides were dominant members of the LAB population on alfalfa, clover, sainfoin, and Indian goosegrass, respectively. The comprehensive 16S rDNA and recA-based approach effectively described the LAB community structure of the relatively abundant LAB species distributed on different forage crops. This is the first report describing the diversity and natural populations of LAB associated with Tibetan forage crops, and most isolates grow well at or below 10°C. The results will be valuable for the future design of appropriate inoculants for silage fermentation in this very cold area.


Degradation of ketoprofen by sulfate radical-based advanced oxidation processes: Kinetics, mechanisms, and effects of natural water matrices.
Ketoprofen (KET) is a mostly used nonsteroidal anti-inflammatory drug that has been frequently detected in wastewater effluents and surface waters. In this study, we investigated the degradation of KET by sulfate radical (SO4-) based advanced oxidation processes (SR-AOPs) in aqueous solution. The degradation kinetics, mechanisms, and effects of natural water matrices on thermally activated persulfate (TAP) oxidation of KET were systematically investigated. Increasing the temperature and persulfate (PS) concentrations greatly enhanced the degradation of KET. KET degradation is pH-dependent with an optimum pH of 5.0. Reactions in the presence of radical quenchers revealed the dominant role of SO4- in oxidizing KET. Water matrix significantly influenced the degradation of KET. The common inorganic anions present in natural waters exhibited inhibitory effect on KET degradation, and the inhibition followed the order of Cl- > CO32- > HCO3- > NO3-; however, no significant inhibition of KET degradation was observed in the presence of Ca2+ and Mg2+ cations. The presence of natural organic matter (NOM) suppressed KET degradation, and the suppression increased as NOM concentration increase. Products identification and mineralization experiments revealed that KET and its degradation intermediates were finally transformed into CO2 and H2O. The results of this study indicated that applying SR-AOPs for the remediation of KET contaminated water matrix is technically possible.


Antenatal interventions to reduce risk of low birth weight related to maternal infections during pregnancy.
Maternal infections during pregnancy have been linked to increased risk of adverse birth outcomes, including low birth weight (LBW), preterm birth (PTB), small for gestational age (SGA), and stillbirth (SB). The purpose of this article was to summarize evidence from published literature on the effect of key interventions targeting maternal infections on adverse birth outcomes. We searched MEDLINE, Embase, Cochrane Database of Systematic Reviews, Cochrane Central Register of Controlled Trials, and CINAHL Complete between March 2020 and May 2020 with an update to cover until August 2022. We included randomized controlled trials (RCTs) and reviews of RCTs of 15 antenatal interventions for pregnant women reporting LBW, PTB, SGA, or SB as outcomes. Of the 15 reviewed interventions, the administration of 3 or more doses of intermittent preventive treatment in pregnancy with sulphadoxine-pyrimethamine [IPTp-SP; RR: 0.80 (95% CI: 0.69, 0.94)] can reduce risk of LBW compared with 2 doses. The provision of insecticide-treated bed nets, periodontal treatment, and screening and treatment of asymptomatic bacteriuria may reduce risk of LBW. Maternal viral influenza vaccination, treatment of bacterial vaginosis, intermittent preventive treatment with dihydroartemisinin-piperaquine compared with IPTp-SP, and intermittent screening and treatment of malaria during pregnancy compared with IPTp were deemed unlikely to reduce the prevalence of adverse birth outcomes. At present, there is limited evidence from RCTs available for some potentially relevant interventions targeting maternal infections, which could be prioritized for future research.


Suppression of stellate cell type I collagen gene expression involves AP-2 transmodulation of nuclear factor-1-dependent gene transcription.
The regulation of collagen gene expression was studied in culture-activated rat hepatic stellate cells, the fibrogenic effector cell involved in hepatic fibrogenesis. Treatment of cells with a 5-lipoxygenase-specific inhibitor caused a reduction in alphaI(I) collagen mRNA transcript abundance, which suggested that leukotriene production was involved in maintaining the activated cell's high level of collagen mRNA production. The underlying mechanism involved a decrease in collagen gene transcription. Suppression of gene transcription was localized to an nuclear factor-1 (NF-1) binding domain in the proximal promoter and an AP-2 binding domain adjacent to it. Gel retardation assays demonstrated that an increase in AP-2 binding adjacent to the NF-1 site was likely to be the transmodulator responsible for the suppression of the NF-1-dependent gene expression. The data suggest that post-translational alterations in AP-2 activity are responsible for this unappreciated mechanism of regulating the collagen gene.


Transdermal Lidocaine for Perioperative Pain: a Systematic Review of the Literature.
The purpose of this review is to provide a summary of the perioperative studies that have examined transdermal lidocaine (lidocaine patch) as an analgesic and put the evidence in context of the likely overall benefit of transdermal lidocaine in the perioperative period. Several randomized controlled trials have been published in the past 4 years that concluded transdermal lidocaine can reduce acute pain associated with laparoscopic trocar or cannula insertion. Transdermal lidocaine may reduce short-term pain after surgery in selected surgery types and has a low risk of toxicity but its overall clinical utility in the perioperative setting is questionable. Transdermal lidocaine does not consistently reduce opioid consumption after surgery and has not been shown to improve patient function.


Corneal ectasia after LASIK in a patient with normal scheimpflug evaluation but a high Ectasia Risk Score.
To present a case of ectasia after LASIK that developed in a patient with normal Scheimpflug imaging but a high Ectasia Risk Score System (ERSS) score. Case report. A 24-year-old woman with no preoperative abnormalities on Scheimpflug evaluation but a cumulative ERSS risk of 4 (high risk) in the right eye and 3 (medium risk) in the left eye developed ectasia in both eyes 27 months after LASIK. She underwent ultraviolet-A corneal collagen cross-linking, with no signs of further progression. In this case, the ERSS was effective at predicting ectasia risk despite normal Scheimpflug imaging and only mild asymmetry on Placido topography.