Folic acid reduces adhesion molecules VCAM-1 expession in aortic of rats with hyperhomocysteinemia.
To investigate effects of supplementation of folic acid on the expression of adhesion molecules VCAM-1 in the aortas of rats with hyperhomocysteinemia. Thirty male SD rats (200 +/- 20 g) were invided into 3 groups (n = 10 for each group): control group(Control), high Met group(Met) and Met plus Folate group(Met + Folate), fed. for 45 days. Plasma Hcy levels were higher with the high-methionine diet (140.68 +/- 36.87 micromol/L vs 6.47 +/- 1.10 micromol/L in control rats) an effect which was reduced by folate. Respectively, the aortic expression of adhesion molecules VCAM-1 at protein and mRNA levels were higher in the Met groups than those in the control groups or the Met + Folate groups. A high methionine diet for 45 days was sufficient to induce hyperhomocysteinemia. Folate supplementation prevented elevation of Hcy levels in the blood, and reduced expression of the adhesion molecule VCAM-1. Hyperhomocysteinemia is now regarded as one of the important risk factors for cardiovascular and cerebralvascular disorders.[Welch GN, Loscalzo J. Homocysteine and atherothrombosis. N Engl J Med 1998; 38(15):1042-50.] Several plausible mechanisms for Hcy-induecd atherosclerosis have been proposed. These include endothelial dysfunction, enhancement of oxidative stress, reduction in NO bioavailability, and augmentation of thrombus formation.[Holven KB, Holm T, Aukrust P, et al. Effect of folic acid treatment on endothelium-dependent vasodilation and nitric oxide-derived end products in hyperhomocysteinemic subjects . Am J Med 2001;110(7):536-42; Guba SC, Fonseca V, Fink LM. Hyperhomocysteinemia and thrombosis. Semin Thromb Hemost 1999;25(3):291-309.] However, the precise molecular mechanism is still unclear. Recent reports have suggested a role for inflammatory processes in the pathogenesis of atherosclerosis.[Gerard C, Rollins BJ. Chemokines and disease. Nat Immunol 2001;2(2):108-15.] Dysfunction of endothelial cells is the key process promoting inflammatory reactions. On injury, endothlial cells are capable of producing various cytokines that participate in inflammatory reactions in the arterial wall. Although results from in vitro studies suggest that Hcy, at pathophysiological concentrations, stimulates chemokine expression in vascular cells, it is unknown whether hyperhomocysteinemia can initiate similar changes, leading to enhanced momocyte adhesion/binding to the vascular endothelium in vivo.[Zeng X, Dai J, Remick DG, Wang X. Homocysteine mediated expression and secretion of monocyte chemoattractant protein-1 and interleukin-8 in human monocytes. Circ Res 2003;93(4):311-20.] On the basis of the potential pathogenic role of chemokines in atherogenesis, the objective of the present study was to investigate that homocsteine may exert its effect in part though adhesion molecules VCAM-1 and that folic acid supplementation may downregulate these inflammatory responses. Male Sprague-Dawley rats (bred from animal centers of Tongji Medical College, Huazhong Science and Technology University) aged 8 weeks were divided into 3 groups(n=10 for each group) and maintained for 45 days on the following diets before the experiments: (1) regular diet; (2) high-metheionine diet, consisting of regular diet plus 1.7% methionine; and (3) high-methionine plus folate -rich diet, consisting of regular diet plus 1.7% methionine and 0.006% folate.[Boisvert WA, Curtiss LK, Terkeltaub RA. Interleukin-8 and its receptor CXCR2 in atherosclerosis. Immunol Res 2000;21(2-3):129-d37.] Plasma and serum samples wee colleced and stored at -80 degrees C after 45 days until analysis. The plasma homocysteine concentration of rats in three groups were determined by high-pressue liquid chromatography. To detect the endothelial expression of adhesion molecules VCAM-1, the thoracic aorta was isolated and dived into segments. These segments were immersion-fixed in 10% neutral-buffered formalin overlight and then embedded in paraffin. Sequential 5 mum paraffin-embedded cross sections were prepared. Immunohistochemical analyisis was performed to detect vascular cell adhesion molecule(VCAM)-1, The fixed cryosections were immediately blcked in 10% horse serum and phosphate baffered saline(PBS) at room temperature for 30 min. Goat polyclonal andibodies against rat VCAM-1(Santa Cruz Biotechnology) were diluted 1:100 in PBS and incubated with the cryosections for 1 h of room temperature. After three washes, the sections were incubated with biotin-conjugated rabbit anti-goat immunoglobulins(Dako) at 1:250 dilution in PBS. After three washes, the samples were mounted in 90% glycerol-PBS. Photographs were taken by use of a light microscope at a mignification of x200.


Chlorpyrifos induced autophagy and mitophagy in common carp livers through AMPK pathway activated by energy metabolism disorder.
Water pollution caused by widely used agricultural pesticide chlorpyrifos (CPF) has aroused extensive public concern. While previous studies have reported on toxic effect of CPF on aquatic animal, little is known about its effect on common carp (Cyprinus carpio L.) livers. In this experiment, we exposed common carp to CPF (11.6 μg/L) for 15, 30, and 45 days to establish a poisoning model. Histological observation, biochemical assay, quantitative real-time polymerase chain reaction (qRT-PCR), Western blot, and integrated biomarker response (IBR) were applied to assess the hepatotoxicity of CPF in common carp. Our results displayed that CPF exposure damaged histostructural integrity and induced liver injury in common carp. Furthermore, we found that CPF-induced liver injury may be associated with mitochondrial dysfunction and autophagy, as evidenced by swollen mitochondria, broken mitochondrial ridges, and increased the number of autophagosomes. Moreover, CPF exposure decreased the activities of ATPase (Na+/K+-ATPase, Ca2+-ATPase, Mg2+-ATPase, and Ca2+Mg2+-ATPase), altered glucose metabolism-related genes (GCK, PCK2, PHKB, GYS2, PGM1, and DLAT), and activated energy-sensing AMPK, indicating that CPF caused energy metabolism disorder. The activation of AMPK further induced mitophagy via AMPK/Drp1 pathway, and induced autophagy via AMPK/mTOR pathway. Additionally, we found that CPF induced oxidative stress (abnormal levels of SOD, GSH, MDA, and H2O2) in common carp livers, which further contributed to the induction of mitophagy and autophagy. Subsequently, we confirmed a time-dependent hepatotoxicity caused by CPF in common carp via IBR assessment. Our findings presented a new insight into molecular mechanism of CPF induced-hepatotoxicity in common carp, and provided a theoretical basis for evaluating CPF toxicity to aquatic organisms.


OSPs and ESPs and ISPs, Oh My! An Update on Sinonasal (Schneiderian) Papillomas.
Sinonasal (Schneiderian) papillomas are benign neoplasms that arise in the sinonasal tract. Since their initial descriptions, sinonasal papillomas have triggered debate regarding their classification, etiology, rate or predictors of malignant transformation, and other issues. While significant strides have been made in recent years, there are still aspects of sinonasal papillomas that remain unclear even now. This review will serve to update the practicing pathologist on the current understanding of sinonasal papillomas.


A Lung Ultrasound-Based Nomogram for the Prediction of Refractory Mycoplasma pneumoniae Pneumonia in Hospitalized Children.
Early diagnosis of refractory Mycoplasma pneumoniae pneumonia (RMPP) is challenging because of the lack of practical diagnostic imaging tools. Lung ultrasound (LUS) is an emerging tool for diagnosing childhood pneumonia. Hence, we evaluated the role of a nomogram combining LUS findings, clinical features, and laboratory indices in the early prediction of RMPP in children. We retrospectively analyzed 225 children with Mycoplasma pneumoniae pneumonia (MPP) admitted to our hospital between Dec 2018 and Aug 2021. Logistic regression analysis incorporated LUS findings and clinical predictors into the nomogram. Ninety patients hospitalized from Sep 2021 to Dec 2021 were used for external validation of the prediction model. Receiver operating characteristics (ROC) and calibration curves were used to evaluate the performance of the nomogram in the early diagnosis of RMPP. Ultimately, Consolidation size /BSA (odds ratio (OR) 1.015, 95% confidence interval (CI) 1.536-2.446), Pleural Effusion (OR 3.551, 95% CI 1.921-15.600), LDH (OR 1.044, 95% CI 1.006-1. 021) and CRP (OR 3.293, 95% CI 1.019-1.098) were independent risk factors for the development of RMPP. The prediction model was represented visually as a nomogram. The area under the ROC curve for the predictive nomogram was 0.955 (95% CI 0.919-0.978) in the training cohort and 0.916 (95% CI 0.838-0.964) in the validation cohort. The calibration curve is close to the diagonal. This is the first-time lung ultrasound was added to the predicted nomogram, which can more comprehensively assess the condition and more accurately predict the occurrence of RMPP early. Therefore, this nomogram can be widely used in the early diagnosis of RMPP, especially in primary care hospitals.


KRAS mutations are associated with solid growth pattern and tumor-infiltrating leukocytes in lung adenocarcinoma.
KRAS mutations define a clinically distinct subgroup of lung adenocarcinoma patients, characterized by smoking history, resistance to EGFR-targeted therapies, and adverse prognosis. Whether KRAS-mutated lung adenocarcinomas also have distinct histopathological features is not well established. We tested 180 resected lung adenocarcinomas for KRAS and EGFR mutations by high-sensitivity mass spectrometry-based genotyping (Sequenom) and PCR-based sizing assays. All tumors were assessed for the proportion of standard histological patterns (lepidic, acinar, papillary, micropapillary, solid, and mucinous), several other histological and clinical parameters, and TTF-1 expression by immunohistochemistry. Among 180 carcinomas, 63 (35%) had KRAS mutations (KRAS+), 35 (19%) had EGFR mutations (EGFR+), and 82 (46%) had neither mutation (KRAS-/EGFR-). Solid growth pattern was significantly over-represented in KRAS+ carcinomas: the mean±s.d. for the amount of solid pattern in KRAS+ carcinomas was 27±34% compared with 3±10% in EGFR+ (P<0.001) and 15±27% in KRAS-/EGFR- (P=0.033) tumors. Furthermore, at least focal (≥20%) solid component was more common in KRAS+ (28/63; 44%) compared with EGFR+ (2/35; 6%; P<0.001) and KRAS-/EGFR- (21/82; 26%; P=0.022) carcinomas. KRAS mutations were also over-represented in mucinous carcinomas and were significantly associated with the presence of tumor-infiltrating leukocytes and heavier smoking history. EGFR mutations were associated with non-mucinous non-solid patterns, particularly lepidic and papillary, lack of necrosis, lack of cytological atypia, hobnail cytology, TTF-1 expression, and never/light smoking history. In conclusion, extended molecular and clinicopathological analysis of lung adenocarcinomas reveals a novel association of KRAS mutations with solid histology and tumor-infiltrating inflammatory cells and expands on several previously recognized morphological and clinical associations of KRAS and EGFR mutations. Solid growth pattern was recently shown to be a strong predictor of aggressive behavior in lung adenocarcinomas, which may underlie the unfavorable prognosis associated with KRAS mutations in these tumors.


Analysis of Pregnancy and Birth Rates Among Black and White Medicaid-Enrolled Teens.
In the United States, black teens overall have higher pregnancy and birth rates than whites, and it is commonly believed that minority race and low income account for this disparity. We examined racial differences in pregnancy and birth rates among teens from low-income households using Medicaid-enrollment as a proxy for low income. This was a retrospective study of Louisiana Medicaid claims data for female teens aged 15-17 years in 2014 (n = 66,069). Pregnancy and pregnancy outcome codes were identified (n = 2,276) and analyzed for differences by black and white race. We conducted validity analyses with different rate definitions and teens' claims status. The cohort was 36% white and 54% black. More black teens than whites lacked any claims data (15.6% vs. 12.6%; p < .001). Rates calculated as events per 1,000 person-years of Medicaid coverage showed no difference in live birth rates between white and black teens (24.6 vs. 25.8; relative incidence ratio, 1.05; 95% confidence interval, .93-1.18; p = .43); however, pregnancy rates for whites were higher than those for blacks (42.7 vs. 36.1; relative incidence ratio, .85; 95% confidence interval, .77-.93; p < .001). In contrast to national trends, which include teens from diverse racial and socioeconomic backgrounds, Louisiana Medicaid-enrolled teens aged 15-17 years had equal birth rates regardless of black or white race, and whites had higher pregnancy rates. Decreased racial disparities in pregnancy and birth rates among these adolescents highlights socioeconomic influences in sexual health behavior and a need to examine the interplay of risk factors contributing to racial disparities seen among adolescents nationally.


Effect of heparin and enoxaparin on platelet interaction with fibrin clots.
It has been recently shown that platelet-rich thrombi are particularly resistant to thrombolysis. Since unfractionated heparin was reported to enhance fibrinogen binding to platelets responsible for the hyperaggregating effect of this drug, the purpose of this work was to determine whether or not heparin could also modulate platelet interaction to whole blood clot. We have therefore investigated the retention to a standard clot of 111In-labelled platelets suspended in plasma in presence of saline (control), heparin or a low molecular weight heparin (Enoxaparin). We have shown that the platelet interaction to the clot was significantly increased by heparin but not by enoxaparin used at the same anti Xa activity. In conclusion, this difference may favor the use of enoxaparin over heparin in clinical situation associated with platelet retention to fibrin clot, such as thrombolysis.


Synthesis of native 60S and 40S ribosomal subunits in Yoshida rat ascites hepatoma AH-130 cells: correlation with the rate of cell growth.
The 60Sn and 40Sn subunit ribosome synthesis declined significantly in Yoshida rat ascites hepatoma AH-130 cells from the log phase to the plateau phase of the in vivo growth. Two main classes of 40Sn particles with protein/RNA ratios of 1.82 (p2) and 1.20 (p3) and a minor "heavy" one with protein/RNA ratio of 0.96 (p1) could be distinguished reproducibly by their ultraviolet absorption after sucrose zone sedimentation. The p2 particles appeared the dominating class in log phase cells. In plateau phase cells a decrease of p2 and an increase of p3 particles was observed. Under these conditions the p1 particles and the peaks corresponding to 60Sn subunits and to 80S ribosomes were also increased. Newly synthetized 40Sn particles banded in the p3 region of the gradient and p2 particles originated from them. These particles entered into the ribosomal cycle and contained poly(A) RNA. Formation of radioactive 80S couples by subunits entering into the ribosomal cycle was markedly stimulated in log phase cells and almost completely blocked in cells at the plateau phase of growth.


Prokaryotic Genome Annotation.
In the last decade, the high-throughput and relatively low cost of short-read sequencing technologies have revolutionized prokaryotic genomics. This has led to an exponential increase in the number of bacterial and archaeal genome sequences available, as well as corresponding increase of genome assembly and annotation tools developed. Together, these hardware and software technologies have given scientists unprecedented options to study their chosen microbial systems without the need for large teams of bioinformaticists or supercomputing facilities. While these analysis tools largely fall into only a few categories, each may have different requirements, caveats and file formats, and some may be rarely updated or even abandoned. And so, despite the apparent ease in sequencing and analyzing a prokaryotic genome, it is no wonder that the budding genomicist may quickly find oneself overwhelmed. Here, we aim to provide the reader with an overview of genome annotation and its most important considerations, as well as an easy-to-follow protocol to get started with annotating a prokaryotic genome.


Public health data collection and sharing using HIPAA messages.
Public health information has significant value for doctors, public health officials, epidemiological researchers, the general public, and government agencies. Unfortunately, these data are difficult to obtain and are typically collected on as-needed basis and maintained locally. This localized process unavoidably limits the access to important public health data by its users. Moreover, the diversity of data transmission standards and collection techniques make the collected data less usable. This paper proposes a new standardized public health information system based on the HIPAA (Health Insurance Portability and Accountability Act) messages, which are the standard transactions between hospitals and insurance companies. In particular, this paper explores the applicability of HIPAA messages as a data source and transmission standard, and proposes a prototype design of a new system to collect and share public health data using HIPAA messages.


Impact of CXCL1 overexpression on growth and invasion of prostate cancer cell.
The role of CXCL1 in prostate cancer (PCa) progression has been poorly investigated. A limitation of previous studies is linked to the use of human PCa cell lines PC3 and DU145, producing CXCL8 at levels strongly exceeding CXCL1 levels. Moreover, in mouse models the sharing of CXCR2 receptor by both ligands makes the phenotype induced by CXCL8 and CXCL1 almost indistinguishable. To overcome this problem we used the murine TRAMP-C2 cell line, not expressing CXCL8 and expressing CXCL1 at low levels. The effect of CXCL1 overexpression was examined by in vivo subcutaneous tumor studies and in vitro functional assays of invasion and adhesion. Biochemical modifications were evaluated by Western blotting and antibody arrays. Our data show that the overexpression of CXCL1 in TRAMP-C2 cells represses tumor establishment and in situ invasion. In vitro, the main action of CXCL1 expression in TRAMP cells is associated with the perturbation of molecules linked to cell adhesion and migration thus explaining in vivo data. Other in vitro findings also suggest that signaling by CXCL1 might activate a secretory network limiting in vivo tumor growth by reinforcing senescence. Immunohistochemical staining of human PCa, BPH, and normal prostate biopsies strengthen our observations on the mouse model: when expressed, CXCL1 is limited to small areas with faint staining and PCa progression does not rely on CXCL1 expression. We could speculate that CXCL1 overexpression acts as a suppressor of malignancy limiting the escape of tumor cells from the primary tumor and reinforcing growth arrest.


Structural basis for inhibition of an archaeal CRISPR-Cas type I-D large subunit by an anti-CRISPR protein.
A hallmark of type I CRISPR-Cas systems is the presence of Cas3, which contains both the nuclease and helicase activities required for DNA cleavage during interference. In subtype I-D systems, however, the histidine-aspartate (HD) nuclease domain is encoded as part of a Cas10-like large effector complex subunit and the helicase activity in a separate Cas3' subunit, but the functional and mechanistic consequences of this organisation are not currently understood. Here we show that the Sulfolobus islandicus type I-D Cas10d large subunit exhibits an unusual domain architecture consisting of a Cas3-like HD nuclease domain fused to a degenerate polymerase fold and a C-terminal domain structurally similar to Cas11. Crystal structures of Cas10d both in isolation and bound to S. islandicus rod-shaped virus 3 AcrID1 reveal that the anti-CRISPR protein sequesters the large subunit in a non-functional state unable to form a cleavage-competent effector complex. The architecture of Cas10d suggests that the type I-D effector complex is similar to those found in type III CRISPR-Cas systems and that this feature is specifically exploited by phages for anti-CRISPR defence.


[Gait analysis and tailored exercise prescription in older adults].
Examining gait characteristics in older adults enhances our understanding of movement control in this population and helps to better target preventive interventions. Walking is a highly automated, regular motor behavior that is mostly controlled by subcortical locomotor brain regions. With increasing age, walking requires higher levels of attention and thus more cortical involvement in motor control. This can affect gait regularity by increasing stride-to-stride variability that is characteristically high among fallers. A growing number of clinical gait analysis systems is now available to determine gait variability and thus the falling risk in older adults. Interventions targeting high gait variability in older adults need to consider basic principles of motor learning. Previously common and automatic gait patterns have to be relearned and again brought up to a highly automated level of motor control. Regular walking exercise, and particularly T'ai Chi with its emphasis on sequenced, slow, and highly controlled movements, might be helpful in this relearning process. Further research is needed to explore other interventions that could favorably affect stride-to-stride variability of older adults.


Characterization of bioresidues for biooil production through pyrolysis.
Biomass is a renewable resource utilized to produce energy, fuels and chemicals. In this study, 25 bioresidues were selected and the physical, chemical, thermal and elemental analyses of the residues were studied as per standard methods. The bioresidues were pyrolyzed at 450°C in a fixed bed reactor to produce biooil. Among the residues, paper (pinfed computer) and Parthenium produced maximum (45%) and minimum biooil (6.33%), respectively. Arecanut stalk, redgram stalk, rice husk, wheat husk, maize cob, coir pith, Cumbu Napier grass Co5, Prosopis wood and paper resulted in a better biooil yield. Models were developed to predict the effect of constituents of bioresidues on the yield of biooil. The volatile matter and cellulose had significant effect on biooil yield. Biooil thus obtained can be used as fuel that may replace considerable fossil fuels.


Plasticity and cytokinetic dynamics of the hair follicle mesenchyme during the hair growth cycle: implications for growth control and hair follicle transformations.
Hair fiber production is the macroscopic end-point of a highly complex set of interactions between the hair follicle's epithelial and mesenchymal components. The nature of this relationship is largely set during hair follicle morphogenesis, but is dramatically revisited in the adult during the unique tissue remodeling events required for hair follicle cycling. Whereas significant attention has focused on the fate of the hair follicle epithelium during these events, associated changes in hair follicle fibroblast subpopulations remain unclear. Here, we present a speculative review that represents a critical and innovative synthesis of the current literature and summarizes a recently submitted original study by the authors, on the nature of hair cycle-dependent fibroblast dynamics and on how perturbations thereof may lead to several clinical manifestations of altered human hair growth.


In vitro studies of serum albumin interaction with poly(D,L-lactide) nanospheres loaded by hydrophobic cargo.
The various polymer-based nanocarriers are very attractive for in vitro and in vivo bioapplications. A new type of a promising drug delivery systems for cancer tissues-poly(D,L-lactide) nanospheres stabilized with Cremophor EL and loaded with hydrophobic cyanines (IR-780 or ZnPc) or curcumin (CUR) were fabricated by the nanoprecipitation method. The Cremophor EL/PLA/water nanospheres demonstrated regular shape, low polydispersity (PdI<0.3) and high entrapment efficiency of selected cargo (over 90%). The size of those nanoconstructs below 130 nm are in the desired nanocarriers size range for tumor delivery. Low level of in vitro drug release from loaded nanospheres after long-time storage indicates their good stability. The half-life of nanocarriers in the circulation, and their biodistribution after parenteral administration are associated with the ability of plasma proteins adsorption. For these reasons the affinity of obtained nanospheres for albumin as a major plasma protein was in vitro investigated. The binding of nanocarrier containing cyanine IR-780 with albumin immobilized in the wells of polystyrene plate occurred with lower efficiency than analogs loaded with ZnPc or CUR. Similar relationships were observed after UV-vis spectra analysis of nanospheres in the presence of albumin at various protein concentrations.


Fluid and electrolyte balance during a cool weather marathon.
Changes in blood, serum, and urine parameters that are usually associated with fluid and electrolyte balance were studied in 45 volunteers who ran the 1987 Pittsburgh Marathon. There were 39 males and 6 females. The mean age was 39.3 years. Their mean fluid intake was 1650 cc and the mean finishing time was 4 hours and 1 minute. The race was run in the rain with a temperature of 46 degrees F. When the prerace and postrace values of the runners were compared, significant increases were noted in the serum sodium, potassium, blood urea nitrogen (BUN), creatinine, uric acid, creatine phosphokinase (CPK), protein, plasma renin, vasopressin, and urinary potassium. Significant decreases were found in weight, blood pressure, and urinary sodium. No significant differences were noted in serum chloride, serum glucose, and hemoglobin/hematocrit. The mean weight loss of 1.9 kg was less than weight losses reported in marathons run under warmer conditions.


Improving the inhibitory activity of arylidenaminoguanidine compounds at the N-methyl-D-aspartate receptor complex from a recursive computational-experimental structure-activity relationship study.
Using a combination of both the partial least squares (PLS) and back-propagation artificial neural network (ANN) pattern recognition methods, several models have been developed to predict the activity of a series of arylidenaminoguanidine analogs as inhibitory modulators of the N-methyl-D-aspartate receptor complex. This was done by correlating structural and physicochemical descriptors obtained from computation software with the experimentally observed [(3)H]MK-801 displacement ability of a small library of synthesized and in vitro screened arylidenaminoguanidines. Results for the generated PLS model were r(2)=0.814, rmsd=0.208, rCV(2)=0.714, loormsd=0.261. The ANN model was created utilizing the eleven descriptors from the PLS model for comparison. The quality of the ANN model (r(2)=0.828, rmsd=0.200, rCV(2)=0.721, loormsd=0.257) is similar to the PLS model, and indicates that the feature between the inputs and the output is majorly linear. These computational models were able to predict inhibition of the NMDA receptor complex by this series of compounds in silico, affording a predictive structure-based 'pre-screening' paradigm for the arylideneaminoguanidine analogs.


Targeted screening for biogenic amine transporters: potential applications for natural products.
The biogenic amine transporters (BATs) are integral membrane proteins that terminate the actions of dopamine (DA), serotonin (5-HT) and norepinephrine (NE) by pumping these substrates from the extracellular space back into the nerve terminal. Numerous drugs and medications target BATs, acting as inhibitors or substrates. This paper will review some of the methods used to measure the activity of test drugs at the BATs. These methods include traditional uptake inhibition assays and transporter binding assays, as well as methods developed in our lab to determine if a test agent is a BAT substrate or inhibitor. Newer methods, developed in our lab, are used to determine the potency of test drugs as BAT substrates in a relatively high throughput manner. The potential application of these methods to characterizing natural products will be discussed in reference to results obtained with "purified" natural products, such as ephedrine stereoisomers.


Translating Inhibitory Fc Receptor Biology into Novel Therapeutic Approaches.
Innate and adaptive immune responses represent well balanced reactions aimed at resolving microbial infections without causing major collateral damage to the host. Disturbances in this system either due to enhanced activating or decreased inhibitory signaling pathways may lead to excessive immune activation resulting in tissue damage, the induction of autoimmune disease and/or chronic inflammation. On the molecular level this balance is achieved by the integration of inhibitory and activating signals, which are delivered by pairs of activating and inhibitory cell surface receptors expressed on innate and adaptive immune cells. The regulation of immunoglobulin G activity through cellular Fc receptors is a prime example for this type of regulation. This is not only relevant for the regulation of antibody-mediated effector functions through innate immune effector cells but also for the regulation of B cell activation and antibody production itself.


Multivariate analysis of primitive reflexes in patients with human immunodeficiency virus type-1 infection and neurocognitive dysfunction.
To prove that primitive reflexes are independent markers of symptomatic human immunodeficiency virus type-1 (HIV-1) infection, a case-control study was carried out in a tertiary care, university teaching hospital. Thirty HIV-1-positive symptomatic cases, 30 seropositive asymptomatic controls and 30 HIV-1 seronegative controls consented to participate and were selected consecutively. A single examiner blinded to serostatus administered the Mini-Mental State Exam and a structured neurological exam to each participant. Up to 45% of cases had cognitive impairment. The occurrence of neurologic signs between seropositive cases and seropositive controls was similar, but the number of primitive reflexes was significantly higher in cases (P < 0.001). By multivariate discriminant analysis, all primitive reflexes but two correctly classified 83.3% of all participants (P = 0.0013). The model had a positive predictive value of 97% when motor, mood, and cognitive symptoms were added (P = 0.0001). Primitive reflexes were independent predictors of HIV-1 serostatus, especially for those with cognitive dysfunction. Primitive reflexes should be included in future case definitions of HIV-1-related neurocognitive disorders.


The effects of thyroxine replacement on the levels of serum asymmetric dimethylarginine (ADMA) and other biochemical cardiovascular risk markers in patients with subclinical hypothyroidism.
The relationship between subclinical hypothyroidism (SH) and cardiovascular disease (CVD) is still under debate. The purpose of the present study was to evaluate plasma total homocysteine (tHcy), high sensitive C-reactive protein (hsCRP), small dense low-density lipoprotein (sdLDL), L-arginine and asymmetric dimethylarginine (ADMA) concentrations and their relationship to nitric oxide (NO) production, measured as plasma nitrite-plus-nitrate (NO(x)) concentration, in patients with SH before and after thyroxine replacement therapy and compared with control group values. Eighty-four women with SH and 33 healthy women as controls matched to the patient group for sex, age and body mass index (BMI), were enrolled in this study. Lipoprotein profile, tHcy, hsCRP, sdLDL, ADMA, L-arginine and NO(x) were measured in pre- and post-treatment blood samples. The pretreatment total cholesterol (TC), LDL-C, hsCRP, ADMA and L-arginine levels were significantly higher and NO(x) levels were lower than in the control group. After treatment, hsCRP, ADMA and L-arginine levels were significantly reduced and sdLDL and NO(x) levels were significantly increased. The present study demonstrated an elevation of hsCRP and ADMA plasma levels of patients with SH associated with a reduction in NO production, which may contribute to some cardiovascular alterations. The elevated ADMA and hsCRP levels were reduced after thyroxine replacement. Also the sdLDL levels of SH patients were found to be lower than the control group values whereas TC and LDL were elevated. Even though we found an elevation in sdLDL levels after treatment, those values were still not higher than in the control group.


Preoperative chemotherapy unmasks underlying Barrett's mucosa in patients with adenocarcinoma of the distal esophagus.
Intestinal metaplasia of the distal esophagus frequently cannot be detected in patients with esophageal adenocarcinoma. It has therefore been questioned whether Barrett's esophagus is the primary precursor lesion of such lesions. We hypothesized that the underlying Barrett's mucosa may be masked by tumor overgrowth in the majority of these patients. The pretherapeutic endoscopy and biopsy records of 79 patients with locally advanced esophageal adenocarcinoma who had undergone preoperative chemotherapy were reviewed and compared to findings on restaging endoscopy/biopsy and subsequent resection and histopathologic analysis of the resected specimen. Pretherapeutic endoscopy and biopsy showed associated Barrett's esophagus in 59/79 patients, whereas there was no evidence of associated intestinal metaplasia in 20/79 patients on extensive biopsies. Following neoadjuvant chemotherapy, Barrett's mucosa was unmasked and later documented by biopsy or histopathologic assessment of the resected specimen in 18 of the latter 20 patients. This resulted in an overall association of Barrett's mucosa with adenocarcinoma in the distal esophagus of 97.4% Underlying Barrett's mucosa is frequently masked by tumor overgrowth in patients with locally advanced adenocarcinoma of the distal esophagus.


Moroccan actinobacteria with promising activity against toxic cyanobacteria Microcystis aeruginosa.
In recent decades, harmful cyanobacterial blooms (HCBs) have become a severe hazard for human health mainly in drinking water resources and are responsible for serious ecological disturbances in freshwater ecosystems. The present study aims to explore the potential of actinobacteria isolated from sediment samples collected from Moroccan salt river to control HCBs mainly through Microcystis aeruginosa lysis. In order to investigate the possible anti-cyanobacterial response mechanisms, the antioxidant enzyme activities of M. aeruginosa cells were analysed. Anti-cyanobacterial activity was tested using the agar cylinder method against the toxic cyanobacteria Microcystis aeruginosa. Amongst the twenty-three isolates tested, only one showed promising anti-cyanobacterial activities with inhibition zone (ZI) equal to 22.00 mm, minimum inhibitory concentration (MIC) equal to 19.53 mg/L and minimum bactericidal concentration MBC equal to 39.06 mg/L. Phylogenetic analysis of the near-complete 16S rRNA gene sequence indicated that the strain DS1R1 belongs to the genus Streptomyces and has the highest similarity (100%) to Streptomyces sp. Indeed, M. aeruginosa growth, chlorophyll-a and protein content were significantly reduced by Streptomyces sp. DS1R1 extract. Superoxide dismutase (SOD), catalase (CAT) and malondialdehyde (MDA) were significantly elevated after treatment with Streptomyces sp. DS1R1 extract. These experimental findings provided insights in the development of a new eco-friendly procedure based on the use of actinobacteria for toxic cyanobacterial bloom bio-control.


Evaluation of mineral content of tooth enamel after application violet led associated with 35% hydrogen peroxide.
The aim of this study was to evaluate the microhardness and quantify the presence of minerals (Calcium and Phosphorous) of tooth enamel submitted to bleaching with violet LED light, either associated with 35% hydrogen peroxide gel, or not. Bovine incisors were selected and divided into 4 Groups according to the bleaching technique used: C- Without bleaching (Control); VL- violet LED; HP- 35% Hydrogen Peroxide; HP+VL- 35% Hydrogen Peroxide + violet LED. The response variables were surface microhardness (n = 12), Energy Dispersive Spectroscopy (EDS) (n = 6) and scanning electron microscopy (SEM) (n = 3). The 2-way ANOVA test with repeated measures showed that there was difference in the microhardness values of enamel in the specimens in all of the Groups when the values measured in the initial and final time intervals were compared, however, without significant differences between the values of the different treatments in both time intervals. The results with reference to EDS were analyzed with the 1-way ANOVA test. There was difference among the Groups only in the Ca%, with Group HP obtaining the lowest values, with significant difference from those of the C Group. SEM images showed a smooth, homogeneous enamel surface and similarity among the Groups, irrespective of the treatment performed. Therefore, it was concluded that the changes caused in enamel by the use of violet LED Light, either associated with 35% hydrogen peroxide, or not, did not differ from those of non-bleached tooth enamel, showing absence of harm to the mineral content of tooth enamel resulting from the use of violet LED.


High density lipoprotein cholesterol and coronary artery occlusion.
The relationship between the levels of high density lipoprotein cholesterol (HDLC) and the extent of coronary artery occlusion (ascertained by arteriography) was studied in four hundred male patients. The group with HDLC levels over 50 mg/dl showed a significantly lower coronary artery occlusion score and lower plasma triglyceride levels than the patients with HDLC levels less than 35 mg/dl. The former group also imbibed more alcohol and had less smoking experience.


Radical retropubic versus laparoscopic prostatectomy: a prospective comparison of functional outcome.
To evaluate and compare prospectively the functional results (urinary continence and erectile function) after retropubic and laparoscopic radical prostatectomy. From 1998 to 2001, 300 patients underwent radical prostatectomy, 70 by the retropubic and 230 by the laparoscopic approach. All of them completed a confidential, self-administered questionnaire regarding urinary continence and erectile function before and 1, 3, 6, and 12 months after surgery. The results were analyzed separately for day and nighttime urinary continence and by neurovascular bundle status and age for erectile function. No differences were found between the two groups except for the duration of bladder catheterization (7.8 days for the retropubic and 5.8 for the laparoscopic approach, P = 0.0006). Before surgery, 296 patients (98.7%) were continent and 290 (96.6%) were potent. One year after surgery, diurnal urinary continence increased progressively to 66.7% and 71.6% (P >0.05), nocturnal continence to 66.7% and 87.1% (P = 0.008), and potency rates to 30% and 41% (P >0.05) for the retropubic and the laparoscopic approaches, respectively. After preservation of one and two neurovascular bundles, the potency rate increased to 27% and 46% and 44% and 53% for the retropubic and laparoscopic approaches, respectively (P >0.05). For patients younger than 60 years old with bilateral neurovascular bundle preservation, the potency rates were 72% and 81% (P >0.05). The two approaches showed similar outcomes regarding continence and erectile function 1 year after surgery. However, the recovery of nocturnal continence seemed to occur earlier after the laparoscopic approach. It is necessary to conduct additional studies to determine the potential benefits of the laparoscopic approach regarding nocturnal continence.


Chromosome 18 breakpoint in t(11;18)(q21;q21) translocation associated with MALT lymphoma is proximal to BCL2 and distal to DCC.
The t(11;18)(q21;q21) translocation has recently been identified as a recurring chromosomal abnormality in a subset of extranodal marginal zone B-cell lymphoma, a low-grade lymphoma of mucosa-associated lymphoid tissue (MALT). Neither the 11q21 nor the 18q21 breakpoints have been characterized by molecular genetic analysis. As a prelude to isolation of the gene(s) involved in this translocation, we have mapped the 18q21 breakpoint region by fluorescence in situ hybridization (FISH) of YAC and PAC clones. We mapped 37 YACs assigned to a 29-cM region within the chromosomal band 18q21. Using nine of these YACs in single- and/or dual-color FISH to analyze three cases of MALT lymphomas with the t(11;18)(q21;q21) translocation, we localized the breakpoints within a 1.6-Mb nonchimeric YAC (938E1). This YAC is useful for the detection of the translocation in metaphase and in interphase cells. A nonchimeric YAC contig of an 8-cM region around the breakpoint comprising nine YACs and a PAC contig of YAC 938E1 were constructed, which enabled the refinement of the breakpoint region in the proximal region of the YAC within a <820-kb segment. This breakpoint is proximal to the BCL2 locus and distal to DCC and DPC4 loci in chromosomal band 18q21.


Persistent nasal symptoms and mediator release after continuous pollen exposure in an environmental challenge chamber.
Immediate- and late-phase reactions are associated with nasal symptoms of patients with allergic rhinitis. To examine the symptoms and mediators released after continuous allergen exposure in an environmental challenge chamber (ECC). Fifteen patients with Japanese cedar pollinosis were enrolled in this study and continuously exposed to cedar pollen at a concentration of 8,000 grains/m(3) for 3 hours in an ECC. Nasal function tests were performed, and nasal secretions were collected before pollen exposure (0 hour), immediately after exiting the ECC (3 hours), and 6 hours after exiting the ECC (9 hours). Symptom scores were recorded every 30 minutes in the ECC and every 3 hours after exiting the ECC. The frequency of sneezing and nose blowing also was monitored. The severity of symptoms in the ECC peaked approximately 2 hours after the beginning of pollen exposure and continued more than 6 hours after leaving the ECC. Concentrations of histamine, tryptase, interleukins 5, 3, 33, and 31, and substance P increased over time, whereas that of nasal fractional exhaled nitric oxide decreased. Various mediators are released during continuous allergen exposure, which subsequently induce persistent nasal symptoms. Effective treatment is required to control the intense inflammation observed after allergen exposure.


Increasing the number of biopsy cores improves the concordance of biopsy Gleason score to prostatectomy Gleason score.
To evaluate taking more biopsy cores for predicting the radical prostatectomy (RP) Gleason score compared with the biopsy Gleason score, as although random sextant biopsies are the standard for a tissue diagnosis of prostate cancer, and taking more biopsies increases the detection rate, it is uncertain whether taking more cores improves the prediction of the RP Gleason score. We analysed retrospectively 404 patients from three centres (Seattle 162, Washington 107 and Chicago 135) who had RP for prostate cancer. Six, eight or 10 biopsies were taken based on the physician's preference and the patient's characteristics. Before RP, 158 (39%) patients had six, 65 (16%) had eight and 181 (45%) had 10 biopsy cores taken. The accuracy of the Gleason sum of the three groups was 65/158 (41%), 26/65 (40%) and 104/181 (57.5%), respectively (P < 0.004, 10-core vs six-core). However, when comparing the Gleason score separately (i.e. 4 + 3 is not equal to 3 + 4), the accuracy of the three groups was 48/158 (30%), 20/65 (31%), and 95/181 (52.5%), respectively (P < 0.001, 10-core vs six core). Taking more biopsy cores improves the accuracy of the biopsy Gleason score in predicting the final Gleason score at RP; the predictive accuracy of the final Gleason score may be increased from 41% to 58% by increasing the number of biopsies from six to 10.


Enhanced resistance to early blight in transgenic tomato lines expressing heterologous plant defense genes.
Genes coding for an iris ribosomal-inactivating protein (I-RIP), a maize beta-glucanase (M-GLU), and a Mirabilis jalapa antimicrobial peptide (Mj-AMP1) were separately introduced into tomato (Lycopersicon esculentum cv. Sweet Chelsea) cotyledons via Agrobacterium tumefaciens-mediated transformation. Transgenic lines carrying each of the transgenes were confirmed for integration into the tomato genome using Southern blot hybridization. Transcription of I-RIP, M-GLU, and Mj-AMP1 genes in various transgenic lines was determined using Northern blot analysis. Plants of selected transgenic lines were inoculated with a 2-3x10(4) conidial spores/ml suspension of the fungal pathogen Alternaria solani, the causal agent of tomato early blight. Compared to control (non-transformed) plants, two transgenic lines carrying either a M-GLU or Mj-AMP1 showed enhanced resistance to early blight disease. None of the four lines carrying the I-RIP transgene showed increased resistance to early blight.


First report on delineation of differentially expressed genes and pathways in milk somatic cells of mastitic and healthy Murrah buffaloes.
Despite immense contribution of buffaloes as dairy species, limited studies have addressed the bubaline mastitis as compared to cattle. This was the first differential transcriptomic study investigating the alterations induced by clinical mastitis in buffalo milk relative to healthy controls. Comparative gene expression profiling of three biological replicates of each group identified 1014 up-regulated and 999 down-regulated genes in the diseased buffaloes (Fold change > 2, FDR < 0.05). Activation of immune and inflammatory responses were the most enriched GO terms in the mastitic animals, with higher transcript abundance of many genes coding for anti-microbial proteins such as β-defensins, perforin, granzymes, granulysin, cathelicidins etc. Analysis of the gene regulatory interactions of the up-regulated DEGs identified many hub genes that govern the cellular and macromolecular metabolic processes (E2F4, E2F1, RBL2, FOXM1, IRF1 and MYB). This study contributes to an insightful understanding of molecular mechanisms governing immune response of buffaloes to mastitis.


Pregnancy and the susceptibility of Lewis rats to experimental allergic encephalomyelitis.
Pregnant Lewis rats challenged with encephalitogen during the second or third week of gestation were afforded a high level of protection against experimental allergic encephalomyelitis, whilst females sensitised during the first week of pregnancy enjoyed only limited protection from clinical signs of disease. When rechallenged with encephalitogen, females which had been sensitised during pregnancy were marginally more susceptible to reinduction of clinical signs of disease than their virgin counterparts. Mothers inoculated during the first week of gestation were the only group to produce abnormal young.


The obesity paradox and hypoglycemia in critically ill patients.
A high body mass index (BMI) has been associated with decreased mortality in critically ill patients. This association may, in part, relate to the impact of BMI on glycemia. We aimed to study the relationship between BMI, glycemia and hospital mortality. We included all patients with a recorded BMI from four large international clinical databases (n = 259,177). We investigated the unadjusted association of BMI with average glucose levels, mortality and hypoglycemia rate. We applied multivariate analysis to investigate the impact of BMI on hypoglycemia rate, after adjusting for glycemia-relevant treatments (insulin, dextrose, corticosteroids, enteral and parenteral nutrition) and key physiological parameters (previous blood glucose level, blood lactate, shock state, SOFA score). We analyzed 5,544,366 glucose measurements. On unadjusted analysis, increasing BMI was associated with increasing glucose levels (average increase of 5 and 10 mg/dL for the 25-30, 30-35 kg/m2 BMI groups compared to normal BMI (18.5-25 kg/m2) patients). Despite greater hyperglycemia, increasing BMI was associated with lower hospital mortality (average decrease of 2% and 3.25% for the 25-30, 30-35 kg/m2 groups compared to normal BMI patients) and lower hypoglycemia rate (average decrease of 2.5% and 3.5% for the 25-30, 30-35 kg/m2 groups compared to normal BMI patients). Increasing BMI was significantly independently associated with reduced hypoglycemia rate, with odds ratio (OR) 0.72 and 0.65, respectively (95% CIs 0.67-0.77 and 0.60-0.71, both p < 0.001) when compared with normal BMI. Low BMI patients showed greater hypoglycemia rate, with OR 1.6 (CI 1.43-1.79, p < 0.001). The association of high BMI and decreased mortality did not apply to diabetic patients. Although diabetic patients had higher rates of hypoglycemia overall and higher glucose variability (p < 0.001), they also had a reduced risk of hypoglycemia with higher BMI levels (p < 0.001). Increasing BMI is independently associated with decreased risk of hypoglycemia. It is also associated with increasing hyperglycemia and yet with lower mortality. Lower risk of hypoglycemia might contribute to decreased mortality and might partly explain the obesity paradox. These associations, however, were markedly modified by the presence of diabetes.


Developmental exposure to polycyclic aromatic hydrocarbons (PAHs): Focus on benzo[a]pyrene neurotoxicity.
Polycyclic Aromatic Hydrocarbons (PAHs) are a class of ubiquitous organic compounds produced during the incomplete combustion or pyrolysis of organic material. Dietary source is the main route for PAH human exposure by environmental contamination, food industrial processing or domestic cooking methods. The most studied PAH is benzo[a]pyrene (B[a]P), due to its harmful and multiple effects on human health: in addition to its well-known carcinogenic effects, emerging evidence indicates that B[a]P also induces neurotoxicity earlier and at lower doses than B[a]P-induced carcinogenicity making B[a]P neurotoxicity relevant to human health risk assessment. Developmental neurotoxicity of B[a]P has indeed received increasing attention: both human and experimental studies provide evidence of detrimental effects of prenatal or early postnatal B[a]P exposure, even at low doses. Indeed, in some of the multi-dose animal studies, maximal adverse effects were observed at lower B[a]P doses, according to a non-monotonic dose-response curve typical of endocrine-disrupting compounds. In substantial agreement with epidemiological studies, both rodents and zebrafish developmentally exposed to B[a]P exhibit long-term changes in multiple behavioural domains, in the absence of overt toxicological effects at birth (e.g. body weight and morphologic abnormalities). Notably, most targeted behavioural responses converge on locomotor activity and emotional profile, often, but not always, leading to a disinhibitory/hyperactive profile.


Early prediction of pathologic complete response of breast cancer after neoadjuvant chemotherapy using longitudinal ultrafast dynamic contrast-enhanced MRI.
The purpose of this study was to evaluate the temporal trends of ultrafast dynamic contrast-enhanced (DCE)-MRI during neoadjuvant chemotherapy (NAC) and to investigate whether the changes in DCE-MRI parameters could early predict pathologic complete response (pCR) of breast cancer. This longitudinal study prospectively recruited consecutive participants with breast cancer who underwent ultrafast DCE-MRI examinations before treatment and after two, four, and six NAC cycles between February 2021 and February 2022. Five ultrafast DCE-MRI parameters (maximum slope [MS], time-to-peak [TTP], time-to-enhancement [TTE], peak enhancement intensity [PEI], and initial area under the curve in 60 s [iAUC]) and tumor size were measured at each timepoint. The changes in parameters between each pair of adjacent timepoints were additionally measured and compared between the pCR and non-pCR groups. Longitudinal data were analyzed using generalized estimating equations. The performance for predicting pCR was assessed using area under the receiver operating characteristic curve (AUC). Sixty-seven women (mean age, 50 ± 8 [standard deviation] years; age range: 25-69 years) were included, 19 of whom achieved pCR. MS, PEI, iAUC, and tumor size decreased, while TTP increased during NAC (all P < 0.001). The AUC (0.92; 95% confidence interval [CI]: 0.83-0.97) of the model incorporating ultrafast DCE-MRI parameter change values (from timepoints 1 to 2) and clinicopathologic characteristics was greater than that of the clinical model (AUC, 0.79; 95% CI: 0.68-0.88) and ultrafast DCE-MRI parameter model at timepoint 2 when combined with clinicopathologic characteristics (AUC, 0.82; 95% CI: 0.71-0.90) (P = 0.01 and 0.02). Early changes in ultrafast DCE-MRI parameters after NAC combined with clinicopathologic characteristics could serve as predictive markers of pCR of breast cancer.


Brain metastases from breast cancer.
Breast cancer (BC) is one of the commonest causes of brain metastases (BM): approximately 10-16 % of patients diagnosed with metastatic breast cancer will eventually develop BM during the course of their disease, however, certain subtypes have a higher risk of this event. The aim of this analysis was therefore to evaluate the prognosis and the pattern and imaging features of BM according to different BC subtypes. We retrospectively reviewed the case records of patients with breast cancer and evidence of brain metastases from the database of IPO Porto between 2014-2018. The data obtained were statistically analysed. We analysed 147 patients with BM from BC. The triple-negative subtype had the shortest overall survival (OS) after BM, besides a short period of time between BC and BM. HER2 overexpressing tumors had the longest OS. The estrogen-receptor positive group had the greatest interval between initial BC diagnosis and diagnosis of BM. Larger lesions showed a heterogeneous contrast enhancement and were heterogeneous pn T2WI sequences; a hyposignal on T2*WI was also associated with larger lesions. Triple-negative BC tended to have more heterogeneous lesions on T1WI. We noticed that the hippocampus is rarely affected by metastatic lesions. Based on the BC subtype it is possible to make a prediction about the prognosis of the disease and some imaging features of the BM, but not about their pattern of distribution. These data support further research concerning prevention, early detection, and treatment of BM from BC.


Scanning and transmission electron microscopy of biodegradable microspheres (DSM) in blood vessels.
A type of biodegradable microsphere (DSM), approximately 45 microns in diameter, made of polymerized potato starch (Pharmacia, Sweden) was intravenously injected into rats to observe the state of DSM in small blood vessels in the kidney and liver at the electron microscopic level. Prior to their digestion with amylase, individual DSM changed their round shape to an irregularly folded one to occupy almost the whole area of the lumen. At the transmission electron microscopic level, DSM were impregnated with colloidal iron and were easily identified. Interaction of the iron labelled DSM with the surface of endothelial cells was unexpectedly loose and no adherence or fusion of this surface was observed. The starch substance was not visible in the pinocytotic vesicles of the endothelium. These findings suggest the independent profile of DSM in situ.


Glycyrrhizic Acid Alleviates 6-Hydroxydopamine and Corticosterone-Induced Neurotoxicity in SH-SY5Y Cells Through Modulating Autophagy.
Recent researches have shown that autophagy is associated with the pathogenesis of neurodegenerative disorders, but there is no paper to investigate the effects of autophagy modulation on Parkinson's disease depression (PDD). In addition, glycyrrhizic acid (GA), the major bioactive ingredient of Radix glycyrrhizae, can induce autophagy and ease rotenone-induced Parkinson's disease (PD). However, there is also no paper to study the action and molecular mechanisms of GA on PDD. In this research, we built the injury model of SH-SY5Y cells through 6-hydroxydopamine (6-OHDA) and corticosterone (CORT). Then, our results showed that GA markedly increased the viability and decreased the apoptosis in SH-SY5Y cells after pre-treating with 6-OHDA and CORT. Moreover, GA notably decreased the expressions of α-Syn and p-S1292-LRRK2 proteins, and significantly increased the levels of CREB and BDNF proteins. Previous papers have suggested that CORT contributed to dopaminergic neurodegeneration via the glucocorticoid (GC)/glucocorticoid receptor (GR) interaction, and our results showed that GA reduced GC level and hypothalamic-pituitary-adrenal (HPA) activity in SH-SY5Y cells by regulating GR signaling pathway. Furthermore, mechanism investigations also showed that GA had the ability to up-regulate the conversion of LC3B II/I and the expression of Beclin-1, and induce autophagy in SH-SY5Y cells, which were reversed by the autophagy inhibitor 3-methyladenine (3-MA). Collectively, these findings proved that GA exerted efficient activity against neurotoxicity in SH-SY5Y cells induced by 6-OHDA and CORT via activation of autophagy, which should be developed as an efficient candidate for treating PDD in the future.


Correlated effects of guanabenz on single-neuron activity in the nucleus reticularis gigantocellularis, systemic arterial pressure and heart rate in the rat.
The simultaneous effects of guanabenz on the activity of single-neurons in the nucleus reticularis gigantocellularis of the medulla oblongata, systemic arterial pressure and heart rate were assessed in Sprague-Dawley rats, anesthetized with pentobarbital sodium (40 mg/kg, i.p.). Of the 35 arterial pressure-related neurons (neurons that temporally altered their spike rhythm subsequent to fluctuations in arterial pressure) which were evaluated in this reticular nucleus, 32 changed their discharge frequencies that exhibited a degree and time-course parallel to the hypotension promoted by the amino-guanidine compound (5, 10 or 20 micrograms/kg, i.v.). More importantly, these alterations neuronal activity preceded the occurrence of the induced vasodepression, signifying a causative relationship between the two events. Four of the 5 non-arterial pressure-related neurons in the gigantocellular reticular nucleus, on the other hand, manifested no basic modification in their spike frequencies in relation to the hypotension induced by guanabenz. These observations provided further support for the idea that the nucleus reticularis gigantocellularis participates actively in the cardiovascular suppressive actions of the aminoguanidine compound.


Three new cases of non-Hodgkin lymphoma with t(9;14)(p13;q32).
The majority of non-Hodgkin lymphomas of B-cell type (B-NHL) exhibit chromosomal abnormalities including many types of reciprocal translocations closely related to specific histopathologic entities. The t(9;14)(p13;q32) has been recognized as a primary genetic event directly involved in the development of lymphoplasmacytic lymphoma. In the 14 published cases, the t(9;14)(p13;32) seems to delineate a variety of low-grade B-cell disorders characterized by a common clinical history and immunopathologic similarities. We report here three new cases presenting a t(9;14)(p13;q32) with other chromosomal abnormalities which have been referred to as B-cell low-grade or high-grade malignant lymphoproliferative disorders. Two of these cases showed diffuse large B cell lymphoma morphology and two patients had a favorable clinical outcome. These data suggest that t(9;14)(p13;q32) is not restricted to low-grade lymphoma.


Short-term ocular tolerability of dorzolamide 2% and brinzolamide 1% vs placebo in primary open-angle glaucoma and ocular hypertension subjects.
To compare ocular tolerability of dorzolamide 2%, brinzolamide 1%, and placebo given three times daily. A prospective, double-masked, three-centre, crossover comparison in which 25 ocular hypertensive or primary-open angle glaucoma subjects were randomized to receive dorzolamide, brinzolamide, or placebo three times daily for 3 days. Intraocular pressure, visual acuity, a visual analogue scale, and ocular and systemic symptom queries were completed at the end of each period. After chronic dosing, there was a significant difference in ocular pain on the visual analogue scale among the groups at the 10-s postinstillation time point with dorzolamide having the highest level (22.5+/-28.9) compared to brinzolamide (5.0+/-8.7) or placebo (3.2+/-10.4) (P=0.0006). No differences between groups were observed preinstillation nor following dosing at 3 or 10-min postinstillation. On the initial instillation, the 10-s postinstillation pain was rated as 43.3+/-77.1, which was significantly higher than after chronic dosing (P=0.017). On the ocular symptom query, dorzolamide had the highest incidence of burning/stinging and redness compared to the other groups, but was generally characterized as mild. There were no significant differences in the visual acuity at any time point. This study suggests that subjects treated with dorzolamide suffer more ocular pain upon instillation compared to brinzolamide or placebo. However, pain symptoms are fewer following chronic dosing and are generally characterized as mild.


Occupational exposure to chemicals and sensory organs: a neglected research field.
The effect of industrial chemicals on the sensory perception of exposed workers has received scant attention from the medical community to date, and the scientific literature is mainly limited to some case-reports or isolated studies. Possible explanations for this include the complexity of sensory perception, and the lack of agreement among researchers on methods for testing large groups of subjects. Nevertheless, some published studies showed that vision, hearing and olfactory function can be affected by various industrial metals and solvents, and some data exist also for touch and taste. This review discusses the main industrial chemicals involved. The pathogenesis of the toxicity of chemicals to sensory perception may be related to an action on receptors, nerve fibers, and/or the brain; probably, different pathogenetic mechanisms are involved. One of the main problems in this research field is that most of the studies to date evaluated the effect of a single industrial chemical on a single sense: as an example, we know that styrene exposure can impair smell and also hearing and vision but we have little idea whether different senses are impaired in the same worker, or whether each impairment is independent. In addition, workers are frequently exposed to different chemicals: co-exposure may have no effect, or result in both an increase or a decrease of the effect, as was observed for hearing loss, but studies on this aspect are largely insufficient. Research shows that both occupational and environmental exposure to industrial chemicals can affect sense organs, and suggests that the decline of perception with age may be, at least partly, related to this exposure. Nevertheless, available evidence is incomplete, and is largely inadequate for an estimation of a "safe" threshold of exposure. Good quality further research in this field is needed. This is certainly complex and demands adequate resources, but is justified by the ultimate result: the possibility to prevent an avoidable part of the decline in sensory function with age.


A comprehensive approach to effectively engage physicians during a hospital closure: using the physician engagement model.
In 2003, St. Vincent's Hospital (SVH) closed its doors. The authors investigate the involvement of the medical staff in the successful closure of SVH using the Physician Engagement (PE) Model. This 10-strategy model is based on engagement, communication, education and support. Results were gathered by surveys, unstructured interviews and meetings. Data suggested that engaging physicians in the process was favourable, particularly by using the PE model. Six recommendations are given to assist administrators/decision-makers in future closures.


Cash transfers and residential demand for electricity: insights from BISP, Pakistan.
We examine the causal impacts of the cash transfer program, namely the Benazir Income Support Program (BISP), on residential demand for electricity among ultra-poor in Pakistan. We also analyze the effects of BISP cash transfers on a household's decision to acquire electrical appliances. The empirical analysis is based on the fuzzy regression discontinuity design (RDD) using primary data collected from 1200 households. We find that BISP cash transfer has a significant positive impact on electricity demand among the target group. The cash transfer positively affects the use of few essential electric appliances, such as a washing machine and refrigerator, but not all electrical appliances. The electricity demand mainly stems from the additional use of existing electrical devices. Therefore, the extra income from BISP may not allow the recipients to move up the electric appliances ladder. The provincial analysis shows that the impact of BISP cash transfers on electricity demand varies across provinces and the development level, signifying the importance of regional heterogeneities, such as electricity supply. The findings suggest that cash transfers may facilitate the transition from traditional to modern energy to overcome the rising pollution problem and protect health. The expansion in the cash transfer program demands continuous investment in the power sector to fulfill the growing need for electricity.


Lymphocyte responses to an aqueous extract from cow hair and dander. Investigations of asthmatics and normal controls with lymphocyte transformation tests.
The proliferative responsiveness to cow dander antigens of lymphocytes from 23 asthmatic patients with allergy to cow hair and dander, from 10 patients with allergic asthma to other antigens and from 31 non-atopic subjects was investigated. Lymphocytes from patients with allergic asthma due to cow epithelium antigens showed a statistically significant decreased responsiveness to an extract from cow dander, compared to that seen with lymphocytes from newborn infants, lymphocytes from normal controls and from asthmatic patients without allergy to cow. The response to the cow dander extract seemed to be an accessory cell dependent T lymphocyte response.


[Effect of angular pyranocoumarin isolated from peucedanum praeruptorum on the proliferation and apoptosis of U266 cells].
To investigate the effects of angular pyranocoumarin (±) -4'-O- acetyl-3'-Oangeloyl- cis- khellactone (APC) extracted from peucedanum praeruptoruon on the proliferation and apoptosis of U266 cells, and to explore its related mechanism. APC was extracted by petroleum ether technique, and its purity was tested by high performance liquid chromatography, and its chemical structure was identified by magnetic resonance spectroscopy. U266 cells were treated with APC in various concentrations (0, 10, 20, 30, 40 μg/ml)for different durations(24 and 48 h). The inhibitive effect of APC on cell growth was detected by CCK-8 method. After U266 cells were incubated with APC(0, 10, 20, 30, 40 μg/ml)for 24 h, the apoptosis of cells were observed by flow cytometry stained with Annexin Ⅴ/PI and Hochest33342; the expression levels of caspase-3, 8, ERK, p-ERK, AKT and p-AKT protein were assayed by Western blot; the expression of hTERT mRNA was measured by RT-PCR. The purity of APC identified by magnetic resonance imaging was 98.8%. The proliferation of U266 cells was inhibited, and the apoptosis was induced in a time- and/or dose- dependent manner after treatment with APC. APC could upregulate the caspase- 8, 3 protein expression and downregulate the p- ERK, p-AKT protein expression along with the increase of APC dose. APC also could downregulate the hTERT mRNA expression. Angular pyranocoumarin APC could inhibit the proliferation and induce the apoptosis of U266 cells. The probable mechanism might be achieved by upregulating caspase-8, 3 protein expression and downregulating p-ERK, P-AKT protein and the hTERT mRNA expression.


31 P MRSI at 7 T enables high-resolution volumetric mapping of the intracellular magnesium ion content in human lower leg muscles.
The non-invasive determination of the free magnesium ion concentration ([Mg2+free ]) using 31 P MRSI in vivo is of interest in research on various pathologies, e.g. diabetes. The purpose of this study was to demonstrate the potential of 31 P MRSI at 7 T to enable volumetric, high-resolution mapping of [Mg2+free ]. 3D 31 P MRSI datasets from the lower leg of three healthy volunteers were acquired at B0 = 7 T with a nominal spatial resolution of (8 × 8 × 16) mm3 in 56 min. Volumetric [Mg2+free ] maps were calculated based on the quantified local chemical shift difference between the α- and β-resonance of adenosine triphosphate (ATP) considering also local pH values. Mean [Mg2+free ] values from three different muscle groups were compared. To demonstrate the potential of reducing the measurement time, the analysis was repeated on the acquired MRSI data retrospectively reconstructed with fewer averages. The generated [Mg2+free ] maps revealed local differences, and mean [Mg2+free ] values of (1.08 ± 0.03) mM were found in the tibialis anterior, (0.91 ± 0.04) mM in the soleus and (0.98 ± 0.03) mM in the gastrocnemius medialis. The time-reduced 28-min scan resulted in comparable [Mg2+free ] maps, and mean values being in agreement with the values from the 56-min scan. 31 P MRSI at 7 T enables volumetric, high-resolution mapping of free magnesium ion content in human lower leg muscles. The measurement time of the 31 P MRSI acquisition can be reduced to 28 min, opening the potential to apply volumetric [Mg2+free ] mapping for the investigation of pathologies with altered magnesium homeostasis.


The secretome of the maize pathogen Ustilago maydis.
Ustilago maydis establishes a biotrophic relationship with its host plant, i.e. plant cells stay alive despite massive fungal growth in infected tissue. The genome sequence has revealed that U. maydis is poorly equipped with plant cell wall degrading enzymes and uses novel secreted protein effectors as crucial determinants for biotrophic development. Many of these effector genes are clustered and differentially regulated during plant colonization. In this review, we analyze the secretome of U. maydis by differentiating between secreted enzymes, likely structural proteins of the fungal cell wall (excluding GPI-anchored proteins) as well as likely effectors with either apoplastic or cytoplasmic function. This classification is based on the presence of functional domains, general domain structure and cysteine pattern. In addition, we discuss possible functions of selected protein classes with a special focus on disease development.


Comparing adsorption performance of microfibers and nanofibers with commercial molecularly imprinted polymers and restricted access media for extraction of bisphenols from milk coupled with liquid chromatography.
Advanced solid phase extraction (SPE) fibrous sorbents including polyethylene, polypropylene poly (hydroxybutyrate), and polyamide 6 nanofibers, polycaprolactone microfibers/nanofibers, polycaprolactone microfibers/polyvinylidene difluoride nanofibers, and poly (hydroxybutyrate) microfibers/polypropylene microfibers composites, as well as commercial molecularly imprinted polymers and restricted access media sorbent were compared in terms of bisphenols extraction from milk and their clean-up efficiency. Three on-line SPE-HPLC methods were completely validated for the extraction and detection of bisphenols A, AF, C, A diglycidyl ether, and F diglycidyl ether in bovine milk. Polycaprolactone composite nanofibers compared favorably to restricted access media, enabled excellent clean-up of bisphenols from the proteinaceous matrix, and yielded recoveries 98.0-124.5% and 93.0-115.0%, respectively, with RSD less than 10%. Total analysis time including on-line SPE step lasted only 12 min, which represents a significant reduction in time compared with previously reported as well as official European Union and AOAC methods defined for the determination of bisphenols in various matrices.


Disposition kinetics and dynamics of nicainoprol, a new antiarrhythmic agent, in humans.
Kinetics and resting and exercise-induced hemodynamic and ECG effects of nicainoprol, a new antiarrhythmic structurally resembling propafenone or propranolol, were investigated in eight healthy male subjects receiving a 1-hour infusion (100 mg) and oral dose (200 mg) in a randomized-crossover fashion. Nicainoprol in plasma and urine was determined by a specific HPLC assay. Plasma concentration-time data were fitted to a triexponential equation. Mean postinfusion kinetic data were: alpha-phase half-life = 3.1 minutes, beta-phase half-life = 106.6 minutes, and gamma-phase half-life = 12.4 hours; volume of central compartment = 0.114 L/kg; steady-state volume of distribution = 0.67 L/kg; total clearance = 3.6 ml/min/kg; and renal clearance = 0.56 ml/min/kg. Absolute bioavailability was approximately 70% and peak plasma drug concentration occurred 2.3 hours after oral administration. Interindividual variability in AUC was 1.6- and 2.4-fold after intravenous and oral administration, respectively. Cumulative fraction excreted unchanged in urine was approximately 15% and 9% of the dose after intravenous and oral administration, respectively. Resting heart rates were increased, whereas exercise-induced heart rates were unchanged after both doses. QRS durations were widened after both doses. PR and QTc intervals were prolonged during intravenous study. The results suggest that nicainoprol is an enzyme-limited or poorly extracted drug suitable for both intravenous and oral administration and devoid of beta-blocking action in humans, at least with doses tested in this study. Its ECG properties appear to be similar to those of class I antiarrhythmics.


Reintegration needs of young women following genitourinary fistula surgery in Uganda.
Genitourinary fistulas (usually arising following prolonged obstructed labor) are particularly devastating for women in low-income counties. Surgical repair is often difficult and delayed. While much attention has been devoted to technical surgical issues, the challenges of returning to normal personal, family, and community life after surgical treatment have received less scrutiny from researchers. We surveyed young Ugandan women recovering from genitourinary fistula surgery to assess their social reintegration needs following surgery. A cross-sectional survey of 61 young women aged 14-24 years was carried out 6 months postoperatively. Interviews were carried out in local languages using a standardized, interviewer-administered, semistructured questionnaire. Data were entered using EpiData and analyzed using SPSS. Ongoing reintegration needs fell into interrelated medical, economic, and psychosocial domains. Although >90% of fistulas were closed successfully, more than half of women had medical comorbidities requiring ongoing treatment. Physical limitations, such as foot drop and pelvic muscle dysfunction impacted their ability to work and resume their marital relationships. Anxieties about living arrangements, income, physical strength, future fertility, spouse/partner fidelity and support, and possible economic exploitation were common. Sexual dysfunction after surgery-including dyspareunia, loss of libido, fear of intercourse, and anxieties about the outcome of future pregnancies-negatively impacted women's relationships and self-esteem. Young women recovering from genitourinary fistula surgery require individualized assessment of their social reintegration needs. Postoperative social reintegration services must be strengthened to do this effectively.


Conformational and functional studies of a cytosolic 90 kDa heat shock protein Hsp90 from sugarcane.
Hsp90s are involved in several cellular processes, such as signaling, proteostasis, epigenetics, differentiation and stress defense. Although Hsp90s from different organisms are highly similar, they usually have small variations in conformation and function. Thus, the characterization of different Hsp90s is important to gain insight into the structure-function relationship that makes these chaperones key regulators in protein homeostasis. This work describes the characterization of a cytosolic Hsp90 from sugarcane and its comparison with Hsp90s from other plants. Previous expressed sequence tag (EST) studies in Saccharum spp. (sugarcane) predicted the presence of an mRNA coding for a cytosolic Hsp90. The corresponding cDNA was cloned, and the recombinant protein was purified and its conformation and function characterized. The structural conformation of Hsp90 was assessed by chemical cross-linking and hydrogen/deuterium exchange using mass spectrometry and hydrodynamic assays, which revealed regions accessible to solvent and that Hsp90 is an elongated dimer in solution. The in vivo expression of Hsp90 in sugarcane leaves was confirmed by western blot, and in vitro functional characterization indicated that sugarcane Hsp90 has strong chaperone activity.


Mapping of macrophage elastase cleavage sites in insoluble human skin elastin.
Macrophage elastase (MMP-12) is a member of the family of matrix metalloproteinases (MMPs) and is active against multiple extracellular protein substrates such as elastin. Its effect on elastin is central to emphysema in the lung and photoaging of skin. Its expression in the skin increases on photodamaged skin and upon aging. Detecting and characterizing peptides cleaved in elastin, therefore, helps to understand such degradative disease processes in the skin and is also needed to assist in the rational design of agents that specifically inhibit the degradation. In this study, cleavage sites of MMP-12 in human skin elastin were extensively investigated. The peptides formed as a result of cleavages by this enzyme in the human skin elastin were characterized using mass spectrometry. A total of 41 peptides ranging from 4 to 41 amino acids were identified and 36 cleavage sites were determined. Amino acids encoded by exons 5, 6, 26, 28-31 were particularly susceptible to cleavages by MMP-12 and none or very few cleavages were detected from domains encoded by the remaining exons. The amino acid preferences of the different subsites on the catalytic domain of MMP-12 were analyzed.


[Evaluation of interest in research among surgically active medical officers in the German Armed Forces].
Research in military medicine and in particular combat surgery is a broad field that has gained international importance during the last decade. In the context of increased NATO missions, this also holds true for the Bundeswehr (German Armed Forces); however, medical officers in surgery must balance research between their clinical work load, missions, civilian and family obligation. To evaluate engagement with and interest in research, a questionnaire was distributed among the doctors of the surgical departments of the Bundeswehr hospitals by the newly founded working group Chirurgische Forschung der Bundeswehr (surgical research of the Bundeswehr). Returned data were recorded from October 2013 to January 2014 and descriptive statistics were performed. Answers were received from 87 out of 193 military surgeons (45 %). Of these 81 % announced a general interest in research with a predominance on clinical research in preference to experimental settings. At the time of the evaluation 32 % of the participants were actively involved in research and 53 % regarded it as difficult to invest time in research activities parallel to clinical work. Potential keys to increase the interest and engagement in research were seen in the implementation of research coordinators and also in a higher amount of free time, for example by research rotation. Research can be regarded as having a firm place in the daily work of medical officers in the surgical departments of the Bundeswehr; however, the engagement is limited by time and structural factors. At the departmental level and in the command structures of the military medical service, more efforts are recommended in the future in order to enhance the engagement with surgical research. This evaluation should be repeated in the coming years as a measuring instrument and data should be compared in an international context.


Neutrophil-to-Lymphocyte Ratio as an Early Predictor of Complication and Mortality Outcomes in Individuals With Acute Pancreatitis at a UK District General Hospital: A Retrospective Analysis.
Aim The purpose of this study was to evaluate the predictive usefulness of the neutrophil-to-lymphocyte ratio (NLR) in patients with acute pancreatitis (AP) and establish a threshold for the prediction of poor outcomes. Methods For this investigation, we looked back at all cases of acute pancreatitis treated at Torbay Hospital in Torquay, UK, between January 1st, 2019, and December 31st, 2020. Those who were found to have chronic pancreatitis or whose baseline laboratory values could not be obtained were not included. Each patient's entire hospital stay was analyzed, including up to 72 hours of medical and laboratory data. Results According to the Glasgow Coma Scale scoring system, 28 of the 314 included patients had severe acute pancreatitis, and 81 patients had pancreatitis with complications. Those with complications had a substantially higher NLR on day 1 (9.43 ± 7.57) than patients who recovered without complications (7.37 ± 5.88) (P-value = 0.028). The NLR on day 0 (>18.71) exhibited a sensitivity of 80%, a specificity of 90.2%, and an accuracy of 83.9% in forecasting the death of patients with pancreatitis. Conclusion Elevated baseline NLR corresponds with pancreatitis with complications and can predict mortality.


Decreased sensitivity to interferon associated with in vitro transformation of X-ray-transformed C3H/10T1/2 cells.
We have compared the interferon sensitivity of parental 10T1/2 cells, having a phenotype characteristic of normal cells, with that of transformed cells isolated after X-irradiation of treatment with methylcholanthrene. Transformed cells were 10- to 20-fold less sensitive than the parental untransformed cells to the antiviral action of interferon and were also less sensitive to the inhibitory effect of interferon on cell multiplication. The decreased sensitivity of these cells to interferon is a useful marker of the transformed phenotype.


RIPK1 Expression Associates With Inflammation in Early Atherosclerosis in Humans and Can Be Therapeutically Silenced to Reduce NF-κB Activation and Atherogenesis in Mice.
Chronic activation of the innate immune system drives inflammation and contributes directly to atherosclerosis. We previously showed that macrophages in the atherogenic plaque undergo RIPK3 (receptor-interacting serine/threonine-protein kinase 3)-MLKL (mixed lineage kinase domain-like protein)-dependent programmed necroptosis in response to sterile ligands such as oxidized low-density lipoprotein and damage-associated molecular patterns and that necroptosis is active in advanced atherosclerotic plaques. Upstream of the RIPK3-MLKL necroptotic machinery lies RIPK1 (receptor-interacting serine/threonine-protein kinase 1), which acts as a master switch that controls whether the cell undergoes NF-κB (nuclear factor κ-light-chain-enhancer of activated B cells)-dependent inflammation, caspase-dependent apoptosis, or necroptosis in response to extracellular stimuli. We therefore set out to investigate the role of RIPK1 in the development of atherosclerosis, which is driven largely by NF-κB-dependent inflammation at early stages. We hypothesize that, unlike RIPK3 and MLKL, RIPK1 primarily drives NF-κB-dependent inflammation in early atherogenic lesions, and knocking down RIPK1 will reduce inflammatory cell activation and protect against the progression of atherosclerosis. We examined expression of RIPK1 protein and mRNA in both human and mouse atherosclerotic lesions, and used loss-of-function approaches in vitro in macrophages and endothelial cells to measure inflammatory responses. We administered weekly injections of RIPK1 antisense oligonucleotides to Apoe-/- mice fed a cholesterol-rich (Western) diet for 8 weeks. We find that RIPK1 expression is abundant in early-stage atherosclerotic lesions in both humans and mice. Treatment with RIPK1 antisense oligonucleotides led to a reduction in aortic sinus and en face lesion areas (47.2% or 58.8% decrease relative to control, P<0.01) and plasma inflammatory cytokines (IL-1α [interleukin 1α], IL-17A [interleukin 17A], P<0.05) in comparison with controls. RIPK1 knockdown in macrophages decreased inflammatory genes (NF-κB, TNFα [tumor necrosis factor α], IL-1α) and in vivo lipopolysaccharide- and atherogenic diet-induced NF-κB activation. In endothelial cells, knockdown of RIPK1 prevented NF-κB translocation to the nucleus in response to TNFα, where accordingly there was a reduction in gene expression of IL1B, E-selectin, and monocyte attachment. We identify RIPK1 as a central driver of inflammation in atherosclerosis by its ability to activate the NF-κB pathway and promote inflammatory cytokine release. Given the high levels of RIPK1 expression in human atherosclerotic lesions, our study suggests RIPK1 as a future therapeutic target to reduce residual inflammation in patients at high risk of coronary artery disease.


In vitro induction of tumor necrosis factor-α by ochratoxin A (OTA) from rat liver: role of Kupffer cells.
Tumor necrosis factor-α (TNF-α) is released from blood-free perfused rat liver by the fungal metabolite ochratoxin A. Here we have identified Kupffer cells as the sole source of OTA-mediated cytokine release. If single cell preparation of Kupffer cells, hepatocytes, or sinusoidal endothelial cells were prepared from rat livers, only Kupffer cells released TNF-α upon incubation with 2.5 μmol/l OTA. OTA failed to induce TNF-α release in the blood-free perfused isolated rat liver when Kupffer cells were blockedin vitro by 15 μmol/l gadolinium chloride. When rats were pretreatedin vivo with the Kupffer cell depleting clodronate liposomes, OTA-mediated TNF-α release was abrogated in the isolated perfused liver model.


Effect of follicle size on hormonal status of follicular fluid, oocyte ATP content, and in vitro embryo production in prepubertal sheep.
The aim of this study was to assess the effect of follicular size on estradiol (E2) and progesterone (P4) levels in intrafollicular fluid, ATP content in oocytes, and the embryo development rate in prepubertal sheep. Slaughterhouse ovaries were dissected to recover the follicles, which were classified according to the follicle diameter as <3 mm (n = 20) and ≥3 mm (n = 17). Then, follicular fluid was obtained and analyzed by radioimmunoassay to determine the E2 and P4 concentrations. Another group of ovaries was used to recover cumulus-oocyte complexes according to follicle size. In vitro maturation (IVM), in vitro fertilization (IVF), and embryo culture were performed using standard procedures, and ATP level was assessed at 0 and 24 h of IVM. Intrafollicular concentrations of E2 and P4 and E2:P4 ratio were higher in ≥3 mm (18.7 ± 5.9 ng/mL, 7.8 ± 1.2 ng/mL, and 3.6 ± 1.3, respectively) than <3 mm (1.8 ± 0.4 ng/mL, 2.6 ± 0.3 ng/mL and 0.9 ± 0.3, respectively) follicles. The rate of ATP increased during IVM and was higher in oocytes from ≥3 mm than <3 mm (22.4 ± 0.7 and 8.6 ± 2.2-fold change; respectively) follicles. After IVF, the blastocyst development was higher in oocytes recovered from ≥3 mm (11.1 ± 0.9%) than from <3 mm (6.5 ± 0.7%) follicles. These results indicate an improvement in the competence and development of oocytes from ≥3 mm follicles with a higher E2:P4 ratio. Thus, this ratio could be used as reference to design IVM medium and to enhance the in vitro embryo production in lambs.


Novel ECG criteria for right ventricular systolic dysfunction in patients with right bundle branch block.
Altered hemodynamics of a failing right ventricle (RV) may place stress on the right bundle branch and Purkinje network, which may be evident as conduction delay on surface electrocardiogram (ECG). We hypothesized that prolonged R' duration in lead V1 would be an indicator of RV dysfunction in patients with RBBB. The Mayo Clinic Arizona echocardiography database was reviewed from 2007 to 2009 to identify patients with RV dysfunction and coexistent right bundle branch block (RBBB). Specific ECG features of RBBB were compared between the RV dysfunction cohort and a randomly selected control population. Features found to be predictive of RV dysfunction were then tested on 100 consecutive patients with RBBB on ECG between January and June 2010. In lead V1, the QRS duration was longer in the RV dysfunction cohort (164 ± 22 ms) compared to controls (148 ± 12 ms), predominantly due to R' prolongation (117 ± 27 ms vs. 87 ± 13 ms, p<.001). Retrospective analysis suggested that V1 R' duration ≥ 100 ms may be 82.3% specific for the presence of RV systolic dysfunction. When applied prospectively, V1 R' duration ≥ 100 ms yielded sensitivity and specificity of 39.0% and 82.9% respectively for detection of abnormal RV systolic function with a positive predictive value of 76.7%. Lead V1 R' duration ≥ 100 ms is predictive of RV systolic dysfunction in patients with RBBB.


Comparison of propofol with thiopental and isoflurane for induction and maintenance of general anesthesia.
Propofol, a phenol compound with a short elimination half-life, was compared with thiopental and isoflurane for induction and maintenance of general anesthesia in 60 consenting ASA I, II, and III patients. The study was randomized and open label in design. Hemodynamically, the propofol patients showed a mean +/- SEM decrease in systolic blood pressure in comparison with the thiopental/isoflurane group at 2 (115.1 +/- 4.9 vs. 136.6 +/- 6.0 mmHg), 3 (125.7 +/- 5.1 vs. 149.4 +/- 5.6 mmHg), and 5 min (126.6 +/- 3.8 vs. 144.4 +/- 6.1 mmHg) postinduction and at intubation (135.2 +/- 4.7 vs. 157.8 +/- 6.0 mmHg) (p less than 0.05). The heart rate was lower in the propofol group throughout the induction period (p less than 0.05). Patients who received propofol were ready for discharge from the recovery room sooner (67.9 +/- 4.0 vs. 80.0 +/- 3.6 min) than the thiopental/isoflurane-treated patients (p less than 0.05). Propofol is as safe and effective for induction and maintenance of general anesthesia as thiopental and isoflurane.


Genesis and evolution of proteinuria in diabetes mellitus.
Approximately 40-45% of insulin-dependent diabetic (IDD) patients will develop, with time, clinical proteinuria, a forerunner of certain renal failure. Before this, however, up to 45% of IDD patients excrete supranormal amounts of protein in the urine, though still undetectable by dipstix test. This microproteinuria appears to be glomerular in origin, consists mainly of albumin and IgG, and is associated with poor glycemic control and marginal elevation of arterial pressure. Glomerular hemodynamic disturbances, and loss of charge selectivity of the glomerular membrane, are probably responsible for this microproteinuria, which appears reversible by correction of hyperglycemia and raised blood pressure. Once the dipstix test becomes positive (i.e. total urinary protein excretion exceeds 0.5 g/24h) and blood pressure rises into the hypertensive range, glomerular filtration rate (GFR) falls relentlessly. By the time GFR is as low as 20 ml/min/1.73 m2, more IgG relative to albumin is being filtered, giving rise to a low selectivity proteinuria, a condition consistent with changes in the size selectivity properties of the glomerular filtre. Glycemic control does not affect the decline in GFR, although blood pressure control and low protein diet can slow it, presumably by altering the self-perpetuating hemodynamic disturbances that occur in surviving glomeruli. The recent demonstration that IDD patients with microalbuminuria in excess of 30 micrograms/min have approximately a 20-fold increase in risk of developing persistent detectable proteinuria has provided a link between these two phases of diabetic nephropathy. The reversibility of the early microalbuminuria heralds a real chance of preventing the later irreversible phase of end-stage renal failure.


"Something Extra on Chromosome 5": Parents' Understanding of Positive Prenatal Chromosomal Microarray Analysis (CMA) Results.
This study aims to explore how couples' understanding of the nature and consequences of positive prenatal chromosomal microarray analysis (CMA) results impacts decision-making and concern about pregnancy. We interviewed 28 women and 12 male partners after receiving positive results and analyzed the transcripts to assess their understanding and level of concern about the expected clinical implications of results. Participant descriptions were compared to the original laboratory interpretation. When diagnosed prenatally, couples' understanding of the nature and consequences of copy number variants (CNVs) impacts decision-making and concern. Findings suggest women, but less so partners, generally understand the nature and clinical implications of prenatal CMA results. Couples feel reassured, perhaps sometimes falsely so, when a CNV is inherited from a "normal" parent and experience considerable uncertainty when a CNV is de novo, frequently precipitating a search for additional information and guidance. Five factors influenced participants' concern including: the pattern of inheritance, type of possible phenotypic involvement, perceived manageability of outcomes, availability and strength of evidence about outcomes associated with the CNV, and provider messages about continuing the pregnancy. A good understanding of results is vital as couples decide whether or not to continue with their pregnancy and seek additional information to assist in pregnancy decision-making.


Diffuse large B cell lymphoma presenting as multiple cavitary lung lesions.
A female patient in her 50s presented to the hospital with a 2 months history of shortness of breath and productive cough. She also had ongoing fatigue and unintentional weight loss. Investigations demonstrated a normal white cell count and elevated lactate dehydrogenase). A CT scan of the chest revealed multiple bilateral cavitary lung masses and pulmonary nodules. Bronchoscopy with transbronchial biopsy and fine needle aspiration of mediastinal lymph node was performed. The histopathology was consistent with the non-germinal centre B cell subtype of diffuse large B cell lymphoma (DLBCL). A regimen of rituximab, cyclophosphamide, doxorubicin, vincristine and prednisone was started, which improved her symptoms. Although a rare presentation, this case highlights the need to consider DLBCL among the differential diagnoses for cavitary lung lesions when the workup fails to elicit an alternate cause. Given its aggressive nature, prompt diagnosis and treatment are critical.


The effects of nifedipine and verapamil on antigen-induced bronchoconstriction in dogs.
The effects of calcium channel blockers, nifedipine and verapamil (i.v. and aerosol), were investigated in beagle dogs natively allergic to Ascaris suum antigen. Control exposures to an aerosol of Ascaris antigen provoked significant bronchopulmonary changes, i.e., increases in pulmonary resistance (RL) and decreases in dynamic lung compliance (CDYN). Pretreatment with either nifedipine or verapamil (200 micrograms/kg, i.v.) provided significant inhibition in the RL responses to Ascaris antigen (P less than 0.015) while neither agent significantly affected CDYN changes. When administered as an aerosol, verapamil (1.0%; 10 breaths) significantly inhibited both the RL and CDYN responses to Ascaris antigen (P less than 0.05), whereas a similar concentration of nifedipine was without effect. Resting basal levels of RL and CDYN were unaffected by either the i.v. route or by aerosols of either nifedipine or verapamil. These results suggest that calcium channel blockers may have beneficial effects against allergen-provoked bronchoconstriction; however, differences appear to exist in the choice of agent, route of administration and site of action.


Ovarian carcinoma as an incidental finding during cesarean section in a preeclamptic woman: case report.
Gynecologic malignancies are rarely associated with pregnancy and ovarian tumors diagnosed during cesarean section are very uncommon. A 38-year-old grandmultipara with no prenatal care was hospitalized at an estimated 28 weeks of gestation for high blood pressure and increased proteinuria but no other symptoms of preeclampsia. We present a rare case of advanced ovarian carcinoma diagnosed during cesarean section.


Fast and ultrafast magnetic resonance imaging in renal lesions.
Our purpose was to analyze and compare the image quality and contrast-to-noise ratio (CNR) of different fast T1- and T2-weighted sequences with conventional spin-echo sequences in renal MRI. Twenty-three patients with focal renal lesions were examined with a T2-weighted ultrafast turbo spin-echo (UTSE) sequence with and without frequency selective fat suppression (SPIR), a combined gradient-and-spin-echo sequence (GraSE), and a conventional spin-echo sequence (SE). In addition, T1-weighted images were obtained pre- and postcontrast, using a fast spin-echo sequence (TSE) with and without SPIR and the conventional SE sequence. Among the T2-weighted images, the highest CNR and the best image quality were obtained with the UTSE sequence, followed by the fat-suppressed UTSE sequence. GraSE and conventional SE sequences showed a significantly lower CNR and image quality (p < 0.05). The T1-weighted sequences did not show significant differences, in either precontrast or postcontrast measurements. T2-weighted UTSE with and without fat suppression combined excellent image quality and high CNR for imaging and detection of renal lesions. The T1-weighted fast sequences provided no alternative to the gradient-echo or to the conventional SE sequences. The results of this systematic study suggest the use of T2-weighted fast techniques for improved diagnostic accuracy of renal MRI.