The gene expression profile of inflammatory, hypoxic and metabolic genes predicts the metastatic spread of human head and neck squamous cell carcinoma.
To assess the prognostic value of the expression profile of the main genes implicated in hypoxia, glucose and lactate metabolism, inflammation, angiogenesis and extracellular matrix interactions for the metastatic spread of head and neck squamous cell carcinoma. Using a high-throughput qRT-PCR, we performed an unsupervised clustering analysis based on the expression of 42 genes for 61 patients. Usual prognostic factors and clustering analysis results were related to metastasis free survival. With a median follow-up of 48months, 19 patients died from a metastatic evolution of their head and neck squamous cell carcinoma and one from a local recurrence. The unsupervised clustering analysis distinguished two groups of genes that were related to metastatic evolution. A capsular rupture (p=0.005) and the "cluster CXCL12 low" (p=0.002) were found to be independent prognostic factors for metastasis free survival. Using a Linear Predictive Score methodology, we established a 9-gene model (VHL, PTGER4, HK1, SLC16A4, DLL4, CXCL12, CXCR4, PTGER3 and CA9) that was capable of classifying the samples into the 2 clusters with 90% accuracy. In this cohort, our clustering analysis underlined the independent prognostic value of the expression of a panel of genes involved in hypoxia and tumor environment. It allowed us to define a 9-gene model which can be applied routinely to classify newly diagnosed head and neck squamous cell carcinoma. If confirmed by an independent prospective study, this approach may help future clinical management of these aggressive tumors.


High-Pressure Nonequilibrium Dynamics on Second-to-Microsecond Time Scales: Application of Time-Resolved X-ray Diffraction and Dynamic Compression in Ice.
The study of nonequilibrium transition dynamics on structural transformation from the second to microsecond regime, a time scale between static and shock compression, is an emerging field of high-pressure research. There are ample opportunities to uncover novel physical phenomena within this time regime. Herein, we briefly review the development and application of a dynamic compression technique based on a diamond anvil cell (DAC) and time-resolved X-ray diffraction (TRXRD) for the study of time-, pressure-, and temperature-dependent structural dynamics. Applications of the techniques are illustrated with our recent investigations on the mechanisms of the interconversions between different high-pressure ice polymorphs. These examples demonstrate that a combination of dynamic compression and TRXRD is a versatile approach capable of providing information on the kinetics and thermodynamic nature associated with structural transformations. Future improvement of rapid compression and TRXRD techniques and potentially interesting research topics in this area are suggested.


Mitochondrial Entry of Cytotoxic Proteases: A New Insight into the Granzyme B Cell Death Pathway.
The mitochondria represent an integration and amplification hub for various death pathways including that mediated by granzyme B (GB), a granule enzyme expressed by cytotoxic lymphocytes. GB activates the proapoptotic B cell CLL/lymphoma 2 (Bcl-2) family member BH3-interacting domain death agonist (BID) to switch on the intrinsic mitochondrial death pathway, leading to Bcl-2-associated X protein (Bax)/Bcl-2 homologous antagonist/killer- (Bak-) dependent mitochondrial outer membrane permeabilization (MOMP), the dissipation of mitochondrial transmembrane potential (ΔΨm), and the production of reactive oxygen species (ROS). GB can also induce mitochondrial damage in the absence of BID, Bax, and Bak, critical for MOMP, indicating that GB targets the mitochondria in other ways. Interestingly, granzyme A (GA), GB, and caspase 3 can all directly target the mitochondrial respiratory chain complex I for ROS-dependent cell death. Studies of ROS biogenesis have revealed that GB must enter the mitochondria for ROS production, making the mitochondrial entry of cytotoxic proteases (MECP) an unexpected critical step in the granzyme death pathway. MECP requires an intact ΔΨm and is mediated though Sam50 and Tim22 channels in a mtHSP70-dependent manner. Preventing MECP severely compromises GB cytotoxicity. In this review, we provide a brief overview of the canonical mitochondrial death pathway in order to put into perspective this new insight into the GB action on the mitochondria to trigger ROS-dependent cell death.


Anemia and undernutrition among children aged 6-23 months in two agroecological zones of rural Ethiopia.
Child malnutrition during the first 1,000 days, commencing at conception, can have lifetime consequences. This study assesses the prevalence of anemia and undernutrition among children aged 6-23 months in midland and lowland agroecological zones of rural Ethiopia. Cross-sectional data examining sociodemographic, anthropometry, hemoglobin levels, and meal frequency indicators were collected from 216 children aged 6-23 months and their mothers randomly selected from eight rural kebele (villages). Of 216 children, 53.7% were anemic, and 39.8%, 26.9%, and 11.6% were stunted, underweight, and wasted, respectively. The prevalence of anemia was higher in the lowland agroecological zone (59.5%) than the midland (47.6%). Among those children who were stunted, underweight, and wasted, 63.5%, 66.7%, and 68.0% were anemic, respectively. Child anemia was significantly associated with the child not achieving minimum meal frequency, sickness during the last 2 weeks before the survey, stunting and low body mass index, and with maternal hemoglobin and handwashing behavior. The prevalence of stunting was higher in the lowland agroecological zone (42.3%) than the midland (36.2%). The predictors of stunting were age and sex of the child, not achieving MMF, maternal body mass index, and age. As maternal height increases, the length for age of the children increases (P=0.003). The overall prevalence of anemia and undernutrition among children aged 6-23 months in these study areas is very high. The prevalence was higher in the lowland agro-ecological zone. Health information strategies focusing on both maternal and children nutrition could be sensible approaches to minimize stunting and anemia.


Electrophysiological and proarrhythmic parameters in transmural canine left-ventricular needle biopsies.
This study was designed to validate the use of small, transmural, left-ventricular biopsies in the dog for investigations of electrophysiological and proarrhythmic properties of the heart. This technique could facilitate pharmacological in vitro testing in remodelled hearts of both man and animal. Small, transmural, semi-cylindrical, left-ventricular biopsies from dogs with normal sinus rhythm (SR) were characterized electrophysiologically and compared with biopsies from electrically remodelled hearts from dogs with chronic, complete AV-block (CAVB). In at least five biopsy segments recordings were made to determine the action potential duration (APD), the transmural gradient of repolarization, the maximal transmural dispersion (deltatM(max)) and presence of early after-depolarizations (EADs) at different pacing cycle lengths (PCLs) in the absence and presence of a class-III agent, ibutilide (10(-6) M). The biopsies showed stable and normal AP characteristics, a conduction velocity of 0.22 +/- 0.05 m/s and normal frequency dependence of the APD. The location of the longest APD varied, thus creating transmural repolarization gradients with differing morphology. Ibutilide prolonged the APD, accentuated repolarization gradients and induced EADs. CAVB biopsies had significantly longer APDs, a larger dispersion of repolarization and showed more EADs in the presence of ibutilide than SR biopsies. We conclude that this biopsy technique provides coherent and valid transmural electrophysiological data in dogs under various conditions.


Proteomic characterization of murid herpesvirus 4 extracellular virions.
Gammaherpesvirinae, such as the human Epstein-Barr virus (EBV) and the Kaposi's sarcoma associated herpesvirus (KSHV) are highly prevalent pathogens that have been associated with several neoplastic diseases. As EBV and KSHV are host-range specific and replicate poorly in vitro, animal counterparts such as Murid herpesvirus-4 (MuHV-4) have been widely used as models. In this study, we used MuHV-4 in order to improve the knowledge about proteins that compose gammaherpesviruses virions. To this end, MuHV-4 extracellular virions were isolated and structural proteins were identified using liquid chromatography tandem mass spectrometry-based proteomic approaches. These analyses allowed the identification of 31 structural proteins encoded by the MuHV-4 genome which were classified as capsid (8), envelope (9), tegument (13) and unclassified (1) structural proteins. In addition, we estimated the relative abundance of the identified proteins in MuHV-4 virions by using exponentially modified protein abundance index analyses. In parallel, several host proteins were found in purified MuHV-4 virions including Annexin A2. Although Annexin A2 has previously been detected in different virions from various families, its role in the virion remains controversial. Interestingly, despite its relatively high abundance in virions, Annexin A2 was not essential for the growth of MuHV-4 in vitro. Altogether, these results extend previous work aimed at determining the composition of gammaherpesvirus virions and provide novel insights for understanding MuHV-4 biology.


Shoulder strength imbalances as injury risk in handball.
This study was conducted to analyze whether internal (IR) and external (ER) rotator shoulder muscles weakness and/or imbalance collected through a preseason assessment could be predictors of subsequent shoulder injury during a season in handball players. In preseason, 16 female elite handball players (HPG) and 14 healthy female nonathletes (CG) underwent isokinetic IR and ER strength test with use of a Con-Trex® dynamometer in a seated position with 45° shoulder abduction in scapular plane, at 60, 120 and 240°/s in concentric and at 60°/s in eccentric, for both sides. An imbalanced muscular strength profile was determined using -statistically selected cut-offs from CG values. For HPG, all newly incurred shoulder injuries were reported during the season. There were significant differences between HPG and CG only for dominant eccentric IR strength, ER/IR ratio at 240°/s and for IRecc/ERcon ratio. In HPG, IR and ER strength was higher, and ER/IR ratios lower for dominant than for nondominant side. The relative risk was 2.57 (95%CI: 1.60-3.54; P<0.05) if handball players had an imbalanced muscular strength profile. In youth female handball players IR and ER muscle strength increases on the dominant side without ER/IR imbalances; and higher injury risk was associated with imbalanced muscular strength profile.


Kazachstania gamospora and Wickerhamomyces subpelliculosus: Two alternative baker's yeasts in the modern bakery.
Saccharomyces cerevisiae, the conventional baker's yeast, remains the most domesticated yeast monopolizing the baking industry. Its rapid consumption of sugars and production of CO2 are the most important attributes required to leaven the dough. New research attempts highlight that these attributes are not unique to S. cerevisiae, but also found in several non-conventional yeast species. A small number of these yeast species with similar properties have been described, but remain poorly studied. They present a vast untapped potential for the use as leavening agents and flavor producers due to their genetic and phylogenetic diversity. We assessed the potential of several non-conventional yeasts as leavening agents and flavor producers in dough-like conditions in the presence of high sugar concentrations and stressful environments mimicking conditions found in flour dough. We tested the capabilities of bread leavening and aroma formation in a microbread platform as well as in a bakery setup. Bread leavened with Kazachstania gamospora and Wickerhamomyces subpelliculosus had better overall results compared to control baker's yeast. In addition, both displayed higher stress tolerance and broader aroma profiles than the control baker's yeast. These attributes are important in bread and other farinaceous products, making K. gamospora and W. subpelliculosus highly applicable as alternative baker's yeasts.


Intracellular acidification and ADP enhance nicorandil induction of ATP sensitive potassium channel current in cardiomyocytes.
Potassium channel openers target ATP sensitive potassium (KATP) channels, and might be beneficial in the treatment of ischaemic cardiac conditions. Ischaemia is associated with an increase in the concentrations of intracellular nucleoside diphosphates and protons, two regulators of KATP channels. Since it is largely unknown whether these factors modulate the action of potassium channel openers, the aim of this study was to define the effects of ADP and intracellular acidification on KATP channel activation by nicorandil, a potassium channel opener currently in clinical use. To measure ionic currents, the whole cell patch clamp technique was employed in guinea pig single ventricular myocytes. Induction of KATP channel current by pinacidil, a potassium channel opener chemically unrelated to nicorandil, was used for comparison. Nicorandil (300 microM) and pinacidil (100 microM) induced a time independent K+ current, which was inhibited by glibenclamide (6 microM), a selective blocker of KATP channels. ADP in the pipette solution was required for the nicorandil induced activation of KATP channels when the pH of the pipette solution was 7.2. In the absence of ADP, lowering the pH of the pipette solution to 6.5 was necessary for nicorandil to induce KATP channel current. For pinacidil to induce KATP channel current neither the addition of intracellular ADP nor acidification was required. Nicorandil may activate KATP channel current more effectively in conditions associated with a change in intracellular proton and ADP concentrations such as cardiac ischaemia.


A ratiometric fluorescent chemosensor for iron: discrimination of Fe2+ and Fe3+ and living cell application.
A newly designed probe, 6-thiophen-2-yl-5,6-dihydrobenzo[4,5]imidazo-[1,2-c] quinazoline (HL(1)) behaves as a highly selective ratiometric fluorescent sensor for Fe(2+) at pH 4.0-5.0 and Fe(3+) at pH 6.5-8.0 in acetonitrile-HEPES buffer (1/4) (v/v) medium. A decrease in fluorescence at 412 nm and increase in fluorescence at 472 nm with an isoemissive point at 436 nm with the addition of Fe(2+) salt solution is due to the formation of mononuclear Fe(2+) complex [Fe(II)(HL)(ClO(4))(2)(CH(3)CN)(2)] (1) in acetonitrile-HEPES buffer (100 mM, 1/4, v/v) at pH 4.5 and a decrease in fluorescence at 412 nm and increase in fluorescence at 482 nm with an isoemissive point at 445 nm during titration by Fe(3+) salt due to the formation of binary Fe(3+) complex, [Fe(III)(L)(2)(ClO(4))(H(2)O)] (2) with co-solvent at biological pH 7.4 have been established. Binding constants (K(a)) in the solution state were calculated to be 3.88 × 10(5) M(-1) for Fe(2+) and 0.21 × 10(3) M(-1/2) for Fe(3+) and ratiometric detection limits for Fe(2+) and Fe(3+) were found to be 2.0 μM and 3.5 μM, respectively. The probe is a "naked eye" chemosensor for two states of iron. Theoretical calculations were studied to establish the configurations of probe-iron complexes. The sensor is efficient for detecting Fe(3+)in vitro by developing a good image of the biological organelles.


Early mucosal responses in blue catfish (Ictalurus furcatus) skin to Aeromonas hydrophila infection.
Bacterial pathogens are well-equipped to detect, adhere to, and initiate infection in their finfish hosts. The mucosal surfaces of fish, such as the skin, function as the front line of defense against such bacterial insults that are routinely encountered in the aquatic environment. While recent progress has been made, and despite the obvious importance of mucosal surfaces, the precise molecular events that occur soon after encountering bacterial pathogens remain unclear. Indeed, these early events are critical in mounting appropriate responses that ultimately determine host survival or death. In the present study, we investigated the transcriptional consequences of a virulent Aeromonas hydrophila challenge in the skin of blue catfish, Ictalurus furcatus. We utilized an 8×60K Agilent microarray to examine gene expression profiles at key early timepoints following challenge (2 h, 12 h, and 24 h). A total of 1155 unique genes were significantly altered during at least one timepoint. We observed dysregulation in a number of genes involved in diverse pathways including those involved in antioxidant responses, apoptosis, cytoskeletal rearrangement, immunity, and extracellular matrix protein diversity and regulation. Taken together, A. hydrophila coordinately modulates mucosal factors across numerous cellular pathways in a manner predicted to enhance its ability to adhere to and infect the blue catfish host.


Management of the pregnant substance abusing woman.
Operation PAR has been involved in prevention and treatment service development for drug-dependent women for nearly 40 years. Management starts with prevention programs. For opiate-addicted pregnant women, methadone is the treatment of choice and should be maintained throughout pregnancy, despite the manageable problem of neonatal opiate withdrawal. Women are actively engaged in innovative educational and therapeutic activities to help moms build skills to enhance their personal health and develop recovery orientation and drug refusal skills. Specific parenting skills are modeled and taught both on an outpatient and inpatient basis as appropriate.


Skin closure after infrainguinal bypass surgery: a prospective randomised study.
wound complications after infrainguinal bypass surgery occur frequently and may jeopardise an underlying graft. The purpose of this study was to investigate the role of suture material in the occurrence of wound complications. prospective randomised trial. 170 consecutive patients underwent femoro-popliteal or femoro-tibial bypass surgery and were randomised between wound closure with continuous polyamide sutures (Ethilon) or with skin staples. ten patients were excluded because of re-intervention or death within 2 weeks after operation. The wound was closed with a continuous polyamide suture (Ethilon) in 77 patients and with metallic skin staples in 83 patients. The groups were comparable in age, use of corticosteroids, occurrence of diabetes mellitus and operation time. Skin closing time was significantly shorter in the staples group (6.4 min versus 2.7 min, p<0.001). Overall, there were no significant differences between continuous polyamide sutures and metallic skin staples in superficial infections (8% versus 2%), deep infections (1% versus 1%), serous leakage or haematoma. we found no significant differences in the incidence of wound complications in infrainguinal bypass surgery comparing continuous polyamide (Ethilon) and skin staples. The time needed for wound closure was significantly reduced using metallic staples, but this had no effect on the total operation time. Choices of closing materials should be guided by surgeon's preference.


Vitamin status in patients with chronic renal failure.
Many factors complicate the effort for a recommendation on individual vitamin requirements in CRF. On the basis of our present incomplete knowledge about the handling of vitamins in uremia, suggestions for appropriate supplementation only of water-soluble vitamins are given. Patients with advanced CRF without dialysis treatment should receive daily supplements of vitamin B6 (5 mg), ascorbic acid (70-100 mg), and the normal recommended daily allowance of the other water-soluble vitamins in addition to the vitamin intake from the diet. We give folic acid only in patients taking antifolate drugs or in combination with iron in iron deficiency state and anemia (1 tablet of Folicombin contains 0.5 mg folic acid and 0.4 g elemental iron). There is still a pressing need for more data on the vitamin status, on vitamin requirements, and on long-term effects of vitamin administration in CRF.


Effect of lipid peroxidation on microtubular protein.
CCl4 and ADP-Fe3+ determine an increased microsomal lipid peroxidation in liver tissue which is related to an impairment of the tubulin molecule in the case of CCl4 while no alteration is evident with ADP-Fe3+. Such difference could be explained in terms of different intermediate or end products towards which the tubulin molecule shows a different susceptibility.


Mononuclear phagocytes in rheumatoid arthritis: Fc-receptor expression by peripheral blood monocytes.
Fc receptor expression by enriched monocytes from rheumatoid arthritis (RA) patients and age and sex matched controls (healthy subjects) was compared by measuring the uptake of IgG on monocytes in a competitive radioassay. The association constant (Ka) between IgG and the monocytes and the number of Fc binding sites per cell was calculated from Scatchard plots of 4 degrees C binding data. RA monocytes had increased expression of Fc receptors as compared with those of controls. This increase was particularly pronounced in those RA patients affected by extra-articular disease. There were significant correlations between the numbers of Fc receptors on monocytes and both C1q binding and anticomplementary activity but none between monocyte Fc receptor numbers and serum rheumatoid factors (IgG and IgM). It is considered that monocyte handling of circulating immune complexes is unimpaired in RA and that monocytes make an adaptive response to increased levels of immune complexes.


Genomic analysis of liver cancer unveils novel driver genes and distinct prognostic features.
Objective: Hepatocellular carcinoma (HCC) is a highly heterogeneous disease with a dismal prognosis. However, driver genes and prognostic markers in HCC remain to be identified. It is hoped that in-depth analysis of HCC genomes in relation to available clinicopathological information will give rise to novel molecular prognostic markers. Methods: We collected genomic data of 1,061 HCC patients from previous studies, and performed integrative analysis to identify significantly mutated genes and molecular prognosticators. We employed three MutSig algorithms (MutSigCV, MutSigCL and MutSigFN) to identify significantly mutated genes. The GISTIC2 algorithm was used to delineate focally amplified and deleted genomic regions. Nonnegative matrix factorization (NMF) was utilized to decipher mutational signatures. Kaplan-Meier survival and Cox regression analyses were used to associate gene mutation and copy number alteration with survival outcome. Logistic regression model was applied to test association between gene mutation and mutational signatures. Results: We discovered 11 novel driver genes, including RNF213, VAV3 and TNRC6B, with mutational prevalence ranging from 1% to 3%. Seven mutational signatures were also identified in HCC, some of which were associated with mutations of classical driver genes (e.g., TP53, TERT) as well as alcohol consumption. Focal amplifications of TERT and other druggable targets, including AURKA, were also revealed. Targeting AURKA by a small-molecule inhibitor potently induced apoptosis in HCC cells. We further demonstrated that HCC patients with TERT amplification displayed shortened overall survival independent of other clinicopathological parameters. In conclusion, our study identified novel cancer driver genes and prognostic markers in HCC, reiterating the translational importance of omics data in the precision medicine era.


Morphine exposure alters Fos expression in a sex-, age-, and brain region-specific manner during adolescence.
Data in both humans and preclinical animal models clearly indicate drug exposure during adolescence, when the "reward" circuitry of the brain develops, increases the risk of substance use and other mental health disorders later in life. Human data indicate that different neural and behavioral sequelae can be observed in early versus late adolescence. However, most studies with rodent models examine a single adolescent age compared to a mature adult age, and often only in males. Herein, we sought to determine whether the acute response to the opioid morphine would also differ across adolescence, and by sex. By quantifying Fos positive cells, a proxy for neural activity, at different stages during adolescence (pre-, early, mid-, and late adolescence) and in multiple reward regions (prefrontal cortex, nucleus accumbens, caudate/putamen), we determined that the neural response to acute morphine is highly dependent on adolescent age, sex, and brain region. These data suggest that heterogeneity in the consequences of adolescent opioid exposure may be due to age- and sex-specific developmental profiles in individual reward processing regions. In future studies, it will be important to add age within adolescence as an independent variable for a holistic view of healthy or abnormal reward-related neural development.


Confocal examination of untreated fresh specimens from basal cell carcinoma: implications for microscopically guided surgery.
To evaluate the diagnostic accuracy of confocal examination of basal cell carcinoma (BCC) in microscopy-guided surgery. Four independent observers with no previous experience in confocal laser scanning (CLS) microscopy received standardized instruction about diagnostic CLS microscopic features. Subsequently, 120 confocal images of fresh excisions from BCCs or normal skin were evaluated by each observer, imaged using a commercially available, near-infrared, reflectance CLS microscope. Logistic regression analysis was performed on a combination of all morphologic features using the forward-stepwise (Wald) method. Reliability (interobserver agreement) data were evaluated by kappa statistic. Department of Dermatology, Medical University of Graz. Twenty patients with histologically verified BCC. Evaluation of fresh BCC excisions by CLS microscopy. Diagnostic accuracy of the method was evaluated by chi2 test. Diagnostic impact and reliability of each morphologic feature were evaluated by logistic regression analysis and kappa statistic, respectively. Overall, high diagnostic accuracy was achieved by the 4 observers. Logistic regression analysis revealed that mainly tumor cell nuclei and tumor nests should be taken into account for diagnostic decisions, whereas disintegration of tumor cells, peripheral palisading, and retraction of stroma were rarely useful. However, most of the features were highly reliable. This diagnostic validation study of CLS microscopy in microscopy-guided surgery yielded promising results and opens avenues for further studies. In the future, CLS microscopy may guide microsurgery of any skin cancer.


A qualitative study to identify reasons for Clostridium difficile testing in pediatric inpatients receiving laxatives or stool softeners.
To understand why clinicians send Clostridium difficile tests from hospitalized children receiving laxatives or stool softeners, we performed a mixed-methods study. We prospectively identified tested patients and surveyed their clinicians by e-mail. Reasons for testing included changes in stooling pattern on baseline bowel regimen, other changes in clinical status, and risk factors for C difficile infection. Education targeting discontinuing bowel medications before C difficile testing could improve the specificity of pediatric C difficile infection diagnosis.


Synthesis and characterization of solution-processable polyelectrolyte complexes and their homogeneous membranes.
Solution-processable polyelectrolyte complexes (PECs) between poly(diallyldimethylammonium chloride) (PDDA) and poly(acrylic acid) (PAA) were synthesized in aqueous NaOH and obtained in their solid forms by protection and deprotection of carboxylic acid groups. Elemental analysis, conductance measurement, and FT-IR showed that the composition and ionic complexation degree (ICD) of the PECs can be controlled effectively by tuning the NaOH concentration in both parent polyelectrolyte solutions. Thermal gravity analysis showed that PECs revealed good thermal stability, and differential scanning calorimetry showed that the glass transition temperature (Tg) of PECs increased with increasing ICD and finally became undetectable when ICD was above 0.16. Viscosity properties of the PEC solutions were well correlated to the ICD of PECs, and it was found that solid PECs could be redissolved in dilute NaOH without breaking the ionic complexation between PDDA and PAA. Homogeneous PEC membranes (HPECMs) were made from their concentrated solutions, and their morphologies were examined by field emission scanning electron microscopy. These novel HPECMs were subjected to dehydration of organics for the first time, and a very promising performance was obtained. Furthermore, another two solution-processable PECs between weak anionic polyelectrolyte and cationic polyelectrolyte were also synthesized by the same method and showed a very high separation performance.


Interleukin 10 polymorphisms differentially influence the risk of gastric cancer in East Asians and Caucasians.
Although a number of association studies of gastric cancer (GC) risk have been conducted worldwide, their results have been inconsistent among different populations. The association between GC incidence and Helicobacter pylori (H. pylori) infection is somewhat of an enigma that has yet to be clearly explained. Geographically-restricted positive selection due to unique environmental pressures often result in large allele frequency differences between populations. Thus, population differences need to be investigated when attempting to identify genes that contribute to phenotypes that differ greatly between populations. We analyzed population differences in 18 polymorphisms of 12 GC-associated or immune response-related genes from 3 ethnic groups comprising 50 Koreans, 46 Indians, and 60 Caucasians; these groups differed in H. pylori seropositivity and susceptibility to GC. An interleukin 10 (IL10) polymorphism demonstrated a significantly different genotype distribution (F(ST)=0.306, P=0.014), indicating a large difference between the Korean and Caucasian populations. The odds ratio of IL10 polymorphism allele between the populations was 38.32 (95% confidence interval, 11.49-127.83). This finding, taken together with previous evidence, provides a possible explanation for previous discrepant association results and supports the idea that IL10 gene polymorphisms can differentially affect GC development among populations.


Process development for degradation of phenol by Pseudomonas putida in hollow-fiber membrane bioreactors.
The degradation of phenol (100-2800 mg/L) by cells Pseudomonas putida CCRC14365 in an extractive hollow-fiber membrane bioreactor (HFMBR) was studied, in which the polypropylene fibers were prewetted with ethanol. The effects of flow velocity, the concentrations of phenol, and the added dispersive agent tetrasodium pyrophosphate on phenol degradation and cell growth were examined. It was shown that about 10% of phenol was sorbed on the fibers at the beginning of the degradation process. The cells P. putida fully degraded 2000 mg/L of phenol within 73 h when the cells were immobilized and separated by the fibers. Even at a level of 2800 mg/L, phenol could be degraded more than 90% after 95-h operation. At low phenol levels (< 400 mg/L) where substrate inhibition was not severe, it was more advantageous to treat the solution in a suspended system. At higher phenol levels (> 1000 mg/L), however, such HFMBR-immobilized cells could degrade phenol to a tolerable concentration with weak substrate-inhibition effect, and the degradation that followed could be completed by suspended cultures due to their larger degradation rate. The process development in an HFMBR system was also discussed.


Translocation, activation and association of protein-tyrosine kinase (p72syk) with phosphatidylinositol 3-kinase are early events during platelet activation.
We have previously reported that a non-receptor-type protein-tyrosine kinase p72syk, exists in both membrane and cytosolic fractions in porcine platelets and is activated after thrombin stimulation. To facilitate the understanding of the function of p72syk, we have investigated the topological features, kinase activities and the interaction with another signal-transducing molecule, namely phosphatidylinositol 3-kinase, during platelet activation. Membrane and cytosolic fractions were separated from thrombin-treated porcine platelets, and the amount of p72syk was quantified by the immunoblot technique or the kinase activity of each fraction was determined by an immunoprecipitation kinase assay. After stimulation by thrombin, cytosolic p72syk rapidly translocated to the membrane fraction within 10 s and there was also a significant increase in the amount of p72syk in the cytoskeletal fraction. The autophosphorylation activity of membrane-associated p72syk significantly increased approximately tenfold and reached a maximum at 10 s; the activity subsequently decreased to almost the basal level within 120 s. For similar time courses, association of p72syk with phosphatidylinositol 3-kinase and tyrosine phosphorylation of p72syk were observed. These results suggest that translocation, activation, and association of p72syk with transducing molecules such as phosphatidylinositol 3-kinase, events which occur during platelet activation, may participate in early signal-transduction events.


Conservation and divergence in cellulosome architecture between two strains of Ruminococcus flavefaciens.
A 17-kb scaffoldin gene cluster in Ruminococcus flavefaciens strain FD-1 was compared with the homologous segment published for strain 17. Although the general design of the cluster is identical in the two strains, significant differences in the modular architecture of the scaffoldin proteins were discovered, implying strain-specific divergence in cellulosome organization.


[Microsurgical tissue transplantation for correction of hand abnormalities].
Established procedures in hand surgery are dominating the management of congenital anomalies of the hand. In complex deficiencies and in certain forms of thumb hypoplasia however, replacement of structures capable of growing is needed, as can be free transplantation of vascularized metatarsal bone or free toe transplantation as substitute for a whole part of a limb. Capabilities of and limitations to microsurgical procedures are presented in selected cases.


Hypothermic machine perfusion alleviates ischemia-reperfusion injury of intestinal transplantation in pigs.
Intestinal transplantation (IT) has become an important procedure for the treatment of irreversible intestinal failure. However, IT is extremely vulnerable to ischemia-reperfusion injury (IRI). Due to the limitations of static cold storage (SCS), hypothermic machine perfusion (HMP) is rapidly gaining popularity. In this study, the intestinal HMP system is established and HMP is compared with SCS. An intestinal HMP system was built. Ten miniature pigs were randomly divided into the HMP and SCS groups, and their intestines were perfused using the HMP device and SCS, respectively, followed by orthotopic auto-transplantation. Analysis was done on the grafts between the two groups. Operation success rates of the surgery were 100% in both groups. The 7-day survival rate was 100% in the HMP group, which was significantly higher than that of the SCS group (20%, P< 0.05). The pathological results showed that fewer injuries of grafts were in the HMP group. Endotoxin (ET), IL-1, IL-6, IFN-γ and TNF-α levels in the HMP group were significantly lower than in the SCS group (P<0.05), whereas IL-10 levels were significantly higher (P<0.05).The intestinal expression levels of ZO-1 and Occludin were higher in the HMP group compared to the SCS group, whereas Toll-like receptor 4 (TLR4), nuclear factor kappa B (NFκB), and caspase-3 were lower. In this study, we established a stable intestinal HMP system and demonstrated that HMP could significantly alleviate intestinal IRI and improve the outcome after IT.


Mapping of contact sites in the caldesmon-calmodulin complex.
The interaction of intact calmodulin and its four tryptic peptides with deletion mutants of caldesmon was analysed by native gel electrophoresis, fluorescence spectroscopy and zero-length cross-linking. Deletion mutants H2 (containing calmodulin-binding sites A and B) and H9 (containing sites B and B') interacted with intact calmodulin to form complexes whose stoichiometries varied from 2:1 to 1:1. The N-terminal peptides of calmodulin (TR1C, residues 1-77, and TR2E, residues 1-90) bound H2 with higher affinity than H9. At the same time H2 was less effective than H9 in binding to the C-terminal peptides of calmodulin TR2C (residues 78-148) and TR3E (residues 107-148). The N-terminal peptides of calmodulin (TR1C and TR2E) could be cross-linked to intact caldesmon and its deletion mutants H2 and H9. The similarity in the primary structures of sites A and B' of caldesmon and our measurements of the affinities of H2 and H9 to calmodulin and its peptides strongly indicate an orientation of the protein complex where sites A and B' interact with the N-terminal domain of calmodulin, whereas site B interacts with the C-terminal domain of calmodulin. The spatial organization of contact sites in the caldesmon-calmodulin complex agrees with the earlier proposed two-dimensional model of interaction of the two proteins [Huber, El-Mezgueldi, Grabarek, Slatter, Levine and Marston (1996) Biochem. J. 316, 413-420].


Omentin-1 exerts bone-sparing effect in ovariectomized mice.
Omentin-1 inhibited osteoblast differentiation in vitro. In co-culture systems of osteoblasts and osteoclast precursors, omentin-1 reduced osteoclast formation by stimulating osteoprotegerin (OPG) and inhibiting receptor activator for nuclear factor κB ligand (RANKL) production in osteoblasts. In vivo, adenovirus-mediated overexpression of omentin-1 suppressed bone turnover and restored bone mineral density (BMD) and bone strength in ovariectomized mice. Omentin-1 (also intelectin-1) is a recently identified visceral adipose tissue-derived cytokine that is highly abundant in plasma. This study was undertaken to investigate the effects of omentin-1 on bone metabolism. Osteoblast differentiation was assessed by measuring alkaline phosphatase activity, osteocalcin production and matrix mineralization. OPG and RANKL protein expression and secretion in osteoblasts were detected by Western blot and ELISA, respectively. The effect of recombinant omentin-1 on osteoclast formation was examined in co-culture systems of osteoblasts and osteoclast precursors. The effects of intravenous administration of adenoviral-delivered omentin-1 on bone mass, bone strength, and bone turnover were also examined in ovariectomized mice. In vitro, omentin-1 inhibited osteoblast differentiation, while it had no direct effect on osteoclast differentiation; it also reduced osteoclast formation in the co-culture systems through stimulating OPG and inhibiting RANKL production in osteoblasts. In vivo, adenovirus-mediated overexpression of omentin-1 partially restored BMD and bone strength in ovariectomized mice, accompanied by decreased levels of plasma osteocalcin and tartrate-resistant acid phosphatase-5b and lower serum RANKL/OPG ratios. The present study suggests that omentin-1 ameliorates bone loss induced by estrogen deficiency via downregulating the RANKL/OPG ratio.


Salivary cortisol and alpha-amylase: Is there consistency between psychosocial stress test and burdensome work shifts?
This study examined the consistency of salivary cortisol and alpha-amylase (sAA) total daily secretion between laboratory and field circumstances. The 95 participants were shift working female health care professionals with high (n = 53) or low (n = 42) psychosocial stress (job strain) measured by the Job Content Questionnaire (JCQ). The Trier Social Stress Test including a 5-min free speech and a mental arithmetic task was conducted with four, and field measurements with three daily saliva samples of cortisol and sAA during circadian rhythm and inter-shift recovery controlled morning shift, night shift, and a day off. The associations of salivary cortisol and sAA area under the curve with respect to ground (AUCg) and area under the curve with respect to increase (AUCi) between laboratory and field were tested using OLS (Ordinary Least Squares) regression. The sAA AUCg output in the laboratory was correlated with the output during all field measurement days and similarly among high and low job strain groups (p < 0.001). SAA AUCi and salivary cortisol AUCg and AUCi were not correlated between laboratory and field measurement, neither in the whole sample nor among the low or high job strain group. In conclusion, a laboratory measure of sAA AUCg output is promising in predicting stress-related output during burdensome work shifts and leisure time, whereas sAA AUCi or salivary cortisol seem not to have this potential.


Phenylnannolones A-C: biosynthesis of new secondary metabolites from the myxobacterium Nannocystis exedens.
Myxobacteria are gliding bacteria that belong to the delta-Proteobacteria and are known for their unique biosynthetic capabilities. Among myxobacteria, Nannocystis spp. are most closely related to marine myxobacteria and their secondary metabolism has hardly been investigated. Phenylnannolones A (1), B (2) and C (3) were obtained from a culture of Nannocystis exedens that was isolated from the intertidal region of Crete. Compound 1 had inhibitory activity toward the ABCB1 gene product P-glycoprotein and reversed daunorubicin resistance in cultured cancer cells. Phenylnannolone A has an unusual structural architecture; it is composed of an ethyl-substituted polyene chain linked to a pyrone moiety on one side and to a phenyl ring on the other. The investigation of the biosynthesis with labelled precursors revealed acetate, butyrate and phenylalanine as building blocks for 1. The labelling pattern suggested novel biochemical reactions for the biosynthesis of the starter unit.


Education and assortative marriage in northern and urban Sudan: 1945-79.
This study examines the extent of assortative mating for education in Northern Sudan and urban Khartoum. More males than females were found at higher levels of education. Increasingly, people tend to marry persons of equal educational level, but the unequal educational opportunities for males and females have led to the emergence of educational exogamy in which members of different educational levels are more likely to marry from the adjacent educational category than from distant categories. The increasing level of education for both sexes, and especially for females, may in part explain the rising trend in age at marriage.


Caring for the carers: safeguarding oncologists' mental health in the time of COVID-19.
Taking care of patients with chronic, terminal diseases presents unique challenges to the mental health of medical oncologists. The current coronavirus disease 2019 (COVID-19) pandemic has exacerbated these mental health risks brought about by isolation and exhaustion. Delegated to be a national COVID-19 referral centre, the University of the Philippines-Philippine General Hospital faced many challenges, including the increased workload in a perilous and anxiety-inducing national crisis which placed the entire healthcare team in an unprecedented situation. To adapt to these challenges, the Division of Medical Oncology employed the following measures to safeguard the mental health of its faculty and fellows: 1) use of psychological support materials; 2) initiation of a psychological intervention programme and 3) establishment of peer support programmes. Caring for the carers through evidence-based interventions ensures the delivery of quality care to our cancer patients despite the challenges during these trying times.


Shoulder mobility after axillary sentinel node biopsy for early infiltrating breast cancer treatment.
It is known that complete axillary lymph node dissection for breast cancer treatment causes more frequent sensitive and motor alterations in the homolateral shoulder and upper limb than sentinel lymph node (SLN) biopsy. However, it is not clear how often patients treated by SLN biopsy suffer from shoulder mobility (SM) restriction, as well as its severity and duration. This study was done aiming to evaluate SM in 38 patients with early infiltrating breast cancer treated by SLN biopsy in whom shoulder movements were assessed before surgery and repeated at one, two and three months later. Shoulder-arm mobility was evaluated by goniometry considering flexion, abduction, aduction, extension, internal rotation and external rotation. An abnormal result for each movement was defined by restriction greater than ten degrees compared to preoperative findings. Significant abnormal results for flexion and abduction were found in all of the patients at the first month evaluation. At the third month assessment no women showed any kind of SM impairment. The average restriction evolution for each of the parameters is presented. It is concluded that there is frequently a slight and transient SM limitation in patients undergoing SLN biopsy. Early postoperative physiotherapeutical assistance should thus be advisable to relieve and shorten disability symptomatology.


Application of a marketing concept to patient-centered care: co-producing health with heart failure patients.
Increasing numbers of patients are being treated for heart failure each year. One out of four of the heart failure patients who receives care in a hospital is readmitted to the hospital within 30 days of discharge. Effective discharge instruction is critical to prevent these patient readmissions. Co-production is a marketing concept whereby the customer is a partner in the delivery of a good or service. For example, a patient and nurse may partner to co-produce a patient-centered health regimen to improve patient outcomes. In this article we review the cost of treating heart failure patients and current strategies to decrease hospital readmissions for these patients along with the role of the nurse and the concept of co-producing health as related to heart failure patients. Next we describe our study assessing the degree to which discharge processes were co-produced on two hospital units having a preponderance of heart failure patients, and present our findings indicating minimal evidence of co-production. A discussion of our findings, along with clinical implications of these findings, recommendations for change, and suggestions for future research are offered. We conclude that standardized discharge plans lead to a mindset of 'one size fits all,' a mindset inconsistent with the recent call for patient-centered care. We offer co-production as a patient-centered strategy for customizing discharge teaching and improving health outcomes for heart failure patients.


Utility of diffusion tensor imaging parameters for diagnosis of hemimegalencephaly.
Hemimegalencephaly is a rare hamartomatous entity characterised by enlargement of all or part of the cerebral hemisphere ipsilaterally with cortical dysgenesis, large lateral ventricle and white matter hypertrophy with or without advanced myelination. Although conventional magnetic resonance imaging (MRI) is useful for detecting these diagnostic features, hemimegalencephaly is not always easily distinguished from other entities, especially when hemimegalencephaly shows blurring between the grey and white matter. Diffusion tensor imaging (DTI) is a functional MRI technique commonly used to assess the integrity of white matter. The usefulness of DTI in assessing hemimegalencephaly has not been fully elucidated. In this study, we clarified the characteristics of hemimegalencephaly with regard to DTI and its parameters including fractional anisotropy and apparent diffusion coefficient. Three patients with hemimegalencephaly underwent MRI including DTI. We first visually compared fractional anisotropy mapping and conventional MRI. Next, we quantitatively measured the fractional anisotropy and apparent diffusion coefficient values in the subcortical white matter of the hemisphere with hemimegalencephaly and corresponding normal-appearing contralateral regions and analysed the values using the Mann-Whitney U test. On fractional anisotropy mapping, we could clearly distinguish the junction of grey and white matter and observed thicker white matter in the hemisphere with hemimegalencephaly, which was unclear on conventional MRI. The white matter in the hemisphere with hemimegalencephaly showed significantly higher fractional anisotropy (P<0.0001) and lower apparent diffusion coefficient (P=0.0022) values than the normal contralateral side. DTI parameters showed salient hemimegalencephaly features and could be useful in its assessment.


Isolation and immunobiochemical characterization of a major allergen (65 kDa) from Fusarium equiseti.
Fusarium equiseti is one of the most important species in the class Deuteromycetes (Fungi Imperfecti). For proper diagnosis and immunotherapy, isolation and characterization of allergens of F. equiseti are necessary. In the present study, culture filtrate (CF) extract of F. equiseti was resolved into 35-37 bands on isoelectric focusing pI (3-9) and SDS-PAGE (mol. wt. 10-100 kDa). Most of them were glycoproteins, as identified by PAS staining. F. equiseti CF revealed 15 allergenic proteins on immunoblot with an allergic serum pool. It was fractionated into nine fractions (I-IX) on a Superose-12 column by FPLC. Fraction IV (65 kDa) and fraction VI (25 kDa) were found to be highly allergenic by IgE ELISA. A 65-kDa protein was observed as a major allergen because it was recognized by most of the patient sera on immunoblot. After elution from SDS-PAGE gel, it gave two bands of pI 7.4 and 6.0. Inhibition in IgE-binding components of F. equiseti CF with CF extracts of F. solani and F. moniliforme by immunoprint inhibition assay indicated the allergenicity shared between the extracts of Fusarium species. Data suggested that the 65-kDa is the major allergen in the Fusarium species and can be used for the treatment of allergic patients.


Basal cell adenocarcinoma of the minor salivary glands involving palate and maxillary sinus.
Basal cell adenocarcinoma (BCAC) is a rare neoplasm accounting for only 2.9% of all salivary gland neoplasms. BCAC involving palatal minor salivary glands are exceedingly rare, and only 10 cases have been reported in the literature. The treatment of choice is surgical excision. Here, we report a case of a 55-year-old male patient with massive BCAC of palatal minor salivary gland extending into the maxillary sinus. This is the first case of BCAC treated by radiotherapy followed by chemotherapy. A follow-up check conducted after 14-months showed good prognosis.


NARP syndrome and adult-onset generalised seizures.
The neurogenic muscle weakness, ataxia and retinitis pigmentosa (NARP) syndrome is a maternally inherited disorder attributable to a heteroplasmic mtDNA point mutation. Catastrophic epilepsy may accompany severe, early onset forms of NARP, but seizures seem to be rare in cases with adolescent and adult onset. We describe a patient who developed clumsiness and visual problems in her teens. She had no clinical seizures but an EEG showed generalized spike and wave discharges. At this time the patient remained without a specific diagnosis. At the age of 21, the patient developed progressive ataxia and she also experienced a tonic-clonic status epilepticus. Further examinations revealed NARP syndrome. EEG abnormalities may precede adult onset seizures in the NARP syndrome.


Vitamin D-inducible calcium transport and gene expression in three Caco-2 cell lines.
The parental cell line (P) of Caco-2 cells and two clones, BBe and TC7, were studied at 11 days postconfluence to test the facilitated diffusion model of vitamin D-mediated intestinal calcium absorption (CaTx). Nuclear vitamin D receptor (nVDR) and calbindin D(9k) (CaBP) were measured by Western blot; 1,25-hydroxyvitamin D(3) 24-hydroxylase (CYP24), CaBP, plasma membrane Ca-ATPase (PMCA), and Ca transport channel (CaT1) mRNA levels were examined by RT-PCR; and net apical-to-basolateral CaTx was examined after treating cells with vehicle or 10 nM calcitriol for 8 (mRNA levels) or 48 h (protein, CaBP mRNA, CaTx). nVDR level was lowest in BBe (38% P value) and directly related to CYP24 induction (TC7 = P, which were 1.56 times greater than BBe). nVDR was inversely related to the vitamin D-induced levels of CaT1 mRNA, CaBP mRNA, PMCA mRNA, and net CaTx, with the highest induction seen in BBe. Basal CaBP mRNA (86 times greater than P) and protein levels were highest in TC7 cells and were not associated with higher net CaTx, suggesting CaBP may not be rate limiting for CaTx in these cells.


Pro-apoptotic role of Cdc25A: activation of cyclin B1/Cdc2 by the Cdc25A C-terminal domain.
Cdc25A is a dual specificity protein phosphatase that activates cyclin/cyclin-dependent protein kinase (Cdk) complexes by removing inhibitory phosphates from conserved threonine and tyrosine in Cdks. To address how Cdc25A promotes apoptosis, Jurkat cells were treated with staurosporine, an apoptosis inducer. Upon staurosporine treatment, a Cdc25A C-terminal 37-kDa fragment, designated C37, was generated by caspase cleavage at Asp-223. Thr-507 in C37 became dephosphorylated, which prevented 14-3-3 binding, as shown previously. C37 exhibited higher phosphatase activity than full-length Cdc25A. C37 with alanine substitution for Thr-507 (C37/T507A) that imitated the cleavage product during staurosporine treatment interacted with Cdc2, Cdk2, cyclin A, and cyclin B1 and markedly activated cyclin B1/Cdc2. The dephosphorylation of Thr-507 might expose the Cdc2/Cdk2-docking site in C37. C37/T507A also induced apoptosis in Jurkat and K562 cells, resulting from activating cyclin B1/Cdc2 but not Cdk2. Thus, this study reveals that Cdc25A is a pro-apoptotic protein that amplifies staurosporine-induced apoptosis through the activation of cyclin B1/Cdc2 by its C-terminal domain.


Effect of beta-adrenergic blockade on lactate turnover in exercising dogs.
Dogs with indwelling catheters in the jugular vein and in the carotid artery ran on the treadmill (slope: 15%, speed: 133 m/min). Lactate turnover and glucose turnover were measured using [U-14C]lactate and [3-3H]glucose as tracers, according to the primed constant-rate infusion method. In addition, the participation of plasma glucose in lactate production (Ra-L) was measured with [U-14C]glucose. Propranolol was given either (A) before exercise (250 micrograms/kg, iv) or (B) in form of a primed infusion administered to the dog running at a steady rate. Measurements of plasma propranolol concentration showed that in type A experiments plasma propranolol fell in 45 min below the lower limit of the complete beta-blockade. In the first 15 min of work Ra-L rose rapidly; then it fell below that of the control (exercise) values. During steady exercise, the elevated Ra-L was decreased by propranolol infusion close to resting values. beta-Blockade doubled the response of glucose production, utilization, and metabolic clearance rate to exercise. In exercising dogs approximately 40-50% of Ra-L arises from plasma glucose. This value was increased by the blockade to 85-90%. It is concluded that glycogenolysis in the working muscle has a dual control: 1) an intracellular control operating at the beginning of exercise, and 2) a hormonal control involving epinephrine and the beta-adrenergic receptors.


[EVALUATION OF CARDIOVASCULAR RISKS WITH THE USE OF MORPHOGENETIC KLOTHO PROTEIN IN PATIENTS WITH CHRONIC RENAL DISEASE].
of the study was to explore the Klotho protein significance in patients with different stages of chronic kidney disease (CKD) and to assess the influence of antihypertensive therapy on Klotho protein serum levels. 130 patients with stage 5 CKD1 were included in the study. Serum PTH, calcium and phosphorus were measured. ELISA was used to determine serum soluble alpha Klotho. Blood pressure including brachial and central (aortic) pressure was measured in all patients together with pulse wave velocity (using a «Sfigmokor» device); in addition, echocardiography (EchoCG), and X-ray examination of the abdominal aorta by Kauppila method were performed. The dynamic study of serum Klotho level showed that it changes with decreasing glomerular filtration rate faster than a rise in phosphate and PTH levels starting from stage 3A of CKD. The two later variables increased at stages 4-5.According to the ROC analysis, the values of serum Klotho below 387 pg /ml suggested enhanced risk of myocardial calcification with 80% sensitivity and 76% specificity. In addition, the highest Klotho serum levels were observed in patients whose target BP values were achieved with angiotensin receptor blockers (ARB) compared to those who used other drugs [р<0,01] or failed to reached target BP levels [p=0,008]. The study showed the possibility of practical use of Klotho protein as an early diagnostic marker of cardiovascular risk. Reduced serum Klotho was less pronounced in patients who used ARB for correction of high blood pressure. Normal Klotho protein levels in serum have been associated with a lower frequency of heart and vessels calcification in CKD patients.


First-principles T-matrix calculations of double-ionization energy spectra of atoms and molecules.
Strong electron correlation plays an important role in the determination of double ionization energy, which is required for removing or adding two electrons, particularly in small-sized systems. Starting from the state-of-the-art GW approximation, we evaluate the particle-particle ladder diagrams up to the infinite order by solving the Bethe-Salpeter equation of the T-matrix theory to calculate the double-ionization energy spectra of atoms and molecules (Be, Mg, Ca, Ne, Ar, Kr, CO, C(2)H(2), Li(2), Na(2), and K(2)) from first principles. The ladder diagrams up to the infinite order are significant to calculations of double-ionization energy spectra. The present results are in good agreement with available experimental data as well as the previous calculations using, e.g., the configuration-interaction method.


Transduction of mitogenic activity of platelet-derived growth factor (PDGF) AB by PDGF-beta receptor without participation of PDGF-alpha receptor in vascular smooth muscle cells.
In cultured rat vascular smooth muscle cells, platelet-derived growth factor (PDGF) beta receptor was expressed at a high level, whereas PDGF-alpha receptor was not detected. PDGF-BB showed a high binding activity at 4 degrees C in the cells and was not displaced by PDGF-AA or -AB. This result indicates that PDGF-AB as well as PDGF-AA does not bind to the cells lacking PDGF-alpha receptor at 4 degrees C. However, at 37 degrees C, PDGF-AB bound to the cells and induced the internalization of PDGF-beta receptor. DNA synthesis was also stimulated potentially by PDGF-AB as well as PDGF-BB in the cells, although PDGF-AA was completely inactive. At 37 degrees C, PDGF-AB caused tyrosine phosphorylation of a group of proteins including PDGF-beta receptor and phospholipase C-gamma 1, but at a slower rate than PDGF-BB. At 4 degrees C, PDGF-AB did not stimulate protein tyrosine phosphorylation, whereas PDGF-BB did. A chemical cross-linking experiment showed that PDGF-beta receptor was dimerized by PDGF-AB as well as PDGF-BB. These results indicate that PDGF-beta receptor binds PDGF-AB without participation of PDGF-alpha receptor at 37 degrees C (but not at 4 degrees C), and PDGF-AB as well as PDGF-BB acts as a potent mitogen in the vascular smooth muscle cells.


HOXA5 and p53 cooperate to suppress lung cancer cell invasion and serve as good prognostic factors in non-small cell lung cancer.
Lung cancer is the leading cause of cancer mortality worldwide and tumor metastasis is the major cause of cancer-related death. Our previous study suggested that Homeobox A5 (HOXA5) could inhibit lung cancer cell invasion via regulating cytoskeletal remodeling and involved in tumor metastasis. Recently, consensus HOX binding sites was found in the p53 gene promoter region. However, whether the HOXA5 could cooperate with p53 and contribute the inhibition of lung cancer cell invasion is still unclear. The aim of the current study is to elucidate the correlation of HOXA5 and p53 in tumor invasion and its prognostic influence in lung cancer patient specimens. Totally 71 cases of primary non-small cell lung cancer (NSCLC) were collected. The median follow-up period is 6.8 years. Immunohistochemical stain for p53 and HOXA5 were performed. Kaplan-Meier plot was done for overall survival analysis. In addition, lung cancer cell lines transfected with wild-type or mutated p53 constructs were overexpressed with HOXA5 for invasion assay. In human specimens, HOXA5 expressed mainly in the cytoplasm (54.1%) rather than nuclei (14.6%) of the NSCLC tumor part. The HOXA5 expression is higher in adenocarcinoma than in squamous cell carcinoma (P < 0.001). In addition, poor prognosis is seen in group with both non-immunoreactive for p53 and HOXA5. HOXA5 and p53 could cooperate to inhibit tumor cell invasion significantly partly by decreasing MMP2 activity in a concentration-dependent manner. Our studies provide new insights into how HOXA5 and p53 cooperate to contribute to the suppression of lung cancer cell invasion and play good prognostic roles in NSCLC.


A Previously Healthy Premature Infant Treated With Thrombolytic Therapy for Life-threatening Pulmonary Artery Thrombosis.
Pulmonary artery thrombosis is rarely reported in preterm neonates. Although treatment of neonatal thrombosis remains controversial, thrombolytic agents must be considered when the thrombosis is life threatening. We herein present a case of a preterm newborn with pulmonary artery thrombosis accompanied by acute-onset respiratory failure and cyanotic congenital heart disease. The thrombosis was successfully treated using tissue plasminogen activator. In conclusion, the thrombolytic therapy should be considered in treatment of patients in whom the thrombosis completely occludes the pulmonary arteries.


Validation of an enzyme immunoassay for detection and semiquantification of cannabinoids in oral fluid.
Oral fluid (OF) is gaining prominence as an alternative matrix for monitoring drugs of abuse in the workplace, criminal justice, and driving under the influence of drugs programs. It is important to characterize assay performance and limitations of screening techniques for Delta(9)-tetrahydrocannabinol (THC) in OF. We collected OF specimens by use of the Quantisal OF collection device from 13 daily cannabis users after controlled oral cannabinoid administration. All specimens were tested with the Immunalysis Sweat/OF THC Direct ELISA and confirmed by 2-dimensional GC-MS. The limit of detection was <1 microg/L THC equivalent, and the assay demonstrated linearity from 1 to 50 microg/L, with semiquantification to 200 microg/L. Intraplate imprecision (n = 7) ranged from 2.9% to 7.7% CV, and interplate imprecision (n = 20) was 3.0%-9.1%. Cross-reactivities at 4 microg/L were as follows: 11-hydroxy-THC, 198%; Delta(8)-tetrahydrocannabinol (Delta(8)-THC), 128%; 11-nor-9-carboxy-THC (THCCOOH), 121%; THC (target), 98%; cannabinol, 87%; THCCOOH-glucuronide, 11%; THC-glucuronide, 10%; and cannabidiol, 2.4%. Of 499 tested OF specimens, 52 confirmed positive (THC 2.0-290 microg/L), with 100% diagnostic sensitivity at the proposed Substance Abuse and Mental Health Services Administration screening cutoff of 4 microg/L cannabinoids and GC-MS cutoff of 2 microg/L THC. Forty-seven specimens screened positive but were not confirmed by 2D-GC-MS, yielding 89.5% diagnostic specificity and 90.6% diagnostic efficiency. Thirty-one of 47 unconfirmed immunoassay positive specimens were from 1 individual and contained >400 ng/L THCCOOH, potentially contributing to cross-reactivity. The Immunalysis Sweat/OF THC Direct ELISA is an effective screening procedure for detecting cannabinoids in OF.


[Carcinoma of the prostate: correlation between local staging and systemic progression].
Management of prostate cancer requires considerable economic and social efforts and may causes some discomfort to patient. To attempt a simplification of the prostate cancer follow-up (dosage of PSA, ultrasound and abdominal and pelvic TC and/or RM, bone scan, X-ray chest and X-ray bone), Authors have review the patients with prostatic cancer who were undergone to a conservative treatment with hormonal and/or radiation therapy. From January 1984 to September 1995, 136 patients have been evaluated in the Urological Department of University of Perugia. Local staging cancer has been made according to TNM system. Ultrasound transrectal follow-up study (TRUS) we made through longitudinal and/or transversal scans. The echographic features have been evaluated are the prostate size, volume and echogenic pattern of cancer, extracapsular extention and, finally, the echographic appearance of surrounding tissues (seminal vesicles, rectum, bladder). In 116 patients (85.3%) Authors found a good correlation of clinical course and the results of the investigations whereas in 20 patients (14.7%) such correlation falls to be demonstrated. Although PSA level remains an important prognostic marker for monitoring patients with prostatic cancer, TRUS is useful to determinate not only the local status but also to predict the subsequent clinical course of these patients. The later could be performed at least twice a year whereas further investigations could be prescribed only in presence of clinical suspicious of metastasis. It can be concluded that TRUS and PSA levels are sufficiently safe indexes on follow-up of prostate cancer which can replace the most expensive examinations avoiding an unnecessary cost to the health care system. Furthermore TRUS can recognize patients with urine flow obstruction who may benefit by treatment with improvement of their quality of life.


The risk of healing complications in primary teeth with intrusive luxation: A retrospective cohort study.
Intrusive luxation is a frequent injury in the primary dentition. Complications such as ankylosis or pulp necrosis (PN) and infection with periapical inflammation may affect the developing permanent tooth if not diagnosed and treated in time. The aim of this study was to report the risk of PN, pulp canal obliteration (PCO), infection-related resorption (IRR), ankylosis-related resorption (ARR) and premature tooth loss (PTL) in primary teeth following intrusive luxation, and to identify possible risk factors for PN and PTL. A retrospective analysis of a cohort comprising 149 patients 194 intruded primary incisors. No treatment was performed. The follow-up programme included examination after 4 weeks, 8 weeks, 6 months, 1 year, and at 6 years of age. The minimum follow-up period was 1 year or until time of tooth loss. The Kaplan-Meier and Aalen-Johansen methods were employed along with Cox regression analysis. The level of significance was 5%. Risks estimated after 3 years: PCO 38.9% (95% CI: 31.8-46.0), PN 24.2% (95% CI: 17.7-30.6), IRR 8.8% (95% CI: 4.5-13.1), ARR 3.6% (95% CI: 1.0-6.2) and PTL 39.4% (95% CI: 31.2-47.5). Most teeth (83.7%) spontaneously re-erupted within the first year. Most complications were diagnosed within the first year. The risk of PN was lowest in patients less than 2 years of age. The degree of intrusion or a concomitant crown fracture did not affect the risk of PN or PTL. Over 80% of the intruded primary teeth re-erupted spontaneously. However, nearly one-third of the teeth showed complications such as pulp infection/periapical inflammation or ankylosis, which could potentially affect the development of the permanent incisor. Therefore, patients should be monitored regularly, especially during the first year after injury, to diagnose and treat complications in time.


An annotated checklist of the Iranian Dasytidae and Rhadalidae (Coleoptera: Cleroidea).
We herein provide an annotated checklist for the Iranian Dasytidae and Rhadalidae (Coleoptera: Cleroidea). A total of 54 species and three subspecies of Dasytidae in 16 genera, four tribes and three subfamilies, and 11 species of Rhadalidae in five genera and three subfamilies are reported and discussed where necessary. Of these, 27 (sub-) species of Dasytidae (48%) and eight species of Rhadalidae (73%) are currently known only from Iran, and are here considered as endemic species. However, some species occurences are doubtful or need confirmation. One species (Dasytes xanthocnemus Kolenati, 1846) is reported from Iran for the first time. Furthermore, Danacea micans astrabadensis Pic, 1922 is resurrected from synonymy and given subspecific status.


Fitness Level Modulates Intraocular Pressure Responses to Strength Exercises.
Purpose/Aim: The execution of strength exercises has demonstrated to increase the intraocular pressure (IOP) levels, and it may have a negative impact on the ocular health. We aimed to explore the influence of fitness level on the acute IOP response to strength exercises performed under different loading conditions, as well as to test whether the IOP responses differ between the bench press and jump squat when performed against the same relative loads. Forty military personnel males were divided in two subgroups (20 high-fit and 20 low-fit) based on their relative to body mass one-repetition maximum (1-RM). Participants performed an incremental loading test in the bench press and jump squat exercises, and IOP was assessed before and after each repetition by rebound tonometry. IOP increased immediately after executing both exercises (p < 0.01 in both cases), being the magnitude of the IOP increment positively and linearly associated with the increment of the load in both groups (i.e., high-fit and low-fit) and in both exercises (R2 range: 0.81-1.00). Higher fitness level attenuated the IOP rise produced by both exercises (p < 0.01 in both cases). The bench press induced higher IOP increments than the jump squat for both groups at relative loads of ~50%1-RM and ~60%1-RM (p < 0.01 in all cases). These data indicate that IOP increases as a consequence of performing strength exercises, being the increment accentuated with the increase of the load and in the bench press compared to the jump squat exercise. Of special importance would be that the IOP responses were significantly reduced in high-fit individuals. These findings should be addressed in glaucoma patients.


[SUCCESSFUL LAPAROSCOPIC TREATMENT OF HETEROTOPIC PREGNANCY AFTER ICSI / FREEZE EMBRYO TRANSFER - A CASE REPORT.]
The coexistence of extrauterine and intrauterine or haeterotopic pregnancy (HP) is a rare condition with reported incidence of 1 in 8000 to 1 in 30 000 pregnancies in natural conceived pregnancies, but with the widespread application of assisted reproductive technologies (ART) the incidence of HP in women with ART has risen to about 0.09% to 1.00%. We have reported a case of a 36 old woman, pregnant in 9 gestation week, gravidity 2, 0 para, who have achieved a pregnancy after in-vitro fertilization - embryo-transfer of two freeze embryos, developed after an ICSI procedure with donor eggs. The patient attended our clinic because of acute abdominal pain. She was evaluated by trans-vaginal sonography and coexistence of ectopic tubal and intrauterine pregnancy has been confirmed. Emergency laparoscopy was carried out. Left tubal ectopic pregnancy and hemoperitoneum (1200 ml) were diagnosed. Left salpingectomy was performed. At present, the patient is at 24 gestation week and the pregnancy is developing in a proper manner. We report such a case due to the increase of the awareness to the condition and the possibility of a successful treatment by a laparoscopic surgery. Intrauterine pregnancy does not exclude extrauterine pregnancy especially in cases after in-vitro fertilization.


Age differences in the association of physical leisure activities with incident disability among community-dwelling older adults.
The relationship between leisure activities (LA) in old age and prevention of disability has not been fully investigated, and age and gender differences of these relationships are unknown. This study aimed to investigate whether physical and cognitive LA predicted incident disability among community-dwelling older adults by age and gender. We prospectively observed 8,275 residents aged 65 or above without disability at baseline for 3 years. Incident disability was defined as a new certification of the public long-term care insurance system. LA were classified into two types: physical LA and cognitive LA. The frequency of LA was categorized into frequent (i.e., once a week or more), moderate (i.e., monthly or yearly), and non-engagement. Covariates included age, gender, family number, education, perceived economic situation, body mass index, chronic medical conditions, alcohol consumption, smoking status, regular dental visits, depression, cognitive functioning, and social participation. Multivariable Poisson regression models were used to estimate adjusted cumulative incidence ratio (CIR) and 95% confidence interval (CI) for incident disability. We performed stratified analyses by age groups (i.e., the young-old aged 65-74 and the old-old aged 75-97) and gender (i.e., men and women). The 3-year cumulative incidence of disability was 7.5%. After adjustment for covariates and mutual adjustment for both types of LA, a significant dose-response relationship between more frequent LA and lower risk of incident disability was found in young-old physical LA (P-trend < 0.001), in old-old cognitive LA (P-trend = 0.012), in male cognitive LA (P-trend = 0.006), and in female physical LA (P-trend = 0.030). Compared with people without LA, adjusted CIR (95% CI) of frequent LA was 0.47 (0.30-0.74) in young-old physical, 0.75 (0.58-0.96) in old-old cognitive, 0.65 (0.46-0.89) in male cognitive, and 0.70 (0.52-0.95) in female physical. Regarding the effect modification according to age and gender, only interaction between age and physical LA significantly prevented incident disability (P for interaction = 0.019). We found age differences in the association of physical LA with incident disability among community-dwelling older adults. An effective measure to prevent long-term care in the community would be to recommend frequent physical LA for the young-old.


[A pregnant woman with acute shoulder pain].
A 34-year-old woman presented with abdominal pain, vaginal blood loss and severe shoulder pain. Ultrasound revealed an ectopic pregnancy. The shoulder pain was due to irritation of the diaphragm caused by bleeding from a ruptured fallopian tube into the abdominal cavity.


Effectiveness of a web-based program on self-care behaviors and glycated hemoglobin in patients with type 2 diabetes: Study protocol of a randomized controlled trial.
Type 2 diabetes (T2DM) decreases the life expectancy and quality of life of diabetics and causes economic and societal problems. For this purpose, diabetes self-management education and support (DSMES) has been designed for many years, which is recently provided through technology-assisted education. Therefore, we developed a web-based program in accordance with DSMES to assess its effect on self-care behaviors and glycated hemoglobin (HbA1c) for patients with T2DM during the coronavirus disease (COVID-19) pandemic, which is described in detail in this paper. This randomized controlled trial (RCT) was performed on 70 diabetic patients in Al-Zahra Hospital for three months. After random allocation, web-based educational content (including videos, lectures, educational motion graphics, text files, educational posters, and podcasts) according to DSMES was provided for the intervention group to improve self-care behaviors and HbA1c levels. The control group received routine educational pamphlets. A diabetes self-management questionnaire (21 questions) with a Likert scale was completed to assess self-care behaviors scores before and after intervention and three months later. Also, HbA1c was determined before and after the intervention. Analysis of variance with repeated measurements will be applied to compare mean scores of self-care behaviors components three times, and an independent t-test analyzed mean differences of HbA1c values. The obtained results of this study might be useful for promoting self-care behaviors and assessing HbA1c in diabetic patients.


Fluid overload in the ICU: evaluation and management.
Fluid overload is frequently found in acute kidney injury patients in critical care units. Recent studies have shown the relationship of fluid overload with adverse outcomes; hence, manage and optimization of fluid balance becomes a central component of the management of critically ill patients. In critically ill patients, in order to restore cardiac output, systemic blood pressure and renal perfusion an adequate fluid resuscitation is essential. Achieving an appropriate level of volume management requires knowledge of the underlying pathophysiology, evaluation of volume status, and selection of appropriate solution for volume repletion, and maintenance and modulation of the tissue perfusion. Numerous recent studies have established a correlation between fluid overload and mortality in critically ill patients. Fluid overload recognition and assessment requires an accurate documentation of intakes and outputs; yet, there is a wide difference in how it is evaluated, reviewed and utilized. Accurate volume status evaluation is essential for appropriate therapy since errors of volume evaluation can result in either in lack of essential treatment or unnecessary fluid administration, and both scenarios are associated with increased mortality. There are several methods to evaluate fluid status; however, most of the tests currently used are fairly inaccurate. Diuretics, especially loop diuretics, remain a valid therapeutic alternative. Fluid overload refractory to medical therapy requires the application of extracorporeal therapies. In critically ill patients, fluid overload is related to increased mortality and also lead to several complications like pulmonary edema, cardiac failure, delayed wound healing, tissue breakdown, and impaired bowel function. Therefore, the evaluation of volume status is crucial in the early management of critically ill patients. Diuretics are frequently used as an initial therapy; however, due to their limited effectiveness the use of continuous renal replacement techniques are often required for fluid overload treatment. Successful fluid overload treatment depends on precise assessment of individual volume status, understanding the principles of fluid management with ultrafiltration, and clear treatment goals.


Impact of Major Depressive Disorder on Prediabetes by Impairing Insulin Sensitivity.
Reports regarding the associations between major depressive disorder (MDD) and diabetes remain heterogeneous. Our aim was to investigate whether glucose homeostasis and insulin sensitivity were impaired in the MDD patients and its mechanisms. A total of 30 patients with MDD and 30 matched controls were recruited. The oral glucose tolerance test and dual-energy X-ray absorptiometry scan were performed in each participant. Insulin signaling in postmortem brain tissues from other depressive patients and controls (obtained from Alabama brain bank) was examined. Insulin sensitivity was reduced substantially in the MDD patients, however, the fasting and 2-h glucose concentrations remained within the normal range through compensatory insulin secretion. Despite increased insulin secretion, 1-h glucose concentrations in the MDD patients were significantly elevated compared with the controls. MDD patients had greater visceral fat mass but lower adiponectin levels compared with the controls. Furthermore, phosphorylated-AKT levels in insulin signaling were decreased in postmortem brain tissues in patients with MDD. These results suggest that MDD patients are at a greater risk for diabetes due to decreased insulin sensitivity, reduced disposition index, and impaired glucose tolerance as manifested by elevated 1-h glucose concentrations following an oral glucose challenge. Mechanistic studies reveal that decreased insulin sensitivity is associated with increased visceral fat mass, lower adiponectin levels and impaired insulin action in postmortem brain tissues in the MDD patients. Our findings emphasize the importance of screening depressive patients to identify susceptible individuals for developing future diabetes with the hope of improving their health outcomes.


"It's just a way of fitting in:" Tobacco use and the lived experience of lesbian, gay, and bisexual appalachians.
Lesbian, gay, and bisexual (LGB) people are affected by multiple health disparities and risk factors, including tobacco use. Few studies to date have examined tobacco use specifically in rural LGB populations, and none has investigated the intersections of identity, rural LGB culture, and tobacco. The purpose of this study was to explore the perspective of Appalachian LGB people regarding tobacco use. . Nineteen LGB-identified Appalachian residents participated in audiotaped, semi-structured interviews. Two authors analyzed and coded transcripts through constant comparison, and determined themes through consensus. Five themes emerged: the convergence of Appalachian and LGB identities, tacit awareness of LGB identity by others, culture and tobacco use, perceived associations with tobacco use, and health beliefs and health care. LGB Appalachians connect stress and culture to tobacco, but seem less aware that partial concealment of their identity might be a source of the stress that could influence their smoking.


Use of Thoracic Catheters to Drain Pelvic Abscesses in Horses: Two Cases.
Pelvic abscesses occur in horses of all ages and breeds. Although conservative management may be successful, surgical drainage may be necessary for larger abscesses or in case of secondary intestinal obstruction. The purpose of this report is to describe two cases of pelvic abscesses that were successfully drained using indwelling chest tubes, repeated lavages and targeted antibiotic treatment.


The biomechanical effect of tunnel placement on ACL graft forces in double-bundle ACL reconstruction - A 3D computational simulation.
The posterolateral (PL) graft experiences a high failure rate in anterior cruciate ligament double-bundle (DB) reconstruction. It is hypothesized that tunnel positions could dramatically affect the graft forces. A validated computational model was used to simulate DB reconstruction with various femoral PL tunnel locations (8-11 mm center-center tunnel spacing). Graft fixation was simulated at both 0° and 30°. Knee biomechanics were examined with the knee under a 134 N anterior load and 400 N quadriceps load at 0°, 30°, 60°, and 90° of flexion. Graft forces, tibial translation, and tibial rotation were calculated. PL graft forces at full extension increased with increasing tunnel spacing under both fixation settings, but the knee kinematics was not dramatically affected. Small changes in the femoral PL tunnel position could result in large changes in graft forces, implying that precise PL tunnel position is an important factor in a successful DB reconstruction.


Lipid metabolism in pregnancy. IV. C Apoprotein changes in very low and intermediate density lipoproteins.
In vitro studies indicate that relative amounts of apolipoproteins CII and CIII in plasma may alter lipoprotein lipase mediated triglyceride removal from very low density lipoprotein. Therefore, we have determined relative amounts of these peptides in pregnant women in late gestation (mean 36 weeks) and 6 and 20 weeks postpartum. Very low density (d less than 1.006) and intermediate density (d 1.006-1.019) lipoproteins have been examined because they are affected similarly in pregnancy. Triglyceride and cholesterol in these two fractions increased 3 1/2 to 4 1/2-fold in pregnancy in keeping with previous reports. On a relative basis, apo CII was decreased and apo CIII1 and CIII2 increased in both VLDL and IDL at 36 weeks gestation. At 6 and 20 weeks postpartum relative amounts of C peptides returned to identical levels and were unaffected by lactation. It remains to be seen if in vivo triglyceride removal in pregnancy is altered by the reduction in apo CII relative to apo CIII.


Myocardial infarction induces expression of midkine, a heparin-binding growth factor with reparative activity.
We examined midkine (MK) expression in the rat heart upon experimental myocardial infarction. Immunohistochemical staining revealed, 6 hours after ligation of the left anterior descending coronary artery, strong MK immunoreactivity in myocytes and endothelial cells of a non-infarcted cardiac region. The myocytes of the infarcted cardiac region destined for death showed only a little immunoreactivity. Northern blot analysis suggested that the increased immunoreactivity was due to increased MK synthesis. The induced MK expression is likely to mimic the expression during embryogenesis: MK was strong lye-expressed in the myocytes of embryonic heart, and the expression decreased during embryogenesis.


Congenital insensitivity to pain with anhidrosis.
A nine-year-old child presented with congenital insensitivity to pain and anhidrosis. Quantitative studies and electron microscopy of the cutaneous branch of the radial nerve revealed almost complete absence of small myelinated and unmyelinated fibers and a disproportionate number of nerve fibers with a diameter of 6-10 micrometers. A grouping of both type 1 and type 2 muscle fibers was also seen. We suggest that this disease entity is not caused by a hereditary sensory neuropathy, but rather that it derives from a developmental defect.


Calbindin content and differential vulnerability of midbrain efferent dopaminergic neurons in macaques.
Calbindin (CB) is a calcium binding protein reported to protect dopaminergic neurons from degeneration. Although a direct link between CB content and differential vulnerability of dopaminergic neurons has long been accepted, factors other than CB have also been suggested, particularly those related to the dopamine transporter. Indeed, several studies have reported that CB levels are not causally related to the differential vulnerability of dopaminergic neurons against neurotoxins. Here we have used dual stains for tyrosine hydroxylase (TH) and CB in 3 control and 3 MPTP-treated monkeys to visualize dopaminergic neurons in the ventral tegmental area (VTA) and in the dorsal and ventral tiers of the substantia nigra pars compacta (SNcd and SNcv) co-expressing TH and CB. In control animals, the highest percentages of co-localization were found in VTA (58.2%), followed by neurons located in the SNcd (34.7%). As expected, SNcv neurons lacked CB expression. In MPTP-treated animals, the percentage of CB-ir/TH-ir neurons in the VTA was similar to control monkeys (62.1%), whereas most of the few surviving neurons in the SNcd were CB-ir/TH-ir (88.6%). Next, we have elucidated the presence of CB within identified nigrostriatal and nigroextrastriatal midbrain dopaminergic projection neurons. For this purpose, two control monkeys received one injection of Fluoro-Gold into the caudate nucleus and one injection of cholera toxin (CTB) into the postcommissural putamen, whereas two more monkeys were injected with CTB into the internal division of the globus pallidus (GPi). As expected, all the nigrocaudate- and nigroputamen-projecting neurons were TH-ir, although surprisingly, all of these nigrostriatal-projecting neurons were negative for CB. Furthermore, all the nigropallidal-projecting neurons co-expressed both TH and CB. In summary, although CB-ir dopaminergic neurons seem to be less prone to MPTP-induced degeneration, our data clearly demonstrated that these neurons are not giving rise to nigrostriatal projections and indeed CB-ir/TH-ir neurons only originate nigroextrastriatal projections.


Tumor-Microenvironment Relaxivity-Changeable Gd-Loaded Poly(L-lysine)/Carboxymethyl Chitosan Nanoparticles as Cancer-Recognizable Magnetic Resonance Imaging Contrast Agents.
Magnetic resonance imaging (MRI) contrast agents with tumor-microenvironment changeable relaxivity are effective to increase the sensitivity and selectivity of MRI in tumor diagnosis. In this study, pH-sensitive Gd-loaded Poly(L-lysine)/ Carboxymethyl Chitosan Nanoparticles (Gd-PCNPs) were developed as relaxivity-changeable MRI contrast agents based on the "on–off" switchable strategy. The "on–off" switchable nano-contrast agents were capable of releasing Gd3+ in response to physical stimulation, with structure transformed. Gd-PCNPs could responsively disassemble in an acidic tumor-microenvironment and increase the exchange of protons between water molecules and Gd3+ ions, thus selectively enhance the relaxivity in tumor area. Gd-PCNPs were self-assembled via electrostatic interaction between poly(L-lysine)-diethylenetriamine pentaacetic acid-gadolinium and pH-sensitive carboxymethyl chitosan (CMCS). Gd-PCNPs exhibited spherical shape with uniform particle size distribution (166.00 ± 1 .71 nm) and negative zeta potential (–13.2 ± 4.7 mV). The relaxivity of Gd-PCNPs increased from 6.618 mM–1 · s–1 to 10.008 mM–1 · s–1 when the pH values decrease from 7.4 to 6.0, which was higher than Magnevist® (3.924 mM–1 · s–1 at both pH 7.4 and 6.0 (p <0 05). The changeable relaxivity of Gd/PCNPs would result in enhanced tumor/normal tissue signal contrast, which was verified by in vivo MRI test. In vivo MRI test showed that the signal of Gd-PCNPs was significantly enhanced with prolonged imaging time in tumor tissue compared to Magnevist® (p <0 05). Furthermore, Gd-PCNPs exhibited unobvious in vitro cytotoxicity under the experimental concentrations in B16 cells. No obvious damage was observed in the different tissues of mice. These results indicated that the relaxivity-changeable Gd-PCNPs exhibited demonstrated sensitivity and selectivity in tumor diagnosis with a great potential as a novel MRI contrast agent.


Viscosity measurements in pulsed magnetic fields by using a quartz-crystal microbalance.
Viscosity measurements in combination with pulsed magnetic fields are developed by use of a quartz-crystal microbalance (QCM). When the QCM is immersed in liquid, the resonant frequency, f0, and the quality factor, Q, of the QCM change depending on (ρη)0.5, where ρ is the mass density and η the viscosity. During the magnetic-field pulse, f0 and Q of the QCM are simultaneously measured by a ringdown technique. The typical resolution of (ρη)0.5 is 0.5%. As a benchmark, the viscosity of liquid oxygen is measured up to 55 T.


Aspiration and the laryngeal mask airway: three cases and a review of the literature.
The primary limitation of the laryngeal mask airway (LMA) is that it does not reliably protect the lungs from regurgitated stomach content. We describe three cases of aspiration associated with the LMA, including the first brain injury, the first death, and the first associated with the intubating LMA, and review the 20 specific case reports of aspiration associated with the LMA that we were able to find described in the literature.


Country-wide seroprevalence studies on Crimean-Congo hemorrhagic fever and hantavirus infections in general population of Bulgaria.
Crimean-Congo hemorrhagic fever (CCHF) and hantavirus infections are the two viral hemorrhagic fevers spread in Europe. To test actual circulation of CCHF virus (CCHFV) and hantaviruses in Bulgaria, we conducted country-wide seroepidemiological studies. Serum samples were collected prospectively from 1500 residents of all 28 districts in Bulgaria. CCHFV seroprevalence of 3.7% was revealed. Anamnesis for tick bites, contact with livestock, age over 40 years and residency in Haskovo district were found as risk factors. The highest CCHFV seroprevalence was observed in the known endemic districts in southeastern Bulgaria: Haskovo (28%) and Yambol (12%). Reactive samples were found in residents of 20 of the 28 districts in Bulgaria. In comparison with the previous studies, the data presented indicate that CCHFV increased substantially its circulation in the endemic regions and was introduced in many new areas. Hantavirus seroprevalence was based on results of the immunoblot and estimated as 3.1%. Surprisingly, contrary to all available data, Puumala virus seroprevalence rate was 2.3% versus 0.8% of Dobrava-Belgrade virus. Evidence for hantavirus IgG seropositivity was found in residents of 23 of the 28 districts in the country. The first hantavirus seroprevalence study in Bulgaria showed that Puumala virus is probably more wide-spread in the country than Dobrava-Belgrade virus.


Tympanic membrane organ culture using cell culture well inserts engrafted with tympanic membrane tissue explants.
Tissue engineering approaches using growth factors and various materials for repairing chronic perforations of the tympanic membrane are being developed, but there are surprisingly few relevant tissue culture models available to test new treatments. Here, we present a simple three-dimensional model system based on micro-dissecting the rat tympanic membrane umbo and grafting it into the membrane of a cell culture well insert. Cell outgrowth from the graft produced sufficient cells to populate a membrane of similar surface area to the human tympanic membrane within 2 weeks. Tissue grafts from the annulus region also showed cell outgrowth but were not as productive. The umbo organoid supported substantial cell proliferation and migration under the influence of keratinocyte growth medium. Cells from umbo grafts were enzymatically harvested from the polyethylene terephthalate (PET) membrane for expansion in routine culture and cells could be harvested consecutively from the same graft over multiple cycles. We used harvested cells to test cell migration properties and to engraft a porous silk scaffold material as proof-of-principle for tissue engineering applications. This model is simple enough to be widely adopted for tympanic membrane regeneration studies and has promise as a tissue-equivalent model alternative to animal testing.


Hepatocyte Concentrations of Imaging Compounds Associated with Transporter Inhibition: Evidence in Perfused Rat Livers.
In the liver, several approaches are used to investigate and predict the complex issue of drug-induced transporter inhibition. These approaches include in vitro assays and pharmacokinetic models that predict how inhibitors modify the systemic and liver concentrations of the victim drugs. Imaging is another approach that shows how inhibitors might alter liver concentrations stronger than systemic concentrations. In perfused rat livers associated with a gamma counter that measures liver concentrations continuously, we previously showed how fluxes across transporters generate the hepatocyte concentrations of two clinical imaging compounds, one with a low extraction ratio [gadobenate dimeglumine (BOPTA)] and one with a high extraction ratio [mebrofenin (MEB)]. BOPTA and MEB are transported by rat organic anion transporting polypeptide and multiple resistance-associated protein 2, which are both inhibited by rifampicin. The aim of the study is to measure how rifampicin modifies the hepatocyte concentrations and membrane clearances of BOPTA and MEB and to determine whether these compounds might be used to investigate transporter-mediated drug-drug interactions in clinical studies. We show that rifampicin coperfusion greatly decreases BOPTA hepatocyte concentrations, but increases those of MEB. Rifampicin strongly decreases BOPTA hepatic clearance. In contrast, rifampicin decreases moderately MEB hepatic clearance and blocks the biliary intrinsic clearance, increasing MEB hepatocyte concentrations. In conclusion, low concentrations prevent the quantification of BOPTA biliary intrinsic clearance, while MEB is a promising imaging probe substrate to evidence transporter-mediated drug-drug interactions when inhibitors act on influx and efflux transporters.


Does computed tomography or positron emission tomography/computed tomography contribute to detection of small focal cancers in the prostate?
Prostate cancer is considered to be a multifocal tumor in the majority of patients. Based on histologic data after prostatectomy, there is a growing insight that a considerable number of men who receive a diagnosis in the contemporary setting of prostate-specific antigen screening have unilateral or unifocal disease. With this, the current concept of whole-gland therapy has come into discussion. The need for improvement of intraprostatic tumor characterization is clear. Molecular imaging is one of the areas of research on this aspect. The clinical indications for positron emission tomography (PET)/CT have increased rapidly in the field of oncology and are largely based on fluorodeoxyglucose (FDG) PET. Both conventional CT and FDG PET, however, cannot detect prostate cancer foci <5 mm within the prostate. Dynamic contrast-enhanced CT involves imaging a region of interest rapidly (usually <10 seconds between images) during a bolus intravenous injection of a contrast agent. Through analysis of the contrast enhancement time curves, it is possible to distinguish tissues with different microvascular properties such as cancer. The technologic aspects of both imaging techniques and the clinical results of 11C-choline PET/CT for intraprostatic tumor characterization are discussed. Based on preliminary studies, dynamic contrast-enhanced (DCE)-CT may be a useful tool for localization of prostate tumors and, perhaps more importantly, quantification of therapeutic response in prostate cancer. Validation work is necessary, however, to define its accuracy and role in therapeutic paradigms such as focal therapies, particularly given the current accuracy of MRI. In the future, combining DCE-CT with CT or (11)C-choline PET/CT may be an alternative to MRI, offering a combination of quantitative parameters that may correlate to tumor prognosis as well as cancer localization for focal therapy.