Effects of Twitch Contraction Induced by Magnetic Stimulation on Expression of Skeletal Muscle Fibrosis Related Genes and Limited Range of Motion in Rats.
We examined the effects of twitch contraction induced by lumbar spinal root magnetic stimulation on immobilization-induced limited range of motion and skeletal muscle fibrosis in rat soleus muscle. The groups included male Wistar rats (controls), rats with both bilateral ankle joints immobilized with plaster casts for 4 wks (immobilization [IM]), and rats in which twitch contraction of the soleus muscle was induced by lumbar magnetic stimulation for 4 wks of immobilization (twitch contraction [TC]). Group differences in ankle dorsiflexion range of motion, collagen content as determined by hydroxyproline assay, and the expression of fibrosis-related genes as determined by reverse transcription-polymerase chain reaction (hypoxia inducible factor 1α, α-smooth muscle actin, and types I and III collagen) in the soleus muscle were examined after the 4-wk-long experiment. Range of motion in the TC group was significantly greater than that in the IM group. The hydroxyproline content and the expressions of fibrosis-related genes decreased significantly in the TC group compared with those in the IM group. No significant differences were seen in the expression of transforming growth factor β mRNA. These results suggest that twitch contraction induced by lumbar spinal root magnetic stimulation may reduce immobilization-induced limited range of motion and skeletal muscle fibrosis.


Strontium ranelate normalizes bone mineral density in osteopenic patients.
To assess the capacity of strontium ranelate to restore normal bone mineral density (WHO definition: T-score >or=-1) in post-menopausal osteopenic women (T-score between -1 and -2.5) at baseline. Post-hoc analysis from SOTI and TROPOS studies of 1428 patients randomly assigned to receive either 2 g of strontium ranelate a day or placebo for three years. Bone mineral density was measured at baseline and each year for three years. Results were analyzed on an intention-to-treat basis. At lumbar spine, after one, two and three years of treatment with strontium ranelate, 26.4, 42.1 and 58.2% respectively of osteopenic patients normalized their bone mineral density, compared with 6.6, 8.9 and 11.9% in the placebo group (all p<0.001). At total hip, the percentage of patients normalizing their bone mineral density was 5.4, 10.0 and 19.6% in the strontium ranelate group and 1.8, 1.4 and 1.6% in the placebo one (all p<0.001). Strontium ranelate is able to normalize bone mineral density in a significant proportion of osteopenic patients after one, two and three years of treatment. The clinical relevance of these results should be confirmed by direct demonstration of the anti-fracture efficacy of strontium ranelate in osteopenic patients.


Autophagy promotes programmed cell death and corpse clearance in specific cell types of the Arabidopsis root cap.
Autophagy is a conserved quality control pathway that mediates the degradation of cellular components by targeting them to the lysosomes or vacuoles.1 Autophagy has been implicated in the regulation of some regulated cell death processes in animal systems.2 However, its function in developmentally controlled programmed cell death (dPCD) in plants remains little studied and controversial.3 Some studies have reported autophagy pro-survival roles,4,5 while others have suggested pro-death functions for autophagy,6,7 calling for further detailed investigations. Here, we investigated the role of autophagy in dPCD using the Arabidopsis root cap as an accessible and genetically tractable model system.8 In Arabidopsis, dPCD is an integral part of root cap differentiation, restricting root cap organ size to the root meristem.9 The root cap consists of two distinct tissues: the proximally positioned columella that is located at the very root tip and the lateral root cap (LRC) that flanks the root meristem up to its distal end at the start of the root elongation zone.10 We show that autophagic flux strongly increased prior to dPCD execution in both root cap tissues and depends on the key autophagy genes ATG2, ATG5, and ATG7. Systemic and organ-specific mutation of these genes shows delayed PCD execution and lack of postmortem corpse clearance in the columella but no defects in dPCD execution or corpse clearance in the distal LRC. Our results reveal a high degree of cell-type specificity in autophagy functions and suggest that autophagy roles in dPCD can considerably diverge between different cell types of the same plant organ.


[Comparative evaluation of the efficacy of pre- and intraoperative reserving of autoblood in surgical treatment of ischemic heart disease and while using a Cell Saver].
A prospective investigation included 142 patients who had undergone operations of coronary bypass under conditions of extracorporeal circulation for ischemic heart disease. Two methods of taking blood were used: one day before operation and at the beginning of operation. The transfusion of autoblood was performed after cessation of the extracorporeal circulation. Cell saver was used during operations on all the patients. The preoperative reserving of autoblood promoted the greater degree of hemodilution (both during extracorporeal circulation and at the early postoperative period). The intraoperative reserving of autoblood led to saving higher indices of content of hemoglobin, number of erythrocytes and hematokrit index at the early postoperative period. However, when using the cell saver, the method of reserving blood (7-8 ml/kg of body mass) was of no substantial influence on the frequency of using the allogenic blood.


Chronopharmacology of strychnine and allylglycine in the mouse.
1. The response of individually caged mice to the lethal actions of allylglycine and strychnine was evaluated in animals previously conditioned on an LD 12:12 (12 h light-12 h darkness) schedule in a controlled environment. 2. These convulsant agents were most toxic at 18.00 hours (during the light phase), and least toxic in the dark phase of the programmed lighting schedule. The relationship is considered of the circadian fluctuations in levels of inhibitory transmitter substances to the time-linked action of convulsant agents.


Blood calcium as a prognostic indicator of success after surgical correction of left displaced abomasum.
Subclinical hypocalcemia, a reduction in blood calcium concentrations without apparent clinical signs of milk fever, occurs in 25 to 50% of early-postpartum dairy cows. Research has shown that these cows are at an increased risk of additional early-lactation diseases, including left displaced abomasum (LDA), compared with their normocalcemic counterparts. However, no work has assessed the association of total calcium (tCa) concentration at the time of LDA correction with subsequent milk yield and survival within the herd. Pending future development of an economical and accurate on-farm test for hypocalcemia, the ability to assess LDA prognosis after surgical correction based on precorrection blood tCa concentration is of interest to practitioners. Therefore, our objective was to determine the association of blood tCa concentration at the time of LDA surgical correction with milk yield and herd removal in the 60 d after correction. Nine bovine practitioners, located in New York State, were involved in sample collection from 17 herds for this prospective cohort study. Immediately following LDA diagnosis and before surgical correction, blood samples were collected from 152 dairy cows within the first 30 d in milk for serum tCa determination. The association of tCa with milk yield (n = 110) and herd removal (n = 127) in the first 60 d after LDA surgical correction was analyzed using repeated-measures ANOVA and a generalized linear mixed model, respectively, controlling for parity and days in milk at LDA surgical correction and the random effect of herd or practitioner. We found no evidence to support an association of tCa at time of LDA diagnosis with subsequent milk yield, and when cows were grouped by median tCa into low and high groups (tCa ≤2.1 and >2.1 mmol/L, respectively), subsequent milk production was similar (39.4 ± 1.6 and 40.1 ± 1.4 kg/d for low and high, respectively). Furthermore, we found no evidence to support an association of tCa at time of LDA diagnosis with the odds of herd removal (odds ratio = 0.6; 95% confidence interval = 0.4-1.7). Therefore, our study provided no clear evidence to support the association of tCa at time of LDA diagnosis and correction with either milk yield or herd removal during the 8 wk after correction. However, it did demonstrate low to extremely low tCa concentration in cows with LDA. Whether calcium supplementation would improve health and production outcomes in these cows is unclear and worth investigating further.


Two distinct phenotypes of asthma in elite athletes identified by latent class analysis.
Clusters of asthma in athletes have been insufficiently studied. Therefore, the present study aimed to characterize asthma phenotypes in elite athletes using latent class analysis (LCA) and to evaluate its association with the type of sport practiced. In the present cross-sectional study, an analysis of athletes' records was carried out in databases of the Portuguese National Anti-Doping Committee and the Norwegian School of Sport Sciences. Athletes with asthma, diagnosed according to criteria given by the International Olympic Committee, were included for LCA. Sports practiced were categorized into water, winter and other sports. Of 324 files screened, 150 files belonged to asthmatic athletes (91 Portuguese; 59 Norwegian). LCA retrieved two clusters: "atopic asthma" defined by allergic sensitization, rhinitis and allergic co-morbidities and increased exhaled nitric oxide levels; and "sports asthma", defined by exercise-induced respiratory symptoms and airway hyperesponsiveness without allergic features. The risk of developing the phenotype "sports asthma" was significantly increased in athletes practicing water (OR = 2.87; 95% CI [1.82-4.51]) and winter (OR = 8.65; 95% CI [2.67-28.03]) sports, when compared with other athletes. Two asthma phenotypes were identified in elite athletes: "atopic asthma" and "sports asthma". The type of sport practiced was associated with different phenotypes: water and winter sport athletes had three- and ninefold increased risk of "sports asthma". Recognizing different phenotypes is clinically relevant as it would lead to distinct targeted treatments.


Modulation of intestinal L-glutamate transport by luminal leptin.
Leptin is secreted into the digestive tract and contributes to the absorption of dietary molecules by regulating transporters activity. Here, we studied the effect of luminal leptin on the intestinal transport of L-glutamate, an important component of human diet. We examined the effect of leptin on L-glutamate uptake in rat intestine in vitro measuring glutamate-induced short-circuit current (Isc) in Ussing chambers and L-[(3)H (U)]-glutamate uptake in jejunal everted rings. Glutamate-induced Isc was only observed in Na(+)-free conditions. This Isc was concentration (1-60 mmol L(-1)) and pH dependent. Luminal leptin increased glutamate Isc (∼100 %). Dose-response curve showed a biphasic pattern, with maximal stimulations observed at 10(-13) and 10(-10) mmol L(-1), that were sensitive to leptin receptor antagonist. In everted rings, two glutamate transport mechanisms were distinguished: a Na(+)-dependent, H(+)-independent, that was inhibited by leptin (∼20 %), and a Na(+)-independent but H(+)-dependent, that was enhanced by leptin (∼20 %), in line with data obtained in Ussing chambers. Altogether, these data reveal original non-monotonic effect of luminal leptin in the intestine and demonstrate a new role for this hormone in the modulation of L-glutamate transport, showing that luminal active gut peptides can influence absorption of amino acids.


Cancer detection and the community pharmacist.
A questionnaire was prepared to asses community pharmacists' perceptions of their abilities to recognize the common signs and symptoms of cancer and their preferred methods for obtaining further education. Of 5,539 questionnaires mailed, 1,187 were returned and analyzed. Seventy percent of the pharmacists reported that at least one patient per month sought advice about possible cancer signs and symptoms. Almost half estimated that between 1% and 25% of the patients who had sought advice for these cancer signs and symptoms had attempted to purchase a medication to treat these symptoms. The vast majority of the pharmacists perceived that they could recognize common signs and symptoms of skin, breast, and colorectal cancers. However, the majority felt they could not identify the symptomatology for six other common cancers. Essentially all pharmacists rated education in cancer symptomatology to be important. There was no consensus as to the type of provider, format, or times for this continuing education. These data suggest that pharmacists can play an important role in the early detection and prevention of cancer. The rate of interaction by pharmacists in this study translates to more than 60,000 patients counseled about cancer symptoms per 1,000 pharmacists per year. Further education of the pharmacist in cancer detection is warranted, and appraisal of the outcome of such educational interventions on patient-referral patterns and cancer-related diagnoses is imperative.


Reassessing triglyceride synthesis in adipose tissue.
The synthesis and breakdown of triglycerides in adipose tissue and muscle is a crucial element of energy metabolism because it ensures that adequate fuel is available during starvation. Triglyceride turnover determines the availability of fatty acids for utilization by mammalian tissues, and any dysfunction in this process can lead to alterations in glucose metabolism, insulin resistance and type 2 diabetes. Our understanding of the reactions involved in triglyceride synthesis is currently being reassessed, primarily because of the recently identified role that re-esterification of fatty acids plays in triglyceride deposition and, thus, in controlling fatty-acid availability. Here, we review recent information on triglyceride synthesis and introduce the pathway of glyceroneogenesis as an important and highly regulated source of glyceride-glycerol in adipose tissue.


On studentising and blocklength selection for the bootstrap on time series.
For independent data, non-parametric bootstrap is realised by resampling the data with replacement. This approach fails for dependent data such as time series. If the data generating process is at least stationary and mixing, the blockwise bootstrap by drawing subsamples or blocks of the data saves the concept. For the blockwise bootstrap a blocklength has to be selected. We propose a method for selecting the optimal blocklength. To improve the finite size properties of the blockwise bootstrap, studentised statistics is considered. If the statistic can be represented as a smooth function model this studentisation can be approximated efficiently. The studentised blockwise bootstrap method is applied for testing hypotheses on medical time series.


Prevention of alcohol-related problems. Introduction to a symposium.
Despite major biomedical advances, the most important health gains of this century have been due to preventive measures. Primary prevention includes: (a) health protection--the use of public direct regulatory activities (e.g. regulations regarding the sale and availability of alcohol, requirements for warning labels); (b) disease prevention--services directed to reducing the occurrence of specific identifiable disorders (e.g. Fetal Alcohol Syndrome) using strategies derived from risk factor research; and (c) health promotion--activities which foster positive behaviors and general good health practices. Although there is a measure of agreement that 'something must be done' to prevent alcohol problems, it is not quite so easy to find agreement on what is the best preventive approach, or even what the focus of prevention should be--problem drinkers, drinking habits or alcohol itself. To change attitudes and practices requires more than token nods towards alcohol education. More attention is now being given to psychologically and socially based prevention programs. The next three papers discuss behavioural approaches to prevention of alcohol-related problems.


Bit1 knockdown contributes to growth suppression as well as the decreases of migration and invasion abilities in esophageal squamous cell carcinoma via suppressing FAK-paxillin pathway.
There is growing evidence that Bit1 exerts different roles in the development and progression of human cancers. Although Bit1 was highly exhibited in ESCC tissues in our previous study, its roles and molecular mechanisms implicated in development and progression of ESCC remain unknown. Bit1 protein expression in ESCC cell lines and normal esophageal epithelial cell was detected by Western blotting. Bit1 protein expression mediated by Bit1 shRNA was investigated by Western blotting. MTT, migration assay, invasion experiment, ELISA and Flow cytometry were utilized to determine the effects of Bit1 knockdown on cell proliferation, migration, invasion and apoptosis, respectively. A xenograft model was used to examine in vivo tumourigenicity, and immunohistochemistry and TUNEL were utilized to evaluate the related protein expression and apoptosis. Gene microarray was determined by Agilent SurePrint G3 Human GE 8 × 60 K Microarray, the interaction of Bit1 and FAK proteins were detected by Immunoprecipitation and the key protein expressions of FAK-paxillin pathway were detected by Western blotting. We found Bit1 expression in all human ESCC cell lines tested was significantly higher than that in normal esophageal epithelial cell Het-1A (P < 0.05), in which EC9706 presented the highest Bit1 level. Bit1 protein level was significantly downregulated at day 1 after transfection with specific shRNA against Bit1 (P < 0.05). At days 2 and 3, Bit1 level reached the lowest value after transfection with Bit1 shRNA. Moreover, Bit1 depletion contributed to growth inhibition in vitro and in vivo, reduced cell migration and invasion abilities, and induced cell apoptosis in EC9706 and TE1 cells. More importantly, Bit1 downregulation significantly lowered Bcl-2 and MMP-2 levels in EC9706 xenografted tumor tissues, meanwhile triggered apoptosis after treatment with different doses of Bit1 shRNA. Further gene microarray revealed that 23 genes in Bit1-RNAi group were markedly downregulated, whereas 16 genes were obviously upregulated. Notably, Bit1 intrinsically interacted with FAK protein in EC9706 cells. Moreover, paxillin was downregulated at mRNA and protein levels in Bit1 shRNA group, coupled with the decreases of FAK mRNA and protein expressions. Bit1 may be an important regulator in cell growth, apoptosis, migration and invasion of ESCC via targeting FAK-paxillin pathway, and thereby combinative manipulation of Bit1 and FAK-paxillin pathway may be the novel and promising therapeutic targets for the patients with ESCC.


The impact of preceptor and student learning styles on experiential performance measures.
To identify preceptors' and students' learning styles to determine how these impact students' performance on pharmacy practice experience assessments. Students and preceptors were asked to complete a validated Pharmacist's Inventory of Learning Styles (PILS) questionnaire to identify dominant and secondary learning styles. The significance of "matched" and "unmatched" learning styles between students and preceptors was evaluated based on performance on both subjective and objective practice experience assessments. Sixty-one percent of 67 preceptors and 57% of 72 students who participated reported "assimilator" as their dominant learning style. No differences were found between student and preceptor performance on evaluations, regardless of learning style match. Determination of learning styles may encourage preceptors to use teaching methods to challenge students during pharmacy practice experiences; however, this does not appear to impact student or preceptor performance.


Different effects of oral conjugated equine estrogens and transdermal estrogen on undercarboxylated osteocalcin concentration in postmenopausal women.
Undercarboxylated osteocalcin (ucOC) is a sensitive marker of vitamin K status, and triglyceride (TG) has been shown to be the main transporter of vitamin K. In the present study, we examined the difference between ucOC concentrations in postmenopausal women receiving hormone therapy (HT) with oral conjugated equine estrogens (CEE) and transdermal estradiol (TE2). We also examined the associations of ucOC concentration with estradiol concentration and TG. Ninety-two postmenopausal women were recruited for this study. Serum concentrations of ucOC, intact osteocalcin, estradiol, and TG were measured before and after 12 months of HT. Forty-six women received oral administration of 0.625 mg of CEE and 2.5 mg of medroxyprogesterone acetate daily, and 46 women received transdermal administration of 50 mug of 17beta-estradiol twice weekly and 2.5 mg of medroxyprogesterone acetate daily. The ucOC concentration in women during HT with oral CEE was significantly (P < 0.01) lower than that in women during HT with TE2. Serum estradiol concentrations during HT with CEE showed a significant inverse correlation with ucOC concentrations and the ratio of ucOC/OC during HT (P < 0.05 and P < 0.01, respectively). In addition, the serum ucOC concentration in women with an increased percentage of change in TG was significantly (P < 0.01) lower than that in women with a decreased percentage of change in TG during HT with oral CEE. The effect of HT with TE2 on ucOC concentration in women is weaker than the effect of HT with oral CEE. Suppression of ucOC concentration in postmenopausal women during HT with oral CEE might be associated with the effect of vitamin K through increased TG induced by oral CEE.


Understanding genomics and the immune environment of penile cancer to improve therapy.
The incidence of penile squamous cell carcinoma (PSCC) has increased in developed countries over the past decades owing to increased human papilloma virus (HPV) exposure. Despite successful surgical treatment of locoregional PSCC, effective treatment options for advanced disease are limited. The prognosis of patients with bulky nodal and metastatic PSCC is dismal and new management approaches are urgently needed. Genomic analyses have provided transformative knowledge on the genomic and molecular landscape and tumour microenvironment of PSCC. Around one-quarter of patients with metastatic PSCC have clinically actionable genomic alterations in mechanistic target of rapamycin, DNA repair and receptor tyrosine kinase pathways. These patients might benefit from combined and sequential targeted therapies. HPV vaccination might be another therapeutic option as PSCC is genetically similar to other HPV-driven cancers. In addition, 40-60% of PSCC tumours show strong PDL1 expression, and the frequency of mutational signatures suggestive of immunotherapy resistance is low, pointing to potential utility of immunotherapy for PSCC. Finally, identification of the composition of the penile microbiota and its biological role might lead to new cancer prevention and treatment strategies.


Associations Between Apparent Diffusion Coefficient Values and the Prognostic Factors of Breast Cancer.
Breast imaging can offer great information about breast cancer heterogeneity. The purpose of this study was to analyze the relationship between apparent diffusion coefficient (ADC) values and various prognostic factors and investigate whether ADC values are useful for breast cancer diagnosis, evaluation of treatment response, and determination of prognosis. A total of 111 cases of breast cancer were included in this study. Magnetic resonance findings were recorded according to the Breast Imaging Reporting and Data System magnetic resonance imaging lexicon. Diffusion-weighted imaging rim sign and minimum, maximum, and difference ADC values (ADCdiff) were also evaluated. ADCdiff was related to all prognostic factors such as histological grade, Ki-67, tumor size, molecular subtype, axillary node metastasis, lymphvascular invasion, internal enhancement pattern, intratumoral high T2 signal, peritumoral edema, and diffusion-weighted imaging rim sign, whereas minimum and maximum ADC values showed variable associations. Apparent diffusion coefficient values were shown to be correlated with many proven or possible prognostic factors of breast cancer. In particular, ADCdiff can reflect tumor heterogeneity and showed higher correlation.


Synthesis and biological activities of transition metal complexes based on acetylsalicylic acid as neo-anticancer agents.
[(μ(4)-η(2))-(Prop-2-ynyl)-2-acetoxybenzoate]dicobalthexacarbonyl (Co-ASS), a derivative of aspirin (ASS), demonstrated high growth-inhibitory potential against various tumor cells with interference in the arachidonic acid cascade as probable mode of action. The significance of the kind of metal and cluster was verified in this structure-activity study: Co(2)(CO)(6) was respectively exchanged by a tetrameric cobalt-, trimeric ruthenium-, or trimeric ironcarbonyl cluster. Furthermore, the metal binding motif was changed from alkyne to 1,3-butadiene. Compounds were evaluated for growth inhibition, antiproliferative effects, and apoptosis induction in breast (MCF-7, MDA-MB 231) and colon cancer (HT-29) cell lines and for COX-1/2 inhibitory effects at isolated isoenzymes. Additionally, the major COX metabolite prostaglandin E2 (PGE(2)) was quantified in arachidonic acid-stimulated MDA-MB 231 breast tumor cells. It was demonstrated that the metal cluster was of minor importance for effects on cellular activity if an alkyne was used as ligand. Generally, no correlation existed between growth inhibition and COX activity. Cellular growth inhibition and antiproliferative activity at higher concentrations of the most active compounds Prop-ASS-Co(4) and Prop-ASS-Ru(3) correlated well with apoptosis induction.


Immunogenic properties of alpha (1----6) dextran, its protein conjugates, and conjugates of its breakdown products in mice.
Mice were immunized with alpha (1-6) dextran, either as such or coupled to protein carriers, and their anti-dextran response was measured by a solid-phase radioimmunoassay and the Farr assay. Like earlier investigators we found that protein-conjugated dextran was more antigenic than plain dextran. Our novel findings were that (1) a standard dose (30 micrograms of dextran per injection) coupled to strongly antigenic protein (chicken serum albumin (CSA) was three times more antigenic than dextran coupled to weakly antigenic bovine serum albumin (BSA); (2) dextrans of low molecular weight (1000-10,000 daltons) coupled to CSA induced at least ten times stronger secondary responses than did a similarly coupled macromolecular dextran (5-40 million daltons); (3) variation of the CHO/protein ratio from 0.3 to 1 had little effect on the antigenicity of the dextran. Increase of the ratio from one appeared to decrease immunogenicity when BSA was the carrier but not when CSA was the carrier.


Ovarian follicle cells in torpedo marmorata synthesize vitellogenin.
The pattern of vitellogenesis is similar in all non-mammalian vertebrates: the liver, under oestrogenic stimulus, synthesizes vitellogenin (VTG) that, via the maternal circulation, is delivered to the oocyte and here internalized by receptor-mediated endocytosis (Wallace, 1985: Development Biology. A comprehensive synthesis. Vol. 1 Oogenesis:127-177; Schneider, 1996: Int Rev Cytol 166:103-134; LaFleur, 1999: Encyclopedia of Reproduction Vol. 4:985-992). The contribution to vitellogenesis of different components of the ovarian follicle has also been reported in amphibians (Wallace, 1985), squamate reptiles (Ghiara and Limatola, 1980: Acta Embryol Morphol Exper 1:5-6; Andreuccetti, 1992: J Morphol 212:1-11), and recently, supporting previous reports (Chieffi and Pierantoni, 1987: Hormones and Reproduction in Fishes, Amphibians and Reptiles Single vol.:117-144), in Torpedo marmorata (Prisco et al., 2001: Perspective in comparative endocrinology: Unity and diversity Single vol.:1197-1201; Prisco et al., 2002b: Gen Comp Endocrinol 128:171-179). The present investigation, performed with immunoblotting, immunohistochemical, and in situ hybridization techniques during different stages of follicular growth in T. marmorata, shows that, as previously supposed (Prisco et al., 2002b), granulosa cells in both previtellogenic and vitellogenic phases actively synthesize VTG. This is the first time among vertebrates that the synthesis of this protein has been found to occur also within the ovarian follicle. The present data also demonstrate that the contribution of granulosa cells becomes particularly evident during vitellogenesis. Indeed, in vitellogenic follicles, small, intermediate, and pyriform-like cells cross-react with an anti-VTG antibody and are positive to a hybridization signal with a VTG mRNA probe. By contrast, in previtellogenesis only the enlarged cells, i.e., intermediate and pyriform-like cells, are involved in VTG synthesis.


Interstitial pneumonitis after treatment with pemetrexed: a rare event?
Pneumonitis after pemetrexed has been rarely reported in conjunction with radiation therapy. Two cases of pemetrexed-induced pneumonitis in different clinical settings and with unequal outcomes are discussed with a review of the literature. Two patients with stage IIIB non-small cell lung cancer developed interstitial lung disease after chemotherapy with pemetrexed. The first patient was previously treated with thoracic radiotherapy, and radiation pneumonitis was initially suspected. He died shortly after pemetrexed reexposition. The second patient developed pemetrexed-induced interstitial lung disease despite no prior radiotherapy. After discontinuation of pemetrexed and administration of steroids, pneumonitis resolved completely. Interstitial lung disease is a rare but potentially fatal side effect of pemetrexed. It occurs more often after radiotherapy but can also be encountered in the absence of radiotherapy. Reexposition to pemetrexed may lead to severe interstitial lung disease and even death and should be strictly avoided.


Hierarchical climate-driven dynamics of the active channel length in temporary streams.
Looking across a landscape, river networks appear deceptively static. However, flowing streams expand and contract following ever-changing hydrological conditions of the surrounding environment. Despite the ecological and biogeochemical value of rivers with discontinuous flow, deciphering the temporary nature of streams and quantifying their extent remains challenging. Using a unique observational dataset spanning diverse geomorphoclimatic settings, we demonstrate the existence of a general hierarchical structuring of river network dynamics. Specifically, temporary stream activation follows a fixed and repeatable sequence, in which the least persistent sections activate only when the most persistent ones are already flowing. This hierarchical phenomenon not only facilitates monitoring activities, but enables the development of a general mathematical framework that elucidates how climate drives temporal variations in the active stream length. As the climate gets drier, the average fraction of the flowing network decreases while its relative variability increases. Our study provides a novel conceptual basis for characterizing temporary streams and quantifying their ecological and biogeochemical impacts.


Activation of AMP-activated protein kinase alleviates homocysteine-mediated neurotoxicity in SH-SY5Y cells.
Mammalian AMP-activated protein kinase (AMPK) acts as a metabolite-sensing protein kinase in multiple tissues. Recent studies have shown that AMPK activation also regulates intracellular signaling pathways involved in cellular survival and apoptosis. Previously, we have reported that AMPK activation alleviates the endoplasmic reticulum (ER) stress-mediated neurotoxicity and tau hyperphosphorylation caused by palmitate. Therefore, we investigated whether AMPK activation alleviates ER stress-mediated neurotoxicity in SH-SY5Y human neuroblastoma cells incubated with homocysteine. Regulation of AMPK activity by isoflavone was also determined to investigate the underlying mechanism of its neuroprotective effect. Treatment of SH-SY5Y human neuroblastoma cells with N (1)-(β-D-ribofuranosyl)-5-aminoimidazole-4-carboxamide (AICAR), a pharmacological activator of AMPK, significantly protected cells against cytotoxicity imposed by tunicamycin and homocysteine. Homocysteine significantly suppressed AMPK activation, which was alleviated by AICAR. We observed a significant inhibition of the unfolded protein response by AICAR in cells incubated with homocysteine, suggesting a protective role of AMPK activation against ER stress-mediated neurotoxicity. AICAR also significantly reduced tau hyperphosphorylation by inactivating glycogen synthase kinase-3β and c-Jun N-terminal kinase in cells incubated with homocysteine. Furthermore, treatment of cells with soy isoflavone, genistein and daidzein significantly activated AMPK, which was repressed by tunicamycin and homocysteine. Therefore, our results suggest that AMPK activation by isoflavone as well as AICAR alleviates homocysteine-mediated neurotoxicity in SH-SY5Y cells.


Vascular endothelial growth factor and its receptor Flk-1 are expressed in the hippocampus following entorhinal deafferentation.
Vascular endothelial growth factor (VEGF) has been thought of as a mitogen that promotes proliferation of endothelial cells and as a neurotrophic factor that stimulates neurogenesis and axonal growth in both peripheral and central nervous systems. To investigate the potential involvement of VEGF in the lesion-induced reorganization in the brain, the expression changes of VEGF and its receptor Flk-1 were analyzed in the mouse hippocampus after transections of the entorhinal afferents. In situ hybridization and immunohistochemistry showed the time-dependent expression upregulation of VEGF mRNA and protein in the entorhinally denervated hippocampal stratum lacunosum-moleculare and dentate outer molecular layer, which initiated by 3 days postlesion, reached its maximum at 7-15 days postlesion, still persisted by 30 days postlesion for protein, and recovered to the normal levels at 30 days postlesion for mRNA and at 60 days postlesion for protein. Double labeling of VEGF and glial fibrillary acidic protein revealed that VEGF-expressing cells in the denervated areas were reactive astrocytes. Semi-quantitative RT-PCR analysis showed that VEGF receptor Flk-1 mRNA was also time-dependently upregulated in the deafferented hippocampus with its maximal elevation at 7-15 days postlesion while the Flt-1 mRNA levels remained unchanged at any time point we examined. Immunohistochemistry analysis also displayed the upregulation of Flk-1 protein in the denervated stratum lacunosum-moleculare and outer molecular layer with a time course similar to that of VEGF mRNA upregulation. Flk-1 receptors were found to be expressed not only by reactive astrocytes but also by neurites, which most likely belong to sprouting axons by 7 days postlesion and regrowing dendrites by 15-30 days postlesion. From these data we suggest that the spatiotemporal upregulation of VEGF and Flk-1 in the hippocampus is induced by entorhinal deafferentation and that VEGF may be involved in the structural reorganization in the deafferented hippocampus via directly or indirectly promoting neurite growth.


Mitochondrial oxygen sensing of acute hypoxia in specialized cells - Is there a unifying mechanism?
Acclimation to acute hypoxia through cardiorespiratory responses is mediated by specialized cells in the carotid body and pulmonary vasculature to optimize systemic arterial oxygenation and thus oxygen supply to the tissues. Acute oxygen sensing by these cells triggers hyperventilation and hypoxic pulmonary vasoconstriction which limits pulmonary blood flow through areas of low alveolar oxygen content. Oxygen sensing of acute hypoxia by specialized cells thus is a fundamental pre-requisite for aerobic life and maintains systemic oxygen supply. However, the primary oxygen sensing mechanism and the question of a common mechanism in different specialized oxygen sensing cells remains unresolved. Recent studies unraveled basic oxygen sensing mechanisms involving the mitochondrial cytochrome c oxidase subunit 4 isoform 2 that is essential for the hypoxia-induced release of mitochondrial reactive oxygen species and subsequent acute hypoxic responses in both, the carotid body and pulmonary vasculature. This review compares basic mitochondrial oxygen sensing mechanisms in the pulmonary vasculature and the carotid body.


Cross-ancestry Genomic Data Reveal New Susceptibility Loci in Lung Cancer.
Five new susceptibility loci were identified in lung cancer using cross-ancestry genome-wide studies.


Structural and genomic DNA analysis of a putative transcription factor SCO5550 from Streptomyces coelicolor A3(2): regulating the expression of gene sco5551 as a transcriptional activator with a novel dimer shape.
SCO5550 from the model actinomycete Streptomyces coelicolor A3(2) was identified as a putative transcriptional regulator, and classified into the MerR family by sequence analysis. Recombined SCO5550 was successfully produced in Rhodococcus erythropolis, which can be used to stably express recombinant protein by optimizing the temperature over a wide range (4-35 °C). Crystal structure analysis showed that the dimerization domain (C-terminal domain) of SCO5550 has a novel fold and forms a new dimer shape, whereas the DNA-binding domain (N-terminal domain) is very similar to those of MerR family members. Such the new dimer form suggests that SCO5550 may define a new subfamily as a new member of the MerR family. Binding DNA sequence analysis of SCO5550 using the genomic systematic evolution of ligands by exponential enrichment (gSELEX) and electrophoretic mobility shift assay (EMSA) indicated that SCO5550 regulates the expression of the immediately upstream gene sco5551 encoding a putative protein, probably as a transcriptional activator.


The Notch locus of Drosophila melanogaster.
The Notch locus appears to correspond to a 37 kb transcription unit. Homologous poly(A)+ RNA is about 11.7 kb. Nine RNA coding regions have been detected and localized within the transcription unit by S1 nuclease analyses. These range in size from 130 to 7250 bp. Sequences within the 3' coding region are variably used. Twenty-four Notch locus "point" mutations have been examined. Seven are associated with DNA insertions. Four insertions are associated with dominant mutations and are located within or very near RNA coding sequences of the 37 kb transcription unit. Three insertions are associated with recessive mutations and fall within intervening sequences. The positions of all insertions agree with the genetic map. It is estimated that, in the Notch region, one map unit corresponds to approximately 275 kb. On this basis, most of the 17 mutations that were not associated with DNA insertions can be placed within coding sequences of the 37 kb transcription unit.


Synthesis of Biotinylated PAMAM G3 Dendrimers Substituted with R-Glycidol and Celecoxib/Simvastatin as Repurposed Drugs and Evaluation of Their Increased Additive Cytotoxicity for Cancer Cell Lines.
Recent achievement in anticancer therapy considers the application of repurposed drugs in optimal combinations with the use of specific carriers for their targeted delivery. As a result, new optimized medications with reduced side effects can be obtained. In this study, two known anticancer drugs, celecoxib and/or simvastatin, were conjugated covalently with PAMAM G3 dendrimer and tested in vitro against human squamous carcinoma (SCC-15-15) and glioblastoma (U-118 MG) cells, as well as normal human fibroblasts (BJ). The obtained conjugates were also substituted with biotin and R-glycidol to increase their affinity for cancer cells and were characterized with NMR spectroscopy and dynamic light scattering technique. Conjugates furnished with two celecoxib and four simvastatin residues revealed the very high effectiveness and dramatically decreased the SCC-15 and U-118 MG cell viability at very low concentrations with IC50 equal to about 3 µM. Its action was 20-50-fold stronger than that of either drug alone or as a mixture. Combined conjugate revealed also additive action since it was 2-8-fold more effective than conjugates with either single drug. The combined conjugate revealed rather low specificity since it was also highly cytotoxic for BJ cells. Despite this, it may be concluded that biotinylated and R-glycidylated PAMAM G3 dendrimers substituted with both celecoxib and simvastatin can be considered as a new perspective anticancer agent, effective in therapy of malignant, incurable glioblastomas.


Suppression of presynaptic calcium currents by hypoxia in hippocampal tissue slices.
We tested the hypothesis that suppression of inward calcium current in presynaptic terminals is the cause of failure of synaptic transmission early during cerebral hypoxia. Postsynaptic responses in CA1 zone of hippocampal tissue slices were blocked either by the combined administration of 6,7-dinitroquinoxaline-2,3-dione (DNQX) and 3-((+-)-2-carboxypiperazine-4-yl)-propyl-1-phosphonic acid (CPP) or by lowering extracellular calcium concentration ([Ca2+]o). Repetitive orthodromic activation of central neurons caused transient decrease of [Ca2+]o (measured by ion selective microelectrodes) in neuropil, attributable to influx of Ca2+ in presynaptic terminals. Presynaptic [Ca2+]o responses were rapidly and reversibly suppressed when oxygen was withdrawn from hippocampal tissue slices. The 'resting' baseline level of [Ca2+]o declined at first gradually, then precipitously as in spreading depression (SD). Presynaptic volleys during high frequency train stimulation were also depressed somewhat before SD began. We conclude that (1) presynaptic Ca2+ currents fail during hypoxia, perhaps because 'resting' intracellular free Ca2+ activity is increased and, in part, also because of partial failure of presynaptic impulse conduction; (2) the influx of Ca2+ into brain cells in hypoxic spreading depression is not mediated by glutamate/aspartate dependent channels.


A novel calcium-dependent soluble inorganic pyrophosphatase from the trypanosomatid Leishmania major.
A single-copy gene IPP encoding a putative soluble inorganic pyrophosphatase (LmsPPase, EC 220.127.116.11) was identified in the genome of the parasite protozoan Leishmania major. The full-length coding sequence (ca. 0.8 kb) was obtained from genomic DNA by polymerase chain reaction (PCR) and cloned into an Escherichia coli expression vector, and was overexpressed for functional protein purification and characterization. The recombinant LmsPPase, purified to electrophoretic homogeneity by a two-step chromatography procedure, exhibited a predicted molecular mass of ca. 30 kDa. The enzyme has an absolute requirement for divalent cations, exhibits a pH optimum of 7.5-8.0 and does not hydrolyze polyphosphates or adenosine triphosphate (ATP). LmsPPase differs from previously studied soluble pyrophosphatases with respect to cation selectivity, Ca(2+) being far more effective than Mg(2+). Comparisons to known sPPases show a short N-terminal extension predicted to be a mitochondrial transit peptide, and changes in active-site residues and the neighboring region. Subcellular fractionation of L. major promastigotes suggests a mitochondrial localization. Molecular phylogenetic analysis indicates that LmsPPase is a highly divergent eukaryotic Family I sPPase, perhaps an ancestral class of eukaryotic sPPases functionally adapted to a calcium-rich, probably mitochondrial, environment.


Conformational stability of Cys45-alkylated and hydrogen peroxide-oxidised glutathione S-transferase.
A highly reactive cysteine residue in class pi glutathione S-transferases enhances their susceptibility to chemical alkylation and oxidative stress. Alkylation of the reactive Cys45 in the porcine class pi enzyme (pGSTP1-1) with either N-iodoacetyl-N'-(5-sulpho-1-naphthyl)ethylenediamine or iodoacetamide results in a loss of enzyme activity and glutathione-binding function. Similarly, oxidation of pGSTP1-1 with hydrogen peroxide (H2O2) also results in a loss of catalytic and glutathione-binding function, but these effects are reversed by the addition of 5 mM glutathione or dithiothreitol. Analysis by SDS-PAGE of the H2O2-oxidised enzyme indicates oxidation-induced formation of disulphide bonds involving Cys45. Equilibrium-unfolding studies with guanidinium chloride indicate that the unfolding of Cys45-alkylated and H2O2-oxidised pGSTP1-1 can be described by a two-state model in which the predominant thermodynamically stable species are the folded dimer and unfolded monomer. Unfolding transition curves suggest that the introduction of a large and bulky AEDANS at Cys45 does not affect the unfolding pathway for pGSTP1-1. H2O2-oxidised pGSTP1-1, on the other hand, appears to follow a different unfolding pathway. This appears not to be a result of the introduction of disulphide bonds since the reduction of these bonds in the oxidised protein with dithiothreitol does not affect the unfolding transition. Furthermore, the conformational stability of the oxidised protein is significantly diminished (delta G(H2O) = 11.6 kcal/mol) when compared with unmodified and AEDANS-alkylated enzyme (delta G(H2O) = 22.5 kcal/mol).


Past history and degree of depression in paraplegic individuals.
The purpose of this study was to attempt to collect sufficient data to substantiate the clinical impression that the degree of depression in the paraplegic and his past history are correlated. It was hypothesised that the poorer the past history, the greater the degree of depression in paraplegic individuals. This study was conducted on ten paraplegic subjects from the Spinal Cord Clinic at Columbia Presbyterian Medical Center, Department of Rehabilitation Medicine, based on data obtained from the administration and scoring of the Depression Scale (D-scale) of the Minnesota Multiphasic Personality Inventory (MMPI) and by the clinical observation by the consulting psychiatrist with the Spinal Injury Clinic. Past history was assumed measurable as 'favourable' to 'poor' by the use of an original questionnaire. The results of this investigation suggested a relationship between a paraplegic individual's past history and the degree of depression. The study was deemed significant in presentation of an original past history questionnaire and in the prediction of future difficulties in the rehabilitation of paraplegics so that preventive measures could be instituted.


Formulation and dosage form design in drug-induced topical irritation of the gastrointestinal tract.
To test drugs for topical effects on gastrointestinal mucosa, a new in situ rabbit colon model was used that permits direct application of drugs in suspensions from gel cups, solutions, or commercially available tablets and capsules onto rabbit colonic mucosa for up to 8 hr. For each agent tested an irritation index was calculated--the product of the area of the mucosa affected by drug exposure and a numerical score for observed effect. Irritation indices ranged from 0 (no effect) to 25.6 (maximal irritation measurable). In general, the immediate release of drug onto tissue elicited the greatest effect, whereas slow or controlled release of drug produced the least response. Topical irritation was found to be a function of (1) the drug, (2) the formulation, (3) the delivery rate, and (4) the concentration. The gastrointestinal therapeutic system (GITS) of potassium chloride and of brompheniramine/pseudoephedrine produced far less irritation than current commercial formulations of these drugs. The rabbit colon model is proposed as a useful screening tool during drug development to aid in selecting the formulation of an oral dosage form that will minimize topical irritation.


Bilateral genital asymmetry in livebearer fishes of the genus Phalloceros Eigenmann, 1907, with description of a new species from coastal drainages of southern Brazil (Cyprinodontiformes: Poeciliidae).
A new species of Phalloceros is described from the Tijucas, Maruim, Aririú, Cubatão do Sul, Rio da Madre, D'Una and Tubarão river basins in southern Brazil. The new species is diagnosed among congeners by the bilateral asymmetry of a hood-like structure located immediately anterior to the urogenital papilla of females. The hood is fused with the urogenital papilla on the right side, but free from the papilla on the left side. This hood is reported for the first time in the genus and is present in six of the 21 valid species of Phalloceros, but its bilateral asymmetry is an exclusive condition of the new species. These species do not constitute a monophyletic group. The new species is likely to be phylogenetically related to P. caudimaculatus, whose females have a symmetrical hood.


Design and early clinical evaluation of selective inhibitors of monoamine oxidase.
1. Selective inhibitors of the monoamine oxidase (MAO) isoenzymes, types A and B, are of potential therapeutic utility. Brain selectivity would overcome the risk of tyramine interactions which have been shown to occur with selective MAO-A but not MAO-B inhibitors. 2. (E)-3-Fluoroallylamines of general structure, FHC = C(R)CH2NH2 have been designed as enzyme-activated, irreversible inhibitors of these enzymes. Two compounds, MDL 72145 (R = 3,4 dimethoxyphenyl) and MDL 72974 (R = 4-fluorophenethyl), are selective and irreversible inhibitors of MAO type B which in vivo show high inhibitory potency against the rat brain enzyme (ED50 0.35 and 0.18 mg/kg p.o., respectively). In animals, these inhibitors do not potentiate the cardiovascular effects of tyramine and have no amphetamine-like effects. However, they do potentiate the central effects of L-Dopa and prevent the neurotoxic effects of MPTP in both mice and monkeys. 3. In early clinical studies, MDL 72145 has been shown to be a potent, long-acting inhibitor of MAO type B. Doses of 16 mg per patient totally inhibit platelet enzyme without potentiating the cardiovascular effects of oral tyramine. Compounds of this type should prove useful in Parkinson's disease. 4. Selective inhibition of brain MAO-A can be achieved by using the bioprecursor amino acid MDL 72394 (E-beta-fluoromethylene-m-tyrosine). This amino acid is decarboxylated by aromatic L-amino acid decarboxylase (AADC) to liberate MDL 72392 (R = 3-hydroxyphenyl), a potent irreversible inhibitor of MAO-A. Combination of MDL 72394 with a peripherally selective inhibitor of AADC (e.g., carbidopa) restricts MAO inhibition to the brain. Consequently, under these conditions, there is a greatly reduced propensity to potentiate the cardiovascular effects of tyramine. 5. This has been confirmed in human volunteers; MDL 72394 (8 mg), combined with carbidopa, substantially decreased urinary MHPG and plasma DHPG concentrations with minimal potentiation of the cardiovascular effects of i.v. tyramine. These results predict that such therapy has potential in the treatment of affective disorders.


Significance of pain in psychiatric hospital patients.
In a consecutive series of 227 psychiatric hospital admissions, data were recorded in respect of the complaint of pain. Eighty-six (38%) had pain. Fourty-four (19%) mentioned it spontaneously and 49 (22%) had no relevant physical cause. Women were affected more often than men (P less than 0.01) and tended to complain more often of severe pain (P less than 0.01). Severe pain was more often reported spontaneously (P less than 0.02). Also, the longer pain lasted the more likely the patient was to report it spontaneously (P less than 0.02). Men more often had a relevant physical diagnosis (P less than 0.05) and the low back was the commonest site of pain in them. Pain was relatively often associated with diagnoses of anxiety and personality disorder and relatively infrequently with schizophrenia, organic brain syndromes and transient situational disturbances. It is concluded that whilst there is a strong association between pain and psychiatric illness, this is less prominent, paradoxically, in some of the more severe psychiatric disturbances.


Are morphologic and topographic alterations of the mandibular fossa after fixed functional treatment detectable on tomograms? Visual classification and morphometric analysis.
The goal was to evaluate if changes in morphology and topography of the mandibular fossa after Functional Mandibular Advancer (FMA) treatment are detectable on tomograms. Furthermore, the suitability of digital tomograms (DT) over magnetic resonance imaging (MRI) for this particular question was investigated. In all, 25 patients (14 female, 11 male) with a skeletal class II malocclusion received treatment with a FMA. DTs were available prior to (T1) and after (T2) FMA treatment. A total of 50 temporomandibular joints were investigated. The mandibular fossae were evaluated metrically and visually regarding treatment-induced alterations. A p < 0.05 was set as the level for statistical significance for all tests. Results were compared to the results of a recent MRI study. Visual inspection of all 50 joints in the DT at T1 and T2 revealed no alterations of the fossa shape in the sagittal plane; 24 patients showed identical morphology of right and left joints. The metrical analysis revealed no significant changes regarding width, depth and ratio thereof between T1 and T2. There also were no bilateral differences. Another 18 different distance measurements between porion, mandibular fossa, articular eminence and pterygoid fossa showed no significant changes. There was no detectable proof of a fossa shift. No changes in the sagittal plane, mandibular fossa, the articular tubercle, or a possible fossa shift were found in the DT of class II patients after FMA treatment. DT and MRI measurements and the visual inspection revealed identical findings; thus, DT appears to be a valuable research tool for sagittal analysis of mandibular fossa changes.


[Ankylosing temporomandibular arthritis following masseter cellulitis. 3 cases].
Treatment of temporomandibular ankylosis complicating masseter muscle cellulitis in three patients, as a result of a dental lesion, required wide excision of bone structure and, in two cases, of adjacent infected soft tissues.


Tofogliflozin long-term effects on atherosclerosis progression and major clinical parameters in patients with type 2 diabetes mellitus lacking a history of cardiovascular disease: a 2-year extension study of the UTOPIA trial.
This study aimed to assess the long-term effects of tofogliflozin, a sodium-glucose cotransporter 2 (SGLT2) inhibitor, on atherosclerosis progression and major clinical parameters in patients with type 2 diabetes lacking an apparent history of cardiovascular disease. This was a prospective observational 2-year extension study of the "Using TOfogliflozin for Possible better Intervention against Atherosclerosis for type 2 diabetes patients (UTOPIA)" trial, a 2-year randomized intervention study. The primary endpoints represented changes in the carotid intima-media thickness (IMT). Secondary endpoints included brachial-ankle pulse wave velocity (baPWV) and biomarkers for glucose metabolism, lipid metabolism, renal function, and cardiovascular risks. The mean IMT of the common carotid artery (IMT-CCA) significantly decreased in both the tofogliflozin (- 0.067 mm, standard error 0.009, p < 0.001) and conventional treatment groups (- 0.080 mm, SE 0.009, p < 0.001) throughout the follow-up period; however, no significant intergroup differences in the changes (0.013 mm, 95% confidence interval (CI) - 0.012 to 0.037, p = 0.32) were observed in a mixed-effects model for repeated measures. baPWV significantly increased in the conventional treatment group (82.7 ± 210.3 cm/s, p = 0.008) but not in the tofogliflozin group (- 17.5 ± 221.3 cm/s, p = 0.54), resulting in a significant intergroup difference in changes (- 100.2 cm/s, 95% CI - 182.8 to - 17.5, p = 0.018). Compared to the conventional treatment group, tofogliflozin significantly improved the hemoglobin A1c and high-density lipoprotein cholesterol levels, body mass index, abdominal circumference, and systolic blood pressure. The frequencies of total and serious adverse events did not vary significantly between the groups. Tofogliflozin was not associated with improved inhibition of carotid wall thickening but exerted long-term positive effects on various cardiovascular risk factors and baPWV while showing a good safety profile.


Unexpected role of stress as a possible resilience mechanism upon mild traumatic brain injury (mTBI) in mice.
Mild traumatic brain injury (mTBI) is common and associated with cognitive impairment. Stress and mTBI are known to modulate the neural function. The present study aims at exploring the effect of prior stress exposure on cognitive function following mTBI. Eight weeks old male ICR mice were subjected to either stress induced by forced swimming stress alone, stress followed by an immediate mTBI, or stress followed by 30 min break and then mTBI. We had two control groups: SHAM group - a control group which was not exposed to stress nor to mTBI and control mTBI group - a control group which was exposed only to TBI with no stress. Mice were weighed prior and at 12, 24 h and 1 week following interventions. Motor evaluation was conducted by rotarod. Behavioral changes were evaluated using open field, Y maze, elevated plus maze and staircase tests, at 12 h and 1 week following interventions. Brain levels of NMDAR subunits (R1, R2A, R2B), GABABR1, glucocorticoid and mineralocorticoid receptors (GR, MR) were evaluated using western blot. Stress alone, mTBI alone, and stress followed by immediate mTBI resulted in a significant weight loss compared to control (p < 0.05). Stress 30 min prior to mTBI had a protective effect on weight (p = 0.14 compared to control). The stress and mTBI alone groups showed reduced time at the center of the open field arena 1 week after intervention (p < 0.05 for both). Time in the novel arm of the Y maze was significantly shorter in the mTBI and stress followed by delayed mTBI (p = 0.02). Immediate stress prior to mTBI had normalized times in the novel arm (p = 0.95 compared to control). Combination of stress and mTBI significantly modified NMDAR subunits levels (increased NMDAR1, p < 0.008, decreased NMDAR2A p = 0.02) as well as increased MR levels (p = 0.04). Exposure to stress prior to mTBI may improve the cognitive consequences of mTBI. These data may point towards a novel, unexpected role of stress as a possible resilience mechanism in the setting of mTBI.


MPTherm-pred: Analysis and Prediction of Thermal Stability Changes upon Mutations in Transmembrane Proteins.
The stability of membrane proteins differs from globular proteins due to the presence of nonpolar membrane-spanning regions. Using a dataset of 929 membrane protein mutations whose effects on thermal stability (ΔTm) were experimentally determined, we found that the average ΔTm due to 190 stabilizing and 232 destabilizing mutations occurring in membrane-spanning regions are 2.43(3.1) °C and -5.48(5.5) °C, respectively. The ΔTm values for mutations occurring in solvent-exposed regions are 2.56(2.82) and - 6.8(7.2) °C. We have systematically analyzed the factors influencing the stability of mutants and observed that changes in hydrophobicity, number of contacts between Cα atoms and frequency of aliphatic residues are important determinants of the stability change induced by mutations occurring in membrane-spanning regions. We have developed structure- and sequence-based machine learning predictors of ΔTm due to mutations specifically for membrane proteins. They showed a correlation and mean absolute error (MAE) of 0.72 and 2.85 °C, respectively, between experimental and predicted ΔTm for mutations in membrane-spanning regions on 10-fold group-wise cross-validation. The average correlation and MAE for mutations in aqueous regions are 0.73 and 3.7 °C, respectively. These MAE values are about 50% lower than standard deviations from the mean ΔTm values. The reliability of the method was affirmed on a test set of mutations occurring in evolutionary independent protein sequences. The developed MPTherm-pred server for predicting thermal stability changes upon mutations in membrane proteins is available at https://web.iitm.ac.in/bioinfo2/mpthermpred/. Our results provide insights into factors influencing the stability of membrane proteins and can aid in designing mutants that are more resistant to thermal stress.


The citrus flavonone naringenin reduces lipopolysaccharide-induced inflammatory pain and leukocyte recruitment by inhibiting NF-κB activation.
Lipopolysaccharide (LPS) is the major structural component of Gram-negative bacteria cell wall and a highly pro-inflammatory toxin. Naringenin is found in Citrus fruits and exhibits antioxidant and anti-inflammatory properties through inhibition of NF-κB activation but its effects in LPS-induced inflammatory pain and leukocyte recruitment were not investigated yet. We investigated the effects of naringenin in mechanical hyperalgesia, thermal hyperalgesia and leukocyte recruitment induced by intraplantar injection of LPS in mice. We found that naringenin reduced hyperalgesia to mechanical and thermal stimuli, myeloperoxidase (MPO, a neutrophil and macrophage marker) and N-acetyl-β-D-glucosaminidase (NAG, a macrophage marker) activities, oxidative stress and cytokine (TNF-α, IL-1β, IL-6, and IL-12) production in the paw skin. In the peritoneal cavity, naringenin reduced neutrophil and mononuclear cell recruitment, and abrogated MPO and NAG activity, cytokine and superoxide anion production, and lipid peroxidation. In vitro, pre-treatment with naringenin inhibited superoxide anion and cytokine (TNF-α, IL-1β, IL-6, and IL-12) production by LPS-stimulated RAW 264.7 macrophages. Finally, we demonstrated that naringenin inhibited NF-κB activation in vitro and in vivo. Therefore, naringenin is a promising compound to treat LPS-induced inflammatory pain and leukocyte recruitment.


Green synthesis of silver nanoparticles using methanolic root extracts of Diospyros paniculata and their antimicrobial activities.
Since the discovery and subsequent widespread use of antibiotics, a variety of bacterial species of human and animal origin have developed numerous mechanisms that render bacteria resistant to some, and in certain cases to nearly all antibiotics, thereby limiting the treatment options and compromising effective therapy. In the present study, the green synthesis of nanoparticles is carried out by the reduction of silver acetate in the presence of crude methanolic root extracts of Diospyros paniculata, a member of family Ebenaceae. The UV-Vis absorption spectrum of the biologically reduced reaction mixture showed the surface plasmon peak at 428 nm, a characteristic peak of silver nanoparticles. X-ray diffraction (XRD) analysis confirmed the face-centered cubic crystalline structure of metallic silver. The average diameter of Ag NPs is about 17 nm from Transmission Electron Microscopy (TEM) which is in good agreement with the average crystallite size (19 nm) calculated from XRD analysis. Further the study has been extended to the antimicrobial activity against test pathogenic Gram (+), Gram (-) bacterial and fungal strains. The biologically synthesized silver nanoparticles showed promising activity against all the tested pathogenic strains and the activity has been enhanced with the increased dose levels.


The obesity epidemic: metabolic imprinting on genetically susceptible neural circuits.
The apparent obesity epidemic in the industrialized world is not explained completely by increased food intake or decreased energy expenditure. Once obesity develops in genetically predisposed individuals, their obese body weight is avidly defended against chronic caloric restriction. In animals genetically predisposed toward obesity, there are multiple abnormalities of neural function that prime them to become obese when dietary caloric density and quantity are raised. Once obesity is fully developed, these abnormalities largely disappear. This suggests that obesity might be the normal state for such individuals. Formation of new neural circuits involved in energy homeostasis might underlie the near permanence of the obese body weight. Such neural plasticity can occur during both nervous system development and in adult life. Maternal diabetes, obesity, and undernutrition have all been associated with obesity in the offspring of such mothers, especially in genetically predisposed individuals. Altered brain neural circuitry and function often accompanies such obesity. This enhanced obesity may then be passed on to subsequent generations in a feed-forward, upward spiral of increasing body weight across generations. Such findings suggest a form of "metabolic imprinting" upon genetically predisposed neural circuits involved in energy homeostasis. Centrally acting drugs used for obesity treatment lower the defended body weight and alter the function of neural pathways involved in energy homeostasis. But they generally have no permanent effect on body weight or neural function. Thus, early identification of obesity-prone mothers, infants, and adults and treatment of early obesity may be the only way to prevent the formation of permanent neural connections that promote and perpetuate obesity in genetically predisposed individuals.


Mongoliitalea daihaiensis sp. nov., isolated from Daihai Lake in Inner Mongolia.
A novel Gram-negative strain, designated X100-76T, was isolated from Daihai Lake in Inner Mongolia, Republic of China. The strain was non-motile, non-spore-forming, long-rod-shaped, oxidase positive and catalase positive. Colonies incubated at 33 °C on 2216 marine agar medium for 3 days were circular, smooth, transparent, convex with clear edges, orange-red in colour and approximately 1.0 mm in diameter. Growth occurred at pH 6.5-12 (optimum pH 7.5), in 0-5.0% (w/v) NaCl (optimum 1.5%, w/v) and at 4-40 °C (optimum 28-33 °C). Phylogenetic analysis based on 16S rRNA gene sequences showed that X100-76T belongs to the genus Mongoliitalea and is most closely related to Mongoliitalea lutea MIM18T with 98.3% sequence similarity. The total genome size of X100-76T was 4,816,617 bp with a G + C content of 39.6%. The digital DNA-DNA hybridization and average nucleotide identity values between strain X100-76T and M. lutea MIM18T were both below the recommended cut-off values. Chemotaxonomic analysis revealed iso-C15:0, Sum Feature 4 (anteiso-C17:1 B and/or iso-C17:1 I), C15:1ω6c, Sum Feature 9 (C16:0 10-methyl and/or iso-C17:1ω9c), C16:0 and iso-C15:1 G as the major fatty acids, menaquinone MK-7 as the major isoprenoid quinone, diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine and phosphatidylinositol as the major cellular polar lipids. The results of polyphasic analysis demonstrated that X100-76T represents a novel species within the genus Mongoliitalea, for which the name Mongoliitalea daihaiensis sp. nov. is proposed. The type strain is X100-76T (= CGMCC 1.18762T = KCTC 82458T).


[Physical activity in the regional health plans in Spain: a review of proposals].
To identify if the regional government health plans in Spain include specific objectives and measures related to physical activity. Institutional websites were scanned to identify the most recent health plan of each regional government. The information included in the health plans on objectives and measures and the population groups targeted by these measures was extracted. The search was completed on 24 January 2022. Sixteen health plans were identified of which six are in force in 2022; 15 of these have incorporated specific objectives and measures on physical activity. Most of the actions are focus on general population, children-adolescents and elderly population. The plans of Cantabria, the Comunitat Valenciana and País Vasco cover physical activity more widely, proposing measures for seven of the eight identified population groups. Most of the health plans include measures related to physical activity and target two or more population groups. The objectives, specific measures and population groups were highly heterogeneous. This study could serve to prompt regional governments to review their health plans, update them and improve them in terms of physical activity. It seems advisable that the different health plans should have a homogeneous framework of proposals related to the promotion of physical activity.


Transfer of conjugative elements from rumen and human Firmicutes bacteria to Roseburia inulinivorans.
Studies on Firmicutes bacteria from the gut are hampered by a lack of gene transfer systems. Here the human colonic anaerobe Roseburia inulinivorans A2-194 was shown to be a transfer recipient for two conjugative transposons, Tn1545 from Eubacterium cellulosolvens and TnK10 from Clostridium saccharolyticum K10.


Molecular analysis of the genus Asparagus based on matK sequences and its application to identify A. racemosus, a medicinally phytoestrogenic species.
The plant Asparagus racemosus is one of the most widely used sources of phytoestrogens because of its high content of the steroidal saponins, shatavarins I-IV, in roots. The dry root of A. racemosus, known as "Rak-Sam-Sip" in Thai, is one of the most popular herbal medicines, used as an anti-inflammatory, an aphrodisiac and a galactagogue. Recently, the interest in plant-derived estrogens has increased tremendously, making A. racemosus particularly important and a possible target for fraudulent labeling. However, the identification of A. racemosus is generally difficult due to its similar morphology to other Asparagus spp. Thus, accurate authentication of A. racemosus is essential. In this study, 1557-bp nucleotide sequences of the maturase K (matK) gene of eight Asparagus taxa were analyzed. A phylogenetic relationship based on the matK gene was also constructed. Ten polymorphic sites of nucleotide substitutions were found within the matK sequences. A. racemosus showed different nucleotide substitutions to the other species. A polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of the matK gene was developed to discriminate A. racemosus from others. Only the 650-bp PCR product from A. racemosus could be digested with BssKI into two fragments of 397 and 253-bp while the products of other species remained undigested. Ten commercially crude drugs were analyzed and revealed that eight samples were derived from A. racemosus while two samples of that were not. Thus, the PCR-RFLP analysis of matK gene was shown to be an effective method for authentication of the medicinally phytoestrogenic species, A. racemosus.


Regional environmental regulation efficiency: spatiotemporal characteristics and influencing factors.
Research regarding the regional environmental regulation efficiency (ERE) and influencing factors can provide theoretical guidance for regions to improve their ERE effectively. By employing a two-step approach, the data envelopment analysis (DEA) CCR-BCC model is built with the inclusion of scale changes, and a Tobit model is developed to explore the influencing factors for the regional ERE, followed by an application to analyze the spatiotemporal variations of ERE in Jiangsu province from 2005 to 2015. It is found that in time dimension, the ERE lies generally in a weak effective interval of [0.5, 1) and displays a shock upward trend. In the spatial dimension, the ERE presents an obvious "bilateral effect", namely, the efficiency is high for both the southern and northern of Jiangsu province but lower for the middle area. Besides, GDP per capital, industrial structure, trade openness, and population growth are among the main influencing factors of ERE. The findings revealed that temporary short-term policies have noticeable impact on the regional ERE, and "matching effect" between the ERE and regional economic development does not present.


Diagnostic value of magnetic resonance imaging in cervical lymph node metastasis of oral squamous cell carcinoma.
The objective of this study was to investigate the correlation between magnetic resonance imaging (MRI) characteristics of cervical lymph nodes and the pathologically confirmed status of cervical lymph node metastasis (LNM) in oral squamous cell carcinoma (OSCC) and to provide imaging evaluation parameters for the clinical diagnosis of cervical lymph node status in OSCC. In a retrospective analysis, 79 patients who were first pathologically diagnosed with OSCC were included. The MRI-derived imaging parameters of the cervical lymph nodes were evaluated and the pathological status of lymph nodes in neck dissection specimens was reviewed. The relationship between the imaging parameters and cervical LNM was analyzed. The MRI-derived imaging parameters of 4419 lymph nodes were evaluated, and the pathological status of 2463 lymph nodes was reviewed. The MRI-derived shortest axial diameter (SAD) and unclear boundary of the cervical lymph node were significantly related to LNM. The cutoff value of SAD that enabled identification of LNM was 3.6 mm, and it was 4.2 and 4.1 mm for the prediction of overall survival and disease-specific survival, respectively. The MRI-derived parameters SAD and unclear boundary of the cervical lymph node correlated with LNM in OSCC. MRI-derived SAD larger than 3 mm warrants simultaneous neck dissection at initial surgery.


Peritonsillar abscess (PTA): clinical characteristics, microbiology, drug exposures and outcomes of a large multicenter cohort survey of 412 patients hospitalized in 13 French university hospitals.
The aim of this study was to describe the epidemiology of hospitalized patients with peritonsillar abscess (PTA). We conducted a multicenter survey in 13 French university hospitals in 2009-2012 describing 412 patients. Median age was 29 year (range, 2-84) and current smoking habit was reported by 177 (43 %) patients. Most of the patients (92 %) had consulted a physician for sore throat within 10 days before admission for PTA diagnosis. Additional symptoms such as visible tonsil abnormalities (83 %), tender cervical adenopathy (57 %) and fever ≥ 38.5 °C (53 %) were also reported. A total of 65 % patients (269/412) reported recent systemic anti-inflammatory agents (AIAs) exposure by medical prescription (70 %), self-medication (22 %), or both (8 %); 61 % and 27 % reported recent exposure to antibiotic and topical treatments for sore throat, respectively. Non-steroidal AIAs were used most often (45 %), particularly arylpropionic derivatives. A rapid diagnosis antigen test (RDT) for Streptococcus pyogenes was performed in 70 (17 %) patients and was positive in 17 (24 %), of whom 9 (53 %) were exposed to AIAs and 14 (82 %) to antibiotics. To treat PTA, antibiotic therapy was given to 392 (95 %) patients. Of 333 antibiotic prescriptions, amoxicillin-clavulanic acid and metronidazole were the most prescribed antibiotics (42 and 17 %, respectively). Surgical drainage of the abscess was performed in 119 (29 %) cases and tonsillectomy in 75 (18 %) cases. The clinical outcome was favorable during the hospital stay in 404 (98 %) patients. In conclusion, patients with sore throat are often exposed to AIAs before PTA diagnosis, and antibiotic prescription was not often based on the RDT positivity.


Occurrence of Rickettsia spp. and Coxiella burnetii in ixodid ticks in Kassena-Nankana, Ghana.
Ticks are arthropods of veterinary and medical importance which spread zoonotic pathogens that link animal and human health. In this study, ticks were collected from 448 livestock between February and December 2020 in the Kassena-Nankana Districts of Ghana and screened for the presence of zoonotic pathogens DNA using PCR and sequencing approaches. In total, 1550 ticks were collected and morphologically identified. Three tick genera were identified with Amblyomma variegatum (63%) as the predominant tick species collected. DNA was extracted from 491 tick pools and screened for the presence of DNA of Rickettsia spp. based on the 115 bp fragment of the 17 kDa surface protein and 639 bp of the Outer membrane protein A (ompA) gene and the 295 bp fragment of the transposase gene of Coxiella burnetii IS1111a element. From the 491 pools screened, the DNA of Rickettsia spp. and C. burnetii was detected in 56.8 and 3.7%, respectively. Coinfections were identified in 2.4% of the tick pools. Characterization of the Rickettsia spp. in this study based on the ompA gene showed that the DNA of Rickettsia africae and Rickettsia aeschlimannii accounted for 39.7 and 14.7%, respectively, and were 100% similar to sequences in GenBank. Most R. africae and C. burnetii infections occurred in ticks collected in the wet season, whereas R. aeschlimannii occurred mostly in the dry season. These pathogens are potential public health threats, thus there is a need to implement control measures to reduce the risk of infections in vulnerable populations.


Preference for oral sildenafil or intracavernosal injection in patients with erectile dysfunction already using intracavernosal injection for > 1 year.
Authors from Seoul describe their experience with patients already on triple therapy by intracavernosal injection who changed to oral sildenafil. Rather surprisingly, they found that patients had had a greater preference than expected for triple therapy, feeling that they had a better quality of erection on intracavernosal injection. The subject of the effect of renal transplantation on sperm quality and sex hormone levels is discussed by authors from Teheran. They found that sperm morphology and density remained unchanged, but there were significant improvements in sperm mobility. There was also an improvement in hormone levels and sexual function. To investigate the efficacy and preference for oral sildenafil or intracavernosal injection (ICI) therapy in patients with erectile dysfunction (ED) already using ICI. In all, 69 patients with ED (mean age 55.1 years, sd 12.3) on ICI therapy with triple solution (papaverine/phentolamine/prostaglandin-E1) for > 1 year were recruited for the study. Their erection quality, adverse reactions and selection rate of oral sildenafil or ICI as treatment, after using sildenafil for 3 months, and the reasons for their preferences, were compared between the regimens, Overall, 52 men (75%) responded to sildenafil; of these men, the erection quality with ICI was better than that with sildenafil in 46 (89%) and 16 (31%) preferred ICI as their treatment. Eighteen patients (35%) used each treatment alternately and 18 (35%) used sildenafil exclusively. The main reason given by patients for choosing ICI was a better quality of erection (74%). More patients with ED and using ICI preferred it as their main treatment than was expected, even though they had a good response to oral sildenafil. A better quality of erection with ICI was the reason why experienced patients chose this method, differing from the choice of patients starting treatment for ED.


Meta-analysis of the cognitive effects of the catechol-O-methyltransferase gene Val158/108Met polymorphism.
Cognitive endophenotypes may further our understanding of the genetic basis of psychiatric disorders, and the catechol-O-methyltransferase (COMT) gene is a promising candidate gene for both cognitive function and disorder. We conducted a meta-analysis of reported associations between the COMT Val158/108Met polymorphism and measures of memory and executive function. The PubMed database was searched for studies relating cognitive functions and the COMT Val158/108Met polymorphism. This enabled meta-analyses of six cognitive phenotypes (Trail Making task, verbal recall, verbal fluency, IQ score, n-back task, and Wisconsin Card Sorting Test). Data were extracted by two reviewers and included cognitive scores by COMT genotype, publication year, diagnostic status, ancestry, proportion of male participants, and whether genotype frequencies were consistent with Hardy-Weinberg equilibrium. We found no association between COMT genotype and the majority of phenotypes. There was evidence of association with IQ score (d = .06), which did not differ significantly by ancestry, sex, average sample age, or patient status. For the n-back task, there was no robust evidence for genetic association, but the effect size was significantly larger in patient (d = .40) than nonpatient (d = -.27) populations, larger in both samples with fewer male subjects, and those of greater average age. There was also evidence of publication bias and decreasing effect sizes with later publication. Despite initially promising results, the COMT Val158/108Met polymorphism appears to have little if any association with cognitive function. Publication bias may hamper attempts to understand the genetic basis of psychological functions and psychiatric disorders.


Interferon with novel characteristics produced by human mononuclear leukocytes.
Treatment of human peripheral blood mononuclear leukocytes with phytohaemmaglutinin and the tumour promoter teleocidin, results in the production of large amounts of interferon-gamma and significant amounts of a novel interferon-like substance which we tentatively class as interferon-delta. This novel interferon type possesses all the important characteristics of classical interferon but, of various cell types tested, has antiviral activity only in trisomy-21 human fibroblasts. It differs decisively from previously identified interferon types in its antigenic, biological and physicochemical properties.


XRD, FTIR and ultrasonic investigations of cadmium lead bismuthate glasses.
Cadmium lead bismuthate glasses in the system xCdO-(1-x)[0.5PbO + 0.5Bi2O3](40 mol% ≤ x ≤ 90 mol%) were successfully prepared by melt-quenching method. The structural and elastic properties have been investigated using XRD, FTIR and ultrasonic pulse-echo techniques. The XRD patterns confirmed the amorphous nature of the samples prepared. Density and ultrasonic velocity data were used to evaluate various elastic properties. Addition of CdO gave rise to decreased density and molar volume and increased elastic moduli, micro-hardness, and Debye temperature. The FTIR analysis revealed that increasing CdO content enhances the BiO6 octahedral sites at the expense of the BiO3 and PbO4 units. This results in the formation of Pb-O-Bi(6) and Bi(3)-O-Bi(6) linkages in the glass network, which stiffen the structure and improve the elastic properties. A correlation between elastic and compositional parameters was achieved on the basis of theories and approaches in the field.


Emergency contraception: Knowledge and practice among women and the spouses seeking termination of pregnancy.
India was one of the first countries to launch a formal family planning program. Initially, the main thrust of the program was on sterilization but subsequently it has got evolved and now the stress is to bring about awareness of contraception and make informed choices. Emergency contraception has been included in its armamentarium. This study was conducted to find out about the awareness among the cases who report for induced abortion. A total of 784 willing cases were enrolled in the study; there were no exclusion criteria except unwillingness. A parallel group was also included consisting of their spouses. Information that was being sought about Emergency Contraception (EC) included its knowledge, details of administration, and availability. Of the 784 cases, a large number, 742 (94.6%), underwent first trimester abortion and only 42 (5.3%) underwent second trimester abortion. 286 (36.4%) patients had not used any contraceptive. A large number had used natural methods (35.3%), like lactation, abstinence, or coitus interruptus, and 25.7% had used barrier contraception inconsistently. A very small percentage in both the groups knew about EC; more number of men knew about EC than women. Awareness about emergency contraception is low, as reported in many other studies, though it is available for many years. Awareness about contraceptives needs to be improved and emergency contraceptive should be advocated as a backup method. More efforts are required to generate awareness about regular use of effective contraception and emergency contraception if required.


Compression use during an exercise intervention and associated changes in breast cancer-related lymphedema.
This study assessed the association between compression use and changes in lymphedema observed in women with breast cancer-related lymphedema who completed a 12-week exercise intervention. This work uses data collected from a 12 week exercise trial, whereby women were randomly allocated into either aerobic-based only (n = 21) or resistance-based only (n = 20) exercise. Compression use during the trial was at the participants discretion. Differences in lymphedema (measured by lymphedema index [L-Dex] score and interlimb circumference difference [%]) and associated symptoms between those who wore, and did not wear compression during the 12-week intervention were assessed. We also explored participants' reasons surrounding compression during exercise. No significant interaction effect between time and compression use for lymphedema was observed. There was no difference between groups over time in the number or severity of lymphedema symptoms. Irrespective of compression use, there were trends for reductions in the proportion of women reporting severe symptoms, but lymphedema status did not change. Individual reasons for the use of compression, or lack thereof, varied markedly. Our findings demonstrated an absence of a positive or negative effect from compression use during exercise on lymphedema. Current and previous findings suggest the clinical recommendation that garments must be worn during exercise is questionable, and its application requires an individualized approach.


Asymmetric tandem conjugate addition and reaction with carbocations on acylimidazole Michael acceptors.
We present here a stereoselective tandem reaction based on the asymmetric conjugate addition of dialkylzinc reagents to unsaturated acylimidazoles followed by trapping of the intermediate zinc enolate with carbocations. The use of a chiral NHC ligand provides chiral zinc enolates in high enantiomeric purities. These enolates are reacted with highly electrophilic onium compounds to afford densely substituted acylimidazoles. DFT calculations helped to understand the reactivity of the zinc enolates derived from acylimidazoles and allowed their comparison with metal enolates obtained by other conjugate addition reactions.


Blackfoot disease in Taiwan: a 30-year follow-up study.
Blackfoot disease is an endemic peripheral vascular disease found among the inhabitants of a limited area on the southwest coast of Taiwan, where artesian well water with a high concentration of arsenic has been used for more than eighty years. The natural history of blackfoot disease, based on a prospective study of 1,300 patients, is presented. The overall male/female ratio was 1.5:1. Although the clinical onset was usually insidious, it may be quite sudden and almost always begins with numbness or coldness in one or more extremities, usually the feet. Ultimately, rest pain develops and progresses to gangrene. In this series, 68% of the patients underwent spontaneous or operative amputation, and the reamputation rate was 23.3%. Lower extremity involvement in blackfoot disease was observed in 97.7% of the cases. The average annual rate for major amputation was 3.81 per 100 patient-years. The factors influencing the prognosis, such as amputation in relation to age and disease onset, are analyzed. The case fatality rate was 66.5% during thirty years; 44% of these were cardiovascular deaths. The annual death rate was 4.84 per 100 patient-years. Other reported case fatality rates for vascular insufficiency are reviewed. A dose-response relationship between blackfoot disease and the duration of water intake was also noted. The survival rates after the onset of blackfoot disease were: five years, 76.0%; ten years, 59.5%; twenty years, 38.2%; thirty years, 28.6%. The 50% survival point was 13.5 years after onset of the disease.


Diagnosis and presentation of fatal myocarditis.
The clinical presentation of myocarditis is highly variable, and histopathology is thus considered to be the cornerstone of diagnosis. We studied how accurately myocarditis was diagnosed in a series of routine autopsies and how fatal myocarditis presents clinically. All death certificates with myocarditis recorded as the underlying cause of death in Finland in 1970 to 1998 were collected retrospectively (N = 639). All cases with cardiac autopsy samples and clinical data available (n = 142; median age, 51 years) were included in this study. The cardiac samples were reexamined for the presence of myocarditis by 3 experienced independent pathologists using the Dallas criteria. The clinical data were evaluated for the presenting signs and symptoms of myocarditis. Histopathologic reanalysis showed that only 32% of the 142 subjects met the Dallas criteria for myocarditis (75% of pediatric and 28% of adult patients, P = .001). Clinicians had suspected myocarditis in only one third of the hospitalized Dallas-positive patients. Dallas-positive patients presented more often with features of myocardial infarction (26% versus 9%, P = .026) or heart failure (35% versus 10%, P = .001) than Dallas-negative subjects. The signs and symptoms of infectious disease were also more common in Dallas-positive patients (61% versus 23%, P < .001). In contrast, Dallas-negative subjects died suddenly or were found dead more frequently (68% versus 39%, P = .004). The most evident cause of death in the Dallas-negative subjects was ischemic heart disease (n = 78, 55% of all cases). Our study provides evidence that myocarditis is overdiagnosed on routine autopsies, particularly in patients who have died suddenly or are found dead. Fatal myocarditis appears to present equally often as heart failure, sudden death, or mimicking myocardial infarction.


[John Hunter (1728-1793): founder of scientific surgery and precursor of urology].
The author briefly presents the life and scientific work of the founder of experimental surgery, John Hunter, who made an enormous contribution to the development of urology by his research and books concerning this discipline, which, up until then, simply constituted a branch of surgery. Several examples of his urological work are presented, emphasizing his oncological concepts concerning testicular tumours.


Case-control study assessing the impact of COVID19 in advanced kidney cancer patients treated with antiangiogenics or immunotherapy: the COVID-REN study.
Cancer is a risk factor for developing severe COVID19. Additionally, SARS-CoV2 has a special tropism for renal cells and complications like thrombosis or cytokine storm could be enhanced by standard treatments in kidney cancer (i.e., antiangiogenics or immunotherapy). Thus, understanding the impact of COVID19 in patients with this tumor is key for their correct management. We designed a retrospective case-control study comparing the outcome of three groups of advanced kidney cancer patients on systemic treatment: cohort A (developed COVID19 while on antiangiogenics), cohort B (developed COVID19 while on immunotherapy) and cohort C (non-infected). Matching factors were age, gender, and treatment. 95 patients were recruited in 16 centers in Spain from September 2020 to May 2021. Finally, 85 were deemed as eligible (23 cohort A, 21 cohort B, 41 cohort C). Patients with COVID required more dose interruptions (25 vs. six) and hospitalizations (10 vs. none) than those without COVID (both p = 0.001). No difference between cohorts A and B was observed regarding hospitalization or length of stay. No ICU admission was registered and one patient in cohort B died due to COVID19. Regarding cancer evolution, three patients in cohort A presented progressive disease after COVID19 compared to two in cohort B. One case in cohort B, initially deemed as stable disease, achieved a partial response after COVID19. Kidney cancer patients who developed COVID19 while on systemic therapy required more treatment interruptions and hospitalizations than those non-infected. However, no significant impact on cancer outcome was observed. Also, no difference was seen between cases on antiangiogenics or immunotherapy.


Lymphokine-activated killer (LAK) cells. Analysis of factors relevant to the immunotherapy of human cancer.
Lymphokine-activated killer (LAK) cells can be generated by incubating fresh peripheral blood lymphocytes (PBL) in Interleukin-2 (IL-2). LAK cells kill fresh autologous and allogeneic human tumor cells in vitro. This study analyzes aspects of LAK cells that make them a promising candidate for the adoptive immunotherapy of human cancer. LAK cells can be generated from PBL of normal individuals and tumor-bearing patients. Pure, recombinant IL-2 generates LAK cells capable of killing a wide variety of tumors including sarcomas and cancers of the colon, pancreas, adrenal gland, and esophagus. Thirty-six of 41 (88%) fresh, noncultured, human tumor cell suspensions prepared from surgical specimens were lysed by LAK cells in a standard 4-hour chromium-release assay. Normal PBL were not killed. LAK cells can be expanded in vitro for periods longer than 2 months, potentially more than 10(20)-fold, while maintaining lytic ability. These results and the demonstrated efficacy of LAK cells in the therapy of murine tumors make LAK cells a candidate for clinical use in the adoptive immunotherapy of human cancer.


RNA bandages for photoregulating in vitro protein synthesis.
'RNA bandages' are composed of two 6-12-mer 2'-OMe RNA strands complementary to a mRNA target that are joined by a photocleavable linker. These tandem oligonucleotides typically exhibit much higher affinity for the mRNA than the individual strands. An RNA bandage with binding arms of different lengths and a 4-base gap blocked translation in vitro of GFP mRNA; subsequent near-UV irradiation restored translation. This provides a general method of photomodulating hybridization for a variety of oligonucleotide-based technologies.


Social presence and the composite face effect.
A robust finding in social psychology research is that performance is modulated by the social nature of a given context, promoting social inhibition or facilitation effects. In the present experiment, we examined if and how social presence impacts holistic face perception processes by asking participants, in the presence of others and alone, to perform the composite face task. Results suggest that completing the task in the presence of others (i.e., mere co-action) is associated with better performance in face recognition (less bias and higher discrimination between presented and non-presented targets) and with a reduction in the composite face effect. These results make clear that social presence impact on the composite face effect does not occur because presence increases reliance on holistic processing as a "dominant" well-learned response, but instead, because it increases monitoring of the interference produced by automatic response.


The effects of the combined administration of gamma-hydroxybutyrate and diazepam on sleep parameters in the rat.
The actions of diazepam and gamma-hydroxybutyrate (GHB) were assessed on the sleep-wakefulness cycle of male Wistar rats. One and 2 mg/kg diazepam and 12.5 mg/kg GHB had no effects on the sleep variables. After 25 mg/kg GHB, slow wave sleep (SWS2) was significantly increased. Following the combined administration of non-effective doses of GHB and diazepam, significantly higher amounts of SWS2 at the expense of wakefulness were obtained. The injection of a subconvulsant dose of bicuculline (2.5 mg/kg) prior to treatments which significantly increased SWS2 prevented this effect to show up. It is suggested that the actions of GHB and diazepam on the sleep-awake cycle are related to the same neurotransmitter system.


Fluorimetric assays for cathepsin B and cathepsin H with methylcoumarylamide substrates.
Benzyloxycarbonyl-phenylalanyl-arginine 4-methyl-7-coumarylamide was found to be an excellent substrate for the fluorimetric assay of cathepsin B, and arginine 4-methyl-7-coumarylamide for cathepsin H. Procedures were developed that are very convenient, and avoid the hazards associated with the use of naphthylamides.