[Genetic collections: problems of formation, conservation and use].
Classification of plant genepool collections are presented, the principal tasks worked on by collections are examined. Databases on genepool collections of the Institute of Cytology and Genetics of Syberian Branch of Russian Academy of Sciences (Russia) and the National Centre for Plant Genetic Resources of Ukraine are described. Principles of formation of genepool collections, their existing problems, the methods of maintenance of their viability and genetical authenticity are discussed. Necessary amount of collection accessions, differences and specificity of their reproduction and multiplication aims and tasks are proved. Gradations of the permissible changes of accessions in connection with the type and the purpose of collections are proposed. The necessity of specialized periodical edition on genepool collecting is emphasized.


Molecular mechanisms for the modulation of blood pressure and potassium homeostasis by the distal convoluted tubule.
Epidemiological and clinical observations have shown that potassium ingestion is inversely correlated with arterial hypertension prevalence and cardiovascular mortality. The higher the dietary potassium, the lower the blood pressure and mortality. This phenomenon is explained, at least in part, by the interaction between salt reabsorption in the distal convoluted tubule (DCT) and potassium secretion in the connecting tubule/collecting duct of the mammalian nephron: In order to achieve adequate K+ secretion levels under certain conditions, salt reabsorption in the DCT must be reduced. Because salt handling by the kidney constitutes the basis for the long-term regulation of blood pressure, losing salt prevents hypertension. Here, we discuss how the study of inherited diseases in which salt reabsorption in the DCT is affected has revealed the molecular players, including membrane transporters and channels, kinases, and ubiquitin ligases that form the potassium sensing mechanism of the DCT and the processes through which the consequent adjustments in salt reabsorption are achieved.


Evolution of the Pseudomonas aeruginosa mutational resistome in an international Cystic Fibrosis clone.
Emergence of epidemic clones and antibiotic resistance development compromises the management of Pseudomonas aeruginosa cystic fibrosis (CF) chronic respiratory infections. Whole genome sequencing (WGS) was used to decipher the phylogeny, interpatient dissemination, WGS mutator genotypes (mutome) and resistome of a widespread clone (CC274), in isolates from two highly-distant countries, Australia and Spain, covering an 18-year period. The coexistence of two divergent CC274 clonal lineages was revealed, but without evident geographical barrier; phylogenetic reconstructions and mutational resistome demonstrated the interpatient transmission of mutators. The extraordinary capacity of P. aeruginosa to develop resistance was evidenced by the emergence of mutations in >100 genes related to antibiotic resistance during the evolution of CC274, catalyzed by mutator phenotypes. While the presence of classical mutational resistance mechanisms was confirmed and correlated with resistance phenotypes, results also showed a major role of unexpected mutations. Among them, PBP3 mutations, shaping up β-lactam resistance, were noteworthy. A high selective pressure for mexZ mutations was evidenced, but we showed for the first time that high-level aminoglycoside resistance in CF is likely driven by mutations in fusA1/fusA2, coding for elongation factor G. Altogether, our results provide valuable information for understanding the evolution of the mutational resistome of CF P. aeruginosa.


Domain-specific physical activity and sedentary behaviour in relation to colon and rectal cancer risk: a systematic review and meta-analysis.
Physical activity is associated with reduced risk of colorectal cancer, but most epidemiological studies have focused on occupational and recreational physical activity. The evidence for other domains of activity, and for sedentary behaviour, is limited. Medline, Embase and Web of Science were searched from inception to December 2015 for studies examining domain-specific physical activity or sedentary behaviour and the risk of colon and/or rectal cancer. We extracted maximally adjusted relative risks (RRs) except when RRs not adjusted for body mass index, were also presented. We used random-effects meta-analysis to compute pooled RRs comparing the highest versus the lowest level of exposure. We used meta-regression to assess sources of heterogeneity in estimates. We identified 17 cohort and 21 case-control studies, of which 17 had occupational data, 23 had recreational data, three each had data on transport and household physical activity domains, and 6 studies had data on occupational sedentary behaviour. The pooled relative risks (RRs) for colon cancer were 0.74 (95% confidence interval (CI): 0.67, 0.82) for occupational activity, 0.80 (95% CI: 0.71, 0.89) for recreational activity, 0.66 (95% CI: 0.45, 0.98) for transport-related physical activity, 0.85 (95% CI: 0.71, 1.02) for household physical activity, and 1.44 (95% CI: 1.28, 1.62) for occupational sedentary behaviour. For rectal cancer, the pooled RRs were 0.88 (95% CI: 0.79, 0.98) for occupational activity, 0.87 (95% CI: 0.75, 1.01) for recreational activity, 0.88 (95% CI: 0.70, 1.12) for transport-related physical activity, 1.01 (95% CI: 0.80, 1.27) for household physical activity, and 1.02 (95% CI: 0.82, 1.28) for occupational sedentary behaviour. In addition to increasing occupational and recreational physical activity, promoting physical activity during transport and reducing sedentary behaviour in the workplace may also be useful colorectal cancer prevention strategies.


HPV DNA Associates With Breast Cancer Malignancy and It Is Transferred to Breast Cancer Stromal Cells by Extracellular Vesicles.
A causal link between Human Papillomavirus (HPV) and breast cancer (BC) remains controversial. In spite of this, the observation that HPV DNA is over-represented in the Triple Negative (TN) BC has been reported. Here we remark the high prevalence of HPV DNA (44.4%) in aggressive BC subtypes (TN and HER2+) in a population of 273 Italian women and we convey the presence of HPV DNA in the epithelial and stromal compartments by in situ hybridization. As previously reported, we also found that serum derived-extracellular vesicles (EVs) from BC affected patients contain HPV DNA. Interestingly, in one TNBC patient, the same HPV DNA type was detected in the serum-derived EVs, cervical and BC tissue samples. Then, we report that HPV DNA can be transferred by EVs to recipient BC stromal cells that show an activated phenotype (e.g., CD44, IL6 expression) and an enhanced capability to sustain mammospheres (MS) formation. These data suggest that HPV DNA vehiculated by EVs is a potential trigger for BC niche aggressiveness.


Additive and heterosis effects on preweaning traits, maternal ability and reproduction from crossing of the Angus and brown Swiss breeds in Florida.
Angus (A), Brown Swiss (S) and A X S reciprocal F1 (AS) dams were mated to A, S and AS (also reciprocal F1) sires resulting in nine breed groups of progeny with varying proportions of Angus and Brown Swiss breeding. Breed group of dam and of sire significantly influenced birth weight, preweaning daily gain, weaning weight, 205-d weight, condition score and frame size. The means for birth weight and weaning weight were 33 and 213 kg, respectively. Brown Swiss bulls sired calves with the heaviest birth and weaning weights. Calves produced by S dams likewise were heavier at birth and weaning. Pregnancy rates were influenced significantly by year, age and breed of dam and averaged 79, 95 and 92% for S, AS and A cows, respectively. Survival rate averaged 97% and was not influenced significantly by any of the effects examined. Because survival rates were similar for all breed groups, the results for weaning rate paralleled those for pregnancy rate. Genetic influences on preweaning growth traits and survival rate were partitioned into additive breed differences (B) and heterosis (H) effects for direct (d) and maternal (m) components. Pregnancy and weaning rates were examined using similar analyses except that genotype of service sire of dam replaced that of the offspring for the direct additive breed and direct heterosis components. The Bd values indicated that the Angus breed was inferior (P less than .01) to the Brown Swiss breed for all preweaning growth traits except for condition score, in which the Angus breed surpassed (P less than .01) the Brown Swiss. The Bm values also showed an advantage for the Brown Swiss breed for all preweaning growth traits. The additive maternal effect (the genotype of the females exposed), Bm, was important for pregnancy rate and weaning rate (P less than .001 and P less than .05) but not for survival rate (P greater than .10). The direct additive breed effect was not important for any reproductive trait. Direct heterosis did not affect any of the preweaning or reproduction traits; however, maternal heterosis (Hm) significantly affected all traits except birth weight, frame score and survival rate. The Hm estimates were 12.0 and 8.4 kg for weaning weight and 205-d weight, respectively. The Hm estimates for pregnancy rate, survival rate and weaning rate were 10, 2 and 13%, respectively.


Non-Hodgkin lymphoma and pesticide exposure in Turkey.
Non-Hodgkin lymphoma (NHL) is a solid tumour of lymphocytes, important elements in the immune system. According to 2006 data, in Turkey the incidence was 6.5 per 100,000 in males, and 4.4 in females. The relationship between the use of pesticides and development of NHL has been extensively investigated in many studies, and it has been demonstrated that the risk of NHL is increased by exposure to such compounds. Antalya is a region of intensive agricultural activity. In this study, the relationship between the incidence of lymphoma in Antalya and the amount of pesticides employed was investigated. The study used data from 1995 to 2010 on the patients from the databank of TR Ministry of Health, Antalya Provincial Health Directorate, Cancer Registration Center and the patients who were histopathologically diagnosed with NHL during these years. The relationship between the amount of pesticide used and the incidence was studied with the Spearman correlation analysis and the p value was found as 0.05. The correlation coefficient was 0.497. An increase in the NHL incidence over the years was identified, with a 2.42-fold increment found from 1995 to 2005 and a 2.77 fold elevation from 1995 to 2010. The use of pesticides increased 1.89 fold over the same period. Our study investigated the relationship of the pesticides used with NHL patients diagnosed during the same year. Since the time elapsing after exposure to pesticides until the development of cancer is not clear, no comparison can be made at present. We believe that the increase in use of pesticides since 1995 may be associated with the increase in the incidence of NHLand therefore that further studies on the issue including measurements of serum pesticide levels, are required.


Impaired antibody-dependent cell-mediated cytotoxic activity in patients with the acquired immunodeficiency syndrome.
Since the acquired immunodeficiency syndrome (AIDS) is characterized by opportunistic infections and malignancies indicative of a profound suppression in cell-mediated immunity, we investigated the antibody-dependent cell-mediated cytotoxicity (ADCC) of peripheral blood mononuclear cells of patients with AIDS against chicken red blood cells (CRBC). A marked decrease in ADCC-CRBC activity was observed from patients with AIDS as compared to healthy controls. Furthermore, suppression in ADCC activity was seen when mononuclear cells from healthy subjects were assayed using media containing 25% or 40% sera from AIDS patients. Two of two patients with AIDS and impaired ADCC-CRBC activity were also found to have in vivo impaired reticuloendothelial system Fc-specific clearance of 51Cr-labelled, anti-Rho (D) IgG-sensitized autologous erythrocytes. These data provide further evidence of monocyte-macrophage dysfunction in AIDS and help explain the widespread occurrence of opportunistic pathogens in AIDS.


Identification of markers linked to disease-resistance genes by bulked segregant analysis: a rapid method to detect markers in specific genomic regions by using segregating populations.
We developed bulked segregant analysis as a method for rapidly identifying markers linked to any specific gene or genomic region. Two bulked DNA samples are generated from a segregating population from a single cross. Each pool, or bulk, contains individuals that are identical for a particular trait or genomic region but arbitrary at all unlinked regions. The two bulks are therefore genetically dissimilar in the selected region but seemingly heterozygous at all other regions. The two bulks can be made for any genomic region and from any segregating population. The bulks are screened for differences using restriction fragment length polymorphism probes or random amplified polymorphic DNA primers. We have used bulked segregant analysis to identify three random amplified polymorphic DNA markers in lettuce linked to a gene for resistance to downy mildew. We showed that markers can be reliably identified in a 25-centimorgan window on either side of the targeted locus. Bulked segregant analysis has several advantages over the use of near-isogenic lines to identify markers in specific regions of the genome. Genetic walking will be possible by multiple rounds of bulked segregation analysis; each new pair of bulks will differ at a locus identified in the previous round of analysis. This approach will have widespread application both in those species where selfing is possible and in those that are obligatorily outbreeding.


Primary DNA damage in human blood lymphocytes exposed in vitro to 2450 MHz radiofrequency radiation.
Human peripheral blood samples collected from three healthy human volunteers were exposed in vitro to pulsed-wave 2450 MHz radiofrequency (RF) radiation for 2 h. The RF radiation was generated with a net forward power of 21 W and transmitted from a standard gain rectangular antenna horn in a vertically downward direction. The average power density at the position of the cells in the flask was 5 mW/cm(2). The mean specific absorption rate, calculated by finite difference time domain analysis, was 2.135 (+/-0.005 SE) W/kg. Aliquots of whole blood that were sham-exposed or exposed in vitro to 50 cGy of ionizing radiation from a (137)Cs gamma-ray source were used as controls. The lymphocytes were examined to determine the extent of primary DNA damage (single-strand breaks and alkali-labile lesions) using the alkaline comet assay with three different slide-processing schedules. The assay was performed on the cells immediately after the exposures and at 4 h after incubation of the exposed blood at 37 +/- 1 degrees C to allow time for rejoining of any strand breaks present immediately after exposure, i.e. to assess the capacity of the lymphocytes to repair this type of DNA damage. At either time, the data indicated no significant differences between RF-radiation- and sham-exposed lymphocytes with respect to the comet tail length, fluorescence intensity of the migrated DNA in the tail, and tail moment. The conclusions were similar for each of the three different comet assay slide-processing schedules examined. In contrast, the response of lymphocytes exposed to ionizing radiation was significantly different from RF-radiation- and sham-exposed cells. Thus, under the experimental conditions tested, there is no evidence for induction of DNA single-strand breaks and alkali-labile lesions in human blood lymphocytes exposed in vitro to pulsed-wave 2450 MHz radiofrequency radiation, either immediately or at 4 h after exposure.


Liposuction over a shielded arteriovenous fistula for hemodialysis access maturation.
Vascular access for hemodialysis by arteriovenous fistula (AVF) is broadly recommended. Vein depth in many obese patients makes cannulation of an otherwise successful AVF difficult or impossible. We reviewed the case of an obese patient where liposuction was used to remove adipose tissue resulting in a superficial and easily accessed AVF. An endoscopic vein harvesting device was used to shield the vein during liposuction.


Etiological factor of hepatocellular carcinoma in Italy.
Cirrhosis from any cause is present in the majority (93.1%) of hepatocellular carcinoma (HCC) cases in Italy, it seems to be the common pathway by which several risk factors extent their carcinogenic effect. The mortality rate of HCC in Italy has progressively increased during the period 1969-1994, reflecting the rising number of persons living with cirrhosis as consequence of the remarkable advances in medical management of such patients. Most HCC develops in cirrhosis caused by known and preventable risk factors (hepatitis B virus, HBV, hepatitis C virus, HCV, alcohol and possibly non-alcoholic steatohepatitis, NASH). Unlike alcohol and NASH, HBV and HCV chronic infections act as a risk factors for HCC both because they induce cirrhosis and because they increase the risk in patients with cirrhosis. Moreover, case-control and prospective studies have shown a synergistic effect on HCC risk, when both viral infections occur. Currently, HCV infection is detected in the majority (76.4%) of HCC cases in Italy, reflecting the large cohort of subjects infected with this virus via the iatrogenic route during the 1950s and 1960s. The low rate of HCV infection in the younger Italian generations may generate a downward trend in the HCC mortality rate in the years to come.


ITGA5 inhibition in pancreatic stellate cells attenuates desmoplasia and potentiates efficacy of chemotherapy in pancreatic cancer.
Abundant desmoplastic stroma is the hallmark for pancreatic ductal adenocarcinoma (PDAC), which not only aggravates the tumor growth but also prevents tumor penetration of chemotherapy, leading to treatment failure. There is an unmet clinical need to develop therapeutic solutions to the tumor penetration problem. In this study, we investigated the therapeutic potential of integrin α5 (ITGA5) receptor in the PDAC stroma. ITGA5 was overexpressed in the tumor stroma from PDAC patient samples, and overexpression was inversely correlated with overall survival. In vitro, knockdown of ITGA5 inhibited differentiation of human pancreatic stellate cells (hPSCs) and reduced desmoplasia in vivo. Our novel peptidomimetic AV3 against ITGA5 inhibited hPSC activation and enhanced the antitumor effect of gemcitabine in a 3D heterospheroid model. In vivo, AV3 showed a strong reduction of desmoplasia, leading to decompression of blood vasculature, enhanced tumor perfusion, and thereby the efficacy of gemcitabine in co-injection and patient-derived xenograft tumor models.


Nebivolol induces, via β3 adrenergic receptor, lipolysis, uncoupling protein 1, and reduction of lipid droplet size in human adipocytes.
Most β-blockers may induce weight gain, dysglycemia, and dyslipidemia. Nebivolol is a third-generation β1-blocker with vasodilating properties mediated by β3 adrenergic receptors (β3AR). We investigated whether nebivolol is able to induce β3AR-mediated lipolysis, uncoupling protein 1 (UCP1), and size-reduction in human adipocytes. Human visceral (n = 28) and subcutaneous adipose tissue (n = 26) samples were used to obtain differentiated subcutaneous and visceral preadipocytes. Adipocytes were used to verify the effects of nebivolol onlipolysis, uncoupling protein 1 (UCP1) and other genes of the thermogenic program. Lipolysis was induced by isoproterenol and specific β3AR agonist, as expected,and also by nebivolol at 100 nmol/l and by its L-enantiomer at 10 nmol/l (P < 0.01). Nebivolol-mediated lipolysis was blocked by SR59230A, a specific β3AR antagonist, suggesting that nebivolol acts through β3AR in human adipocytes. Interestingly, in human adipocytes, nebivolol activated UCP1, PPARγ coactivator-1α (PGC-1α) and cytochrome c (CYCS) gene expression in a p38 MAPK-dependent manner. Using propranolol (β1 and β2 antagonist) together with nebivolol we showed that the induction of these genes was still present suggesting again β3AR activation. Moreover, nebivolol significantly reduced the diameter of lipid droplets in cultured adipocytes. In summary, nebivolol, through β3AR, is able to induce lipolysis and promote thermogenic and mitochondrial genes. The induction of lipolysis and the thermogenic program could explain the reduction of lipid droplets size. In conclusion, the lower dysmetabolic effects of nebivolol in humans may depend on its β3 agonist activity and the consequent induction of thermogenic program in human adipocytes.


Consumption of clarified grapefruit juice ameliorates high-fat diet induced insulin resistance and weight gain in mice.
To determine the metabolic effects of grapefruit juice consumption we established a model in which C57Bl/6 mice drank 25-50% sweetened GFJ, clarified of larger insoluble particles by centrifugation (cGFJ), ad libitum as their sole source of liquid or isocaloric and sweetened water. cGFJ and control groups consumed similar amounts of liquids and calories. Mice fed a high-fat diet and cGFJ experienced a 18.4% decrease in weight, a 13-17% decrease in fasting blood glucose, a three-fold decrease in fasting serum insulin, and a 38% decrease in liver triacylglycerol values, compared to controls. Mice fed a low-fat diet that drank cGFJ experienced a two-fold decrease in fasting insulin, but not the other outcomes observed with the high-fat diet. cGFJ consumption decreased blood glucose to a similar extent as the commonly used anti-diabetic drug metformin. Introduction of cGFJ after onset of diet-induced obesity also reduced weight and blood glucose. A bioactive compound in cGFJ, naringin, reduced blood glucose and improved insulin tolerance, but did not ameliorate weight gain. These data from a well-controlled animal study indicate that GFJ contains more than one health-promoting neutraceutical, and warrant further studies of GFJ effects in the context of obesity and/or the western diet.


Nano-optical imaging of exciton-plasmon polaritons in WSe2/Au heterostructures.
We report a nano-optical imaging study of exciton-plasmon polaritons (EPPs) in WSe2/Au heterostructures with scattering-type scanning near-field optical microscopy (s-SNOM). By mapping the interference fringes of EPPs at various excitation energies, we constructed the dispersion diagram of the EPPs, which shows strong exciton-plasmon coupling with a sizable Rabi splitting energy (∼0.19 eV). Furthermore, we found a sensitive dependence of the polariton wavelength (λp) on WSe2 thickness (d). When d is below 40 nm, λp decreases rapidly with increasing d. As d reaches 50 nm and above, λp drops to 210 nm, which is over 4 times smaller than that of the free-space photons. Our simulations indicate that the high spatial confinement of EPPs is due to the strong localization of the polariton field inside WSe2. Our work uncovers the transport properties of EPPs and paves the way for future applications of these highly confined polaritons in nanophotonics and optoelectronics.


Interesting twists on small RNA themes in Pseudomonas aeruginosa.
While many of the initial studies of regulatory small RNA (sRNA) function focused on Escherichia coli and Salmonella, there is expanding characterization of sRNAs in other bacteria. These studies are revealing unique permutations of the themes found in E. coli and Salmonella. This point is nicely illustrated by the Pseudomonas aeruginosa Hfq-binding RNA, PhrS, which activates the synthesis of the key quorum sensing transcription regulator PqsR as described by Sonnleitner et al. in the current issue of Molecular Microbiology. The PhrS RNA combines features of several previously characterized sRNAs; its expression is induced by low oxygen, it encodes a small protein, and it modulates the expression of a quorum sensing regulator by preventing the formation of an inhibitory secondary structure. However, in contrast to other base pairing sRNAs that activate translation, PhrS does not act directly on pqsR but rather increases the translation of a small upstream open reading frame (uof) whose translation is coupled to pqsR.


Detection of Lactate Dehydrogenase Elevating Virus in a Mouse Vivarium Using an Exhaust Air Dust Health Monitoring Program.
Lactate dehydrogenase elevating virus (LDV) continues to be one of the most common contaminants of cells and cell byproducts. As such, many institutions require that tumor cell lines, blood products, and products derived or passaged in rodent tissues are free of LDV as well as other pathogens that are on institutional exclusion lists prior to their use in rodents. LDV is difficult to detect by using a live-animal sentinel health monitoring program because the virus does not reliably pass to sentinel animals. After switching to an exhaust air dust health monitoring system, our animal resources center was able to detect a presumably long-standing LDV infection in a mouse colony. This health monitoring system uses IVC rack exhaust air dust collection media in conjunction with PCR analysis. Ultimately, the source of the contamination was identified as multiple LDV-positive patient-derived xenografts and multiple LDV-positive breeding animals. This case study is the first to demonstrate the use of environmental PCR testing as a method for detecting LDV infection in a mouse vivarium.


Human mitochondrial tRNAs in health and disease.
The human mitochondrial genome encodes 13 proteins, all subunits of the respiratory chain complexes and thus involved in energy metabolism. These genes are translated by 22 transfer RNAs (tRNAs), also encoded by the mitochondrial genome, which form the minimal set required for reading all codons. Human mitochondrial tRNAs gained interest with the rapid discovery of correlations between point mutations in their genes and various neuromuscular and neurodegenerative disorders. In this review, emerging fundamental knowledge on the structure/function relationships of these particular tRNAs and an overview of the large variety of mechanisms within translation, affected by mutations, are summarized. Also, initial results on wide-ranging molecular consequences of mutations outside the frame of mitochondrial translation are highlighted. While knowledge of mitochondrial tRNAs in both health and disease increases, deciphering the intricate network of events leading different genotypes to the variety of phenotypes requires further investigation using adapted model systems.


Morphogenesis of a cytomegalovirus from an American bison affected with malignant catarrhal fever.
A herpesvirus isolated from several organs of an American bison affected with malignant catarrhal fever was cultured in bovine foetal spleen cells and studied by electron microscopy. The fine structural features of the mature virion and the mode of virus morphogenesis were found to be similar to herpesviruses classified in the subgroup cytomegalovirus. The capsids were granular, hexagonal in shape and contained pleomorphic cores in thin sections. Envelopment of the capsids occurred primarily by budding on cytoplasmic membranes which appeared to be formed as extended vesicles of the Golgi apparatus; budding on nuclear membranes was only rarely observed. Cytoplasmic inclusions consisting of granular threads and amorphous electron-dense material were found in association with virions during the late stages of infection. The formation of cytoplasmic inclusions, the morphogenesis and ultrastructure of the virus are all consistent with classification of this virus as a cytomegalovirus.


The effect of the progression of vitamin A deficiency on glucose, galactose and mannose incorporation into sugar phosphates and sugar nucleotides in hamster liver.
The incorporation of [2-3H]mannose into dolichyl phosphate mannose and glycoproteins is markedly reduced in livers of vitamin A-deficient hamsters. To determine whether vitamin A deficiency selectively alters the level of mannose incorporation into sugar phosphates and sugar nucleotides, we studied the in vivo incorporation of [2-3H]mannose, [5-3H]glucose, and [4,5-3H]galactose into sugar phosphates and sugar nucleotides. Male hamsters fed either a vitamin A-depleted or a retinoic acid-supplemented (3 micrograms/g) diet were used at 4, 6 and 8 wk of age; the animals were killed at various time points after an intraperitoneal injection of the radiolabeled sugar. A two- to threefold increase in the amount of [2-3H]mannose was found in liver of hamsters fed a vitamin A-depleted diet for 4 wk, resulting in enhanced incorporation into mannosyl-phosphate and guanosine diphosphate (GDP) mannose. As deficiency progressed, there was a smaller increase in [2-3H]mannose and a significant decrease in [3H]mannose-phosphate and GDP-[3H]mannose, suggesting a decreased mannose kinase activity. [5-3H]Glucose-labeled livers showed no difference in the total uptake of the label or its incorporation into uridine diphosphate glucose and galactose-phosphate during the 8-wk study. However, the synthesis of glucosyl-phosphate was reduced by 50 to 90% at 6 and 8 wk of deficiency, suggesting an impaired gluco-kinase activity. In hamsters injected with [4,5-3H]galactose only [3H]glucose was found within 5 min in the free sugar fraction. In contrast, as much as 70% of the label in the sugar phosphate and sugar nucleotide fraction remained as [3H]galactose even at 60 min. These effects on sugar, sugar phosphate and sugar nucleotide formation in part may explain the effects of vitamin A deficiency on glycoconjugate biosynthesis.


Production and characterization of monoclonal antibodies to ARVCF.
We have generated the first monoclonal antibodies (MAbs) to Armadillo repeat gene deleted in velo-cardiofacial syndrome (ARVCF), a recently identified Armadillo repeat-containing protein closely related to the catenin p120ctn. Six ARVCF-specific MAbs were characterized for isotype, species cross-reactivity, and utility in assays including immunofluorescence, immunoprecipitation, and Western blotting. All six antibodies were isotyped as IgG1 and several cross-reacted with ARVCF from a variety of species including human, rat, dog, and monkey, but not mouse. Importantly, none of the ARVCF MAbs cross-reacted with p120ctn, despite the high homology between these proteins. MAbs 3B2 and 4B1 were consistently the best in all applications and will provide valuable tools for further study of the role of ARVCF in cells.


Carbon source-dependent capsule thickness regulation in Streptococcus pneumoniae.
The polysaccharide capsule of Streptococcus pneumoniae plays a major role in virulence, adherence to epithelial cells, and overall survival of the bacterium in the human host. Galactose, mannose, and N-acetylglucosamine (GlcNAc) are likely to be relevant for metabolization in the nasopharynx, while glucose is the primary carbon source in the blood. In this study, we aim to further the understanding of the influence of carbon sources on pneumococcal growth, capsule biosynthesis, and subsequent adherence potential. We tested the growth behavior of clinical wild-type and capsule knockout S. pneumoniae strains, using galactose, GlcNAc, mannose, and glucose as carbon source for growth. We measured capsule thickness and quantified capsule precursors by fluorescein isothiocyanate (FITC)-dextran exclusion assays and 31P-nuclear magnetic resonance measurements, respectively. We also performed epithelial adherence assays using Detroit 562 cells and performed a transcriptome analysis (RNA sequencing). We observed a reduced growth in galactose, mannose, and GlcNAc compared to growth in glucose and found capsular size reductions in mannose and GlcNAc compared to galactose and glucose. Additionally, capsular precursor measurements of uridine diphosphate-(UDP)-glucose and UDP-galactose showed less accumulation of precursors in GlcNAc or mannose than in glucose and galactose, indicating a possible link with the received capsular thickness measurements. Epithelial adherence assays showed an increase in adherence potential for a pneumococcal strain, when grown in mannose compared to glucose. Finally, transcriptome analysis of four clinical isolates revealed not only strain specific but also common carbon source-specific gene expression. Our findings may indicate a careful adaption of the lifestyle of S. pneumoniae according to the monosaccharides encountered in the respective human niche.


Sentinel lymph node evaluation in melanoma.
The surgical treatment of patients with cutaneous melanoma has been revolutionized by the development of lymphatic mapping and sentinel lymph node biopsy. This procedure involves injecting a radioactive tracer at the site of the primary melanoma (before wide excision); the tracer then travels via the lymphatics to the first draining or sentinel lymph node. The node is removed and evaluated for the presence of metastatic melanoma. In this way, patients who are most likely to benefit can be selected for regional lymphadenectomy. In addition, accurate and minimally invasive staging allows the surgeon to identify patients who may benefit from adjuvant and investigational immunotherapy. We review the development of lymphatic mapping, the technical details related to the procedure itself, and the published clinical studies using this new procedure. In addition, we discuss the controversial issues that have been raised with the introduction of sentinel lymph node biopsy.


Molecular and morphological analysis of the developing nemertean brain indicates convergent evolution of complex brains in Spiralia.
The brain anatomy in the clade Spiralia can vary from simple, commissural brains (e.g., gastrotrichs, rotifers) to rather complex, partitioned structures (e.g., in cephalopods and annelids). How often and in which lineages complex brains evolved still remains unclear. Nemerteans are a clade of worm-like spiralians, which possess a complex central nervous system (CNS) with a prominent brain, and elaborated chemosensory and neuroglandular cerebral organs, which have been previously suggested as homologs to the annelid mushroom bodies. To understand the developmental and evolutionary origins of the complex brain in nemerteans and spiralians in general, we investigated details of the neuroanatomy and gene expression in the brain and cerebral organs of the juveniles of nemertean Lineus ruber. In the juveniles, the CNS is already composed of all major elements present in the adults, including the brain, paired longitudinal lateral nerve cords, and an unpaired dorsal nerve cord, which suggests that further neural development is mostly related with increase in the size but not in complexity. The ultrastructure of the juvenile cerebral organ revealed that it is composed of several distinct cell types present also in the adults. The 12 transcription factors commonly used as brain cell type markers in bilaterians show region-specific expression in the nemertean brain and divide the entire organ into several molecularly distinct areas, partially overlapping with the morphological compartments. Additionally, several of the mushroom body-specific genes are expressed in the developing cerebral organs. The dissimilar expression of molecular brain markers between L. ruber and the annelid Platynereis dumerilii indicates that the complex brains present in those two species evolved convergently by independent expansions of non-homologous regions of a simpler brain present in their last common ancestor. Although the same genes are expressed in mushroom bodies and cerebral organs, their spatial expression within organs shows apparent differences between annelids and nemerteans, indicating convergent recruitment of the same genes into patterning of non-homologous organs or hint toward a more complicated evolutionary process, in which conserved and novel cell types contribute to the non-homologous structures.


Beliefs about worry in community-dwelling older adults.
This study examines the association between several kinds of beliefs about worry and generalized anxiety disorder (GAD) severity in a sample of older individuals recruited from a community setting (N=142, mean age=71.0 years, SD=6.0, range=55-88). Beliefs about worry were assessed with a 17-item scale designed for older adults, including three dimensions: Positive Beliefs, Negative Beliefs, and Responsibility. All three dimensions distinguished between people endorsing GAD symptoms and those without GAD symptoms, and only Negative Beliefs had a significant independent weight when regressed on GAD severity, even after controlling for level of trait worry. Results of this study suggest that negative beliefs appear to be strongly related to pathological worry in older adults. These results are consistent with empirical findings in younger adults and potentially support the use of particular interventions in clinical work with older adults with GAD.


Increase in Candida krusei infection among patients with bone marrow transplantation and neutropenia treated prophylactically with fluconazole.
In early 1990 fluconazole was introduced as a prophylactic antifungal agent after bone marrow transplantation. During the same year Candida krusei emerged as the chief candida pathogen among patients with bone marrow transplants. To determine whether there was a correlation between the introduction of fluconazole and the increased incidence of C. krusei, we conducted a retrospective study based on the medical, mycologic, and autopsy records of all adult inpatients who had undergone bone marrow transplantation (n = 296) or who had leukemia (n = 167) at the study center during 1989 and 1990. The 84 patients who received antifungal prophylaxis with fluconazole had a sevenfold greater frequency of C. krusei infection than the 335 patients who did not receive fluconazole (8.3 percent vs. 1.2 percent, P = 0.002), despite having a lower frequency of disseminated C. albicans and C. tropicalis infections (0 vs. 6.0 percent, P = 0.02). Ten of the 11 C. krusei infections were controlled by a combination of amphotericin B and flucytosine. Colonization by C. krusei was found in 40.5 percent of the patients who received fluconazole but in only 16.7 percent of those who did not receive it (P less than 0.0001). Colonization was independently associated with the prophylactic use of both fluconazole (odds ratio, 3.50; P less than 0.001) and norfloxacin (odds ratio, 2.53; P = 0.04). C. krusei was not susceptible to fluconazole in vitro. In patients at high risk for disseminated candida infections, suppression of bacterial flora and the more common candida pathogens may permit some less pathogenic, but natively resistant candida species, such as C. krusei, to emerge as systemic pathogens.


Inactivation of Escherichia coli JM109, DH5alpha, and O157:H7 suspended in Butterfield's Phosphate Buffer by gamma irradiation.
Food irradiation is a safe and effective method for inactivation of pathogenic bacteria, including Escherichia coli O157:H7, in meat, leafy greens, and complex ready-to-eat foods without affecting food product quality. Determining the radiation dose needed to inactivate E. coli O157:H7 in foods and the validation of new irradiation technologies are often performed through inoculation of model systems or food products with cocktails of the target bacterium, or use of single well-characterized isolates. In this study, the radiation resistance of 4 E. coli strains, 2 DNA repair deficient strains used for cloning and recombinant DNA technology (JM109 and DH5alpha) and 2 strains of serotype O157:H7 (C9490 and ATCC 35150), were determined. The D-10 values for C9490, ATCC 35150, JM109, and DH5alpha stationary phase cells suspended in Butterfield's Phosphate Buffer and irradiated at 4 degrees C were 229 (+/- 9.00), 257 (+/- 7.00), 61.2 (+/- 10.4), and 51.2 (+/- 8.82) Gy, respectively. The results of this study indicate that the extreme radiation sensitivity of JM109 and DH5alpha makes them unsuitable for use as surrogate microorganisms for pathogenic E. coli in the field of food irradiation research. Use of E. coli JM109 and DH5alpha, which carry mutations of the recA and gyrA genes required for efficient DNA repair and replication, is not appropriate for determination of radiation inactivation kinetics and validation of radiation processing equipment.


The effect of dimerization on the activation and conformational dynamics of adenosine A1 receptor.
The adenosine A1 receptor (A1R) is one of four adenosine receptors in humans, which are involved in the function of the cardiovascular, respiratory and central nervous systems. Experimental results indicate that A1R can form a homodimer and that the protomer-protomer interaction in the A1R dimer is related to certain pharmacological characteristics of A1R activation. In this work, we performed docking, metadynamics simulation, conventional molecular dynamics simulations, Gaussian-accelerated molecular dynamics simulations, potential of mean force calculations, dynamic cross-correlation motions analysis and community network analysis to study the binding mode of 5'-N-ethylcarboxamidoadenosine (NECA) to A1R and the effect of dimerization on the activation of A1R. Our results show that NECA binds to A1R in a similar mode to adenosine in the A1R crystal structure and NECA in the A2AR crystal structure. The A1R homodimer can be activated by one or two agonists with NECA occupying its orthosteric pockets in one (which we call the NECA-A1R system) or both protomers (which we call the dNECA-A1R system). In the NECA-A1R system, activation is predicated in the protomer without NECA bound. In the dNECA-A1R system, only one protomer achieves the active state. These findings suggest an asymmetrical activation mechanism of the homodimer and a negative cooperativity between the two protomers. We envision that our results may further facilitate the drug development of A1R.


Apple peel polyphenols: a key player in the prevention and treatment of experimental inflammatory bowel disease.
Diets rich in fruits and vegetables may reduce oxidative stress (OxS) and inflammation via several mechanisms. These beneficial effects may be due to their high polyphenol content. The aims of the present study are to evaluate the preventive and therapeutic aspects of polyphenols in dried apple peel powder (DAPP) on intestinal inflammation while elucidating the underlying mechanisms and clinical benefits. Induction of intestinal inflammation in mice was performed by oral administration of the inflammatory agent dextran sulfate sodium (DSS) at 2.5% for 10 days. Physiological and supraphysiological doses of DAPP (200 and 400 mg/kg/day respectively) were administered by gavage for 10 days pre- and post-DSS treatment. DSS-mediated inflammation caused weight loss, shortening of the colon, dystrophic detachment of the epithelium, and infiltration of mono- and poly-morphonuclear cells in the colon. DSS induced an increase in lipid peroxidation, a down-regulation of antioxidant enzymes, an augmented expression of myeloperoxidase (MPO) and cyclooxygenase-2 (COX-2), an elevated production of prostaglandin E2 (PGE2) and a shift in mucosa-associated microbial composition. However, DAPP normalized most of these abnormalities in preventive or therapeutic situations in addition to lowering inflammatory cytokines while stimulating antioxidant transcription factors and modulating other potential healing pathways. The supraphysiological dose of DAPP in therapeutic situations also improved mitochondrial dysfunction. Relative abundance of Peptostreptococcaceae and Enterobacteriaceae bacteria was slightly decreased in DAPP-treated mice. In conclusion, DAPP exhibits powerful antioxidant and anti-inflammatory action in the intestine and is associated with the regulation of cellular signalling pathways and changes in microbiota composition. Evaluation of preventive and therapeutic effects of DAPP may be clinically feasible in individuals with intestinal inflammatory bowel diseases.


Histone retention preserves epigenetic marks during heat stress-induced transcriptional memory in plants.
Plants often experience recurrent stressful events, for example, during heat waves. They can be primed by heat stress (HS) to improve the survival of more severe heat stress conditions. At certain genes, sustained expression is induced for several days beyond the initial heat stress. This transcriptional memory is associated with hyper-methylation of histone H3 lysine 4 (H3K4me3), but it is unclear how this is maintained for extended periods. Here, we determined histone turnover by measuring the chromatin association of HS-induced histone H3.3. Genome-wide histone turnover was not homogenous; in particular, H3.3 was retained longer at heat stress memory genes compared to HS-induced non-memory genes during the memory phase. While low nucleosome turnover retained H3K4 methylation, methylation loss did not affect turnover, suggesting that low nucleosome turnover sustains H3K4 methylation, but not vice versa. Together, our results unveil the modulation of histone turnover as a mechanism to retain environmentally mediated epigenetic modifications.


The gut microbiota: a possible factor influencing systemic lupus erythematosus.
Systemic lupus erythematosus (SLE) is an autoimmune disease of unknown cause. In recent years, with the emergence of microbiome research, changes in the gut microbiota composition have been correlated with a variety of autoimmune disorders, and several mechanisms linking these together have been suggested, including the hygiene theory, immune system activation and hormonal effects. It has therefore been suggested that gut microbiota may play a role in SLE. In this review, we summarize recent findings on the SLE-related microbiota compositions in both humans and rodents. Evidence linking microbiome with SLE opens a new avenue in researching the cause of SLE as well as improved future treatments. Although two studies found a lower Firmicutes/Bacteroidetes ratio in SLE patients vs. controls, there were inconsistencies regarding significant differences in the abundance of specific genera or species. Studies of mouse disease models have shown some correlations between microbial compositions and disease states, also indicating differences between males and females. Current data support an association between microbiota composition and SLE. Further research is needed to fully unravel this connection, potentially shedding light on mechanisms in SLE development and on the female bias of the disease, improving diagnosis and treatment.


Flow Cytometry-Based Photodynamic Diagnosis with 5-Aminolevulinic Acid for the Detection of Minimal Residual Disease in Multiple Myeloma.
Multiple myeloma is the cancer of plasma cells. Along with the development of new and effective therapies, improved outcomes in patients with multiple myeloma have increased the interest in minimal residual disease (MRD) monitoring. However, the considerable heterogeneity of immunophenotypic and molecular markers of myeloma cells has limited its clinical application. 5-Aminolevulinic acid (ALA) is a natural compound in the heme biosynthesis pathway. Following ALA treatment, tumor cells preferentially accumulate porphyrins because of the differential activities of aerobic glycolysis, known as Warburg effect. Among various porphyrins, protoporphyrine IX is a strong photosensitizer; thus, ALA-based photodynamic diagnosis has been widely used in various solid cancers. Here, the feasibility of flow cytometry-based photodynamic detection of MRD was tested in multiple myeloma. Among various human cell lines of hematological malignancies, including K562 erythroleukemia, Jurkat T-cell leukemia, Nalm6 pre-B cell leukemia, KG1a myeloid leukemia, and U937 monocytic leukemia, human myeloma cell line, KMS18, and OPM2 abundantly expressed ALA transporters, such as SLC36A1 and SLC15A2, and 1 mM ALA treatment for 24 h resulted in nearly 100% porphyrin fluorescence expression, which could be competitively inhibited by ALA transport with gamma-aminobutyric acid. Titration studies revealed that the lowest ALA concentration required to achieve nearly 100% porphyrin fluorescence in KMS18 cells was 0.25 mM, with an incubation period of 2 h. Under these conditions, incubation of primary peripheral blood mononuclear cells resulted in only 1.8 % of the cells exhibiting porphyrin fluorescence. Therefore, flow cytometry-based photodynamic diagnosis is a promising approach for detecting MRD in multiple myeloma.


A single mutation in the FGA locus responsible for false homozygosities and discrepancies between commercial kits in an unusual paternity test case.
We report an unusual paternity test case showing multiple peculiarities. Using AmpFlSTR Profiler Plus and AmpFlSTR Identifiler PCR Amplification kits, the alleged father and the two children were apparently homozygous at the FGA locus, but using the PowerPlex 16 kit the three individuals were found to be heterozygous. Drop-out was caused by a single mutation event in the presumptive binding site of the reverse primer. In addition, three inconsistencies were detected between the daughter and the alleged father among 18 STR markers. The occurrence of the rare null allele at the FGA locus and case history suggested that the true father was the brother of the alleged father. Furthermore, a single-step repeat maternal mutation was also detected at D16S539. This puzzling case was solved by using multiple analytical approaches, including the use of different primer pairs, the use of a high number of STR markers, and the characterization of the mutation causing the "null allele."


Complementary and alternative treatments of multiple sclerosis: a review of the evidence from 2001 to 2016.
People with multiple sclerosis (PwMS) commonly use complementary and alternative medicines (CAM), but an understanding of their efficacy is lacking. Here, we quantitatively review the class I and class II studies of treatment efficacy for multiple sclerosis from January 2001 to January 2017, in order to assess the modern evidence for CAM use. The 38 studies included in this review are divided across five CAM types (cannabis, diet, exercise, psychological approaches and other). We found little evidence to support CAM efficacy. The studies contained little replication in intervention, primary outcomes or study design. Six of 16 CAMs included in this review were only researched in a single study. Future work in this area should build consensus around study methodologies and primary outcomes.


Effects of applications manual pressure and shotblocker to reduce needle-related pain and fear in children with type 1 diabetes mellitus.
Pain and fear associated with insulin injections can cause children with type 1 diabetes mellitus to avoid insulin injections and skip doses. To evaluate and compare pain and fear levels in children aged 6-12 years receiving subcutaneous insulin injection using the manual pressure and ShotBlocker methods. A randomized controlled study was conducted with 90 children with type 1 diabetes who were allocated using block randomization to the manual pressure, ShotBlocker, and control groups (n = 30 in each group). Fear and pain levels were rated by the children, their parents, and a member of the study team immediately before and after insulin injection using the Children's Fear Scale and Wong-Baker Faces Pain Rating Scale, respectively. All groups had similar self-, parent-, and researcher-reported levels of preprocedural pain and fear (p > 0.05). However, pain and fear scores were lower in the manual pressure and ShotBlocker groups than in the control group after injection (p = 0.0001). There was no significant difference in pain and fear scores between the two intervention groups (p > 0.05). Manual pressure and the ShotBlocker both reduced fear and pain associated with insulin injection in 6- to 12-year-old children with type 1 diabetes. Both the manual pressure and ShotBlocker methods can easily be applied in children receiving insulin injections. As manual pressure is completely cost- and equipment-free, it is a useful option to reduce pain and fear related to insulin injection. National Institutes of Health (NIH), ClinicalTrials.gov, NCT05789810.


Neurocognitive, emotional and neuroendocrine correlates of exposure to sexual assault in women.
Survivors of sexual assault are vulnerable to long-term negative psychological and physical health outcomes, but few studies have investigated changes in cognition, emotional processing and brain function in the early stages after sexual assault. We used a multimodal approach to identify the cognitive and emotional correlates associated with sexual assault in women. Twenty-seven female survivors of sexual assault were included within 4 weeks of the traumatic event, and they were compared with 20 age-matched controls. Participants underwent functional MRI while performing cognitive/emotional tasks (n-back, emotional go/no-go, mental imagery). We also measured diurnal salivary cortisol and conducted neuropsychological assessments of attention and memory abilities. Relative to the control group, the survivor group had lower levels of morning cortisol and showed attentional deficits. We observed no between-group differences in brain activation during the n-back or mental imagery tasks. During the emotional go/no-go task, however, the survivor group showed a lack of deactivation in the dorsal anterior cingulate cortex when processing emotional material, relative to neutral material. Exploratory analyses in the survivor group indicated that symptom severity was negatively associated with cerebellar activation when positive emotional (happy) content interfered with response inhibition, and positively associated with cerebellar activation when thinking of positive (happy) memories. The small sample size was the main limitation of this study. Dysfunctions in the dorsal anterior cingulate cortex and the cerebellum may represent early functional brain modifications that alter higher cognitive processes when emotional material is involved.


The N-terminal sequence of the large proteoglycan of articular cartilage.
A peptide with hyaluronic acid-binding properties was isolated from trypsin digests of bovine articular cartilage proteoglycan aggregate. This peptide originated from the N-terminus of the proteoglycan core protein, retained its function of forming complexes with hyaluronate and link protein and contained at least one keratan sulfate chain. Amino acid sequence data demonstrated that the first six amino acid residues of the N-terminus of bovine articular cartilage proteoglycan core protein differed from the same region from the rat chondrosarcoma proteoglycan. Further sequence data indicate areas of considerable sequence homology in the hyaluronic acid-binding regions of proteoglycans from the two species.


Human papillomavirus types 6 and 11 mRNAs from genital condylomata acuminata.
We have identified and mapped a number of RNA species of human papillomavirus types 6 and 11 from condylomata acuminata by the electron microscopic R-loop technique. Each of the early (E)- and late (L)-region open reading frames (ORFs) deduced from the DNA sequences was represented in one or more transcripts. In addition, RNA species that could encode the modulator of DNA replication and the repressor of transcription, functions recently identified in the genetically similar bovine papillomavirus type 1, were also detected. Some ORFs were 5' proximal in one or more transcripts, whereas others were not 5' proximal in any species, suggesting that internal initiation of translation might be required to gain access to these latter ORFs. Virtually all transcripts had their 5' ends located in the E region and were polyadenylated at one of two sites, i.e., at the end of the E region or at the end of the L region. The great majority of the RNAs were derived from the E region of the genome, with one species approximately 50 to 100 times more abundant than the others. For most of the RNAs, the 5' end mapped near nucleotide 700; minor populations had 5' ends near nucleotide 100 or 1200. By correlating our mapping data with the genomic DNA sequences as well as available RNA structures and cDNA sequences of several papillomaviruses, we predict a number of mRNA splice donor and acceptor sites and suggest that the papillomaviruses have sophisticated usage of ORFs through alternative promoters, mRNA splice sites, and polyadenylation sites.


Diabetic Cardiomyopathy: 2023 Update by the International Multidisciplinary Board of Experts.
Diabetes mellitus (DM) is considered by many the pandemic of the 21st century and is associated with multiple organ damages. Among these, cardiovascular complications are responsible for an incredible burden of mortality and morbidity in Western Countries. The study of the pathological mechanisms responsible for the cardiovascular complications in DM patients is key for the development of new therapeutic strategies. The metabolic disorders caused by hyperglycemia, insulin resistance, and dyslipidemia, results in a cascade of pathomorphological changes favoring the atherosclerotic process and leading to myocardial remodeling. Parallel to this, oxidative stress, calcium overload, mitochondrial dysfunction, activation of protein kinase C signaling pathways, myocardial lipomatosis, and low-grade inflammation of the myocardium - are the main pathways responsible for the diabetic cardiomyopathy development. This review aims to appraise and discuss the pathogenetic mechanisms behind the diabetic cardiomyopathy development.


[The interosseous palmaris muscle of the thumb. Results of an anatomic study of a muscle of the first ray].
In sixteen of twenty adult human cadaveric hands a muscle fascicle was isolated from the adductor pollicis muscle. It had osseous origins from the base of the first metacarpal bone and from the trapezium bone in eleven hands. These origins were connected by a fibrous arch, which was a soft tissue origin itself. In two of the sixteen hands, muscle fibers emerged from the palmar aspect of the first dorsal interosseous muscle near the radial artery. The findings can be explained by the ontogenesis of the human hand. The described muscle is neither a part of the oblique head of the adductor pollicis nor an equivalent to the three palmar interosseous muscles. In our dissections it was a constant muscle. Other studies have to show the exact function of the muscle. We postulate a stabilization of the carpometacarpal joint of the thumb.


Hypoxia-inducible factor 2 regulates hepatic lipid metabolism.
In mammals, the liver integrates nutrient uptake and delivery of carbohydrates and lipids to peripheral tissues to control overall energy balance. Hepatocytes maintain metabolic homeostasis by coordinating gene expression programs in response to dietary and systemic signals. Hepatic tissue oxygenation is an important systemic signal that contributes to normal hepatocyte function as well as disease. Hypoxia-inducible factors 1 and 2 (HIF-1 and HIF-2, respectively) are oxygen-sensitive heterodimeric transcription factors, which act as key mediators of cellular adaptation to low oxygen. Previously, we have shown that HIF-2 plays an important role in both physiologic and pathophysiologic processes in the liver. HIF-2 is essential for normal fetal EPO production and erythropoiesis, while constitutive HIF-2 activity in the adult results in polycythemia and vascular tumorigenesis. Here we report a novel role for HIF-2 in regulating hepatic lipid metabolism. We found that constitutive activation of HIF-2 in the adult results in the development of severe hepatic steatosis associated with impaired fatty acid beta-oxidation, decreased lipogenic gene expression, and increased lipid storage capacity. These findings demonstrate that HIF-2 functions as an important regulator of hepatic lipid metabolism and identify HIF-2 as a potential target for the treatment of fatty liver disease.


Extracorporeal shockwave therapy in musculoskeletal disorders.
The sources of shockwave generation include electrohydraulic, electromagnetic and piezoelectric principles. Electrohydraulic shockwaves are high-energy acoustic waves generated under water explosion with high voltage electrode. Shockwave in urology (lithotripsy) is primarily used to disintegrate urolithiasis, whereas shockwave in orthopedics (orthotripsy) is not used to disintegrate tissues, rather to induce tissue repair and regeneration. The application of extracorporeal shockwave therapy (ESWT) in musculoskeletal disorders has been around for more than a decade and is primarily used in the treatment of sports related over-use tendinopathies such as proximal plantar fasciitis of the heel, lateral epicondylitis of the elbow, calcific or non-calcific tendonitis of the shoulder and patellar tendinopathy etc. The success rate ranged from 65% to 91%, and the complications were low and negligible. ESWT is also utilized in the treatment of non-union of long bone fracture, avascular necrosis of femoral head, chronic diabetic and non-diabetic ulcers and ischemic heart disease. The vast majority of the published papers showed positive and beneficial effects. FDA (USA) first approved ESWT for the treatment of proximal plantar fasciitis in 2000 and lateral epicondylitis in 2002. ESWT is a novel non-invasive therapeutic modality without surgery or surgical risks, and the clinical application of ESWT steadily increases over the years. This article reviews the current status of ESWT in musculoskeletal disorders.


An update review of intradialytic hypotension: concept, risk factors, clinical implications and management.
Intradialytic hypotension (IDH) is a frequent and serious complication of chronic haemodialysis, linked to adverse long-term outcomes including increased cardiovascular and all-cause mortality. IDH is the end result of the interaction between ultrafiltration rate (UFR), cardiac output and arteriolar tone. Thus excessive ultrafiltration may decrease the cardiac output, especially when compensatory mechanisms (heart rate, myocardial contractility, vascular tone and splanchnic flow shifts) fail to be optimally recruited. The repeated disruption of end-organ perfusion in IDH may lead to various adverse clinical outcomes affecting the heart, central nervous system, kidney and gastrointestinal system. Potential interventions to decrease the incidence or severity of IDH include optimization of the dialysis prescription (cool dialysate, UFR, sodium profiling and high-flux haemofiltration), interventions during the dialysis session (midodrine, mannitol, food intake, intradialytic exercise and intermittent pneumatic compression of the lower limbs) and interventions in the interdialysis period (lower interdialytic weight gain and blood pressure-lowering drugs). However, the evidence base for many of these interventions is thin and optimal prevention and management of IDH awaits further clinical investigation. Developing a consensus definition of IDH will facilitate clinical research. We review the most recent findings on risk factors, pathophysiology and management of IDH and, based on this, we call for a new consensus definition of IDH based on clinical outcomes and define a roadmap for IDH research.


Immunization with plasmid DNA encoding hepatitis C virus envelope E2 antigenic domains induces antibodies whose immune reactivity is linked to the injection mode.
Plasmids expressing different domains of the hepatis C virus (HCV) envelope E2 glycoprotein from a genotype 1a isolate were constructed to compare the immunogenic potential of E2 in nucleic acid-based immunizations. One plasmid, pCIE2t, expressed a C-terminally truncated form of E2, while others, pS2.SE2A to pS2.SE2E, encoded the adjacent 60-amino-acid (aa) sequences of E2 (inserts A to E) expressed as a fusion with the hepatitis B virus surface antigen. BALB/c mice were given injections of the plasmids intramuscularly (i.m.) or intraepidermally (i.e.) via a gene gun (biolistic introduction), and induced humoral immune responses were evaluated. The i.e. injections resulted in higher seroconversion rates and antibody titers, up to 100-fold, than did the i.m. injections (P = 0.01 to 0.04). Three restricted immunogenic domains, E2A (aa 384 to 443), E2C (aa 504 to 555), and E2E (aa 609 to 674), that yielded antibody titers ranging from 1:59 to > 1:43,700 could be identified. Subtype 1a- and 1b-derived E2 antigens and synthetic peptides were used in Western blot and enzyme-linked immunosorbent assay analyses, which revealed that the cross-reactivity of the plasmid-induced antibodies was linked both to the type of antigen expressed and to the injection mode. Induced anti-E2 antibodies could immunoprecipitate noncovalent E1E2 complexes believed to exist on the surface of HCV virions. This study allowed us to identify restricted immunogenic domains within E2 and demonstrated that different routes of injection of HCV E2 plasmids can result in quantitatively and qualitatively different humoral immune responses.


[Severe inflammation of the muzzle caused by a nose ring in a breeding bull].
This report describes the findings in a bull with severe inflammation of the muzzle and nose attributable to a nose ring. The most striking finding was that the bull continually licked the right side of the upper lip. The muzzle and right upper lip were swollen, hard, reddened and partially depigmented. Mucopurulent nasal discharge and salivation were also noted, and palpation of the right upper lip was extremely painful. Based on the findings, purulent infection of the right side of the muzzle, right naris and external nasal passage was diagnosed. After removing the nose ring the affected areas were washed daily for four days with a camomile-containing solution after which a chlorhexidine and dexpanthenol salve was applied. The bull also received ceftiofur and ketoprofen. The general condition and appetite of the bull normalised within a few days, and the inflammatory lesions resolved with the exception of the areas of depigmentation. After ten days of treatment, the bull was considered healthy and discharged from the clinic.


Site-directed mutagenesis of human membrane-associated ganglioside sialidase: identification of amino-acid residues contributing to substrate specificity.
Unlike microbial sialidases, mammalian sialidases possess strict substrate specificity, for example the human membrane-associated sialidase, which hydrolyzes only gangliosides. To cast light on the molecular basis of this narrow substrate preference, predicted active site amino-acid residues of the human membrane sialidase were altered by site-directed mutagenesis. When compared with the active site amino-acid residues proposed for Salmonella typhimurium sialidase, only five out of 13 residues were found to be different to the human enzyme, these being located upstream of the putative transmembrane region. Alteration of seven residues, including these five, was followed by transient expression of the mutant enzymes in COS-1 cells and characterization of their kinetic properties using various substrates. Substitution of glutamic acid (at position 51) by aspartic acid and of arginine (at position 114) by glutamine or alanine resulted in retention of good catalytic efficiency toward ganglioside substrates, whereas other substitutions caused a marked reduction. The mutant enzyme E51D exhibited an increase in hydrolytic activity towards GM2 as well as sialyllactose (which are poor substrates for the wild-type) with change to a lower Km and a higher Vmax. R114Q demonstrated a substrate specificity shift in the same direction as E51D, whereas R114A enhanced the preference for gangliosides GD3 and GD1a that are effectively hydrolyzed by the wild-type. The inhibition experiments using 2-deoxy-2,3-didehydro-N-acetylneuraminic acid were consistent with the results in the alteration of substrate specificity. The findings suggest that putative active-site residues of the human membrane sialidase contribute to its substrate specificity.


Stimulatory effect of cytokinins and interaction with IAA on the release of lateral buds of pea plants from apical dominance.
Lateral buds of pea plants can be released from apical dominance and even be transformed into dominant shoots when repeatedly treated with synthetic exogenous cytokinins (CKs). The mechanism of the effect of CKs, however, is not clear. The results in this work showed that the stimulatory effects of CKs on the growth of lateral buds and the increase in their fresh weights in pea plants depended on the structure and concentration of the CKs used. The effect of N-(2-chloro-4-pyridyl)-N'-phenylurea (CPPU) was stronger than that of 6-benzylaminopurine (6-BA). Indoleacetic acid (IAA) concentration in shoot, IAA export out of the treated apex and basipetal transport in stems were markedly increased after the application of CPPU or 6-BA to the apex or the second node of pea plant. This increase was positively correlated with the increased concentration of the applied CKs. These results suggest that the increased IAA synthesis and export induced by CKs application might be responsible for the growth of lateral shoots in intact pea plants.


Development and regulation of glucose-6-phosphatase gene expression in rat liver, intestine, and kidney: in vivo and in vitro studies in cultured fetal hepatocytes.
The mRNA and the activity of glucose-6-phosphatase (Glc-6-Pase) were present in the liver, kidney, and small intestine of 15-day-old suckling rats, but were absent from the stomach, colon, lung, white and brown adipose tissues, muscle, heart, brain, and spleen. The mRNA encoding Glc-6-Pase was present in the liver of 21-day-old fetal rats and increased markedly immediately after birth. From 5 days after birth to the end of the suckling period, it returned to 50% of the level found in the liver of 48-h starved adult rats. When rats were weaned at 21 days onto a high-carbohydrate, low-fat (HCLF) diet, the concentration of liver Glc-6-Pase mRNA was markedly increased. In the fetal rat jejunum, the activity and mRNA of Glc-6-Pase were very low. It increased during the 5 days after birth and then declined to reach very low levels. Neither mRNA nor activity of Glc-6-Pase was present in the fetal kidney. They appeared and increased slowly during the suckling period to reach maximal levels 15 days after birth and then remained constant. Weaning onto the HCLF diet did not change the Glc-6-Pase gene expression, neither in the jejunum nor in the kidney. The regulation of Glc-6-Pase gene expression by hormones and nutrients was studied in cultured hepatocytes from 20-day-old rat fetuses. Bt2cAMP stimulated the Glc-6-Pase gene expression in a dose-dependent manner. This probably resulted from an increased gene transcription since the half-life of the transcript was not affected by dibutyryl cAMP (Bt2cAMP). The Bt2cAMP-induced Glc-6-Pase mRNA accumulation was antagonized by insulin in a dose-dependent manner. Long-chain fatty acids (LCFAs), but not medium-chain fatty acids, induced the accumulation of Glc-6-Pase mRNA and the stabilization of the transcript. The peroxisome proliferator, clofibrate, induced a threefold increase in Glc-6-Pase mRNA concentration. Both stimulation of Glc-6-Pase mRNA by LCFAs and clofibrate were inhibited by insulin. Increasing concentrations of glucose (from 0 to 20 mmol/l) did not affect the Bt2cAMP-induced Glc-6-Pase gene expression. By contrast, high glucose concentration (25 mmol/l) markedly induced the Glc-6-Pase gene expression in fed adult rat hepatocytes. The difference in the response to glucose between fetal and adult rat hepatocytes is discussed. We conclude that the rapid increase in hepatic Glc-6-Pase mRNA levels that accompanies the fetal-to-neonatal transition in the rat is triggered by the reciprocal change in circulating insulin and LCFA concentrations, coupled to the rise in liver cAMP concentration.


Variable selection in strong hierarchical semiparametric models for longitudinal data.
In this paper, we consider the variable selection problem in semiparametric additive partially linear models for longitudinal data. Our goal is to identify relevant main effects and corresponding interactions associated with the response variable. Meanwhile, we enforce the strong hierarchical restriction on the model, that is, an interaction can be included in the model only if both the associated main effects are included. Based on B-splines basis approximation for the nonparametric components, we propose an iterative estimation procedure for the model by penalizing the likelihood with a partial group minimax concave penalty (MCP), and use BIC to select the tuning parameter. To further improve the estimation efficiency, we specify the working covariance matrix by maximum likelihood estimation. Simulation studies indicate that the proposed method tends to consistently select the true model and works efficiently in estimation and prediction with finite samples, especially when the true model obeys the strong hierarchy. Finally, the China Stock Market data are fitted with the proposed model to illustrate its effectiveness.


A qualitative analysis of nonorthogonal CO(2) laserbeam delivery.
Recent studies have been conducted to investigate the effects of nonorthogonal beam delivery on the geometry of the craters formed by laser beams in biological media. The final results, obtained with samples of the quasi-biological solid methacrylate Vedril C, clearly show that major alterations to the classical cone-shaped crater do occur. The results of these first investigations are presented here together with a qualitative discussion of the possible causes of this phenomenon. It is concluded that curved craters should also be expected in vivo.


Stroboscopic vision and sustained attention during coincidence-anticipation.
We compared coincidence-anticipation performance in normal vision and stroboscopic vision as a function of time-on-task. Participants estimated the arrival time of a real object that moved with constant acceleration (-0.7, 0, +0.7 m/s2) in a pseudo-randomised order across 4 blocks of 30 trials in both vision conditions, received in a counter-balanced order. Participants (n = 20) became more errorful (accuracy and variability) in the normal vision condition as a function of time-on-task, whereas performance was maintained in the stroboscopic vision condition. We interpret these data as showing that participants failed to maintain coincidence-anticipation performance in the normal vision condition due to monotony and attentional underload. In contrast, the stroboscopic vision condition placed a greater demand on visual-spatial memory for motion extrapolation, and thus participants did not experience the typical vigilance decrement in performance. While short-term adaptation effects from practicing in stroboscopic vision are promising, future work needs to consider for how long participants can maintain effortful processing, and whether there are negative carry-over effects from cognitive fatigue when transferring to normal vision.


Duration of Untreated Psychosis in First-Episode Psychosis is not Associated With Common Genetic Variants for Major Psychiatric Conditions: Results From the Multi-Center EU-GEI Study.
Duration of untreated psychosis (DUP) is associated with clinical outcomes in people with a diagnosis of first-episode psychosis (FEP), but factors associated with length of DUP are still poorly understood. Aiming to obtain insights into the possible biological impact on DUP, we report genetic analyses of a large multi-center phenotypically well-defined sample encompassing individuals with a diagnosis of FEP recruited from 6 countries spanning 17 research sites, as part of the European Network of National Schizophrenia Networks Studying Gene-Environment Interactions (EU-GEI) study. Genetic propensity was measured using polygenic scores for schizophrenia (SZ-PGS), bipolar disorder (BD-PGS), major depressive disorder (MDD-PGS), and intelligence (IQ-PGS), which were calculated based on the results from the most recent genome-wide association meta-analyses. Following imputation for missing data and log transformation of DUP to handle skewedness, the association between DUP and polygenic scores (PGS), adjusting for important confounders, was investigated with multivariable linear regression models. The sample comprised 619 individuals with a diagnosis of FEP disorders with a median age at first contact of 29.0 years (interquartile range [IQR] = 22.0-38.0). The median length of DUP in the sample was 10.1 weeks (IQR = 3.8-30.8). One SD increases in SZ-PGS, BD-PGS, MDD-PGS or IQ-PGS were not significantly associated with the length of DUP. Our results suggest that genetic variation does not contribute to the DUP in patients with a diagnosis of FEP disorders.


Managing a multi-panel clinic with heterogeneous patients.
Primary care providers (PCPs) are considered the first-line defenders in preventive care. Patients seeking service from the same PCP constitute that physician's panel, which determines the overall supply and demand of the physician. The process of allocating patients to physician panels is called panel design. This study quantifies patient overflow and builds a mathematical model to evaluate the effect of two implementable panel assignments. In specialized panel assignment, patients are assigned based on their medical needs or visit frequency. In equal panel assignment, patients are distributed uniformly to maintain a similar composition across panels. We utilize majorization theory and numerical examples to evaluate the performance of the two designs. The results show that specialized panel assignment outperforms when (1) patient demands and physician capacity are relatively balanced or (2) patients who require frequent visits incur a higher shortage penalty. In a simulation model with actual patient arrival patterns, we also illustrate the robustness of the results and demonstrate the effect of switching panel policy when the patient pool changes over time.


Evaluation of a questionnaire for obtaining owner-perceived, weighted quality-of-life assessments for dogs with spinal cord injuries.
To evaluate a questionnaire for obtaining owner-perceived, weighted quality-oflife assessments for dogs with spinal cord injuries. Evaluation study. Animals-100 dogs with spinal cord injuries and 48 healthy control dogs. The questionnaire was adapted from a questionnaire (the schedule for the evaluation of individual quality of life-direct weighting) used for human patients. Specifically, owners were asked to identify 5 areas or activities they believed had the most influence on their dogs' quality of life, assess their dogs' current status in each of those areas, and provide a weighting for the importance of each area. Results were used to construct a weighted quality-of-life score ranging from 0 to 100 for each dog. Owners were also asked to provide a quality-of-life score with a visual analog scale (VAS). A good correlation was found between weighted and VAS quality-of-life scores. Dogs with spinal cord injuries had weighted quality-of-life scores that were significantly lower than scores for control dogs. Quality-of-life areas and activities provided by owners of dogs with spinal cord injuries were similar to areas and activities provided by owners of healthy control dogs and could mostly be encompassed by 5 broader domains: mobility, play or mental stimulation, health, companionship, and other. Results suggested that the questionnaire could be used to obtain owner-perceived, weighted quality-of-life assessments for dogs with spinal cord injuries. Obtaining owner-perceived quality-of-life assessments for individual dogs should allow veterinarians to better address quality-of-life concerns and expectations of owners.


Tumor Necrosis Factor-α Initiates miRNA-mRNA Signaling Cascades in Obstruction-Induced Bladder Dysfunction.
Bladder outlet obstruction (BOO) and the ensuing clinical lower urinary tract dysfunction are common in elderly patients. BOO is accompanied by urodynamic changes in bladder function and leads to organ fibrosis and ultimately loss of contractility. Comprehensive transcriptome analysis of bladder samples from human patients with different urodynamically defined phenotypes of BOO revealed tumor necrosis factor (TNF)-α as the top upstream signaling pathway regulator. Herein, we validated next-generation sequencing and pathway analysis in cell-based models using bladder smooth muscle and urothelial cells exposed to TNF-α. miRNA profiling and transcriptome analysis of TNF-α-treated bladder smooth muscle cells revealed striking similarities with human BOO. Using a comparative approach, TNF-specific and TNF-independent pathways were delineated in human biopsy specimens. Concomitant down-regulation of smooth muscle cell-specific miRNAs and smooth muscle markers after TNF-α treatment was in accordance with the loss of contractility in humans in advanced obstruction-induced bladder remodeling. The expression levels of four abundant TNF-regulated miRNAs were modulated; the compensatory up-regulation of miR-199a-5p reduced NF-κB signaling. Essential hubs of TNF-α signaling pathways mitogen-activated protein kinase kinase kinase (apoptosis signal-regulating kinase 1) and inhibitor of nuclear factor κ B kinase subunit β (IκB kinase β) were targeted by miR-199a-5p. miR-199a-5p might be part of a negative feedback loop, reducing the impact of TNF, whereas its down-regulation in acontractile bladders from BOO patients advances the disease. The compensatory up-regulation of miR-199a-5p together with TNF-α inhibition may be therapeutically beneficial.


Changes in brain function and heart sound in acute sleep deprivation individuals.
Sleep deprivation (SD) has become a health problem in modern society due to its adverse effects on different aspects. However, the relationship between sleep and cardiovascular system function remains unclear. Here we explored the changes occurring in the brain and the heart sounds after SD. Ninety healthy adult men were recruited and subjected to 36 h of Sleep Deprivation (SD). They participated in a number of tests, including measurements of the heart sound, blood oxygen, and heart rate every 2 h. By using of principal component analysis to reduced the dimensionality of heart sound data. While the ALFF and ReHo indexes were measured via fMRI before and after SD. Correlation and regression analyses were used to reveal the relationship between fMRI and heart sound changes due to SD. In this study, there were no abnormal values in the heart rate and blood oxygen during 36 h of SD, whereas the intensity of heart sounds fluctuated significantly increased and decreased. The ALFF was increased in bilateral pericalcarine(Calcarine), left anterior cuneus, (Precuneus_L), right superior temporal gyrus(Temporal_Sup_R), left supplementary motor area (Supp_Motor_Area_L); However, it was reduced in the right medial superior frontal gyrus (Frontal_Sup_Medial_R), right dorsolateral superior frontal gyrus (Frontal_Sup_R) and left medial frontal gyrus (Frontal_Mid_L). The regression analysis uncovered that the intensity of the heart sound in the systole, s1, and s2 phase could be explained by Calcarine_L changes. Acute sleep deprivation affects cardiac-brain axis and the specific brain regions. Calcarine_L changes during sleep deprivation are involved in regulating heart contractions.


Improving contig binning of metagenomic data using [Formula: see text] oligonucleotide frequency dissimilarity.
Metagenomics sequencing provides deep insights into microbial communities. To investigate their taxonomic structure, binning assembled contigs into discrete clusters is critical. Many binning algorithms have been developed, but their performance is not always satisfactory, especially for complex microbial communities, calling for further development. According to previous studies, relative sequence compositions are similar across different regions of the same genome, but they differ between distinct genomes. Generally, current tools have used the normalized frequency of k-tuples directly, but this represents an absolute, not relative, sequence composition. Therefore, we attempted to model contigs using relative k-tuple composition, followed by measuring dissimilarity between contigs using [Formula: see text]. The [Formula: see text] was designed to measure the dissimilarity between two long sequences or Next-Generation Sequencing data with the Markov models of the background genomes. This method was effective in revealing group and gradient relationships between genomes, metagenomes and metatranscriptomes. With many binning tools available, we do not try to bin contigs from scratch. Instead, we developed [Formula: see text] to adjust contigs among bins based on the output of existing binning tools for a single metagenomic sample. The tool is taxonomy-free and depends only on k-tuples. To evaluate the performance of [Formula: see text], five widely used binning tools with different strategies of sequence composition or the hybrid of sequence composition and abundance were selected to bin six synthetic and real datasets, after which [Formula: see text] was applied to adjust the binning results. Our experiments showed that [Formula: see text] consistently achieves the best performance with tuple length k = 6 under the independent identically distributed (i.i.d.) background model. Using the metrics of recall, precision and ARI (Adjusted Rand Index), [Formula: see text] improves the binning performance in 28 out of 30 testing experiments (6 datasets with 5 binning tools). The [Formula: see text] is available at https://github.com/kunWangkun/d2SBin . Experiments showed that [Formula: see text] accurately measures the dissimilarity between contigs of metagenomic reads and that relative sequence composition is more reasonable to bin the contigs. The [Formula: see text] can be applied to any existing contig-binning tools for single metagenomic samples to obtain better binning results.


The structure of parasite component communities in brackish water fishes of the northeastern Baltic Sea.
We used nestedness analysis to seek non-random patterns in the structure of component communities of metazoan parasites collected from 31 sympatric fish species from the northeastern Bothnian Bay, the most oligohaline area of the Baltic Sea. Only 8 marine parasite species were found among the 63 species recorded, although some marine fish species reproduce in the bay and others occasionally visit the area. Marine parasite species can utilize both freshwater and marine fish species as intermediate or final hosts, and marine fish can harbour freshwater parasite species. This exchange of parasite species between marine and freshwater fish has probably resulted from ecological factors acting over short time scales rather than from evolutionary processes acting over longer time; the key factor probably being the immediate presence of suitable intermediate and definitive hosts. Marine fish were expected to harbour species-poor parasite communities consisting mainly of generalists acquired from the sympatric freshwater fish species, which would result in a nested pattern among the different component communities. However, an anti-nested pattern was found in the component communities of metazoan parasites of fishes from the Bothnian Bay. A likely explanation for the observed pattern is that there are specialist parasite species, the majority of which are cestodes, in some of the freshwater fish species which otherwise have depauperate parasite communities.


[Potential for benz(alpha)pyrene uptake from the soil into potato tubers and grain].
A significant correlation between benz(a)pyrene concentration in soil and potato tubers was established for its soil level of 0.6-50.0 mg/kg. This points to potato uptake of the carcinogen even at low levels of pollution. Absence of correlation between benz(a)pyrene concentrations in grain (husk) and soil even for relatively high levels of soil pollution (up to 320 micrograms/kg) shows that wheat and millet grains do not absorb the carcinogen.


[Synthesis of the tobacco mosaic virus in blocked spreading of infection].
The possibility of infection of tobacco upper leaves with tobacco mosaic virus (TMV) was examined in experiments where the inoculum was imbibed through the cut stem. The inoculum used were: a) a preparation of a virus-specific informosome-like ribonucleoproteins (vRNP) isolated from TMV-infected plants; b) a TMV preparation; or c) a mixture of TMV and vRNP. Multiplication of TMV in upper leaves was observed in neither of the variants; nevertheless in the vascular tissue and/or probably in adjoining parenchymal cells, two kinds of RNA were synthesized: of mol. w. (1.1--1.3) X 10(6) and (0.6--0.8) X 10(6). These RNA were not found in healthy plants in the presence of actinomycin D. The synthesis of genomic TMV RNA is suppressed under these conditions. Thus, some kind of abortive TMV infection takes place under the condition of experimental inoculation of plants through a cut stem. Molecular hybridization with the DNA of recombinant plasmid containing a nucleotide sequence complementary to the 3'-portion of genomic TMV RNA proves that short RNAs synthesized under the abortive infection conditions are TMV-specific. The experiments with differential temperature treatment of N-gene-containing plants under abortive infection conditions suggest that necrotization is not necessarily induced by genomic TMV RNA synthesis.


Shock absorption during transtibial amputee gait: Does longitudinal prosthetic stiffness play a role?
Reduced-stiffness components are often prescribed in lower-limb prostheses, but their efficacy in augmenting shock absorption has been inconclusive. To perform a systematic variation of longitudinal prosthetic stiffness over a wide range of values and to evaluate its effect on shock absorption during gait. Repeated-measures crossover experiment. Twelve subjects with a unilateral transtibial amputation walked at normal and fast self-selected speeds. Longitudinal prosthetic stiffness was modified by springs within a shock-absorbing pylon: normal (manufacturer recommended), 75% of normal (medium), 50% of normal (soft), and rigid (displacement blocked). The variables of interest were kinematic (stance-phase knee flexion and pelvic obliquity) and kinetic (prosthetic-side ground reaction force loading peak magnitude and timing). No changes were observed in kinematic measures during gait. A significant difference in peak ground reaction force magnitudes between medium and normal ( p = 0.001) during freely selected walking was attributed to modified walking speed ( p = 0.008). Ground reaction force peaks were found to be statistically different during fast walking, but only between isolated stiffness conditions. Thus, altering longitudinal prosthesis stiffness produced no appreciable change in gait biomechanics. Prosthesis stiffness does not appear to substantially influence shock absorption in transtibial prosthesis users. Clinical relevance Varying the level of longitudinal prosthesis stiffness did not meaningfully influence gait biomechanics at self-selected walking speeds. Thus, as currently prescribed within a transtibial prosthesis, adding longitudinal stiffness in isolation may not provide the anticipated shock absorption benefits. Further research into residual limb properties and compensatory mechanisms is needed.


The integrated stress response regulates BMP signalling through effects on translation.
Developmental pathways must be responsive to the environment. Phosphorylation of eIF2α enables a family of stress-sensing kinases to trigger the integrated stress response (ISR), which has pro-survival and developmental consequences. Bone morphogenetic proteins (BMPs) regulate multiple developmental processes in organisms from insects to mammals. Here we show in Drosophila that GCN2 antagonises BMP signalling through direct effects on translation and indirectly via the transcription factor crc (dATF4). Expression of a constitutively active GCN2 or loss of the eIF2α phosphatase dPPP1R15 impairs developmental BMP signalling in flies. In cells, inhibition of translation by GCN2 blocks downstream BMP signalling. Moreover, loss of d4E-BP, a target of crc, augments BMP signalling in vitro and rescues tissue development in vivo. These results identify a novel mechanism by which the ISR modulates BMP signalling during development.


[Ketamine attenuates the contractile response to vasoconstrictors in isolated coronary artery rings].
Ketamine was shown to increase coronary blood flow. It was the aim of this study to answer the question whether ketamine directly dilates coronary arteries. Using the model of isolated vessel rings we studied the effects of ketamine (2.5, 25, and 250 microg ml(-1)) on the contractile response to three vasoconstrictors, acetylcholine, histamine, and serotonin in porcine coronary artery segments. Other rings were contracted with KCl or PGF (2a) and then treated with ketamine (5 up to 500 microg ml(-1) added cumulatively). Ketamine dose-dependently dilated coronary arteries in concentrations beyond those used in clinical practice. In intact rings ketamine racemate (250 microg ml(-1)) attenuated contractions mediated by acetylcholine by 38.8 +/- 2.8%, histamine by 33.0 +/- 4.4% and serotonin by 42.1 +/- 3.7% (p < 0.05). There were no differences between intact and denuded rings (acetylcholine 38.5 +/- 2.8%, histamine 26.6 +/- 4.7%, serotonin 30.0 +/- 3.2%). With low concentrations of ketamine (2.5 microg ml(-1)) a slight tendency towards a contraction was recorded (n. s.). In rings precontracted with either KCl or PGF (2a) ketamine caused a small enhancement of contraction (KCl: 101.4 +/- 0.4%, PGF (2a): 101.3 +/- 1.4%) when administered in low concentration (5 microg ml(-1)), but almost complete relaxation (KCl: 0.4 +/- 1.3%, PGF (2a): 0.0 +/- 5.4%) in high concentration (500 microg ml(-1)). It is concluded that ketamine dose-dependently dilates porcine coronary arteries in concentrations beyond those used in clinical practice and that this effect is independent of endothelial function.


Neural substrates of specific and general autobiographical memory retrieval in younger and older adults.
Healthy aging is associated with a shift away from the retrieval of specific episodic autobiographical memories (AMs), towards more general and semanticized memories. Younger adults modulate activity in the default mode network according to the episodic specificity of AM retrieval. However, little is known about whether aging disrupts this neural modulation. In the current study we examine age-related changes in the modulation of whole-brain networks in response to three tasks falling along a gradient of episodic specificity. Younger and older adults retrieved specific (unique) AMs, general (routine) AMs, and semantic (general knowledge) memories. We found that both younger and older adults modulated default mode regions in response to varying episodic specificity. In addition, younger adults upregulated activity in several default mode regions with increasing episodic specificity, while older adults either did not modulate these regions, or downregulated activity in these regions. In contrast, older adults upregulated activity in the left temporal pole for tasks with higher episodic specificity. These brain activation patterns converge with prior findings that specific AMs are diminished in episodic richness with age, but are supplemented with conceptual and general information. Age-related reductions in the modulation of default mode regions might contribute to the shift away from episodic retrieval and towards semantic retrieval, resulting in reduced episodic specificity of personal memories.


Fibrin depletion decreases inflammation and delays the onset of demyelination in a tumor necrosis factor transgenic mouse model for multiple sclerosis.
In multiple sclerosis, in which brain tissue becomes permeable to blood proteins, extravascular fibrin deposition correlates with sites of inflammatory demyelination and axonal damage. To examine the role of fibrin in neuroinflammatory demyelination, we depleted fibrin in two tumor necrosis factor transgenic mouse models of multiple sclerosis, transgenic lines TgK21 and Tg6074. In a genetic analysis, we crossed TgK21 mice into a fibrin-deficient background. TgK21fib(-/-) mice had decreased inflammation and expression of major histocompatibility complex class I antigens, reduced demyelination, and a lengthened lifespan compared with TgK21 mice. In a pharmacologic analysis, fibrin depletion, by using the snake venom ancrod, in Tg6074 mice also delayed the onset of inflammatory demyelination. Overall, these results indicate that fibrin regulates the inflammatory response in neuroinflammatory diseases. Design of therapeutic strategies based on fibrin depletion could potentially benefit the clinical course of demyelinating diseases such as multiple sclerosis.


Hourly Relationship between Reference Evapotranspiration and Shoot Shrinkage in Walnut Trees and Pomegranate under Deficit Irrigation.
Diameter fluctuations of branches, shoots, or fruits are related to plant transpiration and water potential. In the past, several models have related dendrometric variables and evapotranspiration on a daily scale. However, trunk-branch shrinkage occurs only between dawn and midday, while evapotranspiration occurs most of the day from sunrise to sunset. Previous models have failed to incorporate this key fact. The objective of the present study was to assess the relationship of hourly daily shrinkage (HDS) between dawn and the next 4 h to the hourly reference evapotranspiration (EToh) of the same period in walnut trees and pomegranate plants under different irrigation regimes. Our data show that the relationship between EToh and HDS is much better than several previous models that included maximum daily shrinkage (MDS) and reference evapotranspiration (ETo). The novel slope analysis of the relationship between HDS versus time used here corresponds to the velocity at which the HDS occurs, which depends on the ETo intensity at that moment. This new method of analyzing this type of data calls for a revision of these models and sets a new baseline for future analysis.