Efficacy of Gemcitabine as Salvage Therapy for Relapsed and Refractory Aggressive Non-Hodgkin Lymphoma.
Gemcitabine-based salvage therapy is considered an effective treatment for relapsed and refractory Non-Hodgkin's lymphoma (NHL). We analyzed the outcome of 41 consecutive NHL patients treated with gemcitabine-based regimens between January 2007 and October 2015. Twenty-eight males and 13 females (median age 66.4 years) were included. The median follow-up from gemcitabine initiation was 7.3 months. Thirty patients (73%) had B-cell, and eleven (27%) had T-cell, lymphoma. All patients received a median of 2 prior regimens, of which at least 1 was anthracycline based. Twenty-eight patients (78%) received full-dose while 9 (22%) received reduced-dose regimens. The overall response rate was 37%, with 24% (n = 10) complete response, 12% (n = 5) partial response, and 63% (n = 22) progressive disease or stable disease. The median progression-free survival (PFS) was 47 days (range 12-1,318), the median overall survival (OS) was 1.9 years. Twenty patients (49%) died during follow-up. Grade 3-4 hematological toxicity was reported in 21 patients (51%). Relapsed vs. refractory disease, as well as a response to gemcitabine, predicted better PFS and OS. Use of a full-dose regimen predicted a better OS. Compared to previously published data, we observed less favorable outcomes. The administration of gemcitabine-based therapy as a salvage regimen for patients with relapsed or refractory NHL had limited success. Innovative therapies for these patients are an unmet need.


Cough augmentation with mechanical insufflation/exsufflation in patients with neuromuscular weakness.
Adults and children with neuromuscular disease exhibit weak cough and are susceptible to recurrent chest infections, a major cause of morbidity and mortality. Mechanical insufflation/exsufflation may improve cough efficacy by increasing peak cough flow. It was hypothesised that mechanical insufflation/exsufflation would produce a greater increase in peak cough flow than other modes of cough augmentation. The acceptability of these interventions was also compared. Twenty-two patients aged 10-56 yrs (median 21 yrs) with neuromuscular disease and 19 age-matched controls were studied. Spirometry was performed and respiratory muscle strength measured. Peak cough flow was recorded during maximal unassisted coughs, followed in random order by coughs assisted by physiotherapy, noninvasive ventilation, insufflation and exsufflation, and exsufflation alone. Subjects rated strength of cough, distress and comfort on a visual analogue scale. In the neuromuscular disease group, mean +/- SD forced expiratory volume in one second was 0.8 +/- 0.6 L x s(-1), forced vital capacity 0.9 +/- 0.8 L, maximum inspiratory pressure 25 +/- 16 cmH2O, maximum expiratory pressure 26 +/- 22 cmH2O and unassisted peak cough flow 169 +/- 90 L x min(-1). The greatest increase in peak cough flow was observed with mechanical insufflation/exsufflation at 235 +/- 111 L x min(-1) (p<0.01). All techniques showed similar patient acceptability. Mechanical insufflation/exsufflation produces a greater increase in peak cough flow than other standard cough augmentation techniques in adults and children with neuromuscular disease.


Water, sanitation and hygiene (WASH) behaviour change research: why an analysis of contingencies of reinforcement is needed.
Diarrheal disease associated with poor water, sanitation and hygiene (WASH) kills more than one million people every year. Safe WASH practices have the potential to greatly reduce these statistics but behaviour change interventions in the field have yielded little success to date. Currently, there is an emphasis on addressing cognitive processes to bring about changes in behaviour. In this review, a case is made for the benefits of a contingency-based perspective, focusing on the contextual antecedents and consequences of behaviour. The role of contingencies of reinforcement, not explored in previous WASH literature, is discussed as an explanatory framework for designing behaviour change strategies. A proper use of contrived reinforcers is recommended to counterbalance the natural reinforcers of convenience associated with risk practices. Recognising the role of consequences in the acquisition and maintenance of behaviour is an important step in the search for the answers urgently needed in the WASH field.


Effects of dexamethasone on ovarian activity in rats.
Rats with 5-day ovarian cycles were treated with dexamethasone (0.025 to 0.4 mg per rat per day) to examine its effects on ovarian activity. All doses of dexamethasone markedly suppressed endogenous secretion of ACTH as evidenced by decrease in weight of the adrenal glands after 7-23 days of treatment. Treatment with 0.4 mg of dexamethasone from the day of ovulation appeared to induce pseudopregnancy in more than 90% of the rats studied: pseudopregnancies lasted for 14-22 days. Induction of pseudopregnancy occurred less, in a dose-dependent fashion, when the lower doses of dexamethasone were administered. However, prolonged treatment with these lower doses resulted also in pseudopregnancy albeit after 1 or more cycles of 5 to 7 days. The effects of large doses of glucocorticoids on ovarian activity in rats, and possibly in humans as well, may be mediated predominantly through a stimulatory action of such drugs on prolactin secretion. Treatment with 0.4 mg per day of dexamethasone from the day before pro-oestrus did not interfere with ovulation and pseudopregnancy ensued this ovulation in all rats studied. Occasionally, unexpectedly large numbers of eggs per ovulating animal were found during treatment with dexamethasone. The data indicate failure of large doses of dexamethasone to interfere with ovulation and, possibly, induction of an increase in numbers of maturing follicles: this latter action deserves further analysis.


Neural inhibition enables selection during language processing.
Whether grocery shopping or choosing words to express a thought, selecting between options can be challenging, especially for people with anxiety. We investigate the neural mechanisms supporting selection during language processing and its breakdown in anxiety. Our neural network simulations demonstrate a critical role for competitive, inhibitory dynamics supported by GABAergic interneurons. As predicted by our model, we find that anxiety (associated with reduced neural inhibition) impairs selection among options and associated prefrontal cortical activity, even in a simple, nonaffective verb-generation task, and the GABA agonist midazolam (which increases neural inhibition) improves selection, whereas retrieval from semantic memory is unaffected when selection demands are low. Neural inhibition is key to choosing our words.


[X-ray guided punch biopsy of the spinal column].
Roentgenographically visible lesions of the skeleton are subjects for microscopy if the nature of the lesion is uncertain. In particular, the vertebral column, as it lies so deeply, needs a major surgical intervention if an excision has to be performed. The radiologically guided trephine biopsy, in contrast, is a great simplification for the patient. Using the Jamshidi-needle, 57 bone biopsies were performed. In 53 cases a reliable pathologic diagnosis was obtained by microscopy. Bacteriology, however, was positive in only 5 out of 23 cases with spondylitis or spondylodiscitis. No complications were observed.


Phosphatidate phosphohydrolase activity as a marker for surfactant synthesis in organotypic cultures of type II alveolar pneumonocytes.
The specific activity of phosphatidate phosphohydrolase (PAPase) (EC 126.96.36.199) has been assayed in organotypic cultures of foetal rat lung type II alveolar pneumonocytes and in L2 cells derived from type II cells of the adult rat lung. This enzyme catalyses a critical step in the synthesis of phosphatidylcholine, the major lipid component of pulmonary surfactant. Surfactant is produced by the mature type II cell in culture as well as in vivo. The specific activity of PAPase in organotypic cultures prepared from foetal rat lung starting at 16 days of gestation increased four- to fivefold during the first 7 days in culture. The specific activity of this enzyme was further increased through the 21 days of culture. In parallel with the increase in PAPase specific activity in the cultures were morphological changes in the type II cells such as the appearance of increased numbers of lamellar bodies. The specific activities of PAPase samples derived from non-type II cell cultures maintained under identical conditions were compared. Organotypic cultures and L2 cells, a culture system that also exhibits type II cell characteristics, show PAPase specific activities five to six times greater than cultures that do not contain type II cells. PAPase activity in the type II cell cultures parallels the development of mature patterns of pulmonary surfactant synthesis storage and secretion.


Postpartum hemorrhage revisited: new challenges and solutions.
Postpartum hemorrhage (PPH) is a leading cause of maternal morbidity and mortality in the United States, and worldwide. Recognition of PPH is challenging, but once hemorrhage is recognized, management needs to focus on achieving adequate uterine tone and maintaining maternal hemodynamic stability. There have been several advances in the management of postpartum hemorrhage, many of which can be implemented at the labor and delivery unit level. There have been many advances in the understanding of at-risk parturients, and the use of hemorrhage protocols and safety bundles have been shown to improve patient outcomes. There are many new advances in transfusion management (e.g. fibrinogen concentrate, prothrombin complex concentrate, tranexamic acid) that can compliment traditional component therapy. Consideration should be given to transferring women at high risk for complications (e.g. invasive placentation) to a higher level facility for delivery. Although postpartum hemorrhage itself may not be preventable, early identification of blood loss, and mobilization of resources may prevent adverse outcomes. Multidisciplinary planning at the system level, ensuring that hemorrhage protocols exist, as well as for management of high-risk patients is important for improving patient outcomes.


Network geometry and market instability.
The complexity of financial markets arise from the strategic interactions among agents trading stocks, which manifest in the form of vibrant correlation patterns among stock prices. Over the past few decades, complex financial markets have often been represented as networks whose interacting pairs of nodes are stocks, connected by edges that signify the correlation strengths. However, we often have interactions that occur in groups of three or more nodes, and these cannot be described simply by pairwise interactions but we also need to take the relations between these interactions into account. Only recently, researchers have started devoting attention to the higher-order architecture of complex financial systems, that can significantly enhance our ability to estimate systemic risk as well as measure the robustness of financial systems in terms of market efficiency. Geometry-inspired network measures, such as the Ollivier-Ricci curvature and Forman-Ricci curvature, can be used to capture the network fragility and continuously monitor financial dynamics. Here, we explore the utility of such discrete Ricci curvatures in characterizing the structure of financial systems, and further, evaluate them as generic indicators of the market instability. For this purpose, we examine the daily returns from a set of stocks comprising the USA S&P-500 and the Japanese Nikkei-225 over a 32-year period, and monitor the changes in the edge-centric network curvatures. We find that the different geometric measures capture well the system-level features of the market and hence we can distinguish between the normal or 'business-as-usual' periods and all the major market crashes. This can be very useful in strategic designing of financial systems and regulating the markets in order to tackle financial instabilities.


Enhancing hypericin production of Hypericum perforatum cell suspension culture by ozone exposure.
Accumulation of secondary metabolites is one of the common reactions of plants to ozone exposure in nature. To investigate the effect of ozone on the production of desired compounds of plant cell cultures, we assayed hypericin production of Hypericum perforatum suspension cell cultures treated with different doses of ozone at different culture phases. The results show that hypericin contents of the cells treated with 60 to 180 nL L(-1) ozone are significantly higher than those of the control, showing that ozone exposure may stimulate hypericin synthesis. Hypericin production of the cells treated with ozone at exponential phase is higher than that of lag and stationary phase, which suggests that exponential phase cell cultures are more responsive to ozone exposure than lag and stationary phase cells. The highest hypericin production is obtained by the cells exposed to 90 nL L(-1) ozone at late exponential phase for 3 h, being about fourfold of the control.


Epilepsy in children with attention-deficit/hyperactivity disorder.
Previous studies have suggested a higher incidence of symptoms of attention-deficit/hyperactivity disorder (ADHD) in children with epilepsy, but few have investigated epilepsy in children with ADHD. The objective here was to compare the incidence and characteristics of epilepsy among population-based, research identified cohorts of children with (n = 358) and without ADHD (n = 728), based on medical record review to age 20 years. Data abstracted included characteristics of seizures, testing, and treatment. Cases were 2.7 times more likely than controls to have epilepsy (95% CI = 0.94-7.76; P = 0.066), had earlier seizure onset (median age, 5.5 vs 15 years; P = 0.020), and exhibited a trend toward more frequent seizures (more than monthly, 63% vs 17%). Among children who met the research criteria for ADHD, those with epilepsy tended to be less likely to have received a clinical diagnosis of ADHD (63% vs 89%; P = 0.052) or to be treated with stimulants (50% vs 85%; P = 0.025). The findings suggest a strong trend toward a higher incidence of epilepsy among children with ADHD than among children without ADHD. Epilepsy in children with ADHD appears to be more severe than in those without. Finally, there appears to be a reluctance to diagnose and initiate treatment for ADHD in children with epilepsy.


Adolescents in Primary Care with Sub-Threshold Depressed Mood Screened for Participation in a Depression Prevention Study: Co-Morbidity and Factors Associated with Depressive Symptoms.
Adolescents in primary care with sub-threshold depression symptoms may be candidates for early intervention to prevent onset of full major depressive disorder. Little is known about this population. We screened consecutive adolescents (ages 14-21) in 13 primary care sites for presence of depression symptoms for "at least a few days" or "nearly every day" in the last two weeks for possible enrollment in a primary care/Internet-based depression prevention intervention (CATCH-IT). We report illness severity, prevalence of self-harm ideation, prevalence correlates (automatic negative thoughts, generalized self-efficacy, perceived social support from family and friends) and co-occurring symptoms of other mental disorders with depressed mood. Twenty-one percent (N=293) of those screened reported a core symptom of depressive disorder of which 83 enrolled in the study and were analyzed. The sample was 40% ethnic minority with a mean zip code household income of $40,249 (SD=$14,500). Reporting at least one symptom of another mental disorder was common for anxiety (48%, N=40), substance abuse (31%, N=15), conduct disorder (71%, N=53), as were self-harm ideation (16%, N=12) and reporting school impairment (100%, N=83). Prevalence correlates for current depressive symptoms included low self-efficacy, automatic negative thoughts, perceived low peer acceptance, and school impairment. Adolescents with sub-threshold depressed mood have frequent co-morbid symptoms that may need to be considered in developing prevention interventions. Early intervention targets to reduce depressed mood include pessimistic thinking, low self-efficacy, low peer acceptance, and school impairment.


[Progress in genetic testing and prevention of hereditary hearing disorders].
The article reviews the latest achievements of molecular-genetic technologies in diagnosis of hereditary hypoacusis, statistics on hereditary hypoacusis morbidity in different countries. Arguments for genetic testing, diagnostic algorithm of congenital hypoacusis and trends in prophylaxis are considered. Presently 65 genes are identified which are related with syndromes and non-syndrome forms of neurosensory hypoacusis. In foreign countries about 200 mutations in 8 genes are diagnosed now. Pathological changes of the majority of genes lead to peripheral neurosensory hypoacusis as all these genes code proteins of the internal ear. In view of this, genetic testing makes topic diagnosis of the disease of the organ of hearing. Most of genetically confirmed hearing problems are indication for cochlear implantation. Prophylactic activity should be focused on explanation of the role and possibilities of genetic counseling and DNA diagnostic tests, promotion of knowledge of etiological structure of congenital hearing disorders. Active measures are necessary for early detection of genetic alterations among risk groups, early diagnosis of mutation carriers among couples of reproductive age with normal hearing.


Overexpression of a pea DNA helicase 45 in bacteria confers salinity stress tolerance.
Salinity stress is one of the major factors negatively affecting growth and productivity in living organisms including plants and bacteria resulting in significant losses worldwide. Therefore, it would be fruitful to develop salinity stress tolerant useful species and also to understand the mechanism of stress tolerance. The pea DNA helicase 45 (PDH45) is a DNA and RNA helicase, homologous to eukaryotic translation initiation factor 4A (eIF-4A) and is involved in various processes including protein synthesis, maintaining the basic activities of the cell, upregulation of topoisomerase I activity and salinity stress tolerance in plant, but its role in salinity stress tolerance in bacteria has not heretofore been studied. This study provides an evidence for a novel function of the PDH45 gene in high salinity (NaCl) stress tolerance in bacteria (Eschericia coli, BL21 cells) also. Furthermore, it has been shown that the functionally active PDH45 gene is required to show the stress tolerance in bacteria because the single mutants (E183G or R363Q) and the double mutant (E183G + R363Q) of the gene could not confer the same function. The response was specific to Na+ ions as the bacteria could not grow in presence of LiCl. This study suggests that the cellular response to high salinity stress across prokaryotes and plant kingdom is conserved and also helps in our better understanding of mechanism of stress tolerance in bacteria and plants. It could also be very useful in developing high salinity stress tolerant useful bacteria of agronomic importance. Overall, this study provides an evidence for a novel function of the PDH45 gene in high salinity stress tolerance in bacteria.


Animal model in the study of colorectal carcinogenesis.
Experimental animal models of neoplastic diseases are important in understanding etiological and pathophysiological processes also in humans. In order to investigate whether the mechanism of genomic instability is associated with chemically induced colorectal tumorigenesis in rat we performed the following study: One hundred and fifty Wistar rats (males 220-280 g and females 140-180 g) were used in the study. Colorectal tumors were induced by means of 15 s.c. applications (20 mg/kg) of 1,2-dimethylhydrazine (DMH). On autopsy, all intestinal lesions were assessed by histological criteria used in human pathology. Forty five tumors were found in the large intestine--30 of these in males and 15 in females, i.e. in 27% of all animals. In four animals multiple primary tumors were found. Histologically 24 tumours were adenocarcinomas, 14 signet-cell carcinoma and 7 adenomas. DNA was extracted from rat neoplastic lesions and adjoining microscopically normal tissues from the same slide and amplified by PCR, using 10 different microsatellite markers from chromosomes 1, 3, 5, 7 and 8. PCR amplicon were analyzed for microsatellite instability with non-isotopic method. In 13 adenocarcinomas (29%) microsatellite instability was found at a minimum of 1 locus. Seven tumors (15.5%) showed microsatellite instability at multiple loci. The results of our experiment suggest that genomic instability is an important molecular event in the pathophysiology of DMH induced colorectal carcinogenesis in rats.


Nucleotide structure and characterization of the murine blood coagulation factor VII gene.
The gene encoding murine coagulation factor VII (fVII) has been cloned. Seven introns and eight exons are present, with the introns positioned as splice junctions between the major domain units of the protein. A total of 11,748 bp of the gene was sequenced, and included 1,077 bp of a 5'-flanking region, in which several high probability binding sites for liver transcription factors were present, as well as a CCAAT sequence and possible GC boxes. Primer extension analysis revealed that the major transcription start site was positioned only 9 residues upstream of the ATG initiation codon, thus providing a very short 5'-untranslated region of the gene. The sequence of the CAP site in the murine fVII gene matched exactly the consensus eukaryotic sequence. A total of 1,484 bp of 3'-flanking nucleotides included a probable polyadenylation site (ATTAAA) and an appropriately positioned downstream consensus sequence (AGTGTTTC) for the efficient formation of a 3' terminus of mRNA. These results indicate that all elements are present for liver-based transcription of the gene for murine factor VII. The sequence and restriction endonuclease map of this gene will facilitate construction of fVII deficient mice and mice containing mutant fVII genes.


Biphasic firing response of nucleus accumbens neurons elicited by THPB-18 and its correlation with DA receptor subtypes.
To investigate the possibility whether THPB-18 (l-12-shloroscoulerine) possesses the D1 agonist-D2 antagonist action on meso-accumbens-mPFC DA system. Single unit spontaneous firing activity was recorded in the nucleus accumbens (NAc) neurons of naive and unilateral-6-hydroxydopamine (6-OHDA)-lesioned Sprague-Dawley rats. The effects of drugs applied intravenously or iontophoretically were determined by the change of firing rates. Under normal conditions, the systemic administration of THPB-18 produced a decrease-increase biphasic firing pattern in the NAc neurons during cumulative doses. High dose of THPB-18 was capable of reversing the inhibition induced by both D2 agonist LY171555 and D1/D2 agonist APO on NAc firing activity. Spiperone pretreatment could not block the high dose of THPB-18-induced firing rate increase, which was reversed by the D1 selective antagonist SCH23390. The tested NAc neurons were effectively inhibited by iontophoretically applied THPB-18 in 90 % of 6-OHDA-lesioned rats, while THPB-18 caused variable effects on the firing of NAc neurons in the neurons of unlesioned rats. The inhibitory effect of THPB-18 was blocked by iontophoretic application of SCH23390, but not D2 antagonist spiperone. Similar to L-stepholidine, THPB-18 also possesses the D1 agonistic-D2 antagonistic dual action on the VTA-NAc DA system.


Alkylresorcinol metabolite concentrations in spot urine samples correlated with whole grain and cereal fiber intake but showed low to modest reproducibility over one to three years in U.S. women.
Two alkylresorcinol (AR) metabolites, 3, 5-dihydroxybenzoic acid (DHBA) and 3-(3,5-dihydroxyphenyl)-1-propanoic acid (DHPPA), in urine have been suggested as biomarkers of whole grain (WG) and cereal fiber intake but the long-term reproducibility and correlation with habitual intake has not been determined. Therefore, we evaluated the long-term reproducibility of AR metabolites in spot urine samples and investigated their correlation with habitual WG and cereal fiber intake in U.S. women. AR metabolites were analyzed in 104 women participating in the Nurses' Health Study II and WG and fiber intakes were assessed using a FFQ. Long-term reproducibility was assessed by calculating the intra-class correlation coefficients (ICC) using samples taken 1-3 y (mean 1.8 y) apart. The observed Spearman correlation coefficients (r(s)) and r(s) adjusted for within-participant variation in the biomarker were calculated between WG and fiber intake and biomarkers. The long-term reproducibility was poor for DHBA [ICC = 0.17 (95% CI: 0.05, 0.43)] and modest for DHPPA [ICC = 0.31 (95% CI: 0.17, 0.51)]. The correlation between WG intake in 1995 and DHPPA measured 2 y later was 0.37 (P < 0.0001); the adjusted correlation was 0.60 (95% CI: 0.37, 0.76). Cereal fiber and WG intake were similarly correlated to the biomarkers. DHPPA in spot urine samples reflected WG intake despite relatively low intake of food sources of AR. The poor to modest reproducibility may limit the use of single measurements of these biomarkers in cohort studies in the US, where WG intake is relatively low and has changed over time. But DHPPA in repeated samples may be useful for validating WG intake and assessing compliance in WG intervention studies.


The evidence for dietary prevention and treatment of cardiovascular disease.
During the past few decades numerous studies have reported the atherogenic potential of saturated fatty acids, trans-fatty acids, and cholesterol, and beneficial effects of fiber, phytostanols/phytosterols, n-3 fatty acids, a Mediterranean diet, and other plant-based approaches. The purpose of this article is to provide a comprehensive and systematic review of the evidence associated with key dietary factors and risk of cardiovascular disease-an umbrella term encompassing diseases that affect the heart and blood vessels, including coronary heart disease, coronary artery disease, dyslipidemia, and hypertension-in conjunction with the work of the American Dietetic Association Evidence Analysis Library review on diet and lipids, updated with new evidence from the past 2 years. The criteria used and results cited provide scientific rationale for food and nutrition professionals and other health professionals for counseling patients. Details of these searches are available within the American Dietetic Association Evidence Analysis Library online (http://adaevidencelibrary.com). Potential mechanisms and needs for future research are summarized for each relevant nutrient, food, or food component.


How soft is a single protein? The stress-strain curve of antibody pentamers with 5 pN and 50 pm resolutions.
Understanding the mechanical functionalities of complex biological systems requires the measurement of the mechanical compliance of their smallest components. Here, we develop a force microscopy method to quantify the softness of a single antibody pentamer by measuring the stress-strain curve with force and deformation resolutions, respectively, of 5 pN and 50 pm. The curve shows three distinctive regions. For ultrasmall compressive forces (5-75 pN), the protein's central region shows that the strain and stress are proportional (elastic regime). This region has an average Young's modulus of 2.5 MPa. For forces between 80 and 220 pN, the stress is roughly proportional to the strain with a Young's modulus of 9 MPa. Higher forces lead to irreversible deformations (plastic regime). Full elastic recovery could reach deformations amounting to 40% of the protein height. The existence of two different elastic regions is explained in terms of the structure of the antibody central region. The stress-strain curve explains the capability of the antibody to sustain multiple collisions without any loss of biological functionality.


Exploring evolution and diversity of Chinese Dipterocarpaceae using next-generation sequencing.
Tropical forests, a key-category of land ecosystems, are faced with the world's highest levels of habitat conversion and associated biodiversity loss. In tropical Asia, Dipterocarpaceae are one of the economically and ecologically most important tree families, but their genomic diversity and evolution remain understudied, hampered by a lack of available genetic resources. Southern China represents the northern limit for Dipterocarpaceae, and thus changes in habitat ecology, community composition and adaptability to climatic conditions are of particular interest in this group. Phylogenomics is a tool for exploring both biodiversity and evolutionary relationships through space and time using plastome, nuclear and mitochondrial genome. We generated full plastome and Nuclear Ribosomal Cistron (NRC) data for Chinese Dipterocarpaceae species as a first step to improve our understanding of their ecology and evolutionary relationships. We generated the plastome of Dipterocarpus turbinatus, the species with the widest distribution using it as a baseline for comparisons with other taxa. Results showed low level of genomic diversity among analysed range-edge species, and different evolutionary history of the incongruent NRC and plastome data. Genomic resources provided in this study will serve as a starting point for future studies on conservation and sustainable use of these dominant forest taxa, phylogenomics and evolutionary studies.


Alternative processing of hepatitis delta virus antigenomic RNA transcripts.
Intrinsic to the life cycle of hepatitis delta virus (HDV) is the fact that its RNAs undergo different forms of posttranscriptional RNA processing. Transcripts of both the genomic RNA and its exact complement, the antigenomic RNA, undergo ribozyme cleavage and RNA ligation. In addition, antigenomic RNA transcripts can undergo 5' capping, 3' polyadenylation, and even RNA editing by an adenosine deaminase. This study focused on the processing of antigenomic RNA transcripts. Two approaches were used to study the relationship between the events of polyadenylation, ribozyme cleavage, and RNA ligation. The first represented an examination under more controlled conditions of mutations in the poly(A) signal, AAUAAA, which is essential for this processing. We found that when a separate stable source of deltaAg-S, the small delta protein, was provided, the replication ability of the mutated RNA was restored. The second approach involved an examination of the processing in transfected cells of specific Pol II DNA-directed transcripts of HDV antigenomic sequences. The DNA constructs used were such that the RNA transcripts were antigenomic and began at the same 5' site as the mRNA produced during RNA-directed HDV genome replication. A series of such constructs was assembled in order to test the relative abilities of the transcripts to undergo processing by polyadenylation or ribozyme cleavage at sites further 3' on a multimer of HDV sequences. The findings from the two experimental approaches led to significant modifications in the rolling-circle model of HDV genome replication.


Can pentoxifylline prevent renal disease?
The characteristics of pentoxifylline(Trental) suggest that it is an ideal agent to be used as adjunct in the therapy of chronic proliferative glomerulonephritides. Theoretical considerations suggest that pentoxifylline should also protect against and even ameliorate tubulo-interstitial fibrosis of affected kidneys.


Infection of Zebrafish Larvae with Aspergillus Spores for Analysis of Host-Pathogen Interactions.
Invasive aspergillosis (IA) is one of the most common fungal infections among immunocompromised individuals. Despite the availability of antifungal drugs, IA can cause >50% mortality in infected immunocompromised patients. It is crucial to determine both host and pathogen factors that contribute to infection susceptibility and low survival rates in infected patients in order to develop novel therapeutics. Innate immune responses play a pivotal role in recognition and clearance of Aspergillus spores, though little is known about the exact cellular and molecular mechanisms. Reliable models are required to investigate detailed mechanistic interactions between the host and pathogen. The optical clarity and genetic tractability of zebrafish larvae make them an intriguing model to study host-pathogen interactions of multiple human bacterial and fungal infections in a live and intact host. This protocol describes a larval zebrafish Aspergillus infection model. First, Aspergillus spores are isolated and injected into the zebrafish hindbrain ventricle via microinjection. Then, chemical inhibitors such as immunosuppressive drugs are added directly to the larval water. Two methods to monitor the infection in injected larvae are described, including the 1) homogenization of larvae for colony forming unit (CFU) enumeration and 2) a repeated, daily live imaging setup. Overall, these techniques can be used to mechanistically analyze the progression of Aspergillus infection in vivo and can be applied to different host backgrounds and Aspergillus strains to interrogate host-pathogen interactions.


[Cutaneous lesions and blood count changes in a 9-month old girl with glutaric aciduria type I].
Non-specific cutaneous lesions are common in patients suffering from acute myeloid leukemia (AML). Leukemic skin infiltrates are present in about 30% of cases of monoblastic or myelomonocytic leukemia. The appearance of specific skin lesions can precede bone marrow involvement. We report the case of a 9-month-old girl with acute myelogenous leukemia (FAB M5) and glutaric aciduria type I which initially presented with cutaneous lesions, anemia and leukopenia.


Involvement of Runx3 in the basal transcriptional activation of the mouse angiotensin II type 1 receptor-associated protein gene.
We previously cloned a molecule that interacts with angiotensin II type 1 (AT1) receptor to exert an inhibitory function on AT1 receptor signaling that we named ATRAP/Agtrap (for AT1 receptor-associated protein). In the present study we examined the regulation of basal ATRAP gene expression using renal distal convoluted tubule cells. We found that serum starvation upregulated basal expression of ATRAP gene, a response that required de novo mRNA and protein synthesis. Luciferase assay revealed that the proximal promoter region directs transcription and that a putative binding site of runt-related transcription factors (RBE) is important for transcriptional activation. The results of RBE-decoy transfection and endogenous knockdown by small interference RNA showed that the runt-related transcription factor Runx3 is involved in ATRAP gene expression. Chromatin immunoprecipitation assay also supported the binding of Runx3 to the ATRAP promoter in renal distal convoluted tubule cells. Immunohistochemistry demonstrated the expression of Runx3 and ATRAP proteins in the distal convoluted and connecting tubules of the kidney in consecutive sections. Furthermore, the Runx3 immunostaining was decreased together with a concomitant suppression of ATRAP expression in the affected kidney after 7 days of unilateral ureteral obstruction. These findings indicate that Runx3 plays a role in ATRAP gene expression in renal distal tubular cells both in vitro and in vivo.


Surface Topography of PDMS Replica Transferred from Various Decellularized Aortic Lumens Affects Cellular Orientation.
Cells sense and respond to various surface topographies of substrates. Many types of topographical architectures have been developed for understanding cell-extracellular matrix (ECM) interactions and for their application in biomaterials. In the present study, as a topographical surface similar to native tissue, we developed a PDMS replica prepared using the transferring method of the decellularized aorta, which is an ECM assembly, and its cellular behaviors, such as orientation and elongation on it. Decellularized aortas were prepared by high hydrostatic pressure (HHP) and sodium dodecyl sulfate (SDS) methods for use as templates. Scanning electron microscopic observation of the SDS replica showed a randomly rough surface. Further, microscaled linear structures along the direction of the aortic longitudinal axis were observed on the HHP replica. These results indicated that the topographical surface of the HHP and SDS decellularized aorta could be replicated to their replicas at a microscale. Fibroblasts (NIH3T3) and endothelial cells (HUVECs) were cultured on their surfaces. Although they were randomly aligned on the SDS replica and flat surface, the high cellular alignment along with the direction of the aortic longitudinal axis was shown in the HHP replica and HHP decellularized aorta. These results suggest that the topographical structure similar to a native aorta could effectively induce the cell alignment, which is important to regulate cellular functions, and could provide important methodologies and knowledge for vascular biomaterials or culture substrates.


Cardiac lesions and initial laboratory data in Kawasaki disease: a nationwide survey in Japan.
Cardiac lesions, such as coronary dilatation, aneurysms, narrowing, myocardial infarction, and valvular lesions, sometimes occur in Kawasaki disease, but most studies have only evaluated cardiac lesions in the later phase of the disease. This study was undertaken to clarify the related factors between cardiac lesions and laboratory data in the initial phase of Kawasaki disease. We conducted a cross-sectional study using data for 26 691 patients from the 22nd nationwide survey of Kawasaki disease in Japan, the observation period of which was from January 2011 through December 2012. We excluded patients with recurrent Kawasaki disease and who were more than seven days from the start of symptoms at admission. We analyzed 23 155 cases (13 353 boys; mean age: 923 ± 734 days) with available laboratory data for white blood cell count, platelet count, serum albumin, and C-reactive protein (CRP). Cardiac lesions were detected in 984 cases (656 boys and 328 girls); lesions were classified as coronary dilatation (764 cases), coronary aneurysm (40), giant coronary aneurysm (6), coronary narrowing (3), and valvular lesions (204). The significant related factors of initial coronary dilatation were male sex (odds ratio [OR] 1.73), older age (OR per 100 days increase 1.03), higher platelet count (OR per 10 000 cells/µL increase 1.006), lower albumin (OR per 1 g/dL increase 0.66), and higher CRP (OR per 1 mg/dL increase 1.02). The factors related to coronary aneurysm were higher platelet count (OR 1.01) and lower albumin (OR 0.34). No factors were significantly related to giant coronary aneurysm. The related factors of valvular lesions were age (OR 0.98), and higher CRP (OR 1.05). Clinicians should consider male sex, older age, higher platelet count, lower albumin levels, and higher CRP levels when assessing risk of cardiac lesions in the initial phase of Kawasaki disease.


Circulating Heparan Sulfate Fragments Attenuate Histone-Induced Lung Injury Independently of Histone Binding.
Extracellular histones are cationic damage-associated molecular pattern molecules capable of directly inducing cellular injury via charge-mediated interactions with plasma membranes. Accordingly, histones released into the plasma during critical illness are known to contribute to the onset and propagation of lung injury. Vascular injury (with consequent degradation of the endothelial glycocalyx) simultaneously releases anionic heparan sulfate fragments (hexa- to octasaccharides in size) into the plasma. It is unknown whether this endogenous release of heparan sulfate fragments modulates charge-dependent histone cytotoxicity, or if exogenous heparan sulfate fragments could therapeutically attenuate histone-induced lung injury. Using isothermic calorimetry, we found that extracellular histones only bind to heparan sulfate fragments ≥ 10 saccharides in size, suggesting that glycocalyx-derived heparan sulfate hexa/octasaccharides are incapable of intercepting/neutralizing circulating histones. However, we found that even heparan sulfate fragments incapable of histone binding (e.g., tetrasaccharides) attenuated histone-induced lung injury in vivo, suggesting a direct, size-independent protective effect of heparan sulfate. We found that histones had no effect on human neutrophils ex vivo but exerted toll-like receptor-independent cytotoxicity on human pulmonary microvascular endothelial cells in vitro. This cytotoxicity could be prevented by either the addition of negatively charged (i.e., highly sulfated) heparan sulfate tetrasaccharides (incapable of binding histones) or decasaccharides (capable of binding histones). Taken together, our findings suggest that heparan sulfate oligosaccharides may directly exert pulmonary endothelial-protective effects that attenuate histone-mediated lung injury.


Antisocial Personality Disorder Subscale (Chinese Version) of the Structured Clinical Interview for the DSM-IV Axis II disorders: validation study in Cantonese-speaking Hong Kong Chinese.
OBJECTIVE. Antisocial personality disorder (ASPD) is a risk factor for violence and is associated with poor treatment response when it is a co-morbid condition with substance abuse. It is an under-recognised clinical entity in the local Hong Kong setting, for which there are only a few available Chinese-language diagnostic instruments. None has been tested for its psychometric properties in the Cantonese-speaking population in Hong Kong. This study therefore aimed to assess the reliability and validity of the Chinese version of the ASPD subscale of the Structured Clinical Interview for the DSM-IV Axis II Disorders (SCID-II) in Hong Kong Chinese. METHODS. This assessment tool was modified according to dialectal differences between Mainland China and Hong Kong. Inpatients in Castle Peak Hospital, Hong Kong, who were designated for priority follow-up based on their assessed propensity for violence and who fulfilled the inclusion criteria for the study, were recruited. To assess the level of agreement, best-estimate diagnosis made by a multidisciplinary team was compared with diagnostic status determined by the SCID-II ASPD subscale. The internal consistency, sensitivity, and specificity of the subscale were also calculated. RESULTS. The internal consistency of the subscale was acceptable at 0.79, whereas the test-retest reliability and inter-rater reliability showed an excellent and good agreement of 0.90 and 0.86, respectively. Best-estimate clinical diagnosis-SCID diagnosis agreement was acceptable at 0.76. The sensitivity, specificity, positive and negative predictive values were 0.91, 0.86, 0.83, and 0.93, respectively. CONCLUSION. The Chinese version of the SCID-II ASPD subscale is reliable and valid for diagnosing ASPD in a Cantonese-speaking clinical population.


Symmetrized correlation function for liquid para-hydrogen using complex-time pair-product propagators.
We present a simple and efficient method for calculating symmetrized time correlation functions of neat quantum fluids. Using the pair-product approximation to each complex-time quantum mechanical propagator, symmetrized correlation functions are written in terms of a double integral for each degree of freedom with a purely positive integrand. At moderate temperatures and densities, where the pair-product approximation to the Boltzmann operator is sufficiently accurate, the method leads to quantitative results for the early time part of the correlation function. The method is tested extensively on liquid para-hydrogen at 25 K and used to obtain accurate quantum mechanical results for the initial 0.2 ps segment of the symmetrized velocity autocorrelation function of this system, as well as the incoherent dynamic structure factor at certain momentum transfer values.


An initial experience with screening for colon polyps using spiral CT with and without CT colography (virtual colonoscopy)
Computed tomographic (CT) colography (virtual colonoscopy) is a new imaging method for detection of colon polyps and cancer. To evaluate the sensitivity of CT colography for polyp detection in a population without symptoms that included persons without colon neoplasia and with radiologists blinded to colonoscopic findings. Forty-six persons without symptoms underwent spiral CT followed by same-day colonoscopy with subsequent inspection of two-dimensional axial CT images, interactive multiplanar images, and surfaced and volume-rendered images of the colon (three-dimensional CT colography). Three-dimensional CT colography was superior to two-dimensional axial imaging for detection of colon polyps. Three-dimensional CT colography depicted 1 of 4 (25%) adenomas 2 cm in diameter or larger, 6 of 10 (60%) adenomas 1 to 1.9 cm, 6 of 14 (43%) 6 to 9 mm, and 7 of 65 (11%) 5 mm in diameter or smaller. Three-dimensional CT colography showed a polyp that might have led to colonoscopy in 3 of 4 (75%) patients whose largest adenoma was 2 cm or larger, 5 of 6 (83%) patients with largest adenoma 1 to 1.9 cm, 3 of 7 (43%) patients with largest adenoma 6 to 9 mm, and 4 of 16 patients (25%) with largest adenoma 5 mm or smaller. Large, flat adenomas of the right colon were difficult to identify with three-dimensional CT colography. The specificity of three-dimensional CT colography for patients with adenomas 1 cm in diameter or larger was 89%. Examination of patients with missed adenomas after unblinding indicated that meticulous bowel preparation and adequate distention are critical to accurate interpretation. Perceptual errors were common. CT colography as performed in this study is not adequate as a colorectal cancer screening test. Several technical factors that appear critical to accurate performance of CT colography are defined.


Hearing Preservation in Pediatric Cochlear Implantation.
Currently, there is a paucity of literature evaluating hearing preservation outcomes in children following cochlear implantation. The objective of the current study is to report pediatric hearing preservation results following cochlear implantation with conventional full-length electrodes. Retrospective review (2000-2016). Tertiary referral center. All pediatric patients with a ≤ 75 dB preoperative low-frequency pure tone average (LFPTA; 250-500 Hz average), who underwent cochlear implantation with a conventional length electrode. Cochlear implantation. Complete, partial, minimal, or no hearing preservation following cochlear implantation (Skarzynski et al., 2013); maintenance of functional low frequency hearing (≤85 dB LFPTA). A total of 43 ears, in 35 pediatric patients, met inclusion criteria. The mean age at time of implantation was 8.6 years (range, 1.4-17.8 yr), 20 (57.1%) patients were female, and 25 (58.1%) cases were left-sided.The mean preoperative ipsilateral low frequency PTA and conventional four-frequency PTA (500, 1000, 2000, 3000 Hz average) were 54.2 dB (range, 15-75 dB) and 82.2 dB (range, 25-102.5 dB), respectively. The mean low frequency PTA and conventional four-frequency PTA shifts comparing the pre- and first postoperative audiogram were Δ25.2 dB (range, -5 to 92.5 dB) and Δ18.3 dB (range, -8.8 to 100 dB), respectively. Overall, 17 (39.5%) ears demonstrated complete hearing preservation, 19 (44.2%) ears partial hearing preservation, 0 minimal hearing preservation, and 7 (16.3%) exhibited no measurable acoustic hearing after surgery. In total, 28 (65.1%) ears maintained functional low-frequency hearing (i.e., ≤85 dB LFPTA) based on the initial postoperative audiogram. There was no statistically significant difference in the initial low frequency PTA shift comparing lateral wall and perimodiolar electrodes (Δ22.2 versus Δ28.1 respectively; p = 0.44), cochleostomy and round window insertions (Δ25.2 vs. Δ24.7 respectively; p = 0.95), or statistically significant association between age at implantation and low frequency PTA shift (r = 0.174; p = 0.26).In total, 22 ears in 19 patients had serial audiometric data available for review. Over a mean duration of 43.8 months (range, 2.6-108.3 mo) following surgery, the mean low frequency PTA and conventional four-frequency PTA shift comparing the initial postoperative and most recent postoperative audiogram was Δ9.7 dB (range, -27.5 to 57.5 dB) and Δ8.1 dB (range, -18.8 to 31.9 dB), respectively. Varying levels of hearing preservation with conventional length electrodes can be achieved in most pediatric subjects. In the current study, 82% of patients maintained detectable hearing thresholds and 65% maintained functional low-frequency acoustic hearing. These data may be used to guide preoperative counseling in pediatric patients with residual acoustic hearing. Additionally, the favorable rates of hearing preservation achieved in children provide further evidence for the expansion of pediatric cochlear implant candidacy to include patients with greater degrees of residual hearing.


Role of hematopoietic growth factors/flt3 ligand in expansion and regulation of dendritic cells.
Dendritic cells (DCs) are hematopoietic cells that initiate immune responses by presenting antigen to T cells. Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a primary growth factor for DCs in vitro, but recently it was recognized that other factors including flt3 ligand (FL) and G-CSF expand various DC subsets in vivo. DCs undergo a complex series of maturation and activation steps after they acquire antigen and before they can activate resting T cells. In addition, they must traffic to T-cell-rich areas of lymph nodes (LN) to achieve this. Each of these steps is tightly regulated, and in the last year progress has been made in identifying some of the key molecules involved in each of these steps. This progress will further the efforts underway to develop DCs as vaccine adjuvants.


Effect of Telephone-Delivered Collaborative Goal Setting and Behavioral Activation vs Enhanced Usual Care for Depression Among Adults With Uncontrolled Diabetes: A Randomized Clinical Trial.
Depression symptoms are present in one-third of patients with diabetes, contributing to significant adverse consequences. Population screening of high-risk patients coupled with telephone delivery of evidence-based therapies for comorbid diabetes may address barriers to care. To evaluate the effectiveness of proactive population screening plus telephone delivery of a collaborative goal-setting intervention among high-risk patients with uncontrolled diabetes and depression. In this randomized clinical trial, 225 participants (intervention [n = 136] and control [n = 89]) were enrolled from a regional Veterans Healthcare System serving Southeast Texas from November 1, 2012, through June 24, 2016. Data were gathered at baseline and 6 and 12 months after intervention. Patients selected had uncontrolled diabetes (hemoglobin A1c [HbA1c] >7.5%]) and clinically significant depression (Patient Health Questionnaire-9 scores [PHQ-9] ≥10) and were living more than 20 miles from the Veterans Affairs medical center. Data collection was completed on December 6, 2016, and final analyses were completed by January 25, 2018. All analyses were intent to treat. Healthy Outcomes Through Patient Empowerment (HOPE) included 9 telephone sessions with 24 trained health care professionals using collaborative goal-setting and behavioral activation methods. The control group received enhanced usual care (EUC) and notification of high-risk status. Change in depression symptoms using PHQ-9 and glycemic control using HbA1c from baseline to 6 months and to 12 months. Secondary analyses evaluated clinically significant responses for these measures. Among 225 participants, 202 (89.8%) were men, the mean (SD) age was 61.9 (8.3) years, 145 (64.4%) were married, and 156 (69.3%) had some education beyond high school. For the overall study, 38 participants (16.9%) were lost to follow-up or withdrew at 6 months and another 21 (9.3%) were lost to follow-up or withdrew at 12 months. Repeated-measures analysis with multiple imputation for missing data assessing the interaction of treatment group (HOPE vs EUC) and time (baseline, 6 months, and 12 months) found no significant improvement in PHQ-9 (β, 1.56; 95% CI, -0.68 to 3.81; P = .17) or HbA1c (β, -0.005; 95% CI, -0.73 to 0.72; P = .82). Analyses using t test for change from baseline to 12 months showed a HOPE vs EUC between-group mean difference for PHQ-9 of 2.14 (95% CI, 0.18 to 4.10; P = .03) and for HbA1c of -0.06% (95% CI, -0.61% to 0.50%; P = .83). A secondary analysis of patients experiencing a clinical response found that 52.1% of HOPE participants had clinically significant responses in PHQ-9 at 12 months vs 32.9% in EUC (difference, 0.19; 95% CI, 0.04-0.33; P = .01). Telephone-delivered, collaborative goal setting produced clinically significant reductions in depression symptoms but not glycemic control among patients who remained engaged at 12 months compared with EUC among a population screened sample of high-risk patients with diabetes and depression. Although the intervention created some lasting effect for depression, additional strategies are needed to maintain engagement of this high-risk population within an interprofessional team approach to primary care. ClinicalTrials.gov identifier: NCT01572389.


Risk factors in autism: Thinking outside the brain.
Autism spectrum disorders (ASD) are complex neurodevelopmental conditions that have been rising markedly in prevalence for the past 30 years, now thought to affect 1 in 68 in the United States. This has prompted the search for possible explanations, and has even resulted in some controversy regarding the "true" prevalence of autism. ASD are influenced by a variety of genetic, environmental, and possibly immunological factors that act during critical periods to alter key developmental processes. This can affect multiple systems and manifests as the social and behavioral deficits that define these disorders. The interaction of environmental exposures in the context of an individual's genetic susceptibilities manifests differently in each case, leading to heterogeneous phenotypes and varied comorbid symptoms within the disorder. This has also made it very difficult to elucidate underlying genes and exposure profiles, but progress is being made in this area. Some pharmaceutical drugs, toxicants, and metabolic and nutritional factors have been identified in epidemiological studies as increasing autism risk, especially during the prenatal period. Immunologic risk factors, including maternal infection during pregnancy, autoantibodies to fetal brain proteins, and familial autoimmune disease, have consistently been observed across multiple studies, as have immune abnormalities in individuals with ASD. Mechanistic research using animal models and patient-derived stem cells will help researchers to understand the complex etiology of these neurodevelopmental disorders, which will lead to more effective therapies and preventative strategies. Proposed therapies that need more investigation include special diets, probiotics, immune modulation, oxytocin, and personalized pharmacogenomic targets. The ongoing search for biomarkers and better treatments will result in earlier identification of ASD and provide much needed help and relief for afflicted families.


Pediatric multiple sclerosis.
Multiple sclerosis (MS) onset in childhood occurs in a small proportion of individuals with the disease, although the precise incidence of pediatric MS is unknown. It may be difficult to distinguish the initial attack of pediatric MS from acute disseminated encephalomyelitis, particularly in very young children. Environmental and genetic factors that appear to increase the risk of pediatric MS include prior infection with Epstein-Barr virus, exposure to cigarette smoke, and HLA-DRB1*15 haplotype. Children may have more posterior fossa involvement at onset and a higher relapse rate than adults with MS. Although time to disability may be longer than in adults, pediatric MS is associated with an earlier age of disability. Off-label use of standard disease-modifying therapies for adult MS is common, although data regarding the efficacy and safety for these medications are limited.


Development and deployment of a supply-chain digital tool to predict fluid milk spoilage due to psychrotolerant sporeformers.
Psychrotolerant sporeformers pose a challenge to maintaining fluid milk quality. Dynamic temperature changes along the supply chain can favor the germination and growth of these bacteria and lead to fluid milk spoilage. In this study, we aim to expand on our previous work on predicting milk spoilage due to psychrotolerant sporeformers. The key model innovations include (i) the ability to account for changing temperatures along the supply chain, and (ii) a deployed user-friendly interface to allow easy access to the model. Using the frequencies and concentrations of 8 Bacillales subtypes specific to fluid milk collected in New York, the model simulated sporeformer growth in half-gallons of high-temperature short-time (HTST) pasteurized fluid milk transported from processing facility to retail store and then to consumer. The Monte Carlo simulations predicted that 44.3% of half-gallons of milk were spoiled (defined as having a bacterial concentration > 20,000 cfu/mL, a conservative estimate that represents the Pasteurized Milk Ordinance [PMO] regulatory limit) after 21 d of refrigerated storage at consumer's home. Model validations showed that the model was the most accurate in predicting the mean sporeformer concentration at low temperatures (i.e., at 3 and 4°C; compared with higher temperatures at 6 and 10°C) within the first 21 d of consumer storage, with a root mean square error of 0.29 and 0.34 log10cfu/mL, respectively. Global sensitivity analyses indicated that home storage temperature, facility-to-retail transportation temperature, and initial spore concentration were the 3 most influential factors for predicting milk spoilage on d 21 of shelf-life. What-if scenarios indicated that microfiltration was predicted to be the most effective strategy to reduce spoilage. The implementation of this strategy (assumed to reduce initial spore concentration by 2.2 log10cfu/mL) was predicted to reduce the percentage of spoiled milk by 17.0 percentage points on d 21 of storage and could delay the date by which 50% of half-gallons of milk were spoiled from d 25 to d 35. Overall, the model is readily deployed as a digital tool for assessing fluid milk spoilage along the supply chain and evaluating the effectiveness of intervention strategies, including those that target storage temperatures at different supply chain stages.


MicroRNA-26a induced by hypoxia targets HDAC6 in myogenic differentiation of embryonic stem cells.
The importance of epigenetic regulation for maintenance of embryonic stem cell (ESC) pluripotency or for initiation of differentiation is widely accepted. However, the molecular mechanisms are poorly understood. We recently reported that a hypoxic microenvironment induces ESC differentiation. In the present study, we found that hypoxia-responsive histone deacetylase 6 (HDAC6) performs an essential signaling function for myogenic differentiation of ESCs. HDAC6 was downregulated in hypoxic ESCs or during differentiation. A knock-down of HDAC6 in ESCs resulted in induction of myogenic markers, including Pax7. Suppression of HDAC6 increased acetylation of core histones H3 and H4, leading to enhanced binding of RNA polymerase II to the Pax7 promoter. Transplantation of HDAC6 knock-down cells facilitated muscle regeneration in vivo. Importantly, the downregulation of HDAC6 by hypoxia was not mediated by HIF1α or HIF2α, master transcription regulators under hypoxia, but by induction of microRNA-26a that directly targeted the 3'-untranslated region (3'-UTR) of HDAC6. A point mutation of the microRNA-26a-binding sequence in the HDAC6 3'-UTR diminished the luciferase reporter activity. Taken together, these results suggest that environmental cues of differentiation modulate the epigenetic machinery and guide stem cells to commit to a specific lineage.


Targeting of interleukin-2 to the periplasm of Escherichia coli.
A synthetic gene coding for interleukin-2 (IL-2) was used to produce large amounts of recombinant IL-2 (met-IL-2) in Escherichia coli. Met-IL-2 was found to accumulate in the cytoplasm in an insoluble, aggregated form. Inclusion bodies located at the pole caps of cells were detected using immunogold labelling. Constructs were designed to fuse the IL-2 gene to DNA fragments encoding signal peptides for an outer-membrane protein (OmpA) or for a periplasmic protein (PhoA) of E. coli. No significant maturation was observed with these fusion proteins which were found in an insoluble form in the cytoplasm. The influence of charge disposition at the N-terminus of the mature portion of the protein was investigated by replacing positively charged amino acids with glutamic acid. None of the introduced substitutions had any effect. Various factors that might affect expression, secretion and folding were examined in an attempt to obtain secretion. By fusing IL-2 to the precursor maltose-binding protein (preMBP) a large fraction of the preMBP-IL-2 protein was correctly processed and transported to the periplasmic space. IL-2 derived from MBP-IL-2 after FXa cleavage possessed similar specific activity to recombinant IL-2 produced in Chinese Hamster ovary cells.


Characterization of a cloned ultraviolet radiation (UV)-induced suppressor T cell line that is capable of inhibiting anti-UV tumor-immune responses.
Ultraviolet radiation (UV) is a potent carcinogen for the induction of skin tumors. In this regard, UV represents a unique carcinogenic agent, in that depending on the dosage and conditions of administration it can function as either a complete carcinogen, a carcinogenic promoting agent, or an immunologic modulator of anti-tumor rejection responses. The immunologic modulatory activity of UV has been demonstrated in numerous studies. These studies have shown that subcarcinogenic doses of UV induce a population of suppressor T lymphocytes (Ts cells) that allow for the emergence and progression of UV-induced tumors. Although the phenotypic and functional properties of these cells have been established, it was unclear as to whether the UV-induced Ts cell population consisted of multiple Ts cell clones able to recognize a range of unique tumor antigens or a limited number of Ts cell clones with functional specificity directed toward a common tumor-associated antigen (TAA). To address this question, an interleukin 2-dependent, UV-induced cloned Ts cell line was derived, by limiting dilution without exogenous antigen stimulation, from the splenic T cell population of a C3H mouse that had been exposed to a subcarcinogenic dose of UV. This Ts cell line, designated UV2.10, was selected for its ability to suppress the in vitro differentiation of cytotoxic T cells from the draining lymph nodes of UV-induced tumor-immune mice. When transferred into non-UV-irradiated syngeneic mice, which normally reject a UV-induced tumor implant, the UV2.10 cells rendered their hosts susceptible to the growth of a battery of UV-induced tumors. Although capable of suppressing in vitro and in vivo UV-induced tumor-immune responses, UV2.10 cells did not inhibit the elicitation of contact hypersensitivity responses, the rejection of allogeneic skin grafts, responses, the rejection of allogeneic skin grafts, or the rejection of allogeneic UV-induced tumors. These data suggest that the cloned UV2.10 Ts cell line possesses functional antigenic specificity that may be limited to the regulation of immune responses that are directed toward the TAA expressed by syngeneic UV-induced tumors. Employing monoclonal antibodies and FACS analysis, the cell surface phenotype of the UV2.10 cell line was determined to be: Thy-1.2+, Lyt-1-, Lyt-2+/- (dim), L3T4a-, I-A/E-, and I-J+. This cell surface phenotype is indicative of a suppressor T cell. These data lend further support to the hypothesis that the UV-induced Ts cell population is clonal in nature and functions through its ability to recognize a common TAA(s) that appears to be expressed by virtually all UV-induced tumors.


Pancreatic cancer: updates on translational research and future applications.
Pancreatic cancer is one of the most lethal malignancies with a mortality rate almost equal to its incidence. It is ranked as the fourth leading cause of cancer-related deaths in the United States, and despite intensive basic and clinical research over the last few years, the survival benefit for the majority of patients with pancreatic cancer is still disappointing. Due to the absence of specific symptoms and the lack of early detection tests, pancreatic cancer is usually diagnosed at an advanced inoperrable stage and palliative chemotherapy with the purine analogue gemcitabine in combination with the targeted agent erlotinib, remains the mainstay method in the management of these patients. Therefore, there is an imperative need for new findings in the translational research field with prognostic, predictive and therapeutic value. In this paper we summarize five most interesting research abstracts as presented at the 2013 American Society of Clinical Oncology (ASCO) Gastrointestinal Cancers Symposium. In particular, we focus on Abstract #141 which investigates the interaction between liver and pancreatic organ damage in patients with pancreatic cancer and the potential contribution of the patatin-like phospholipase domain containing 3 (PNPLA3) gene variation in pancreatic cancer development and on Abstract #149, in which, the prognostic and predictive role of SWI/SNF complex, a chromatin-remodeling complex, is examined. The key role of pharmacogenomics, in terms of predicting response and resistance to chemotherapy in pancreatic cancer patients, is analyzed in Abstract #142 and the contribution of circulating tumor cell detection in the early diagnosis of pancreatic cancer, allowing the avoidance of more invasive procedures like EUS-FNA, is discussed in Abstract #157. Lastly, in Abstract #164, the diagnostic utility of YKL-40 and IL-6 in pancreatic cancer patients is investigated.


Endovascular Cooling Catheter for Selective Brain Hypothermia: An Animal Feasibility Study of Cooling Performance.
Therapeutic hypothermia represents a promising neuroprotective treatment in acute ischemic stroke. Selective cerebral hypothermia applied early, prior to and during endovascular mechanical recanalization therapy, may be beneficial in the critical phase of reperfusion. We aimed to assess the feasibility of a new intracarotid cooling catheter in an animal model. Nine adult sheep were included. Temperature probes were introduced into the frontal and temporal brain cortices bilaterally. The cooling catheter system was introduced into a common carotid artery. Selective blood cooling was applied for 180 minutes. Systemic and local brain temperatures were measured during cooling and rewarming. Common carotid artery diameters and flow were measured angiographically and by Doppler sonography. The common carotid artery diameter was between 6.7 and 7.3 mm. Common carotid artery blood flow velocities increased moderately during cooling and after catheter removal. Maximum cerebral cooling in the ipsilateral temporal cortex was -4.7°C (95% CI, -5.1 to -4.0°C). Ipsilateral brain temperatures dropped significantly faster and became lower compared with the contralateral cortex with maximum temperature difference of -1.3°C (95% CI, -1.5 to -1.0°C; P < .0001) and compared with systemic temperature (-1.4°C; 95% CI, -1.7 to -1.0°C; P < .0001). Sheep proved a feasible animal model for the intracarotid cooling catheter. Fast induction of selective mild hypothermia was achieved within the cooled cerebral hemisphere, with stable temperature gradients in the contralateral brain and systemic blood. Further studies are required to demonstrate any therapeutic benefit of selective cerebral cooling in a stroke model.


Inconsistent echocardiographic grading of aortic stenosis: is the left ventricular outflow tract important?
Discrepancy in the echocardiographic severity grading of aortic stenosis (AS) based on current guidelines has been reported. We sought to investigate the left ventricular outflow tract diameter (LVOTd) as a source of inconsistencies, and to explore hypothetical alternatives for discrepancy improvement. Retrospective echocardiographic cross-sectional analysis. From 2000 to 2010, we identified all AS patients with left ventricular EF ≥50%, mean gradient (MG) ≥20 mm Hg, aortic valve area (AVA) ≤2.5 cm(2), <moderate (2+) aortic regurgitation; and divided them into three groups: patients with 'small ' LVOTd 1.7-1.9 cm, 'average' LVOTd 2.0-2.2 cm and 'large' LVOTd ≥2.3 cm. In each group, inconsistency of data for classification of severity of AS was assessed and alternative thresholds explored. Of 9488 total patients, 58% were men, LVOTd 2.18±0.19 cm, peak velocity (Vmax) 3.9±0.8 m/s, MG 37±16 mm Hg, and AVA 1.09±0.34 cm(2). Small LVOTd patients were older women (91%) with worse systemic haemodynamics and more prevalent paradoxical low-flow, compared with average and large LVOTd patients (all parameters p <0.001). Despite clinically similar MG and Vmax across all groups, mean AVA ranged from 0.88 to 1.25 cm(2) (p <0.001), classifying small LVOTd patients as severe, average LVOTd as moderate-severe and large LVOTd as moderate. For patients with large, average and small LVOTd, an AVA of 1 cm(2) corresponded to MG of 42, 35 and 29 mm Hg, Vmax of 4.1, 3.8 and 3.5 m/s and dimensionless index (DI) of 0.22, 0.29 and 0.36, respectively. An AVA cut-off of 0.8 cm(2) reduced severe AS inconsistency from 48% to 26% for small LVOTd patients. An AVA cut-off of 0.9 cm(2) reduced severe AS inconsistency from 37% to 26% for average LVOTd patients. The current AVA cut-off of 1 cm(2) was consistent for large LVOTd patients. The LVOTd is associated with significant inconsistencies in AS assessment by current guidelines. For patients with normal EF and normal flow, current guideline definition of severe AS is most consistent for patients with large LVOTd, but not so for patients with average or small LVOTd in whom lower AVA cut-offs should be further studied. The DI cut-off for severe AS is highly variable depending on the LVOTd and guideline revision of this threshold should be considered.


A review of varicocele repair for pain.
A dilation of the pampiniform venous plexus in the scrotum above the testicle, called a varicocele, affects approximately 15% of the general male population. While the majority is asymptomatic, pain results in up to 10% of cases of varicoceles. The pain associated with varicoceles is typically mild and is described as heavy, achy, or dull-and is usually isolated to the testicle or spermatic cord. Guidelines clearly recommend varicocele repair in males with varicoceles, infertility, and an abnormal semen analysis. While chronic, severe pain is an additional indication for repair, a careful evaluation to rule out other etiologies in addition to a period of conservative management are necessary prior to surgical treatment because of the high incidental prevalence of varicoceles in the general population. Several techniques for varicocele repair have been described, including retroperitoneal, laparoscopic, inguinal, and subinguinal. Additionally, recent adjuncts to improve visualization and identification of critical structures including the operating microscope and microvascular Doppler ultrasound have improved success and complication rates. With careful patient selection, outcomes of varicocele repair with regard to pain are excellent, with over 90% of patients experiencing symptomatic relief. After failure of conservative treatments, a varicocele associated with pain should be considered for repair, and the microsurgical subinguinal approach is the gold standard surgical treatment, offering excellent outcomes while minimizing risk of complications.


Knowledge, Attitude, and Practice Regarding Contraception among Women with Schizophrenia: An Observational Study from South India.
Women with schizophrenia have needs beyond their mental health needs, such as those arising out of their gender, sexual, and reproductive functions. Very little is known about the knowledge, attitude, and practice regarding contraception among women with schizophrenia from India. Study among women with schizophrenia (in reproductive age group, having at least one living child, and currently staying with husband) from south India explored their knowledge, attitude, and practice of contraception. Adhering to observational design and ethical principles, data were collected using a semi-structured questionnaire. Modified National Family Health Survey-3 questionnaire and Positive and Negative Symptom Scale of Schizophrenia were also used. Ninety-six women with schizophrenia participated. The mean age was 33.5 years [standard deviation (SD): 6.8 years], and the mean age of onset of schizophrenia was 29.2 years (SD: 6.2 years). Although nearly 90% had knowledge on at least one method of contraception, the mean total number of methods known was mere two. Out of 65 women who were practising contraception, 86.2% adopted female sterilization. The common reasons for not using contraception were wish for another child/son, lack of awareness, and fear of side effects. Unmet need for family planning was 14%. Informed choice of contraception was below 3%. There was statistically significant association between those who were currently using contraception and variables such as age 31 years and above, undifferentiated subtype of schizophrenia, and greater severity of schizophrenia. Although the majority had some knowledge about contraception, decision-making largely rested with others, and informed choice regarding contraception was poor. These could pose an obstetric risk on women with schizophrenia. Sociocultural and illness-related factors influencing contraception need to be explored.


Conformational Flexibility Drives Cold Adaptation in Pseudoalteromonas haloplanktis TAC125 Globins.
Significance: Temperature is one of the most important drivers in shaping protein adaptations. Many biochemical and physiological processes are influenced by temperature. Proteins and enzymes from organisms living at low temperature are less stable in comparison to high-temperature adapted proteins. The lower stability is generally due to greater conformational flexibility. Recent Advances: Adaptive changes in the structure of cold-adapted proteins may occur at subunit interfaces, distant from the active site, thus producing energy changes associated with conformational transitions transmitted to the active site by allosteric modulation, valid also for monomeric proteins in which tertiary structural changes may play an essential role. Critical Issues: Despite efforts, the current experimental and computational methods still fail to produce general principles on protein evolution, since many changes are protein and species dependent. Environmental constraints or other biological cellular signals may override the ancestral information included in the structure of the protein, thus introducing inaccuracy in estimates and predictions on the evolutionary adaptations of proteins in response to cold adaptation. Future Directions: In this review, we describe the studies and approaches used to investigate stability and flexibility in the cold-adapted globins of the Antarctic marine bacterium Pseudoalteromonas haloplanktis TAC125. In fact, future research directions will be prescient on more detailed investigation of cold-adapted proteins and the role of fluctuations between different conformational states.


Bearing of the diencephalon on the taxonomic status of the Tupaioidea.
As the taxonomic status of the Tupaioidea within the Order Primates is still unsettled, an attempt is being made to throw some light on this problem through investigations on the nuclear configuration and fibre connections of the diencephalon of the tree-shrew. Several tree-shrew species, particularly Tupaia glis and Urogale everetti have been involved in this qualitative study, and comparisons of diencephalic structures among the tree-shrews species, and with the Insectivores and Prosimii have been made. Thalamic and hypothalamic nuclei which show more advanced development than others, are nuclei anteroventralis, mediodorsalis, centrum medianum, lateralis posterior, pulvinaris, geniculati lateralis and medialis, subthalamicus and mamillaris medialis. The nucleus mediodorsalis becomes much enlarged and more clearly differentiated into two or three regions. The ventrolateral thalamic nuclear group, though comparably small, is clearly definable into anterior, lateral and posterior parts. The pulvinar makes its first definitive appearance as a separate entity. The nuclei pretectalis and thalamicus posterior are larger and more prominent than those in the Prosimii. The dorsal part of the lateral geniculate body, termed nucleus geniculatus lateralis, show a clear stratification of its cells into six layers that correspond well to the primate pattern. The nucleus geniculatus medialis is well developed and differentiated into magnocellular and parvocellular parts. The epithalamus, subthalamus and hypothalamus are not much different from those of other mammalian species. This study has shown that the thalamus and metathalamus show more advanced than primitive phylogenetic features than are found in the Insectivora. These may compare favorably with most of the homologues in the diencephalon of the Lemuroidea.


Influence of age, sex and body weight on renal osteodystrophy in predialysis patients with chronic renal failure.
The aim of the present investigation was to examine the influence of age, sex and body weight on osseous changes in pre-dialysis patients with chronic renal failure (CRF). 87 patients (44 males and 43 females) aged 18-60 years with CRF were studied. The levels of serum creatinine, total and ionized calcium, phosphorus, alkaline phosphatase, intact parathormone and serum osteocalcine were followed up. Body weight is presented as BMI. 47 of the patients were subjected to double X-ray absorptiometry of lumbar vertebra (Lunar) and 40 patients were examined by computed tomography osteometry. No reliable differences in the levels of biochemical parameters in male and female patients with the same degree of CRF were established. A tendency towards an increase in the level of intact parathormone and serum osteocalcine in women with both initial and advanced CRF was recorded. The BMI in patients with advanced CRF was lower as compared to those with initial CRF. Different stages of osseous changes were observed in 29 males (74.35%) and in 25 females (60.97%). A tendency for a higher frequency and severity of osseous changes in men aged up to 40 years was observed. After this age males and females were equally affected. A high positive correlation (r = 0.50) between BMI and the percentage of the normal Bone Mineral Density/Bone Mineral Content in females with CRF stage II and III was noticed. No significant difference in the frequency and severity of osseous changes in male and female uremic patients was observed. Bone changes were more frequent and pronounced in males up to 40 years of age, while this tendency reversed after the menopause. The higher body weight was beneficial for the osseous changes only in females with advanced CRF, while in all other patients no correlation with densitometric parameters was noticed.


Genetics and molecular mapping of genes for race-specific all-stage resistance and non-race-specific high-temperature adult-plant resistance to stripe rust in spring wheat cultivar Alpowa.
Stripe rust, caused by Puccinia striiformis f. sp. tritici, is one of the most widespread and destructive wheat diseases worldwide. Growing resistant cultivars is the preferred control of the disease. The spring wheat cultivar 'Alpowa' has both race-specific, all-stage resistance and non-race-specific, high-temperature adult-plant (HTAP) resistances to stripe rust. To identify genes for the stripe rust resistances, Alpowa was crossed with 'Avocet Susceptible' (AVS). Seedlings of the parents, and F(1), F(2) and F(3) progeny were tested with races PST-1 and PST-21 of P. striiformis f. sp. tritici under controlled greenhouse conditions. Alpowa has a single partially dominant gene, designated as YrAlp, conferring all-stage resistance. Resistance gene analog polymorphism (RGAP) and simple sequence repeat (SSR) techniques were used to identify molecular markers linked to YrAlp. A linkage group of five RGAP markers and two SSR markers was constructed for YrAlp using 136 F(3) lines. Amplification of a set of nulli-tetrasomic Chinese Spring lines with RGAP markers Xwgp47 and Xwgp48 and the two SSR markers indicated that YrAlp is located on the short arm of chromosome 1B. To map quantitative trait loci (QTLs) for the non-race-specific HTAP resistance, the parents and 136 F(3) lines were tested at two sites near Pullman and one site near Mount Vernon, Washington, under naturally infected conditions. A major HTAP QTL was consistently detected across environments and was located on chromosome 7BL. Because of its chromosomal location and the non-race-specific nature of the HTAP resistance, this gene is different from previously described genes for adult-plant resistance, and is therefore designated Yr39. The gene contributed to 64.2% of the total variation of relative area under disease progress curve (AUDPC) data and 59.1% of the total variation of infection type data recorded at the heading-flowering stages. Two RGAP markers, Xwgp36 and Xwgp45 with the highest R (2) values were closely linked to Yr39, should be useful for incorporation of the non-race-specific resistance gene into new cultivars and for combining Yr39 with other genes for durable and high-level resistance.


A framework to describe, analyze and generate interactive motor behaviors.
While motor interaction between a robot and a human, or between humans, has important implications for society as well as promising applications, little research has been devoted to its investigation. In particular, it is important to understand the different ways two agents can interact and generate suitable interactive behaviors. Towards this end, this paper introduces a framework for the description and implementation of interactive behaviors of two agents performing a joint motor task. A taxonomy of interactive behaviors is introduced, which can classify tasks and cost functions that represent the way each agent interacts. The role of an agent interacting during a motor task can be directly explained from the cost function this agent is minimizing and the task constraints. The novel framework is used to interpret and classify previous works on human-robot motor interaction. Its implementation power is demonstrated by simulating representative interactions of two humans. It also enables us to interpret and explain the role distribution and switching between roles when performing joint motor tasks.


High short-term mortality in hospitalized patients with advanced dementia: lack of benefit of tube feeding.
The influence of tube feeding on survival in hospitalized patients with advanced dementia is controversial. To assess long-term survival in an inception cohort, incident tube feeding placement during the index hospitalization, and the influence of tube feeding on survival in this group of patients. Ninety-nine hospitalized patients with advanced dementia and an available surrogate decision maker were followed up through and after the index hospitalization for mortality and placement of a feeding tube. Other variables measured included advance directive status, presence of a long-term primary care physician, level of involvement of the surrogate decision maker, admitting diagnosis, prior hospitalizations, comorbidities, and diagnosis related group diagnostic category. A new feeding tube was placed in 50% (51/99) of the study patients during the index hospitalization, 31% (31/99) left the hospital without a feeding tube, and 17% (17/99) were admitted with a feeding tube already in place. By stepwise logistic regression analysis, predictors of new feeding tube placement included African American ethnicity (odds ratio, 9.43; 95% confidence interval, 2.1-43.2) and residence in a nursing home (odds ratio, 4.9; 95% confidence interval, 1.02-2.5). Median survival of the 99 patients was 175 days. Eighty-five (85%) survived the index hospitalization, and 28 (28%) were still alive at last follow-up, a range of 1.3 to 4.2 years after enrollment in the study. Tube feeding was not associated with survival (P =.90). An admitting diagnosis of infection was associated with higher mortality (odds ratio, 1.9; 95% confidence interval, 1.01-3.6). In this cohort of hospitalized patients with advanced dementia, risk of receiving a new feeding tube is high, associated with African American ethnicity, and prior residence in a nursing home, and has no measurable influence on survival. With or without a feeding tube, these patients have a 50% six-month median mortality.


3-min whole body cryotherapy/cryostimulation after training in the evening improves sleep quality in physically active men.
Exercise training during evening may disturb sleep patterns and hinder recovery process. The present study aimed to examine the effect of whole body cryotherapy (WBC) exposure after training in the evening on sleep quality and night heart rate variability (HRV). A total of 22 physically active men were randomized to undergo either WBC (3-min at -40°C, wind speed of 2.3 m s-1) or passive recovery (control) following an evening training consisting of 25 min of continuous running at 65% of the maximal aerobic speed (MAS) followed by intermittent running at 85% of the MAS. Each night following the training, the number of movements and HRV during sleeping time were recorded. The next morning, subjective sleep quality and perceived pain were assessed using Spiegel questionnaire and a visual analogue scale, respectively. The number of movements during the night following WBC was significantly reduced (p < 0.05) compared with the control condition. Subjective sleep quality following WBC was significantly better than the control group (p < 0.05). During the estimated slow-wave sleep (SWS), the high frequency power (HF) was higher in the WBC group than the control group (p < 0.05), and the low frequency power (LF) and the LF/HF ratio were lower than the control group (p < 0.05). Pain was significantly reduced following WBC compared to the control (p < 0.01). In conclusion, the use of 3-min WBC after training in the evening improves subjective and objective sleep quality in physically active subjects, which may be due to greater pain relief and improved parasympathetic nervous activity during the SWS period.


Are you good enough for your patients? The European certification model in laparoscopic surgery.
Quality control, training and education in gynaecological surgery are challenged and urgent measures are emerging. The implementation of a structured and validated program for training and quality control seems the most urgent measurement to be taken. The European Academy of Gynaecological Surgery has made a first attempt to do so. Through a practical and theoretical tests system, the skills of an individual surgeon is measured and the conditions to enter the different level of expertise are clearly defined. This certification system based on the best possible level of scientific evidence provides a first practical tool, universally implementable for a decent quality control and structured training program in Gynaecological laparoscopic surgery.


Use of tumor suppressor genes of naked mole rats for human cancer treatment.
Cancer not only has a significant prevalence in the human population, but is also leading cause of death in animals. Despite a long history, our battle against cancer continues. Cross-species comparative genomics offers insight into shared genes and pathways by analyzing genomic data across species, enhancing our understanding of cancer mechanisms, evolutionary processes, and possible therapeutic targets. However, no previous study has demonstrated the inhibitory effects of tumor suppressor genes from one species on tumor cells from another. The naked mole rat is the only mammal yet to be found with cancer that is attributed to its tumor suppressor genes. In this study, we constructed phylogenetic trees and assessed the anti-tumor activity of two suppressor genes, programmed cell death molecule 5 (PDCD5) and dickkopf 3 (DKK3), from rats, mice, and humans. DKK3 robustly inhibited the proliferation of breast cancer cells within and across species due to its highly conserved protein sequence. However, the cross-species inhibitory effect of PDCD5 on breast cancer cells was inconsistent due to significant sequence variations. Intriguingly, PDCD5 from the naked mole rat demonstrated potent anti-tumor activity against breast cancer cells from mice, rats, and humans, surpassing that of PDCD5 from parental species. Our results demonstrate that the suppressor genes from the naked mole rat have a strong inhibitory effect on cancer cells, indicating that the powerful anti-cancer functions of the naked mole rat may be useful for human tumor treatment.


Preparing and Presenting Journal Club Content: An Essential Component of Homeopathic Learning.
Homeopathic education has passed through significant incremental changes in the past few years, where especially postgraduate education has become increasingly slanted toward advanced knowledge of clinical work and research methods. Among many educational activities, a great source of learning is from presenting at or attending a journal club meeting, which is a gathering of people to learn and to critically appraise a journal article or other study material. There has been little previous guidance in homeopathy regarding how to prepare and present journal club content. Selection of a suitable topic is one of the critical prerequisites. Each and every step, from preparation to presentation, needs to be carefully planned and considered. For the meeting to be successful, the final discussion phase requires the active participation and critical insight of all those attending.


dbVar structural variant cluster set for data analysis and variant comparison.
dbVar houses over 3 million submitted structural variants (SSV) from 120 human studies including copy number variations (CNV), insertions, deletions, inversions, translocations, and complex chromosomal rearrangements. Users can submit multiple SSVs to dbVAR that are presumably identical, but were ascertained by different platforms and samples, to calculate whether the variant is rare or common in the population and allow for cross validation. However, because SSV genomic location reporting can vary - including fuzzy locations where the start and/or end points are not precisely known - analysis, comparison, annotation, and reporting of SSVs across studies can be difficult. This project was initiated by the Structural Variant Comparison Group for the purpose of generating a non-redundant set of genomic regions defined by counts of concordance for all human SSVs placed on RefSeq assembly GRCh38 (RefSeq accession GCF_000001405.26). We intend that the availability of these regions, called structural variant clusters (SVCs), will facilitate the analysis, annotation, and exchange of SV data and allow for simplified display in genomic sequence viewers for improved variant interpretation. Sets of SVCs were generated by variant type for each of the 120 studies as well as for a combined set across all studies. Starting from 3.64 million SSVs, 2.5 million and 3.4 million non-redundant SVCs with count >=1 were generated by variant type for each study and across all studies, respectively. In addition, we have developed utilities for annotating, searching, and filtering SVC data in GVF format for computing summary statistics, exporting data for genomic viewers, and annotating the SVC using external data sources.


Fibrinogen induces endothelial cell adhesion and spreading via the release of endogenous matrix proteins and the recruitment of more than one integrin receptor.
We have previously shown that fibrinogen (fg) acts as a subendothelial matrix protein in promoting human endothelial cell (EC) adhesion and cytoskeletal organization. In this study we report that EC spreading on fg, at variance with other matrix proteins, requires endogenous matrix protein synthesis and secretion. ECs, upon seeding on fg, promptly released and organized a fibronectin (fn) matrix. Fg was more effective than vitronectin (vn) in promoting the deposition of this protein. ECs treated with monensin to block matrix protein secretion still adhered to fg but did not properly organize their cytoskeleton and adhesion structures. In contrast, monensin did not affect EC spreading either on vn or on fn. Using antibodies to the alpha and beta chains of fn (alpha 5 beta 1) and vn (alpha v beta 3) receptors, it was found that ECs adherent to fg show clustering and organization in adhesion structures of both type of receptors. A faint staining of adhesion structures with alpha 2 but not alpha 3 and alpha 6 antibodies was also observed. Antibodies either to vn or fn receptors were able to disrupt the EC monolayer and to induce EC retraction and detachment, thus indicating that both receptors are important in maintaining a sustained EC adhesion to fg. However, when ECs were treated with monensin only the vn receptor was organized in adhesion structures while the fn receptor was diffusely distributed. This suggests that clustering of the fn receptor is mediated by the release of endogenous matrix proteins induced by the exposure to fg. In conclusion, fg has a peculiar and complex type of interaction with ECs since it requires endogenous matrix protein release and the recruitment of more than one adhesive receptor. This suggests a specific way of response of ECs to each extracellular matrix component.


Achievements and gaps in projection studies on the temperature-attributable health burden: Where should we be headed?
Future projection of the temperature-related health burden, including mortality and hospital admissions, is a growing field of research. These studies aim to provide crucial information for decision-makers considering existing health policies as well as integrating targeted adaptation strategies to evade the health burden. However, this field of research is still overshadowed by large uncertainties. These uncertainties exist to an extent in the future climate and population models used by such studies but largely in the disparities in underlying assumptions. Existing studies differ in the factors incorporated for projection and strategies for considering the future adaptation of the population to temperature. These differences exist to a great degree because of a lack of robust evidence as well as gaps in the field of climate epidemiology that still require extensive input from the research community. This narrative review summarizes the current status of projection studies of temperature-attributable health burden, the guiding assumptions behind them, the common grounds, as well as the differences. Overall, the review aims to highlight existing evidence and knowledge gaps as a basis for designing future studies on temperature-attributable health burden estimation. Finding a robust methodology for projecting the future health burden could be a milestone for climate epidemiologists as this would largely benefit the world when applying this technique to project the climate-attributable cause-specific health burden and adapt our existing health policies accordingly.


Effects of oocyte age, cumulus cells and injection methods on in vitro development of intracytoplasmic sperm injection rabbit embryos.
This study assessed the effects of oocyte age, cumulus cells and injection methods on in vitro development of intracytoplasmic sperm injection (ICSI) rabbit embryos. Oocytes were recovered from female rabbits superovulated with PMSG and hCG, and epididymal sperm were collected from a fertile male rabbit. The oocyte was positioned with the first polar body at 12 o'clock position, and a microinjection needle containing a sperm was inserted into the oocyte at 3 o'clock. Oolemma breakage was achieved by aspirating ooplasm, and the aspirated ooplasm and sperm were re-injected into the oocyte. The injected oocytes were cultured in M199 medium containing 10% fetal calf serum at 38 degrees C with 5% CO2 in air. The results showed that oocytes injected at 1 h post-collection produced a higher (p < 0.05) fertilization rate than those injected at 4 or 7 h post-collection. Blastocyst rate in the 1 h group was higher (p < 0.05) than in the 7 h group. Denuded oocytes (group A) and oocytes with cumulus cells (group B) were injected, respectively. Rates of fertilization and development of ICSI embryos were not significantly different (p > 0.05) between the two groups. Four ICSI methods were applied in this experiment. In methods 1 and 2, the needle tip was pushed across half the diameter of the oocyte, and oolemma breakage was achieved by either a single aspiration (method 1) or repeated aspiration and expulsion (method 2) of ooplasm. In methods 3 and 4, the needle tip was pushed to the oocyte periphery opposite the puncture site, and oolemma breakage was achieved by either a single aspiration (method 3) or repeated aspiration and expulsion (method 4) of ooplasm. Fertilization rate in method 2 was significantly higher (p < 0.05) than in methods 1 and 3. Blastocyst rates were not significantly different (p > 0.05) among methods 1, 3 and 4, but method 2 produced a higher (p < 0.05) blastocyst rate than method 3.