Dietary Proteins, Brown Fat, and Adiposity.
High protein diets have become popular for body weight maintenance and weight loss despite controversies regarding efficacy and safety. Although both weight gain and weight loss are determined by energy consumption and expenditure, data from rodent trials consistently demonstrate that the protein:carbohydrate ratio in high fat diets strongly influences body and fat mass gain per calorie eaten. Here, we review data from rodent trials examining how high protein diets may modulate energy metabolism and the mechanisms by which energy may be dissipated. We discuss the possible role of activating brown and so-called beige/BRITE adipocytes including non-canonical UCP1-independent thermogenesis and futile cycles, where two opposing metabolic pathways are operating simultaneously. We further review data on how the gut microbiota may affect energy expenditure. Results from human and rodent trials demonstrate that human trials are less consistent than rodent trials, where casein is used almost exclusively as the protein source. The lack of consistency in results from human trials may relate to the specific design of human trials, the possible distinct impact of different protein sources, and/or the differences in the efficiency of high protein diets to attenuate obesity development in lean subjects vs. promoting weight loss in obese subjects.


Multiple metals exposure and neurotoxic risk in bald eagles (Haliaeetus leucocephalus) from two Great Lakes states.
In the present study, the authors determined concentrations of several elements (As, Cd, Co, Cu, Cr, Mn, Pb, Sb, Zn) in the brains and livers of 46 bald eagles (Haliaeetus leucocephalus) from two Great Lakes states, Michigan and Minnesota. To explore whether exposures are of neurological concern, the authors assessed their associations with neurochemical receptors (N-methyl-D-aspartate [NMDA] and γ-aminobutyric acid A [GABA(A)]) and enzymes (glutamine synthetase [GS] and glutamic acid decarboxylase [GAD]) that play critical roles in vertebrate neurobehavior and reproduction. For most elements, levels in the livers and brains did not differ between region and gender. Hepatic Pb levels averaged 33.1 ppm (dry wt), 30.4% of all carcasses exceeded proposed avian Pb thresholds (>26.4 ppm), and in 30.8% of the birds examined evidence of Pb pellets or fragments was found. Significant changes in the activities of GS and GAD were related to brain concentrations of several metals (Pb, Cd, Co, Cu, Zn). No relationships were found among any of the nine elements and NMDA or GABA(A) receptor levels. When combined with the authors' previous study on these same eagles that showed Hg-associated alterations in GS, GAD, and NMDA receptor levels, the present research suggests that bald eagles are exposed to various elements, especially Pb and Hg, that are capable of causing changes in GABAergic and glutamatergic neurotransmission. The functional significance of these neurochemical changes warrants attention.


Rheumatoid factor and the pathogenesis of rheumatoid arthritis.
In analogy with the two categories of reactants which are used in the serological tests for the unusual category of macroglobulins called rheumatoid factor, two fluorescent reactants have been prepared for the detection of rheumatoid factor in situ in tissue sections: fluorescent antigen-rabbit antibody (immune) complex, in the present study, and fluorescent aggregated human gamma-globulin, in previous work. Plasma cells in the synovial membrane and germinal center cells and internodular plasma cells in lymph nodes are the sites of origin of rheumatoid factor in active rheumatoid arthritis, whether occurring in adults or children. Plasma cells and germinal center cells which form rheumatoid factor detectable with fluorescent immune complex are less numerous than those which contain factor demonstrable with fluorescent aggregate. In the same tissues, plasma cells and germinal center cells which contain macroglobulin (19S human gamma-globulin) detectable with fluorescent antibody-but not showing the reactivity of rheumatoid factor-are more abundant than those containing rheumatoid factor. While macroglobulin and rheumatoid factor are almost exclusively formed in the cytoplasm, these proteins are also detectable in the nucleus of an occasional plasma cell. Normal and pathological synovial and capsular tissues, lymph nodes, and connective tissues obtained from individuals without rheumatoid arthritis are not stained with fluorescent immune complex or, except for an unusual example of Waldenstrom's macroglobulinemia, with fluorescent aggregate. The cellular origin, as well as certain chemical and immunological attributes, of rheumatoid factor suggests an antibody-like nature and function. The observations cited are consistent with the behavior anticipated for cellular rheumatoid factor, were it primarily an antibody direct to an altered human gamma-globulin and cross-reacting with rabbit gamma-globulin. However, it is also possible that there are two or more cellular rheumatoid factors. Lesion-associated protein precipitates having the composition anticipated for rheumatoid factor-antigen complex are localized in the amyloid depositions in kidney and spleen of an individual who died with amyloidosis secondary to rheumatoid arthritis.


Usefulness and applicability of femorofemoral crossover bypass grafting.
We examined the usefulness of femorofemoral crossover bypass grafting (FFC) and factors influencing its outcome by retrospectively analyzing all FFCs performed in our hospital over a 5-year period, focusing on both patency rates and clinical efficacy. For 95 patients Kaplan-Meier patency rates were calculated (follow-up 40.4 +/- 3.0 months). Clinical outcome was defined according to Rutherford's standardized categories. The influence of cardiovascular risk factors and technical characteristics on outcome was determined. Clinical status of the limb remained improved in 89%. One- and 5-year primary, primary assisted, and secondary patency rates were 88.2% and 57.3%, 90.6% and 62.4%, and 92.6% and 68.1%, respectively. Clinical outcome of the limb was better in patients with <50% stenosis in the femoral arteries preoperatively (p = 0.033). No predictors for patency rates were identified. FFCs are effective in the medium long term for patients in all age categories independently of cardiovascular risk factors. The best predictor of clinical outcome is the preoperative degree of stenosis, with a better outcome for patients affected by <50% stenosis. Success of FFC cannot be reliably measured by graft patency alone but should be assessed by combining patency rates and clinical outcome according to standardized categories.


Association Between Celiac Disease and Mortality Risk in a Swedish Population.
Celiac disease may be associated with a modest but persistent increased long-term mortality risk. It is uncertain whether this risk has changed in the era of wider diagnosis rates, less severe clinical disease, and more widespread availability of gluten-free food. To evaluate the association between celiac disease and mortality risk in a population-based cohort in Sweden. All individuals in Sweden with celiac disease diagnosed between 1969 and 2017 were identified through the Epidemiology Strengthened by histoPathology Reports in Sweden (ESPRESSO) cohort. Participants (n = 49 829) were observed starting on the day of the biopsy. The final date of follow-up was December 31, 2017. Celiac disease was defined by the presence of small intestinal villus atrophy on histopathology specimens during the years 1969-2017 from Sweden's 28 pathology departments. Each individual was matched with as many as 5 control participants in the general population by age, sex, county, and calendar period. The primary outcome was all-cause mortality, and the secondary outcome was cause-specific mortality. Patients with celiac disease were compared with controls using stratified Cox proportional modeling, stratifying by year of diagnosis. There were 49 829 patients with celiac disease, including 24% who were diagnosed between the years 2010 and 2017. The mean (SD) age at diagnosis was 32.2 (25.2) years and 62.4% were women. During a median follow-up time of 12.5 years, 13.2% (n = 6596) died. Compared with controls (n = 246 426), overall mortality was increased in those with celiac disease (9.7 vs 8.6 deaths per 1000 person-years; absolute difference, 1.2 per 1000 person-years; hazard ratio [HR], 1.21 [95% CI, 1.17-1.25]). The relative increase in mortality risk was present in all age groups and was greatest in those diagnosed in the age range of 18 to 39 years (1.9 vs 1.1 per 1000 person-years; HR, 1.69 [95% CI, 1.47-1.94]; P values for heterogeneity comparing 18-39 years with 40-59 years and with ≥60 years were both <.001). Individuals with celiac disease were at increased risk of death from cardiovascular disease (3.5 vs 3.4 per 1000 person-years; HR, 1.08 [95% CI, 1.02-1.13]), cancer (2.7 vs 2.2 per 1000 person-years; HR, 1.29 [95% CI, 1.22-1.36]), and respiratory disease (0.6 vs 0.5 per 1000 person-years; HR, 1.21 [95% CI, 1.08-1.37]). When compared with controls, the overall mortality risk was greatest in the first year after diagnosis (15.3 vs 6.5 per 1000 person-years; HR, 2.34 [95% CI, 2.14-2.55]) but persisted beyond 10 years after diagnosis (10.5 vs 10.1 per 1000 person-years; HR, 1.15 [95% CI, 1.10-1.20]). The mortality risk was likewise present for patients diagnosed during the years 2010-2017 (7.5 vs 5.5 per 1000 person-years; HR, 1.35 [95% CI, 1.21-1.51]). In a Swedish population studied between 1969 and 2017, a diagnosis of celiac disease compared with the general population was associated with a small but statistically significant increased mortality risk.


Genetic engineering in the improvement of plants for phytoremediation of metal polluted soils.
Metal concentrations in soils are locally quite high, and are still increasing due to many human activities, leading to elevated risk for health and the environment. Phytoremediation may offer a viable solution to this problem, and the approach is gaining increasing interest. Improvement of plants by genetic engineering, i.e. by modifying characteristics like metal uptake, transport and accumulation as well as metal tolerance, opens up new possibilities for phytoremediation. So far, only a few cases have been reported where one or more of these characteristics have been successfully altered; e.g. mercuric ion reduction causing improved resistance and phytoextraction, and metallothionein causing enhanced cadmium tolerance. These, together with other approaches and potentially promising genes for transformation of target plants are discussed.


Effects of desipramine treatment on the biliary, fecal and urinary excretion of methadone in the rat.
The effects of desipramine (DMI) given i.p. 1 hour before administration of 14C-methadone (5 mg/kg s.c.) on the excretion of methadone in bile, feces and urine were studied. Rats with cannulated bile ducts excreted 52% of administered 14C into bile within 4 hours, 60% of the excretion as 2-ethylidine-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP) and 30% as water-soluble metabolite (WSM). The percentages of 14C excreted within 48 hours into the feces and urine of rats without biliary fistula were 60.6 and 17.4%, respectively. The excretion of 14C in feces consisted mostly of EDDP. Urinary excretion consisted of unchanged methadone, EDDP and WSM in the first 12 hours and consisted mostly of WSM thereafter. DMI treatment increased biliary flow but decreased the biliary excretion of 14C, mainly by a decrease in WSM. The fecal output was greatly decreased by DMI, thus decreasing fecal excretion of 14C in the first 12 hours. DMI treatment markedly decreased urinary volume but did not change the urine pH. The decreased urinary excretion of WSM in DMI-treated rats during the first 12-hour urine sample accounted for most of the decreased elimination of 14C. It is suggested that DMI-induced inhibition of methadone metabolism in the liver is a major factor responsible for the observed decreases in the biliary and urinary excretion of WSM...


Thromboembolic prophylaxis in 3575 hospitalized patients with atrial fibrillation. The Clinical Quality Improvement Network (CQIN) Investigators.
To define contemporary utilization patterns of anticoagulant and antiplatelet therapy for thromboembolic prophylaxis in atrial fibrillation (AF). Retrospective medical records audit of patients admitted in 1993 and 1994. Twelve Canadian hospitals. Three thousand, three hundred and seventy-five consecutive patients with AF; 1570 females and 2005 males. The mean age was 72 years; 1353 patients were younger than 70 years and 2222 were aged 70 years and older. Overall, 1188 (33%) of the 3575 patients received no prophylaxis, 852 (24%) were treated with warfarin alone, 1247 (35%) received acetylsalicylic acid (ASA) alone and 288 (8%) received both drugs. The pattern of medication use did not change appreciably when possible contraindications to warfarin or ASA therapy were considered. Among the 331 AF patients with valvular heart disease and no contraindications to thromboembolic prophylaxis, 65 (20%) received neither treatment, 181 (55%) received warfarin therapy alone, 46 (14%) received ASA alone and 39 (12%) received both. Among the 2199 AF patients with nonvalvular heart disease and no contraindications, 823 (37%) did not receive either therapy, 677 (31%) received ASA alone, 504 (23%) received warfarin alone and 195 (9%) received both. Elderly and female patients were less likely to receive thromboembolic prophylaxis. Anticoagulation and antiplatelet prophylaxis in AF appears to be less than optimal. Although concerns about bleeding may be one reason thromboembolic prophylaxis is so unevenly and incompletely applied, it will be important to determine the reasons for this practice and to develop effective strategies in order to enhance the process of care and patient outcomes.


DNA sequence analysis and restriction fragment length polymorphism (RFLP) typing of the HLA-DQw2 alleles associated with dermatitis herpetiformis.
Dermatitis herpetiformis (DH) is a blistering autoimmune skin disease associated with a 95-100% incidence of the HLA class II antigen HLA-DQw2. Although the precise role of this antigen in the pathogenesis of DH is unclear, one theory proposes that patients with DH possess a molecularly unique subtype of the HLA-DQw2 antigen that causes immune abnormalities eventuating in the clinical manifestations of DH. To test this hypothesis, we performed DNA sequence analysis on the highly polymorphic HLA-DQB1 and HLA-DQA1 loci of eight patients with dermatitis herpetiformis. All DQB1 alleles sequenced were identical to the previously described HLA-DQB*0201 allele from HLA-DQw2 normal subjects. In addition, DQA1 alleles sequenced were identical to those alleles previously associated with HLA-DQw2 (DQA*0201, DQA*0501). These data document that although HLA-DQw2 appears to be a necessary element in the pathogenesis of DH, the development of DH is not dependent on the presence of a unique HLA-DQw2 antigen. HLA-DQ allelic typing by restriction fragment length polymorphism analysis of PCR-amplified HLA-DQA1 and HLA-DQB1 fragments was also performed in ten patients with DH to determine the allelic distribution among both HLA-DR3 (eight patients) and non-DR3 (two patients) DH patients. At the HLA-DQ beta chain locus, all patients possessed the DQB1*0201 allele. At the HLA-DQ alpha chain locus, all HLA-DR3 patients and one non-DR3 patient displayed a pattern consistent with the DQA1*0501 allele, whereas one non-DR3 patient displayed a pattern consistent with the DQA1*0201 allele. These data document that patients with DH do not express a unique HLA-DQw2 heterodimer, that the HLA-DQw2 molecules present in patients with DH have no DNA sequence differences from those found in normal HLA-DQw2 subjects and therefore that susceptibility to DH is not due to a unique HLA-DQw2 molecule.


[Reactions of neurons of the secondary somatosensory cortex of awake cats to electrocutaneous and acoustic stimulation].
In chronic experiments on alert cats the discharges of neurons in the secondary projection zone of the somato-sensory cortex to click and electrical stimulation of contralateral paw were investigated. 57% of neurons responded to "specific" stimulation by increasing the discharge frequency, 18%--by weakening or stopping the background activity, while 25% did not react at all. 30% of neurons responded by activation to the "non-specific" stimulation (click), 25%--by inhibition and 45% were unresponsive. An investigation of intersensory convergence by sending signals from different analyzer systems showed that bimodal responses are peculiar to most neurons (55%) of this zone. 18% of neurons with a response only to the "specific" or "non-specific" stimuli were found. It is suggested that polysensory convergence on the neurones of the somato-sensory cortex is important for the cortical area in the process of closing of conditioned connections.


An open letter to Magic Johnson.
The last installment in this series dealt with post-test counseling of a person whose HIV test was negative. The plan was to provide counseling for an HIV-infected person in this installment. NBA basketball star Magic Johnson's surprise announcement on November 7, 1991, of his infection with HIV has provided an unexpected opportunity to counsel a well-known person through an "open letter."


trans activation of an immediate-early frog virus 3 promoter by a virion protein.
We investigated the protein and DNA sequence requirements for the expression of an immediate-early frog virus 3 (FV3) gene, infected-cell RNA (ICR) 169. We used a plasmid containing the 78 nucleotides 5' to the transcription start site of ICR-169 placed upstream from the coding sequence for the bacterial enzyme chloramphenicol acetyltransferase (CAT). This construction, when introduced by CaPO4-mediated transfection into various eucaryotic cell lines, promoted CAT synthesis only if the transfected cells were subsequently infected with FV3. Dot-blot hybridization of RNA extracted from transfected, FV3-infected cells with a radioactive CAT probe showed that the induction of CAT synthesis by FV3 was at the level of transcription. When transfected cells were infected with FV3 in the presence of cycloheximide, induction of CAT-specific RNA still occurred, demonstrating that a virion protein was responsible for the trans activation. FV3-induced CAT synthesis was inhibited by alpha-amanitin in wild-type Chinese hamster ovary (CHO) cells but not in CHO cells with an alpha-amanitin-resistant RNA polymerase II. The results suggest that a virion protein alters either the DNA template or the host polymerase to allow transcription from immediate-early FV3 promoters.


cDermo-1 expression indicates a role in avian skin development.
Basic helix-loop-helix (bHLH) transcription factors have been shown to be important regulatory proteins for tissue determination and differentiation. We cloned the chicken homologue of the gene of the murine Twist-related bHLH protein Dermo-1, which we named cDermo-1, and analyzed its sequence and embryonic expression. Our sequence data suggest a decisive role of Dermo-1 proteins in the evolution of amniote skin. We present a detailed analysis of cDermo-1 expression during avian embryonic development. cDermo-1 is first expressed in a variety of mesodermal tissues of the chick embryo including the limb buds, but later becomes restricted to the subectodermal mesenchyme of the integument and the developing feather buds, indicating a role of cDermo-1 during avian skin and feather development.


Molecular toxicity of cerium oxide nanoparticles to the freshwater alga Chlamydomonas reinhardtii is associated with supra-environmental exposure concentrations.
Ceria nanoparticles (NPs) are widely used as fuel catalysts and consequently are likely to enter the environment. Their potential impacts on. biota at environmentally relevant concentrations, including uptake and toxicity, remain to be elucidated and quantitative data on which to assess risk are sparse. Therefore, a definitive assessment of the molecular and phenotypic effects of ceria NPs was undertaken, using well-characterised mono-dispersed NPs as their toxicity is likely to be higher, enabling a conservative hazard assessment. Unbiased transcriptomics and metabolomics approaches were used to investigate the potential toxicity of tightly constrained 4-5 nm ceria NPs to the unicellular green alga, Chlamydomonas reinhardtii, a sentinel freshwater species. A wide range of exposure concentrations were investigated from predicted environmental levels, to support hazard assessment, to supra-environmental levels to provide insight into molecular toxicity pathways. Ceria NPs were internalised into intracellular vesicles within C. reinhardtii, yet caused no significant effect on algal growth at any exposure concentration. Molecular perturbations were only detected at supra-environmental ceria NP-concentrations, primarily down-regulation of photosynthesis and carbon fixation with associated effects on energy metabolism. For acute exposures to small mono-dispersed particles, it can be concluded there should be little concern regarding their dispersal into the environment for this trophic level.


Endoscopic removal of daughter Echinococcus cysts from the common bile duct.
A case report is presented of a patient in whom daughter cysts of Echinococcus granulosus were removed endoscopically from the common bile duct. We would like to suggest in this paper that in cases of operated hydatid disease of the liver with retained cysts in the common bile duct, endoscopic removal instead of surgery is possible; and in particular that anaphylactic reaction need not be expected as the cysts content is drained into the gut.


Dorsal Muscle Attenuation May Predict Failure to Respond to Interleukin-2 Therapy in Metastatic Renal Cell Carcinoma.
To explore whether the sarcopenia body type can help predict response to interleukin-2 (IL-2) therapy in metastatic renal cell carcinoma (RCC). Institutional review board approval was obtained for this Health Insurance Portability and Accountability Act-compliant retrospective cohort study of 75 subjects with metastatic RCC who underwent pretreatment contrast-enhanced computed tomography within 1 year of initiating IL-2 therapy. Cross-sectional area and attenuation of normal-density (31-100 Hounsfield units [HU]) and low-density (0-30 HU) dorsal muscles were obtained at the T11 vertebral level. The primary outcome was partial or complete response to IL-2 using RECIST 1.1 criteria at 6 weeks. A conditional inference tree was used to determine an optimal HU cutoff for predicting outcome. Bonferroni-adjusted multivariate logistic regression was conducted to investigate the independent associations between imaging features and response after controlling for demographics, doses of IL-2, and RCC prognostic scales (eg, Heng and the Memorial Sloan Kettering Cancer Center [MSKCC]). Most subjects had intermediate prognosis by Heng (65% [49 of 75]) and the MSKCC (63% [47 of 75]) criteria; 7% had complete response and 12% had partial response. Mean attenuation of low-density dorsal muscles was a significant univariate predictor of IL-2 response after Bonferroni correction (P = 0.03). The odds of responding to treatment were 5.8 times higher for subjects with higher-attenuation low-density dorsal muscles (optimal cutoff: 18.1 HU). This persisted in multivariate analysis (P = 0.02). Body mass index (P = 0.67) and the Heng (P = 0.22) and MSKCC (P = 0.08) clinical prognostic scales were not significant predictors of response. Mean cross-sectional attenuation of low-density dorsal muscles (ie, sarcopenia) may predict IL-2 response in metastatic RCC. Clinical variables are poor predictors of response.


Site-specific recognition of bacteriophage T4 DNA by T4 type II DNA topoisomerase and Escherichia coli DNA gyrase.
The site specificity of bacteriophage T4-induced type II DNA topoisomerase action on double-stranded DNA has been explored by studying the sites where DNA cleavages are induced by the enzyme. Oxolinic acid addition increases the frequency at which phi X174 duplex DNA is cut by the enzyme by about 100-fold, to the point where nearly every topoisomerase molecule causes a double-stranded DNA cleavage event. The effect of oxolinic acid on the enzyme is very similar to its effect on another type II DNA topoisomerase, the Escherichia coli DNA gyrase. A filter-binding method was developed that allows efficient purification of topoisomerase-cleaved DNA fragments by selecting for the covalent attachment of this DNA to the enzyme. Using this method, T4 topoisomerase recognition of mutant cytosine-containing T4 DNA was found to be relatively nonspecific, whereas quite specific recognition sites were observed on native T4 DNA, which contains glucosylated hydroxymethylcytosine residues. The increased specificity of native T4 DNA recognition seems to be due entirely to the glucose modification. In contrast, E. coli DNA gyrase shows a high level of specificity for both the mutant cytosine-containing DNA and native T4 DNA, recognizing about five strong cleavage sites on both substrates. An unexpected feature of DNA recognition by the T4 topoisomerase is that the addition of the cofactor ATP strongly stimulates the topoisomerase-induced cleavage of native T4 DNA, but has only a slight effect on cleavage of cytosine-containing T4 DNA.


Evaluating care farming as a means to care for those in trauma and grief.
The interrelationships between nature, health, and wellbeing are increasingly recognized and incorporated into therapeutic interventions. Care farming, the concept of utilizing agricultural places and practices for providing care, therapy, and rehabilitation, is a paradigmatic example of this shift. This mixed method study empirically evaluates the efficacy of care farming as an intervention for individuals affected by traumatic grief, a complex experiential condition. Both quantitative and qualitative results suggest this care farm intervention was beneficial, yielding significant reductions in subjective distress to grief intensity. The study's findings add to the growing body of evidence on care farming and support green care as a therapeutic potential for individuals affected by traumatic grief.


Changes in cortical thickness across the lifespan in major depressive disorder.
Neurobiological mechanisms underlying the development of major depressive disorder (MDD) may differ depending on age-of-onset. Our aim was to compare patients who differ in age-of-onset, while controlling for illness duration, and number of depressive episodes. By directly comparing early-(EOD) and late-onset (LOD) patients, we examined whether age-of-onset is associated with changes in the extent or spatial pattern of cortical thickness. Cross-sectional comparison of cortical thickness in EOD vs. LOD. Age-of-onset was determined based on self-report, with EOD defined as onset prior to age 25. Reduced cortical thickness in the dorsal-lateral prefrontal cortex (DLPFC), pre- and postcentral gyrus, and the lingual gyrus were found in EOD compared to healthy controls (p<0.001). In linear regression models controlling for number of episodes, illness duration, severity, and sex, differences (at p<0.001) were found between EOD and LOD in the bilateral posterior cingulate, parahippocampal gyri, right precuneus, lingual, and fusiform gyri, but not the DLPFC. EOD is associated with greater disturbances in cortical thickness than LOD, even when duration of illness and other factors are controlled. These results provide novel insights on how development of depression is differentiated by age.


G-CSF induces elevation of circulating CA 15-3 in breast carcinoma patients treated in an adjuvant setting.
Cancer antigen 15-3 (CA 15-3), a circulating marker that determines secreted products of the polymorphic MUC1 gene, has been established as a convenient tool for monitoring breast carcinoma patients. The authors investigated alterations of soluble CA 15-3 in 57 postoperative breast carcinoma patients while they were receiving intensified adjuvant chemotherapy with granulocyte colony stimulating factor (G-CSF) support; 26 patients had American Joint Committee on Cancer (AJCC) Stage II, and 31 patients had AJCC Stage III breast carcinoma. Serial CA 15-3 values recorded thoughout the treatment were compared with baseline values, analyzed for correlation with hematologic and biochemical parameters, and compared with clinicopathologic characteristics and patient outcome. At a median follow-up time of 32 months, 47 of these patients remained relapse-free. A twofold increase of CA 15-3 was detected at the end of the second week of treatment, remained significantly elevated in most patients at above the cutoff level of 30 U/mL throughout the treatment period (P < 0.0001), and subsided to pretreatment values 1-2 months after treatment cessation. CA 15-3 values were found to be associated strongly with absolute neutrophil count, serum lactate dehydrogenase, and alkaline phosphatase. The median values and the kinetics of tumor markers did not differ over time in regard to hormonal receptor status and disease recurrence. These data provide strong evidence that G-CSF administration can induce elevation of CA 15-3 and indicate that false-positive results should be considered when evaluating CA 15-3 in patients who are receiving G-CSF. It is speculated that this phenomenon occurs through the induction of MUC1 antigen of unknown origin by G-CSF. Experimental investigation of this clinical observation is warranted.


Polarized compartmentalization of organelles in growth cones from developing optic tectum.
We have used computer-assisted reconstructions of continuous serial sections to study the cytoplasmic organization of growth cones in vivo. Optic tecta from 6.25-6.5-d-old chicken embryos were quick-frozen and then freeze-substituted in acetone-osmium tetroxide or, for comparison, prepared by conventional fixation. Images of eight freeze-substituted and two conventionally fixed growth cones were reconstructed from aligned serial micrographs. After freeze-substitution, numerous lumenless membrane-bound sacs arrayed in multilamellar stacks appear to replace the abundant smooth endoplasmic reticulum found after chemical fixation. Microtubule fascicles progressively diverge from their typical fascicular organization in the initial segment of the growth cone and are absent in the varicosity and the more distal segment. Mitochondria, in contrast, are concentrated in the proximal segment of the varicosity; multilamellar stacks and endosome-like vacuoles are in the distal segment; and coated pits and vesicles are concentrated near the terminal filopodium, which is the most distal and organelle-poor domain of the growth cone. These observations suggest that dilation and fusion of the lumenless, membrane-bound sacs that occurs during chemical fixation give rise to the network of smooth endoplasmic reticulum. The three-dimensional reconstructions show that the cytoplasmic components of growth cones, including the membrane-bound sacs and multilamellar stacks revealed by freeze substitution, are polarized along the axis of these growth cones, which suggests that they have a role in recycling of membrane during elongation of the growth cone.


An exploratory investigation of predictors of outcome in face-to-face and online cognitive-behavioural therapy for bulimia nervosa.
Cognitive-behavioural therapy (CBT) delivered face-to-face and via the internet reduces bulimia nervosa (BN) symptoms. However, our empirical understanding of factors affecting patient outcomes is limited. Using data from a randomised, controlled trial comparing internet-based (CBT4BN, n = 78) with face-to-face (CBTF2F, n = 71) group CBT (97% female, M = 28 years), we examined general treatment (across conditions) and modality-specific predictors of end-treatment and 1-year outcomes (abstinence, binge-eating frequency, purging frequency). Improved eating disorder-related quality of life (EDQOL) during treatment and follow-up predicted abstinence at end-treatment and 1-year assessments. Improved EDQOL, disordered eating cognitions, and anxiety symptoms predicted less frequent binge eating and purging. Previous CBT and being employed predicted more frequent binge eating and purging at both assessments. Higher self-transcendence and self-directedness predicted less frequent binge eating. More severe binge eating and purging at baseline and end-treatment predicted more frequent binge eating and purging at subsequent assessments. Improved EDQOL was more strongly associated with positive outcome in CBT4BN; improved depressive symptoms and health-related QOL predicted positive outcome in CBT4BN but not CBTF2F. Symptom improvement and certain character traits predicted positive outcome, whereas more severe presentation and prior CBT experience predicted poorer outcome. Consideration of intreatment symptom improvement may facilitate care recommendations, particularly for internet-based modalities.


Inhibited coupling guiding hollow fibers for label-free DNA detection.
The potentialities in using hollow core tube lattice fibers based on inhibited coupling wave-guiding for label-free DNA detection are numerically investigated and discussed here. The proposed sensing approach does not require any additional transducer component such as Bragg gratings, amplifying techniques such as nanoparticles nor coherent sources. It simply consists of the measurement of the transmittance of a piece of fiber some ten centimeters long. In case of matching DNA sequence, an additional bio-layer is laid down the dielectric-air interface causing a red shift of the transmission spectrum of the fiber. Results show a spectral sensitivity on the bio-layer with shift as high as 42 nm for every 10 nm of bio-layer and robustness against imperfect fiber coupling. The proposed approach can be easily applied to sensing of other complex molecular structures where the presence/absence of analyte can generate or not an additional layer.


Addressing violence against children: A case review in the state of Qatar.
Violence against children (VAC) is a critical public health issue that affects billions of children worldwide. The combination of its prevalence and severity of effects on children creates an urgent need for effective interventions. Multiple studies associate VAC with lifelong implications that affect children through adulthood. In Qatar, multiple approaches such as legislation are being used to protect children from all forms of violence. Despite the gravity of the issue, there is still low readiness for the prevention of VAC in Qatar. This review aimed to map approaches to addressing VAC in Qatar from the panelists' perspectives on current approaches to addressing VAC. The review obtained data from a recorded video entitled "A Public Health Approach to Addressing Violence Against Children." The panel discussion in this video clip was organized as a side event of the WISH virtual summit by UNICEF and WISH on World Children's Day, held in Qatar in November 2020. The video was transcribed and analyzed using thematic analysis. It shows the importance of both global and national level interventions in addressing VAC. The review uses the socioecological model to show relationships among different levels of interventions addressing VAC in Qatar. The findings highlight the national approaches to addressing VAC using public health, and legislative and policy approaches. The interventions addressing VAC at different levels in Qatar are interconnected. Delineating each level is key to the formation of holistic interventions that leverage global, regional, national, communal, familial, and individual factors that support interventions to address VAC.


Identification of Novel Urinary Biomarkers for Predicting Renal Prognosis in Patients With Type 2 Diabetes by Glycan Profiling in a Multicenter Prospective Cohort Study: U-CARE Study 1.
Because quantifying glycans with complex structures is technically challenging, little is known about the association of glycosylation profiles with the renal prognosis in diabetic kidney disease (DKD). In 675 patients with type 2 diabetes, we assessed the baseline urinary glycan signals binding to 45 lectins with different specificities. The end point was a decrease of estimated glomerular filtration rate (eGFR) by ≥30% from baseline or dialysis for end-stage renal disease. During a median follow-up of 4.0 years, 63 patients reached the end point. Cox proportional hazards analysis revealed that urinary levels of glycans binding to six lectins were significantly associated with the outcome after adjustment for known indicators of DKD, although these urinary glycans, except that for DBA, were highly correlated with baseline albuminuria and eGFR. Hazard ratios for these lectins were (+1 SD for the glycan index) as follows: SNA (recognizing glycan Siaα2-6Gal/GalNAc), 1.42 (95% CI 1.14-1.76); RCA120 (Galβ4GlcNAc), 1.28 (1.01-1.64); DBA (GalNAcα3GalNAc), 0.80 (0.64-0.997); ABA (Galβ3GalNAc), 1.29 (1.02-1.64); Jacalin (Galβ3GalNAc), 1.30 (1.02-1.67); and ACA (Galβ3GalNAc), 1.32 (1.04-1.67). Adding these glycan indexes to a model containing known indicators of progression improved prediction of the outcome (net reclassification improvement increased by 0.51 [0.22-0.80], relative integrated discrimination improvement increased by 0.18 [0.01-0.35], and the Akaike information criterion decreased from 296 to 287). The urinary glycan profile identified in this study may be useful for predicting renal prognosis in patients with type 2 diabetes. Additional investigation of glycosylation changes and urinary glycan excretion in DKD is needed.


Acute suppression of circulating sCD40L during hyperglycemia and euglycemic-hyperinsulinemia in healthy young males.
sCD40L is a proatherogenic cytokine, part of the tumor necrosis factor (TNF) superfamily and consistently associated with obesity, diabetes, and increased cardiovascular risk. Although the role of sCD40L in the onset/progression of cardiovascular complications of dysmetabolic diseases may be modulated by acute and/or chronic fluctuations of plasma insulin and glucose, very little has been done to clarify this interaction. The kinetic profile of sCD40L (and, in an exploratory manner, of several immunomodulatory factors), were measured during hyperglycemia and euglycemic-hyperinsulinemia in a group of 10 healthy young males (26.8 +/- 1.4 years). After an overnight fast, intravenous (iv) catheters were placed in antecubital veins of both arms for blood drawing and dextrose/insulin iv infusions. Procedures lasted 240 minutes including baseline (t = 0-60), hyperglycemia (t = 60-150; plasma glucose approximately 220 mg/dL via iv dextrose infusion), and euglycemic-hyperinsulinemia (t = 150-240; glucose infusion continued to clamp glycemic levels between 80 and 110 mg/dL; constant insulin infusion at 1.5 mU/kg/minute).Plasma for cytokine assays was sampled at 12 separate time-points. Plasma levels of sCD40L were significantly reduced (P < 0.01) during hyperglycemia and euglycemic-hyperinsulinemia, paralleling the kinetic profiles of free fatty acids and ketone bodies. This pattern was also observed in other immunomodulatory factors (notably cortisol and epidermal growth factor), while (interleukin [IL]-1alpha, IL-4, IL-6, IL-9, IL-10, TNF-alpha, Eotaxin) did not change significantly. Significant reductions of the proatherogenic cytokine sCD40L were observed during endogenous and exogenous hyperinsulinemia, independent of prevailing glucose concentration, in young healthy males. Our data suggest a mechanism by which correct insulin action may exert a beneficial protective role against inflammation, independent of its immediate glucose-lowering effect.


Efficacy of aerial forward-looking infrared surveys for detecting polar bear maternal dens.
Denned polar bears (Ursus maritimus) are invisible under the snow, therefore winter-time petroleum exploration and development activities in northern Alaska have potential to disturb maternal polar bears and their cubs. Previous research determined forward looking infrared (FLIR) imagery could detect many polar bear maternal dens under the snow, but also identified limitations of FLIR imagery. We evaluated the efficacy of FLIR-surveys conducted by oil-field operators from 2004-2016. Aerial FLIR surveys detected 15 of 33 (45%) and missed 18 (55%) of the dens known to be within surveyed areas. While greater adherence to previously recommended protocols may improve FLIR detection rates, the physical characteristics of polar bear maternal dens, increasing frequencies of weather unsuitable for FLIR detections-caused by global warming, and competing false positives are likely to prevent FLIR surveys from detecting maternal dens reliably enough to afford protections consonant with increasing global threats to polar bear welfare.


Peste des petits ruminants infection among cattle and wildlife in northern Tanzania.
We investigated peste des petits ruminants (PPR) infection in cattle and wildlife in northern Tanzania. No wildlife from protected ecosystems were seropositive. However, cattle from villages where an outbreak had occurred among small ruminants showed high PPR seropositivity, indicating that spillover infection affects cattle. Thus, cattle could be of value for PPR serosurveillance.


Renal renin responses to changes in volume status and perfusion pressure.
The effects on juxtaglomerular apparatus (JGA) renin activity of maneuvers known to acutely alter renin secretory rates were studied in rats. Five groups of rats were studied: group I: control; group II: 1% volume expansion; group III: 10% volume expansion; group IV: hypotensive hemorrhage; and group V: aortic clamping. Superficial (S) JGA renin activity was decreased in group II and further decreased in group III. Deep (D) JGA renin activity did not change. After hypotensive hemorrhage both S- and D-JGA renin activity increased. Similar results were seen after aortic clamping, although the increase in D-JGA renin activity was significantly less. These results indicate that S- and D-JGA renin activity may be under the control of different stimuli and may vary independently. They also support the concept that acute changes in renin secretion may be related to changes in renin synthesis.


Case Report: HTLV-1-Induced Adult T-Cell Leukemia and Tropical Spastic Paresis.
Human T-lymphotropic virus type 1 (HTLV-1) is a retrovirus endemic in many areas around the world. HTLV-1 can induce the development of adult T-cell leukemia (ATL) or myelopathy/tropical spastic paraparesis (HAM/TSP). We report a patient who presented to our outpatient clinic with massive splenomegaly, weight loss, urinary retention, and lower extremity weakness for the previous 3 years. The patient was found to have positive HTLV-1 by ELISA and Western blot from peripheral blood. Evaluation of the spleen demonstrated T-cell large granular lymphocyte leukemia consistent with ATL. In addition to progressive lower extremity weakness, hyperreflexia and clonus, cerebral spinal fluid was positive for HTLV-1 by ELISA and had a reversed CD4-to-CD8 ratio consistent with HAM/TSP. These findings suggest HTLV-1 induced ATL and HAM/TSP presenting simultaneously in the same patient.


Serologic studies in patients with keratoconjunctivitis sicca.
Thirty-two patients with keratoconjunctivitis sicca (KCS) were screened for the presence of antinuclear antibodies, rheumatoid factor, and autoantibodies associated with Sjögren's syndrome (designated SS-A and SS-B). None of these patients had or were found to have clinical evidence of connective-tissue disease. The conditions of 19 (59%) patients were antinuclear-antibody-positive and 18 (56%) were rheumatoid-factor-positive. We found SS-A and/or SS-B autoantibodies in ten (31%) patients. There seems to be a high incidence of serologic abnormalities in patients with KCS, even when those patients with connective-tissue disease are excluded. Serologic testing seems to be a useful adjunct in the early diagnosis of primary Sjögren's syndrome. The presence of SS-A and SS-B autoantibodies correlated well with the clinical diagnosis of Sjögren's syndrome and seemed to identify the conditions of patients who may have a higher incidence of systemic complications with KCS.


In-vitro schedule-dependent interaction between oxaliplatin and 5-fluorouracil in human gastric cancer cell lines.
In order to define the most effective combination schedule of oxaliplatin (L-OHP) and 5-fluorouracil (5-FU), we investigated the in vitro interaction between these drugs in a panel of four human gastric adenocarcinoma cell lines (MKN-1, NUGC-3, NUGC-5 and AZ-521). Cytotoxic activity was determined by the WST-1 assay. Different schedules of the two drugs were compared and evaluated for synergism, additivity or antagonism with a quantitative method based on the median-effect principle of Chou and Talalay. Cell cycle perturbation and apoptosis were evaluated by flow cytometry. Simultaneous and sequential treatments of L-OHP followed by 5-FU exhibited synergistic effects in all four cell lines, whereas the reverse sequence yielded a clear antagonism. 5-FU exclusively arrested cells at the G0/G1 phase, and L-OHP at the G0/G1 and G2/M phases. Apoptosis was most prominent when cells were treated simultaneously or in a sequence of L-OHP followed by 5-FU, producing apoptosis in the majority of treated cells (55.5-61.5%). In contrast, the reverse sequence yielded only 20% induction of apoptosis, the rate being not significantly different from those induced by each drug singly. Moreover, this sequence dependence was further confirmed by the experiment which compared the total number of NUGC-3 cells 7 days after these combination schedules. These findings suggest that the interaction of 5-FU and L-OHP could be highly schedule dependent, with the most efficacious interaction observed in simultaneous combination and that 5-FU followed by L-OHP would not be recommended in clinical trials for patients with advanced gastric cancer.


Age-related factors in the relationship between foot measurements and living stature and body weight.
The measurements of feet and footprints are especially important in forensic identification, as they have been used to predict the body height and weight of victims or suspects. It can be observed that the subjects of forensic-oriented studies are generally young adults. That is to say, researchers rarely take into consideration the body's proportional changes with age. Hence, the aim of this study is to generate equations which take age and sex into consideration, when stature and body weight are estimated from foot and footprints dimensions. With this aim in mind, we measured the stature, body weight, foot length and breadth, heel breadth, footprint length and breadth, and footprint heel breadth of 516 volunteers (253 males and 263 females) aged between 17.6 and 82.9 years using standard measurement techniques. The sample population was divided randomly into two groups. Group 1, the study group, consisted of 80% of the sample (n = 406); the remaining 20% were assigned to the cross-validation group or Group 2 (n = 110). In the first stage of the study, we produced equations for estimating stature and weight using a stepwise regression technique. Then, their reliability was tested on Group 2 members. Statistical analyses showed that the ratios of foot dimensions to stature and body weight change considerably with age and sex. Consequently, the regression equations which include these variables yielded more reliable results. Our results indicated that age and sex should be taken into consideration when predicting human body height and weight for forensic purposes.


Drug use and phylogenetic clustering of hepatitis C virus infection among people who use drugs in Vancouver, Canada: A latent class analysis approach.
This study estimated latent classes (ie, unobserved subgroups in a population) of people who use drugs in Vancouver, Canada, and examined how these classes relate to phylogenetic clustering of hepatitis C virus (HCV) infection. HCV antibody-positive people who use drugs from two cohorts in Vancouver, Canada (1996-2012), with a Core-E2 sequence were included. Time-stamped phylogenetic trees were inferred, and phylogenetic clustering was determined by time to most common recent ancestor. Latent classes were estimated, and the association with the phylogenetic clustering outcome was assessed using an inclusive classify/analyse approach. Among 699 HCV RNA-positive participants (26% female, 24% HIV+), recent drug use included injecting cocaine (80%), injecting heroin (70%), injecting cocaine/heroin (ie, speedball, 38%) and crack cocaine smoking (28%). Latent class analysis identified four distinct subgroups of drug use typologies: (i) cocaine injecting, (ii) opioid and cocaine injecting, (iii) crack cocaine smoking and (iv) heroin injecting and currently receiving opioid substitution therapy. After adjusting for age and HIV infection, compared to the group defined by heroin injecting and currently receiving opioid substitution therapy, the odds of phylogenetic cluster membership was greater in the cocaine injecting group (adjusted OR [aOR]: 3.06; 95% CI: 1.73, 5.42) and lower in the crack cocaine smoking group (aOR: 0.06; 95% CI: 0.01, 0.48). Combining latent class and phylogenetic clustering analyses provides novel insights into the complex dynamics of HCV transmission. Incorporating differing risk profiles associated with drug use may provide opportunities to further optimize and target HCV treatment and prevention strategies.


Bullous lesions in Kaposi's sarcoma: case report.
Bullous lesions have been only rarely described in Kaposi's sarcoma (KS), and their histopathologic features have never been described in detail. We report a case of bullous lesions of KS in an 82-year-old Italian woman. The patient had typical smooth pale reddish-grey slightly-raised KS plaques on the legs, present for at least 10 years. Several dull grayish-pink blisters (0.5 to 2 cm in diameter) affected both dorsa of her feet and ankles symmetrically. Two punch biopsies were taken, one from an infiltrated KS plaque on the right buttock and the other from a bullous lesion on the right foot. Histopathologically, the late KS plaque on the buttock showed typical features of KS, with an increased number of spindle cells arranged in short bundles and extravasation of erythrocytes. The bullous lesion on the foot showed a full-thickness vascular neoplasm involving the upper and lower dermis and the subcutaneous fat. The upper portion of the lesion contained many newly formed, highly-dilated blood vessels, touching the overlying epidermis and separated from it by a narrow band of collagen and endothelial cells; wide, empty spaces characterized the superficial dermis, in which preexisting venules and bands of collagen associated with non-atypical endothelial cells floated. All these findings would suggest a lymphangiomatous lesion, if the presence of specific diagnostic criteria of KS were not recognizable at a deeper level of the lesion. Various criteria actually suggest that the bullous lesion may be regarded as an epiphenomenon of a KS plaque lesion: (a) full-thickness involvement of the reticular dermis and, in this case, also of the subcutaneous fat; (b) dense and patchy lymphoplasmocytic infiltrate typical of plaque lesions and, much less frequently, of patch lesions; (c) presence of ectatic blood vessels, filled with plasma and erythrocytes (pseudoangiomatous findings), a nonpathognomonic but highly characteristic finding of the plaque lesion; and (d) as in the KS plaque lesions, in the bullous lesion as well the reticular dermis was characterized by an increased number of anastomosing bizarrely shaped vascular spaces lined by non-atypical endothelial cells. We hypothesize that the prevalence of lymphangiomatous differentiation in the upper dermis represents one of the many features of KS lesions. When present, it may correlate with the clinical feature of a blister.


[Relation of pre-operative ecography to laparoscopic cholecystectomy difficulty at the Central Military Hospital].
This study included 62 patients diagnosed with known vesicular disease, who underwent laparoscopic cholecystectomy between August 1999 and March 2000, at the Central Military Hospital. Those patients showing alterations in the hepatic function tests and dilatation of the biliary tract in the pre-operative echography were excluded from the study. The patients selected were subjected to evaluation of the echographic parameters one day before the surgery in order to determine the vesicular volume and ejection fraction. The maximum ejection fraction was calculated as the difference between the fasting gastric volume and the residual volume, in the fasting volume percentage. We evaluated the above-mentioned operative parameters one day following the laparoscopic cholecystectomy, using a visual analog scale (VAS) for evaluation of the surgical difficulty and bleeding during surgery. The surgery duration was measured in minutes. Subsequently, the pre-operative echographic parameters, excluding the ballstone parameter in the main biliary tract, were related to the surgery parameters, obtaining the following results: -The pre-operative echographic parameters, thickness of the vesicular wall and vesicular ejection fraction, are the best indicators of surgical difficulty in laparoscopic cholecystectomy. -The pre-operative echographic parameter, vesicular ejection fraction <50% and vesicular wall thickness = 3 mm are indicators of larger bleeding during laparoscopic cholecystectomy. -The pre-operative echographic parameter, vesicular ejection fraction in gallbladder indicates prolongation of surgery duration in laparoscopic cholecystectomy. -Finally, our study supports the use of echography as a useful diagnosis mean in patients undergoing laparoscopic cholecystectomy, and which can predict, through some of its echographic parameters, cases with technical difficulty, bleeding during surgery and prolongation of surgery time in laparoscopic cholecystectomy.


Regulation of muscle protein synthesis and the effects of catabolic states.
Protein synthesis and degradation are dynamically regulated processes that act in concert to control the accretion or loss of muscle mass. The present article focuses on the mechanisms involved in the impairment of protein synthesis that are associated with skeletal muscle atrophy. The vast majority of mechanisms known to regulate protein synthesis involve modulation of the initiation phase of mRNA translation, which comprises a series of reactions that result in the binding of initiator methionyl-tRNAi and mRNA to the 40S ribosomal subunit. The function of the proteins involved in both events has been shown to be repressed under atrophic conditions such as sepsis, cachexia, chronic kidney disease, sarcopenia, and disuse atrophy. The basis for the inhibition of protein synthesis under such conditions is likely to be multifactorial and includes insulin/insulin-like growth factor 1 resistance, pro-inflammatory cytokine expression, malnutrition, corticosteroids, and/or physical inactivity. The present article provides an overview of the existing literature regarding mechanisms and signaling pathways involved in the regulation of mRNA translation as they apply to skeletal muscle wasting, as well as the efficacy of potential clinical interventions such as nutrition and exercise in the maintenance of skeletal muscle protein synthesis under atrophic conditions. This article is part of a Directed Issue entitled: Molecular basis of muscle wasting.


Default network and intelligence difference.
In the last few years, many studies in the cognitive and system neuroscience found that a consistent network of brain regions, referred to as the default network, showed high levels of activity when no explicit task was performed. Some scientists believed that the resting state activity might reflect some neural functions that consolidate the past, stabilize brain ensembles and prepare us for the future. Here, we modeled default network as undirected weighted graph and then used graph theory to investigate the topological properties of the default network of the two groups of people with different intelligence levels. We found that, in both groups, the posterior cingulate cortex showed the greatest degree in comparison to the other brain regions in the default network, and that the medial temporal lobes and cerebellar tonsils were topologically separations from the other brain regions in the default network. More importantly, we found that the strength of some functional connectivities and the global efficiency of default network were significantly different between the superior intelligence group and the average intelligence group, which indicates that the functional integration of the default network might be related to the individual intelligent performance.


Paroxysmal atrial fibrillation: why patients experience different symptoms from the same arrhythmia?
A careful evaluation and interpretation of the symptoms of the patient with atrial fibrillation should be the first step in selecting a treatment modality. Arrhythmia burden is not equal to symptoms. Some patients will have and continue to have symptoms not related to the occurrence of arrhythmias. Recognition of these patients is important in the treatment of atrial fibrillation.


[Perspective of genetic improvement of inflorescences structure in oil crops of Brassicaceae family].
The theoretical substantiation and new scientific decision of ability to genetic construction of agriculturally valuable inflorescence structure in oil crops of Brassicaceae family are proposed on the examples of A. thaliana genes AP1, BP1 and TFL1. New types of plants are more suitable to machine seed harvesting as a result of decreasing non-simultaneous ripeness of seeds within inflorescences. It has been determined that A. thaliana forms inflorescences with new combinations of untypical for family Brassicaceae features by the effects of recessive alleles ap1-1, bp-1 and tfl1-2 expression.


Consumption of dairy products in relation to the presence of clinical knee osteoarthritis: The Maastricht Study.
Observational studies showed inverse associations between milk consumption and knee osteoarthritis (knee OA). There is lack of information on the role of specific dairy product categories. We explored the association between dairy consumption and the presence of knee osteoarthritis in 3010 individuals aged 40-75 years participating in The Maastricht Study. The presence of knee OA was defined according to a slightly modified version of the American College of Rheumatology (ACR) clinical classification criteria. Data on dairy consumption were appraised by a 253-item FFQ covering 47 dairy products with categorization on fat content, fermentation or dairy type. Multivariable logistic regression analyses were performed to estimate odd ratios (ORs) and 95% confidence intervals (95%CI), while correcting for relevant factors. 427 (14%) participants were classified as having knee OA. Significant inverse associations were observed between the presence of knee OA and intake of full-fat dairy and Dutch, primarily semi-hard, cheese, with OR for the highest compared to the lowest tertile of intake of 0.68 (95%CI 0.50-0.92) for full-fat dairy, and 0.75 (95%CI 0.56-0.99) for Dutch cheese. No significant associations were found for other dairy product categories. In this Dutch population, higher intake of full-fat dairy and Dutch cheese, but not milk, was cross-sectionally associated with the lower presence of knee OA. Prospective studies need to assess the relationship between dairy consumption, and in particular semi-hard cheeses, with incident knee OA.


Functionally redundant RXLR effectors from Phytophthora infestans act at different steps to suppress early flg22-triggered immunity.
Genome sequences of several economically important phytopathogenic oomycetes have revealed the presence of large families of so-called RXLR effectors. Functional screens have identified RXLR effector repertoires that either compromise or induce plant defense responses. However, limited information is available about the molecular mechanisms underlying the modes of action of these effectors in planta. The perception of highly conserved pathogen- or microbe-associated molecular patterns (PAMPs/MAMPs), such as flg22, triggers converging signaling pathways recruiting MAP kinase cascades and inducing transcriptional re-programming, yielding a generic anti-microbial response. We used a highly synchronizable, pathogen-free protoplast-based assay to identify a set of RXLR effectors from Phytophthora infestans (PiRXLRs), the causal agent of potato and tomato light blight that manipulate early stages of flg22-triggered signaling. Of thirty-three tested PiRXLR effector candidates, eight, called Suppressor of early Flg22-induced Immune response (SFI), significantly suppressed flg22-dependent activation of a reporter gene under control of a typical MAMP-inducible promoter (pFRK1-Luc) in tomato protoplasts. We extended our analysis to Arabidopsis thaliana, a non-host plant species of P. infestans. From the aforementioned eight SFI effectors, three appeared to share similar functions in both Arabidopsis and tomato by suppressing transcriptional activation of flg22-induced marker genes downstream of post-translational MAP kinase activation. A further three effectors interfere with MAMP signaling at, or upstream of, the MAP kinase cascade in tomato, but not in Arabidopsis. Transient expression of the SFI effectors in Nicotiana benthamiana enhances susceptibility to P. infestans and, for the most potent effector, SFI1, nuclear localization is required for both suppression of MAMP signaling and virulence function. The present study provides a framework to decipher the molecular mechanisms underlying the manipulation of host MAMP-triggered immunity (MTI) by P. infestans and to understand the basis of host versus non-host resistance in plants towards P. infestans.


Inactivation of Escherichia coli O157:H7 on Romaine Lettuce When Inoculated in a Fecal Slurry Matrix.
A field trial was conducted in July 2011 to quantify the inactivation rate of Escherichia coli O157:H7 when mixed with fecal slurry and applied to romaine lettuce leaves. Lettuce was grown under commercial conditions in Salinas Valley, CA. One-half milliliter of rabbit fecal slurry, containing 6.3 × 107 CFU of E. coli O157:H7, was inoculated onto the upper (adaxial) surface of a lower leaf on 240 heads of lettuce within 30 min after a 2.5-h irrigation event. Forty-eight romaine lettuce heads were collected per event at 2.5 h (day 0.1), 19.75 h (day 0.8), 43.25 h (day 1.8), 67.25 h (day 2.8), and 91.75 h (day 3.8) postinoculation and were analyzed for the concentration of E. coli O157:H7 (Ct). E. coli O157:H7 was detected on 100% of collected heads in concentrations ranging from 340 to 3.40 × 1010 most probable number (MPN) per head. Enumeration data indicate substantial growth of E. coli O157:H7 postinoculation (2.5 h), leading to elevated concentrations, 1 to 3 log above the starting inoculum concentration (Co). By the end of the 92-h trial, we observed a net 0.8-log mean reduction of E. coli O157:H7 compared with Co; however, after accounting for the substantial bacterial growth, there was an overall 2.3-log reduction by the final sampling event (92 h). On the basis of two different regression models that used either the raw data for Ct or log-transformed values of Ct/Co during the period 2.5 to 91.75 h postinoculation, there was an estimated 76 to 80% reduction per day in bacterial counts; however, more accurate predictions of MPN per head of lettuce were generated by using non-log-transformed values of Ct. This study provides insight into the survival of E. coli O157:H7 transferred via splash from a contaminated fecal source onto produce during irrigation. Moreover, these findings can help generate inactivation times following a potential contamination incident.


A broad spectrum Kunitz type serine protease inhibitor secreted by the hookworm Ancylostoma ceylanicum.
Although blood-feeding hookworms infect over a billion people worldwide, little is known about the molecular mechanisms through which these parasitic nematodes cause gastrointestinal hemorrhage and iron deficiency anemia. A cDNA corresponding to a secreted Kunitz type serine protease inhibitor has been cloned from adult Ancylostoma ceylanicum hookworm RNA. The translated sequence of the A. ceylanicum Kunitz type inhibitor 1 (AceKI-1) cDNA predicts a 16-amino acid secretory signal sequence, followed by a 68-amino acid mature protein with a molecular mass of 7889 daltons. Recombinant protein (rAceKI-1) was purified from induced lysates of Escherichia coli transformed with the rAceKI-1/pET 28a plasmid, and in vitro studies demonstrate that rAceKI-1 is a tight binding inhibitor of the serine proteases chymotrypsin, pancreatic elastase, neutrophil elastase, and trypsin. AceKI-1 inhibitory activity is present in soluble protein extracts and excretory/secretory products of adult hookworms but not the infective third stage larvae. The native AceKI-1 inhibitor has been purified to homogeneity from soluble extracts of adult A. ceylanicum using size exclusion and reverse-phase high pressure liquid chromatography. As a potent inhibitor of mammalian intestinal proteases, AceKI-1 may play a role in parasite survival and the pathogenesis of hookworm anemia.


Occult cerebrovascular malformations after irradiation.
It has recently been found that patients receiving cerebral irradiation can develop hemorrhagic dysangiogeneses simulating occult vascular malformations. To analyze this connection, we report on five patients with occult cerebrovascular malformations occurring after "standard" or focused irradiation performed for brain tumors in four patients and for a deep-seated cavernous angioma in one patient. All lesions were within the radiation ports. The time interval between irradiation and the detection of the occult vascular malformations varied from 3 to 9 years; the ratio of female to male patients was 4:1. Four patients were < 15 years old when first irradiated. Four patients presented with acute symptoms (headache, vomiting, focal signs) and one was asymptomatic when the lesions were first detected. Serial magnetic resonance imaging scans were available in four patients and a computed tomographic scan in the other patient. The initial appearance was that of a hypointense T1-T2 focus; magnetic resonance imaging then revealed focal or multifocal T1 hyperintensity and T2 mixed signal intensity followed by a late ring of decreased signal intensity. Four patients were operated on and one was under neuroradiological monitoring. Histological features of these lesions included clusters of closely packed vascular spaces resembling cavernous malformations sometimes associated with a thrombosed thick-walled vein with intense hemosiderin deposition and fibroblastic proliferation; telangiectasic changes were also seen in the adjacent brain. Increased awareness of occult cerebrovascular malformations is necessary, because their occurrence is not infrequent and they have hemorrhagic potential. Children receiving cerebral irradiation are at greater risk of this complication.


Plasma biomarker discovery for early chronic kidney disease diagnosis based on chemometric approaches using LC-QTOF targeted metabolomics data.
Chronic kidney disease (CKD) is a progressive pathological condition in which renal function deteriorates in time. The first diagnosis of CKD is often carried out in general care attention by general practitioners by means of serum creatinine (CNN) levels. However, it lacks sensitivity and thus, there is a need for new robust biomarkers to allow the detection of kidney damage particularly in early stages. Multivariate data analysis of plasma concentrations obtained from LC-QTOF targeted metabolomics method may reveal metabolites suspicious of being either up-regulated or down-regulated from urea cycle, arginine methylation and arginine-creatine metabolic pathways in CKD pediatrics and controls. The results show that citrulline (CIT), symmetric dimethylarginine (SDMA) and S-adenosylmethionine (SAM) are interesting biomarkers to support diagnosis by CNN: early CKD samples and controls were classified with an increase in classification accuracy of 18% when using these 4 metabolites compared to CNN alone. These metabolites together allow classification of the samples into a definite stage of the disease with an accuracy of 74%, being the 90% of the misclassifications one level above or below the CKD stage set by the nephrologists. Finally, sex-related, age-related and treatment-related effects were studied, to evaluate whether changes in metabolite concentration could be attributable to these factors, and to correct them in case a new equation is developed with these potential biomarkers for the diagnosis and monitoring of pediatric CKD.


Matrix metalloproteinase inhibition modifies left ventricular remodeling after myocardial infarction in pigs.
Global and regional shape changes that occur within the left ventricular wall after myocardial infarction have been termed infarct expansion. A potential mechanism for this postinfarction remodeling is activation of the matrix metalloproteinases. Accordingly, the present study examined the effects of matrix metalloproteinase inhibition on left ventricular global geometry after myocardial infarction in pigs. Myocardial infarction was created in pigs by means of occlusion of the first and second obtuse marginal branches of the circumflex coronary artery, resulting in a uniform left ventricular free wall infarct size of 21% +/- 2%. At 5 days after infarction, the pigs were randomized to undergo broad-spectrum matrix metalloproteinase inhibition (n = 9; PD166793, 20 mg. kg(-1). d(-1) by mouth) or myocardial infarction alone (n = 8). Ten pigs served as noninfarction control animals. Left ventricular end-diastolic area, determined by means of echocardiography, was measured 8 weeks after infarction. Left ventricular end-diastolic area increased in both the myocardial infarction plus broad-spectrum matrix metalloproteinase inhibition and myocardial infarction only groups compared to reference control animals (3.7 +/- 0.2 cm(2)), but was reduced with broad-spectrum matrix metalloproteinase inhibition compared to myocardial infarction alone (4.5 +/- 0.2 vs 4.9 +/- 0.2 cm(2), respectively; P <.05). Regional radial stress within the infarct region increased in both infarction groups when compared to values obtained from reference control animals (599 +/- 152 g/cm(2)), but was attenuated in the myocardial infarction plus broad-spectrum matrix metalloproteinase inhibition group compared to the myocardial infarction alone group (663 +/- 108 vs 1242 +/- 251 g/cm(2), respectively; P <.05). Similarly, regional myocardial stiffness increased in both the myocardial infarction plus broad-spectrum matrix metalloproteinase inhibition and the myocardial infarction only groups compared with that observed in reference control animals (14 +/- 1 rkm, P <.05) but was lower with broad-spectrum matrix metalloproteinase inhibition than with myocardial infarction alone (42 +/- 6 vs 68 +/- 10 rkm, respectively; P <.05). Matrix metalloproteinase inhibition reduced postinfarction left ventricular dilation, reduced regional myocardial wall stress, and modified myocardial material properties. These unique findings suggest that increased myocardial matrix metalloproteinase activation after infarction contributes directly to the left ventricular remodeling process.


Management in non-traumatic arm, neck and shoulder complaints: differences between diagnostic groups.
Arm, neck and/or shoulder complaints are common in western societies. In the Netherlands, general practice guidelines are issued on shoulder pain and epicondylitis only. Little is known about actual management of the total range of diagnoses. The objectives of the study are: to determine management in patients consulting the GP with a new episode of non-traumatic arm neck and shoulder complaints up to 6 months after the first consultation. To evaluate differences in management between patients with specific diagnoses versus non-specific diagnoses and between specific diagnostic groups. In a prospective cohort study in general practice. We recruited 682 eligible patients. Data on diagnosis, management, patient- and complaint-characteristics were collected. Co-occurrence of treatment options was presented in scaled rectangles. After 6 months, additional diagnostic tests had been performed in 18% of the patients, mainly radiographic examination (14%). Further, 49% had been referred for physiotherapy and 12% to the medical specialist. Patients with specific diagnoses were more frequently referred for specialist treatment, and patients with non-specific diagnoses for physiotherapy. Corticosteroid injections (17%) were mainly applied specific diagnoses (e.g. impingement syndrome, frozen shoulder, carpal tunnel and M. Quervain). Frequencies of prescribed medication (51%) did not differ between specific and non-specific diagnoses. In 19% of the patients no referral, prescribed analgesics or injection was applied. Braces (4%) were mainly prescribed in epicondylitis. Overall, management most frequently consisted of prescribed analgesics and referral for physiotherapy. Specific and non-specific diagnostic subgroups differed in the frequency corticosteroid injections were applied, and referrals to physiotherapy and to a medical specialist.


Developmental stages of the third molar in Israeli children.
The purpose of this study was to examine the developmental stages of the third molar in Israeli children, compare its development in boys and girls, in the maxilla and mandible, right to left sides in various age groups, and relate the developmental stage to chronological age. Panoramic radiographs of 693 healthy children and adolescents (328 boys and 365 girls) ages 7 to 16 were analyzed using a modified 6-stage method. Since the 16-year-old group was too small for statistical analysis, it was later excluded from the study. Radiographs were obtained from patient files. There were no significant differences in the development of the third molar between boys and girls and between right and left sides. Slight differences were found between the maxilla and mandible. The first appearance of a radiolucent bud was at age 8.7 in the mandible and 9.3 in the maxilla. Crown completion was observed on average at age 11.7 in the maxilla and 11.8 in the mandible. There was a high correlation between third molar development and chronological age. Agenesis can be determined conclusively if no radiolucent bud is present beyond age 14. The modified 6-stage method allowed more accurate determination of developmental stages.


Cell Line Models for Human Cytomegalovirus Latency Faithfully Mimic Viral Entry by Macropinocytosis and Endocytosis.
Human cytomegalovirus (HCMV) enters primary CD34+ hematopoietic progenitor cells by macropinocytosis, where it establishes latency in part because its tegument-transactivating protein, pp71, remains associated with endosomes and is therefore unable to initiate productive, lytic replication. Here we show that multiple HCMV strains also enter cell line models used to study latency by macropinocytosis and endocytosis. In all latency models tested, tegument-delivered pp71 was found to be colocalized with endosomal markers and was not associated with the seven other cytoplasmic localization markers tested. Like the capsid-associated pp150 tegument protein, we initially detected capsid proteins in association with endosomes but later detected them in the nucleus. Inhibitors of macropinocytosis and endocytosis reduced latent viral gene expression and precluded reactivation. Importantly, we utilized electron microscopy to observe entry by macropinocytosis and endocytosis, providing additional visual corroboration of the findings of our functional studies. Our demonstration that HCMV enters cell line models for latency in a manner indistinguishable from that of its entry into primary cells illustrates the utility of these cell lines for probing the mechanisms, host genetics, and small-molecule-mediated inhibition of HCMV entry into the cell types where it establishes latency.IMPORTANCE Primary cells cultured in vitro currently provide the highest available relevance for examining molecular and genetic requirements for the establishment, maintenance, and reactivation of HCMV latency. However, their expense, heterogeneity, and intransigence to both long-term culture and molecular or genetic modification create rigor and reproducibility challenges for HCMV latency studies. There are several cell line models for latency not obstructed by deficiencies inherent in primary cells. However, many researchers view cell line studies of latency to be physiologically irrelevant because of the perception that these models display numerous and significant differences from primary cells. Here, we show that the very first step in a latent HCMV infection, entry of the virus into cells, occurs in cell line models in a manner indistinguishable from that in which it occurs in primary CD34+ hematopoietic progenitor cells. Our data argue that experimental HCMV latency is much more similar than it is different in cell lines and primary cells.


Semantator: annotating clinical narratives with semantic web ontologies.
To facilitate clinical research, clinical data needs to be stored in a machine processable and understandable way. Manual annotating clinical data is time consuming. Automatic approaches (e.g., Natural Language Processing systems) have been adopted to convert such data into structured formats; however, the quality of such automatically extracted data may not always be satisfying. In this paper, we propose Semantator, a semi-automatic tool for document annotation with Semantic Web ontologies. With a loaded free text document and an ontology, Semantator supports the creation/deletion of ontology instances for any document fragment, linking/disconnecting instances with the properties in the ontology, and also enables automatic annotation by connecting to the NCBO annotator and cTAKES. By representing annotations in Semantic Web standards, Semantator supports reasoning based upon the underlying semantics of the owl:disjointWith and owl:equivalentClass predicates. We present discussions based on user experiences of using Semantator.


Functional evaluation of the role of reticuloendotheliosis virus long terminal repeat (LTR) integrated into the genome of a field strain of Marek's disease virus.
MDV-GX0101 is a field strain of Marek's disease virus with a naturally occurring insertion of the reticuloendotheliosis virus (REV) LTR fragment. In order to study the biological properties of REV-LTR insertion in the MDV genome, we constructed a full-length infectious BAC clone of MDV-GX0101 strain and deleted the LTR sequences by BAC mutagenesis. The pathogenic properties of the LTR-deleted virus were evaluated in infected SPF birds. The study demonstrated that the LTR-deleted virus had a stronger inhibitory effect on the growth rates of the infected birds and induced stronger immunosuppressive effects. Surprisingly, however, the ability for horizontal transmission of the LTR-deleted virus appeared to be significantly weaker than its parental LTR-intact virus. Even though the precise molecular mechanisms are still not clear, the results of our studies demonstrate that the retention of the REV-LTR in the MDV genome decreases its pathogenic effects but increases its potential for horizontal transmission.


[The use of human insulin in cases of insulin allergy].
The authors treated 14 insulin-allergic diabetic patients in 1985-87. Previous short-term therapy with conventional insulins could be verified in 12 cases. The "presensitizing" effect of this transient treatment seems highly probable. Five of the 14 patients were allergic to all animal insulins incl. monospecies porcine monocomponent (MC) preparations. Surprisingly, all these patients proved to be allergic to biosynthetic human insulin (Huminsulin Normal and Huminsulin Basal) preparations too, but two of them could be treated with semisynthetic human insulins (Actrapid HM and Monotard HM). Another patient was able to receive Monotard HM exclusively. The remaining two patients proved to be allergic to all human insulins used. One of them was desensitized but the insulin therapy had to be supplemented with steroid drugs for two years. The fifth patient continued the oral treatment but her metabolism is poorly controlled. These results show that some diabetics with allergy to animal insulins are allergic to human insulins as well. As a prevention, transient therapy with conventional preparations should be avoided. If, however, there is an absolute indication for it (e.g. gestational diabetes or surgical intervention), it should be carried out with human insulin, or, if that is not available, with monospecies porcine MC insulin.


More (G-proteins) please! Identification of an elaborate network of G-proteins in soybean.
The studies performed in model plants Arabidopsis and rice have revealed a significantly simple repertoire of canonical G-protein components in plants, with the presence of only two possible heterotrimers versus hundreds of possible trimeric combinations in animal systems. Since a number of plant species are polyploid, we assessed if genome duplication events have resulted into multiplicity of G-protein components in such plants and whether the duplicated gene pairs have specific expression patterns or biochemical properties. Our analysis of soybean genome has identified four Gα, four Gβ and two Gγ proteins, predicting thirty-two possible heterotrimeric combinations. All ten G-protein genes are retained in soybean genome and ubiquitously expressed. The G-protein genes have interesting expression profiles during seed developments and germination. The four Gα proteins form two distinct groups based on their GTPase activity. Yeast-based interaction analyses predict that the proteins interact in most but not all of the possible combinations, and there is some degree of interaction specificity between duplicated gene pairs. This research, thus, identifies the most elaborate heterotrimeric G-protein network known to date in plants.


Japanese morning glory dusky mutants displaying reddish-brown or purplish-gray flowers are deficient in a novel glycosylation enzyme for anthocyanin biosynthesis, UDP-glucose:anthocyanidin 3-O-glucoside-2''-O-glucosyltransferase, due to 4-bp insertions in the gene.
Bright blue or red flowers in the Japanese morning glory (Ipomoea nil) contain anthocyanidin 3-O-sophoroside derivatives, whereas the reddish-brown or purplish-gray petals in its dusky mutants accumulate anthocyanidin 3-O-glucoside derivatives. The Dusky gene was found to encode a novel glucosyltransferase, UDP-glucose:anthocyanidin 3-O-glucoside-2''-O-glucosyltransferase (3GGT), which mediates the glucosylation of anthocyanidin 3-O-glucosides to yield anthocyanidin 3-O-sophorosides. Ipomoea nil carries one copy of the 3GGT gene that contains no intron and produces 1.6-kbp transcripts mainly in the petals and tubes of flower buds at around 24 h before flower opening. The gene products of both In3GGT in I. nil and Ip3GGT in the common morning glory (Ipomoea purpurea) comprise 459 amino acids and showed a close relationship to the petunia UDP-rhamnose:anthocyanidin 3-O-glucoside-6''-O-rhamnosyltransferase (3RT), which controls the addition of a rhamnose molecule to anthocyanidin 3-O-glucosides for conversion into anthocyanidin 3-O-rutinosides. All of the 30 dusky mutants tested were found to carry the 4-bp insertion mutations GGAT or CGAT at an identical position near the 3' end of the gene, and the insertions caused frameshift mutations. The expected 3GGT enzymatic activities were found in the crude extracts of Escherichia coli, in which the 3GGT cDNA of I. nil or I. purpurea was expressed, while no such activity was detected in the extracts expressed with the dusky mutant cDNAs containing 4-bp insertions. Moreover, the introduced Ip3GGT cDNA efficiently produced 3GGT that converted cyanidin 3-O-glucoside into cyanidin 3-O-sophoroside in transgenic petunia plants.


Reflections on DSM classification and its utility in primary care: case studies in "mental disorders".
This case series was conducted to demonstrate the limitations of the DSM system in primary care patients. Sixty family health center patients free of mental disorders according to DSM-IV criteria completed monthly quantitative interviews, using multiple rating instruments, concerning the levels of psychiatric symptoms, presence of distress and/or a mental disorder, functional status, support, and stressors. In addition, a purposive sample of 16 subjects completed an in-depth qualitative interview concerning their situation at the time they crossed a DSM threshold. Data were collected from April 2000 to March 2001. Overall, there were 14 subjects with distress alone, 6 subjects with subthreshold disorders, and 3 subjects with known transient (< 2 months' duration) threshold disorders. Thus, even with the incomplete longitudinal data in this study, the clinical needs of 23 (38%) of the original 60 subjects were inadequately met by the DSM criteria. From the 10 subjects who crossed a DSM threshold and completed a qualitative interview, we selected 5 case studies with the most complete and complementary quantitative and qualitative data to illustrate several findings regarding the utility of the DSM classification in primary care. First, these cases show that psychological symptoms and DSM disorders vary considerably. Second, distress and subthreshold disorders are often seen in primary care patients. Third, the crossing of a DSM threshold corresponds to extreme levels of psychological symptoms and may therefore represent symptom severity. Fourth, psychological symptoms are often linked to physical illness. Finally, the context in which these symptoms and disorders develop often produces complex dynamic patterns. The current DSM system failed to adequately reflect the spectrum and context of mental illness in patients from a predominantly low-income, Hispanic primary care population.


"Just Join Them": Improv and Dementia Care.
Improvisational (improv) theatre skill development holds promise for improving the dementia capability of care partners. In this report, we present analysis of data from an ongoing study on meaningful engagement and quality of life among assisted living (AL) residents with dementia. Using ethnographic methods, we collected data from persons with dementia (n = 59) and their care partners (n = 165) in six diverse AL communities each studied for one year. Building cumulatively on past work and existing literature, we demonstrate the potential benefits of training care partners to use improv skills. We discuss implications, including the need for intervention research.


Stem cells and germ cells: microRNA and gene expression signatures.
The study of primordial germ cell development in vivo is hampered by their low numbers and inaccessibility. Recent research has shown the ability of embryonic and adult stem cells to differentiate into primordial germ cells and more mature gametes and this generation of germ cells in vitro may be an attractive model for their study. One of the biggest challenges facing in vitro differentiation of stem cells into primordial germ cells is the lack of markers to clearly distinguish the two. As both cell types originate early in embryonic development they share many pluripotent markers such as OCT4, VASA, FRAGILIS, and NANOG. Genome wide microarray profiling has been used to identify transcriptome patterns unique to primordial germ cells. A more thorough analysis of the temporal and quantitative expression of a panel of genes may be more robust in distinguishing these two cell populations. MicroRNAs, short RNA molecules that have been shown to regulate translation through interactions with mRNA transcripts, have also recently come under investigation for the role they may play in pluripotency. Attempts to elucidate key microRNAs responsible for both stem cell and primordial germ cell characteristics have recently been undertaken. Unique microRNAs, either individually or as global profiles, may also help to distinguish differentiated primordial germ cells from stem cells in vitro. This review will examine gene expression and microRNA signatures in stem cells and germ cells as ways to distinguish these closely related cell types.


Visual outcome at 5 years of newborn infants at risk of cerebral visual impairment.
Visual development at 5 years of age was tested in a group of 39 children who had shown severe neonatal encephalopathy or perinatal brain lesions, documented by medical history, cranial ultrasound, or MRI. In all children, grating acuity was tested during the first 2 years of life. The assessment protocol at 5 years included various visual functions (grating and resolution acuity, visual field size, depth perception, optokinetic nystagmus, and ocular motility), and neurological and cognitive development. The majority of the children showed visual disorders of different type and degree, which were not due to ophthalmological abnormalities. Visual defects correlated well with the results of early visual assessment and of neuroimaging. Visual outcome could be predicted by grating acuity at 1 to 2 years in 27 of the 39 children, by neonatal cranial ultrasound in 26 of the 32 cases examined by this technique, and by later MRI in 23 out of 27. Moreover, a significant correlation was found between visual, motor, and cognitive impairment.


Targeted memory reactivation of face-name learning depends on ample and undisturbed slow-wave sleep.
Face memory, including the ability to recall a person's name, is of major importance in social contexts. Like many other memory functions, it may rely on sleep. We investigated whether targeted memory reactivation during sleep could improve associative and perceptual aspects of face memory. Participants studied 80 face-name pairs, and then a subset of spoken names with associated background music was presented unobtrusively during a daytime nap. This manipulation preferentially improved name recall and face recognition for those reactivated face-name pairs, as modulated by two factors related to sleep quality; memory benefits were positively correlated with the duration of stage N3 sleep (slow-wave sleep) and negatively correlated with measures of sleep disruption. We conclude that (a) reactivation of specific face-name memories during sleep can strengthen these associations and the constituent memories, and that (b) the effectiveness of this reactivation depends on uninterrupted N3 sleep.


An annotated checklist of freshwater Copepoda (Crustacea, Hexanauplia) from continental Ecuador and the Galapagos Archipelago.
An annotated checklist of the free-living freshwater Copepoda recorded in different regions in Ecuador (including the Amazon, the Andes, the coastal region, and the Galapagos Islands) is here provided. We revised all published records, critically evaluated the validity of each taxon and provided short taxonomic and biogeographical remarks for each one. A total of 27 taxa have been reported, including species and records at the generic level only. The species and taxa identified only up to the generic level belong to five families and 14 genera. The Cyclopoida is the most diverse group with 16 records belonging to species (or identified to the generic level only) and eight genera, followed by the Harpacticoida with six species, one identification to the generic level only, and four genera, and Calanoida with four species belonging to two genera. A total of 18 taxa are recorded for the Andes. Six have been recorded in the Amazon, two are recorded for the coastal region, and six for the Galapagos. One species is shared between the Amazon and the Andes. One species is shared between the coastal region and the Amazon. Seventeen are only reported from the Andes and four are only reported from the Amazon. At the current status of the knowledge, any attempt to analyze and generalize distributional patterns of copepods in Ecuador is premature due to the scarcity of available information, and evidently there is an urgent need for more extensive field collections. A few working hypothesis for future studies are identified.


Diagnostic imaging in patients with retinitis pigmentosa.
Retinitis pigmentosa (RP) is a progressive inherited retinal disease, and patients with RP have reduced visual function caused by a degeneration of the photoreceptors and retinal pigment epithelium (RPE). At the end stage of RP, the degeneration of the photoreceptors in the fovea reduces central vision, and RP is one of the main causes of acquired blindness in developed countries. Therefore, morphological and functional assessments of the photoreceptors in the macula area can be useful in estimating the residual retinal function in RP patients. Optical coherence tomography (OCT) is a well-established method of examining the retinal architecture in situ. The photoreceptor inner/outer segment (IS/OS) junction is observed as a distinct, highly reflective line by OCT. The presence of the IS/OS junction in the OCT images is essential for normal visual function. Fundus autofluorescence (FAF) results from the accumulation of lipofuscin in the RPE cells and has been used to investigate RPE and retinal function. More than one-half of RP patients have an abnormally high density parafoveal FAF ring (AF ring). The AF ring represents the border between functional and dysfunctional retina. In this review, we shall summarize recent progress on diagnostic imaging in eyes with RP.


Custirsen (OGX-011) combined with cabazitaxel and prednisone versus cabazitaxel and prednisone alone in patients with metastatic castration-resistant prostate cancer previously treated with docetaxel (AFFINITY): a randomised, open-label, international, phase 3 trial.
Docetaxel and cabazitaxel improve overall survival compared with mitoxantrone in patients with metastatic castration-resistant prostate cancer. Custirsen (OGX011) is a second generation highly specific antisense oligonucleotide that inhibits the production of clusterin, an antiapoptotic protein that is upregulated in response to chemotherapy and that confers treatment resistance. We aimed to assess whether custirsen in combination with cabazitaxel and prednisone increases overall survival in patients with metastatic castration-resistant prostate cancer previously treated with docetaxel. In this randomised, open-label, international, phase 3 trial, men with radiographically documented metastatic castration-resistant prostate cancer that had progressed after docetaxel treatment with a Karnofsky performance status of more than 70% and who were fit for chemotherapy, were recruited from 95 cancer treatment centres in eight countries. Patients were randomly assigned (1:1) centrally using permuted blocks (block size 8) to receive cabazitaxel plus prednisone (cabazitaxel 25 mg/m2 intravenously every 21 days plus oral prednisone 10 mg daily) with or without custirsen (640 mg intravenously on days 1, 8, and 15, plus three previous loading doses) until disease progression, unacceptable toxicity, or the completion of ten treatment cycles. Randomisation was stratified by use of opioids for prostate cancer-related pain at screening, disease progression following first-line docetaxel treatment established by radiographic evidence, and previous treatment with abiraterone or enzalutamide. The co-primary endpoints were overall survival in all randomly assigned patients and in a poor-prognosis subgroup. All analyses were intention to treat with the exception of safety, which was reported for patients who received any assigned treatment. The trial has been completed and the results presented here are the final analysis. This trial is registered with Clinicaltrials.gov, number NCT01578655. Between Sept 9, 2012, and Sept 29, 2014, 795 patients were screened for enrolment. 635 men were eligible for inclusion and were randomly assigned (n=317 in the cabazitaxel and prednisone plus custirsen group and n=318 in the cabazitaxel and prednisone group). Median follow up was 28·3 months (IQR 24·4-34·5) for the custirsen group and 29·8 months (IQR 25·3-35·2) for the control group. Median overall survival in all randomly assigned patients did not differ between the two groups (14·1 months [95% CI 12·7-15·9] in the curtisen group vs 13·4 months [12·1-14·9] in the control group; hazard ratio [HR] 0·95 [95% CI 0·80-1·12]; log-rank p=0·53). In the poor prognosis subgroup, median overall survival also did not differ between the two treatment groups (11·0 months [95% CI 9·3-13·3] in the custursin group vs 10·9 months [8·2-12·4] in the control group; HR 0·97 [95% CI 0·80-1·21]; two-sided p=0·80). The most frequently reported grade 3 or worse adverse events in the custirsen versus control groups were neutropenia (70 [22%] of 315 vs 61 [20%] of 312), anaemia (68 [22%] vs 49 [16%]), fatigue (23 [7%] vs 18 [6%]), asthenia (16 [5%] vs 8 [3%]), bone pain (16 [5%] vs 5 [2%]), and febrile neutropenia (16 [5%] vs 9 [3%]). Serious adverse events were reported in 155 (49%) versus 132 (42%). 27 patients died within 30 days of treatment in the cabazitaxel and prednisone plus custirsen group, seven of which were deemed to be treatment related, versus 17 in the cabazitaxel and prednisone group, eight of which were deemed to be treatment related. Of the 21 deaths reported, 15 were reported as complications related to study treatment, either chemotherapy (eight and three, respectively) or study drug (none and four, respectively). We noted no survival benefit in men with metastatic castration-resistant prostate cancer with the addition of custirsen to cabazitaxel and prednisone treatment. Cabazitaxel and prednisone remains the standard of care for patients with metastatic castration-resistant prostate cancer progressing after docetaxel chemotherapy. OncoGenex Pharmaceuticals.