[Survival in stage T2-T3A bladder cancer treated with radical cystectomy].
The optimal treatment for patients with localized muscle-infiltrating urothelial carcinoma (Jewett stage B or T2-T3a of the TNM classification, UICC 1992) continues to be a controversy. The present study analyzed the survival rate in patients with stage T2-T3a bladder cancer who had been treated by radical cystectomy. The records of 50 patients with T2-T3a NO tumor, submitted to pelvic lymphadenectomy and radical cystoprostatectomy, were reviewed to determine the prognosis in this group of patients. Seventeen patients (34%) received three courses of systemic chemotherapy (CMV) prior to cystectomy. The overall 5-year survival rate was 73%; 76% for those with T2 (n = 30) and 67% for those with T3a (n = 20) (log-rank, p = 0.27). No statistically significant differences were observed for age (less than or over 65 years), tumor growth pattern (papillary or flat), tumor size (less or greater than 5 cms) or treatment (with or without induction CMV). However, patients with G1-2 tumor had a better survival rate (94% at 5 years) than those with G3 tumor (51%), a difference with statistical significance (log-rank, p = 0.047). The Cox regression analysis showed no independent variable of prognostic significance. Muscle-infiltrating urothelial carcinoma is highly curable by radical surgery. Some authors believe it is unnecessary to distinguish T2-T3a lesions; therefore a critical review of the TNM classification appears to be warranted. We are unable to distinguish patients with a better prognosis that might benefit from less aggressive therapeutic options. Similarly, the therapeutic benefits of induction chemotherapy prior to cystectomy in patients with stage T2-T3a tumor could not be demonstrated.


Real-Time Estimation of the Risk of Death from Novel Coronavirus (COVID-19) Infection: Inference Using Exported Cases.
The exported cases of 2019 novel coronavirus (COVID-19) infection that were confirmed outside China provide an opportunity to estimate the cumulative incidence and confirmed case fatality risk (cCFR) in mainland China. Knowledge of the cCFR is critical to characterize the severity and understand the pandemic potential of COVID-19 in the early stage of the epidemic. Using the exponential growth rate of the incidence, the present study statistically estimated the cCFR and the basic reproduction number-the average number of secondary cases generated by a single primary case in a naïve population. We modeled epidemic growth either from a single index case with illness onset on 8 December, 2019 (Scenario 1), or using the growth rate fitted along with the other parameters (Scenario 2) based on data from 20 exported cases reported by 24 January 2020. The cumulative incidence in China by 24 January was estimated at 6924 cases (95% confidence interval [CI]: 4885, 9211) and 19,289 cases (95% CI: 10,901, 30,158), respectively. The latest estimated values of the cCFR were 5.3% (95% CI: 3.5%, 7.5%) for Scenario 1 and 8.4% (95% CI: 5.3%, 12.3%) for Scenario 2. The basic reproduction number was estimated to be 2.1 (95% CI: 2.0, 2.2) and 3.2 (95% CI: 2.7, 3.7) for Scenarios 1 and 2, respectively. Based on these results, we argued that the current COVID-19 epidemic has a substantial potential for causing a pandemic. The proposed approach provides insights in early risk assessment using publicly available data.


Incremental direct expenditure of treating asthma in the United States.
There is a wide range in the estimates of cost of asthma that are available in the literature. Given the growing prevalence of asthma and its associated healthcare resource use in the United States (U.S.), it is important to obtain current and precise cost estimates attributable to asthma treatment. The objectives of this study were to estimate the incremental direct expenditures associated with asthma in the U.S. Retrospective analysis was conducted using the 2004 Medical Expenditure Panel Survey (MEPS) data that are representative of the civilian non-institutionalized population of the U.S. Asthma respondents were identified as those with International Classification of Diseases-9-Clinical Modification (ICD-9-CM) diagnosis codes for asthma in 2004 or those who had a self-report of having asthma in 2004. Incremental total expenditures and expenditures for various categories of resource use including physician office visits, emergency room visits, outpatient visits, inpatient visits, medications, and other medical visits associated with asthma were estimated separately in children (age < 18 years) and in adults (age > or = 18 years) using generalized linear regression models. The models were adjusted for covariates including age, gender, race, ethnicity, education, marital status (for age group > or = 18 years), geographic region, insurance status, and comorbidities. The prevalence of asthma among children and adults in 2004 was estimated at 8.7% (6.4 million persons) and 6.72% (14.8 million persons), respectively. The annual adjusted mean incremental total expenditure associated with asthma was $1,004.6 (SE: $326.1; p = 0.002) per person among children and was $2,077.5 (SE: $544.5; p < 0.0001) per person among adults, after adjusting for covariates. Prescription medications and physician office visits were the major drivers of total expenditures and constituted approximately 38% and 49% of the total incremental expenditures for asthma in children and adults, respectively. Inpatient visit expenditures were high in both age groups but were not significantly different from zero. Given the prevalence of asthma among U.S. children and adults and its associated incremental expenditures, the annual direct medical expenditure attributable to asthma treatment is estimated at approximately $37.2 billion in 2007 U.S. dollars representing a significant portion of healthcare resource use in the U.S.


Crystal structures of phosphodiesterases and implications on substrate specificity and inhibitor selectivity.
Crystal structures of seven phosphodiesterase families (PDE1-5, 7, 9) show a conserved core catalytic domain that contains about 300 amino acids and fourteen alpha-helices. The catalytic domains of the PDE families 1-4, 7, and 9 have a uniform conformation. However, the H-loop at the active site of PDE5 shows four different conformations upon binding of inhibitors, probably implying a special mechanism for recognition of substrates and inhibitors by PDE5. The active site of all PDE families contains two divalent metal ions: zinc and probably magnesium. The PDE4-AMP and PDE5-GMP structures reveal the conserved interactions of the phosphate groups of the products AMP and GMP, and thus suggest a universal mechanism of nucleophilic attack for all PDE families. The substrate specificity has not been well understood. This review will comment on the early proposal, "glutamine switch", on basis of the recent biochemical and structural information. The PDE-inhibitor structures have identified a common subpocket for non-selective binding of all inhibitors and potential elements for recognition of family-selective inhibitors. The kinetic analysis on the mutations of PDE7 to PDE4 suggests that the multiple elements must work together to define inhibitor selectivity.


Charge-based particle separation in microfluidic devices using combined hydrodynamic and electrokinetic effects.
A new microfluidic approach for charge-based particle separation using combined hydrodynamic and electrokinetic effects is presented. A recirculating flow pattern is employed, generated through application of bi-directional flow in a narrow glass microchannel incorporating diverging or converging segments at both ends. The bi-directional flow in turn is a result of opposing pressure-driven flow and electro-osmotic flow in the device. Trapping and preconcentration of charged particles is observed in the recirculating flow, under conditions where the average net velocity of the particles themselves approaches zero. This phenomenon is termed flow-induced electrokinetic trapping (FIET). Importantly, the electrophoretic mobility (zeta potential) of the particles determines the flow conditions required for trapping. In this paper, we exploit FIET for the first time to perform particle separations. Using a non-uniform channel, one type of particle can be trapped according to its zeta-potential, while particles with higher or lower zeta-potentials are flushed away with the pressure-driven or electro-osmotic components, respectively, of the flow. This was demonstrated using simple mixtures of two polystyrene bead types having approximately the same size (3 microm) but different zeta potentials (differences were in the order of 25 to 40 mV). To gain more insight into the separation mechanism, particle separations in straight, 3 cm-long microchannels with uniform cross-section were also studied under conditions of bi-directional flow without trapping. A thorough theoretical analysis confirmed that trapping occurs when electrokinetic and pressure-driven particle velocities are equal and opposite throughout the diverging segment. This makes it possible to predict the pressure and electric field conditions required to separate particles having defined zeta potentials.


Genome-wide association study identifies NRG1 as a susceptibility locus for Hirschsprung's disease.
Hirschsprung's disease (HSCR), or aganglionic megacolon, is a congenital disorder characterized by the absence of enteric ganglia in variable portions of the distal intestine. RET is a well-established susceptibility locus, although existing evidence strongly suggests additional loci contributing to sporadic HSCR. To identify these additional genetic loci, we carried out a genome-wide association study using the Affymetrix 500K marker set. We successfully genotyped 293,836 SNPs in 181 Chinese subjects with sporadic HSCR and 346 ethnically matched control subjects. The SNPs most associated with HSCR were genotyped in an independent set of 190 HSCR and 510 control subjects. Aside from SNPs in RET, the strongest overall associations in plausible candidate genes were found for 2 SNPs located in intron 1 of the neuregulin1 gene (NRG1) on 8p12, with rs16879552 and rs7835688 yielding odds ratios of 1.68 [CI(95%):(1.40, 2.00), P = 1.80 x 10(-8)] and 1.98 [CI(95%):(1.59, 2.47), P = 1.12 x 10(-9)], respectively, for the heterozygous risk genotypes under an additive model. There was also a significant interaction between RET and NRG1 (P = 0.0095), increasing the odds ratio 2.3-fold to 19.53 for the RET rs2435357 risk genotype (TT) in the presence of the NRG1 rs7835688 heterozygote, indicating that NRG1 is a modifier of HSRC penetrance. Our highly significant association findings are backed-up by the important role of NRG1 as regulator of the development of the enteric ganglia precursors. The identification of NRG1 as an additional HSCR susceptibility locus not only opens unique fields of investigation into the mechanisms underlying the HSCR pathology, but also the mechanisms by which a discrete number of loci interact with each other to cause disease.


Genome-wide association analysis in dogs implicates 99 loci as risk variants for anterior cruciate ligament rupture.
Anterior cruciate ligament (ACL) rupture is a common condition that can be devastating and life changing, particularly in young adults. A non-contact mechanism is typical. Second ACL ruptures through rupture of the contralateral ACL or rupture of a graft repair is also common. Risk of rupture is increased in females. ACL rupture is also common in dogs. Disease prevalence exceeds 5% in several dog breeds, ~100 fold higher than human beings. We provide insight into the genetic etiology of ACL rupture by genome-wide association study (GWAS) in a high-risk breed using 98 case and 139 control Labrador Retrievers. We identified 129 single nucleotide polymorphisms (SNPs) within 99 risk loci. Associated loci (P<5E-04) explained approximately half of phenotypic variance in the ACL rupture trait. Two of these loci were located in uncharacterized or non-coding regions of the genome. A chromosome 24 locus containing nine genes with diverse functions met genome-wide significance (P = 3.63E-0.6). GWAS pathways were enriched for c-type lectins, a gene set that includes aggrecan, a gene set encoding antimicrobial proteins, and a gene set encoding membrane transport proteins with a variety of physiological functions. Genotypic risk estimated for each dog based on the risk contributed by each GWAS locus showed clear separation of ACL rupture cases and controls. Power analysis of the GWAS data set estimated that ~172 loci explain the genetic contribution to ACL rupture in the Labrador Retriever. Heritability was estimated at 0.48. We conclude ACL rupture is a moderately heritable highly polygenic complex trait. Our results implicate c-type lectin pathways in ACL homeostasis.


The effect of icotinib or apatinib on the pharmacokinetic profile of oxycodone in rats and the underlying mechanism.
This study aimed to investigate the interactions between icotinib/apatinib and oxycodone in rats and to unveil the underlying mechanism. An ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed and validated to determine oxycodone and its demethylated metabolite simultaneously. In vivo, Sprague-Dawley (SD) male rats were administered oxycodone with or without icotinib or apatinib. Blood samples were collected and subjected to UPLC-MS/MS analysis. An enzyme incubation assay was performed to investigate the mechanism of drug-drug interaction using both rat and human liver microsomes (RLM and HLM). The results showed that icotinib markedly increased the AUC(0-t) and AUC(0-∞) of oxycodone but decreased the CLz/F. The Cmax of oxycodone increased significantly upon co-administration of apatinib. In vitro, the Km value of oxycodone metabolism was 101.7 ± 5.40 μM and 529.6 ± 19.60 μM in RLMs and HLMs, respectively. Icotinib and apatinib inhibited the disposition of oxycodone, with a mixed mechanism in RLM (IC50 = 3.29 ± 0.090 μM and 0.95 ± 0.88 μM, respectively) and a competitive and mixed mechanism in HLM (IC50 = 22.34 ± 0.81 μM and 0.48 ± 0.05 μM, respectively). In conclusion, both icotinib and apatinib inhibit the metabolism of oxycodone in vitro and in vivo. Therefore, the dose of oxycodone should be reconsidered when co-administered with icotinib or apatinib.


Educational resources for vascular laboratory education in vascular surgery residencies and fellowships: Survey of Vascular Surgery Program Directors.
The Registered Physician in Vascular Interpretation (RPVI) credential is a prerequisite for certification by the Vascular Surgery Board of the American Board of Surgery. Of concern, as more current trainees and recent program graduates take the Physician Vascular Interpretation (PVI) examination, vascular surgery trainee pass rates have decreased. Residents and fellows have a lower PVI examination pass rates than practicing vascular surgeons. The purpose of this study was to assess current vascular laboratory (VL) training for vascular surgery residents and fellows and to identify gaps that residency and fellowship programs might address. Program directors (PDs) of Accreditation Council for Graduate Medical Education-accredited vascular surgery programs (107 fellowships, 53 integrated residency programs) were surveyed using a web-based tool. Responses were submitted anonymously. Data collected included information about the program, the PD, accreditation status of the VL, and the curriculum used to meet the PVI prerequisites. Concurrent data (June 2017) on the credentials of all PDs were obtained from the Alliance for Physician Certification and Advancement (APCA). Sixty-one of 117 PDs participated in the survey (52% response rate). Of these, 44 individuals (72% of responders) reported they held the RPVI and/or Registered Vascular Technologist credential. Records from APCA indicated that 51 of 117 PDs of accredited vascular surgery residencies and fellowships (44%) had an RPVI/Registered Vascular Technologist credential. Ninety-four percent reported that their VL was accredited. Practical VL experience for trainees was reported to be 20 hours or less by 62% of respondents. The use of a structured curriculum for practical experience was reported by only 15 programs. Programs with fellowships established for more than 10 years were more likely to have a structured program for didactic instruction (P = .03). Only 23 programs reported a dedicated VL rotation. Didactic instruction provided was 20 hours or less for 75% of the cohort. In the absence of a standardized VL curriculum, there is variation in the VL instruction provided to trainees. Fellowship programs with longer histories have more structured instruction, but time allocated to VL education is substantially less than the 30 hours of didactic and 40 hours of practical experience recommended by the APCA. Programs and learners may benefit from the development of VL training guidelines and curriculum resources.


Ultraviolet A-induced oxidation in cornea: Characterization of the early oxidation-related events.
Exposure to sunlight ultraviolet-A (UVA), the main component of solar UV reaching the eyes, is suspected to play an important part in the onset of ocular pathologies. UVA primary biological deleterious effects arise from the photo-induction of oxidative stress in cells. However, the molecular bases linking UVA-induced oxidation to UVA toxicity in eyes remain poorly understood, especially with regards to the cornea. To shed some light on this issue, we have investigated the susceptibility and response potential of the different corneal cellular layers (epithelium, stroma and endothelium) to UVA-induced oxidation. We have monitored UVA-induced immediate effects on cellular redox balance, on mitochondrial membrane potential, on 8-Hydroxy-2'-deoxyguanosine (8-OHdG) accumulation in cellular DNA and on S-glutathionylated proteins (PSSG) levels along whole rabbit corneas. Higher redox imbalance was observed in the posterior part of the cornea following irradiation. Conversely, UVA-altered mitochondrial membrane potentials were observed only in anterior portions of the cornea. UVA-induced 8-OHdG were found in nuclear DNA of epithelia, while they were found in both nuclear and mitochondrial DNA in stromal and endothelial cells. Finally, significantly higher levels of cytosolic PSSG were measured in epithelia and endothelia immediately after UVA exposure, but not in stromas. Taken together, our findings indicate that while corneal epithelial cells are subjected to important modifications in response to UVA exposure, they efficiently limit the early manifestations of UVA-induced toxicity. On the other hand, the corneal endothelium is more susceptible to UVA-induced oxidation-related toxicity.


Protective effects of SSRI, Citalopram in mutant APP and mutant Tau expressed dorsal raphe neurons in Alzheimer's disease.
Depression is among the most common neuropsychiatric comorbidities in Alzheimer's disease (AD) and other Tauopathies. Apart from its anti-depressive and anxiolytic effects, selective serotonin reuptake inhibitor (SSRI) treatment also offers intracellular modifications that may help to improve neurogenesis, reduce amyloid burden & Tau pathologies, and neuroinflammation in AD. Despite its multifaceted impact in the brain, the exact physiological and molecular mechanism by which SSRIs such as Citalopram improve neurogenesis and synaptogenesis in dementia is poorly understood. In the current study, we investigated the protective role of SSRI, Citalopram, in serotonergic, medullary raphe neurons (RN46A-B14). RN46A-B14 cells were transfected with wild-type and mutant APP and Tau cDNAs for 24 h and then treated with 20 μM Cit for 24 h. We then assessed mRNA and protein levels of pTau, total Tau, serotonin related proteins such as TPH2, SERT, and 5HTR1a, synaptic proteins and the cytoskeletal structure. We also assessed cell survival, mitochondrial respiration and mitochondrial morphology. The mutant APP and Tau transfected cells showed increased levels of serotonin related proteins and mRNA, while the mRNA and protein levels of synaptic proteins were downregulated. Citalopram treatment significantly reduced pathologically pTau level along with the serotonin related protein levels. On the other hand, there was a significant increase in the mRNA and protein levels of synaptic genes and cytoskeletal structure in the treated groups. Further, Citalopram also improved cell survival, mitochondrial respiration and mitochondrial morphology in the treated cells that express mAPP and mTau. Taken together these findings suggest Citalopram could not only be a promising therapeutic drug for treating patients with depression, but also for AD patients.


Role of lipopolysaccharide in regulating colony-stimulating factor-dependent macrophage proliferation in vitro.
Bacterial lipopolysaccharides (LPS) enhance both production of colony-stimulating factors (CSF) and proliferation of mononuclear phagocytes in vivo. The present study was undertaken to determine whether the effects of LPS on CSF-dependent monopoiesis are due solely to enhanced production of CSF or also to direct effects of LPS on the responding progenitor cell. Addition of LPS to CSF-stimulated macrophage populations had different effects, depending upon the concentration of CSF in the cultures. In the presence of optimal to supraoptimal concentrations of CSF, LPS at doses >/=0.01 mug/ml inhibited macrophage colony formation. This inhibitory activity was not due to cytotoxicity of the LPS and was not mediated through prostaglandin synthesis. In the presence of suboptimal concentrations of CSF, minute concentrations of LPS (10(-7) mug/ml) significantly enhanced macrophage colony formation. Both effects of LPS (inhibition and enhancement) appeared to be properties of lipid A since neither effect was noted with cells from LPS-resistant C3H/HeJ mice, whereas both effects could be neutralized by the addition of the antibiotic polymyxin B, which binds to the lipid A portion of LPS. These results suggest that the effects of LPS on monopoiesis in vivo may not be due solely to its capacity to stimulate production of CSF. Rather, LPS may be involved in stimulating monopoiesis both indirectly through stimulation of CSF production and by its effects on the CSF-responsive progenitor cell.


Failure to Recover from Proactive Semantic Interference and Abnormal Limbic Connectivity in Asymptomatic, Middle-Aged Offspring of Patients with Late-Onset Alzheimer's Disease.
We have obtained previous evidence of limbic dysfunction in middle-aged, asymptomatic offspring of late-onset Alzheimer's disease (LOAD) patients, and failure to recover from proactive semantic interference has been shown to be a sensitive cognitive test in other groups at risk for LOAD. To assess the effects of specific proactive semantic interference deficits as they relate to functional magnetic resonance imaging (fMRI) neocortical and limbic functional connectivity in middle aged offspring of individuals with LOAD (O-LOAD) and age-equivalent controls. We examined 21 O-LOAD and 20 controls without family history of neurodegenerative disorders (CS) on traditional measures of cognitive functioning and the LASSI-L, a novel semantic interference test uniquely sensitive to the failure to recover from proactive interference (frPSI). Cognitive tests then were correlated to fMRI connectivity of seeds located in entorhinal cortex and anterodorsal thalamic nuclei among O-LOAD and CS participants. Relative to CS, O-LOAD participants evidenced lower connectivity between entorhinal cortex and orbitofrontal, anterior cingulate, and anterior temporal cortex. In the offspring of LOAD patients, LASSI-L measures of frPSI were inversely associated with connectivity between anterodorsal thalamus and contralateral posterior cingulate. Intrusions on the task related to frPSI were inversely correlated with a widespread connectivity network involving hippocampal, insular, posterior cingulate, and dorsolateral prefrontal cortices, along with precunei and anterior thalamus in this group. Different patterns of connectivity associated with frPSI were observed among controls. The present results suggest that both semantic interference deficits and connectivity abnormalities might reflect limbic circuit dysfunction as a very early clinical signature of LOAD pathology, as previously demonstrated for other limbic phenotypes, such as sleep and circadian alterations.


Occurrence of extended spectrum beta-lactamases producing alpha hemolytic Escherichia coli in neonatal diarrhea.
E. coli, often highly pathogenic in neonatal and immuno-compromised patients, are usually supposed susceptible to a variety of chemotherapeutic agents, however with the time and also due to the extensive use of chemotherapeutic agents irrespective of their susceptibility, have evolved drug resistant strains. Moreover, the production of extended spectrum beta-lactamases producing enteric pathogens is a serious issue. In this context the present study was conducted to find out occurrence of extended spectrum Beta-lactamases producing alpha hemolytic Escherichia coli in neonates, of less than one month of age, suffering from diarrhea. Fecal samples were collected from various private hospitals in Mathura and Agra districts of Uttar Pradesh, India. With the help of hospital nursing staffs sterilized stool samples were collected and processed for isolation of E. coli. The double disk diffusion method was applied to assess the ESBL production. E. coli organisms were isolated from 39 kids out of 120 samples. The assessment of isolates revealed alpha hemolytic nature of 23 isolates on 5% sheep blood agar. As usual when drug sensitivity was performed that revealed their multi drug resistance pattern which on further examination with double disk method showed 17 of them to be extended spectrum beta-lactamases producing E. coli. The presence of enterohemorrhagic extended spectrum beta-lactamases producing Escherichia coli in kids is a matter of concern and public health importance as it may be due to the transmission from hospital itself during the birth time or post birth admission period.


Mucins secreted by a transformed cell line derived from human tracheal gland cells.
High-molecular-mass glycoconjugates are secreted by the continuous cell line MM-39, which has been obtained from cultured human tracheal gland cells transformed by simian virus 40. They were purified on Sepharose(R) CL-4B and then by two steps of density-gradient centrifugation. High-molecular-mass glycoproteins resistant to digestion by hyaluronidase, chondroitin ABC lyase and heparitinase were obtained, in addition to hyaluronic acid and proteoglycans. They were susceptible to beta-elimination. They contained polylactosaminoglycan chains as well as carbohydrate chains with a terminal sialic acid in the NeuAc alpha2-3 sequence. Most of them have a buoyant density of 1.45 g/ml in CsCl-density-gradient centrifugation, except for MUC1. The MM-39 cells were also characterized by a high expression of MUC1 and MUC4 genes, but they did not express MUC2, MUC3, MUC5B and MUC5AC. Therefore the MM-39 cells synthesized mucin-like glycoproteins as well as lysozyme and mucous proteinase inhibitor [Merten, Kammouni, Renaud, Birg, Mattéi and Figarella (1996) Am. J. Respir. Cell. Mol. Biol. 15, 520-528]; they should be considered as having a mixed, both serous and mucous, phenotype.


Circulating levels of tumor necrosis factor-alpha receptor 2 are increased in heart failure with preserved ejection fraction relative to heart failure with reduced ejection fraction: evidence for a divergence in pathophysiology.
Various pathways have been implicated in the pathogenesis of heart failure (HF) with preserved ejection fraction (HFPEF). Inflammation in response to comorbid conditions, such as hypertension and diabetes, may play a proportionally larger role in HFPEF as compared to HF with reduced ejection fraction (HFREF). This study investigated inflammation mediated by the tumor necrosis factor-alpha (TNFα) axis in community-based cohorts of HFPEF patients (n = 100), HFREF patients (n = 100) and healthy controls (n = 50). Enzyme-linked immunosorbent assays were used to investigate levels of TNFα, its two receptors (TNFR1 and TNFR2), and a non-TNFα cytokine, interleukin-6 (IL-6), in plasma derived from peripheral blood samples. Plasma levels of TNFα and TNFR1 were significantly elevated in HFPEF relative to controls, while levels of TNFR2 were significantly higher in HFPEF than both controls and HFREF. TNFα, TNFR1 and TNFR2 were each significantly associated with at least two of the following: age, estimated glomerular filtration rate, hypertension, diabetes, smoking, peripheral vascular disease or history of atrial fibrillation. TNFR2 levels were also significantly associated with increasing grade of diastolic dysfunction and severity of symptoms in HFPEF. Inflammation mediated through TNFα and its receptors, TNFR1 and TNFR2, may represent an important component of a comorbidity-induced inflammatory response that partially drives the pathophysiology of HFPEF.


Heptakis(2,6-O-dimethyl)beta-cyclodextrin: a novel growth stimulant for Bordetella pertussis phase I.
The effect of cyclodextrins on the growth of Bordetella pertussis Tohama phase I in synthetic medium was evaluated. The addition of cyclodextrins, especially heptakis(2,6-O-dimethyl)beta-cyclodextrin (Me beta CD), to a complete synthetic medium such as Stainer-Scholte medium gave the same number of individual colonies and growth rates as those on Bordet-Gengou medium. Furthermore, with the addition of Me beta CD, growth inhibition by fatty acids such as oleic or palmitic acid was overcome and normal cell growth was observed. This modified Stainer-Scholte medium, designated as cyclodextrin solid medium (CSM), supported excellent growth of 20 lyophilized clinical isolates. Serotypes of the organisms after 10 passages on this CSM plate were not changed. These results suggest that Me beta CD is a significant growth stimulant and CSM is one of the most suitable synthetic media for culture of B. pertussis phase I.


In vitro and in vivo antitumor efficacy of docetaxel and sorafenib combination in human pancreatic cancer cells.
The response of pancreatic cancer to treatments remains unsatisfactory, highlighting the need for more effective therapeutic regimens. Sorafenib, an orally available multikinase inhibitor, is active against different tumors, including pancreatic cancer. We studied the antitumor efficacy of sorafenib in combination with different antitumor drugs currently used in clinical practice in in vitro and in vivo experimental models of human pancreatic cancer. The cytotoxic effect of sorafenib and conventional antitumor drug combinations was evaluated in vitro in human pancreatic cancer cell lines and the efficacy of the most active combination was tested on tumor-bearing mice. Flow cytometric, Western blot and immunohistochemistry analyses were performed to investigate the mechanisms involved in the activity of single drugs and in their interaction when used in combination. Sorafenib showed a strong sequence-dependent synergistic interaction in vitro with docetaxel, which was highly dependent on the drug sequence employed. In vivo, human pancreatic cancer-xenografted mice treated with docetaxel followed by sorafenib reduced and delayed tumor growth, with complete tumor regression observed in half of the mice. This marked antitumor effect resulted in an overall increase in mouse survival of about 70% and in a complete cure in 3 of the 8 treated mice. The strong activity was also accompanied by marked apoptosis induction, inhibition of tumor angiogenesis and downregulation of ERK signalling. Our results show that the docetaxel and sorafenib combination exerts high therapeutic efficacy in experimental models of human pancreatic cancer, indicating a promising antitumor strategy for clinical use.


Impact of childhood life events and childhood trauma on the onset and recurrence of depressive and anxiety disorders.
To investigate the effect of childhood life events and childhood trauma on the onset and recurrence of depressive and/or anxiety disorders over a 2-year period in participants without current psychopathology at baseline. Longitudinal data in a large sample of participants without baseline DSM-IV depressive or anxiety disorders (n = 1,167, aged 18 to 65 years; assessed between 2004-2007) were collected in the Netherlands Study of Depression and Anxiety (NESDA). Childhood life events and childhood trauma were assessed at baseline with a semistructured interview. The Composite International Diagnostic Interview, based on DSM-IV criteria, was used to diagnose first onset or recurrent depressive and/or anxiety disorders over a 2-year period. At baseline, 172 participants (14.7%) reported at least 1 childhood life event, and 412 (35.3%) reported any childhood trauma. During 2 years of follow-up, 226 participants (19.4%) developed a new (n = 58) or recurrent (n = 168) episode of a depressive and/or anxiety disorder. Childhood life events did not predict the onset and recurrence of depressive or anxiety disorders. Emotional neglect and psychological, physical, and sexual abuse were all associated with an increased risk of first onset and recurrence of either depressive or comorbid disorders (P < .001), but not of anxiety disorders. In multivariate models, emotional neglect was the only significant independent predictor of first onset and recurrence of any depressive or comorbid disorder (P = .002). These effects were primarily mediated by the severity of (subclinical) depressive symptoms at baseline and, to a lesser extent, by a prior lifetime diagnosis of a depressive and/or anxiety disorder. Childhood maltreatment is a key environmental risk factor, inducing vulnerability to develop new and recurrent depressive and comorbid anxiety and depressive episodes.


Protein expression and purification of G-protein coupled receptor kinase 6 (GRK6), toward structure-based drug design and discovery for multiple myeloma.
Human G-protein coupled receptor kinase 6 (GRK6) belongs to the GRK4 kinase subfamily of the G protein-coupled receptor kinase family which comprises of GRK1, GRK2, and GRK4. These kinases phosphorylate ligand-activated G-protein coupled receptors (GPCRs), driving heterotrimeric G protein coupling, desensitization of GPCR, and β-arrestin recruitment. This reaction series mediates cellular signal pathways for cell survival, proliferation, migration and chemotaxis. GRK6 is a kinase target in multiple myeloma since it is highly expressed in myeloma cells compared to epithelial cells and has a significant role in mediating the chemotactic responses of T and B-lymphocytes. To support structure-based drug design, we describe three human GRK6 constructs, GRK6, GRK6His/EK, and GRK6His/TEV, designed for protein expression in Spodoptera frugiperda Sf9 insect cells. The first construct did not contain any purification tag whereas the other two constructs contained the His10 affinity tag, which increased purification yields. We report here that all three constructs of GRK6 were overexpressed in Sf9 insect cells and purified to homogeneity at levels that were suitable for co-crystallization of GRK6 with potential inhibitors. The yields of purified GRK6, GRK6His/EK, and GRK6His/TEV were 0.3 mg, 0.8 mg and 0.7 mg per liter of cell culture, respectively. In addition, we have shown that GRK6His/TEV with the His10 tag removed was highly homogeneous and monodisperse as observed by dynamic light scattering measurement and actively folded as exhibited by circular dichroism spectroscopy. The described methods will support the structure-based development of additional therapeutics against multiple myeloma.


Lesion-symptom mapping of language impairments in patients suffering from left perisylvian gliomas.
Brain tumors cause local structural impairments of the cerebral network. Moreover, brain tumors can also affect functional brain networks more distant from the lesion. In this study, we analyzed the impact of glioma WHO grade II-IV tumors on grey and white matter in relation to impaired language function. In a retrospective analysis of 60 patients, 14 aphasic and 46 non-aphasic, voxel-based lesion-symptom mapping (VLSM) was used to identify tumor induced lesions in grey (GM) and white matter (WM) related to patients' performance in subtests of the Aachen Aphasia Test (AAT). Significant clusters were analyzed for atlas-based grey and white matter involvements in relation to different linguistic modalities. VLSM analysis indicated significant contribution of a posterior perisylvian cluster covering WM and GM to AAT performance averaged across subtests. When considering individual AAT subtests, a substantial overlap between significant clusters for analysis of the token test, picture naming and language comprehension results could be observed. The WM-cluster intersections reflect the overall importance of the perisylvian area in language function, similarly to GM participations. Especially the constant high percentages of Heschl's gyrus, superior temporal gyrus, inferior longitudinal and middle longitudinal fascicles, but also arcuate and inferior fronto-occipital fascicles highlight the importance of the posterior perisylvian area for language function.


Health content analysis of organ donation and transplantation news on Turkish television channels and in Turkish print media.
The media affects individuals' behaviors, especially by means of news and advertisements. In this study, we evaluated health content of organ donation and transplantation news in the printed media and on television programs for a 1-year period in Turkey. We examined 2449 news items in 230 newspapers and magazines; 1179 news programs on 45 television channels, all concerning organ donation and transplantation. The news obtained from the Media Pursuit Center were transferred to an electronic file to evaluate the format and content of the news. Nine variables were examined about the scope and the formal characteristics of the news: the publication name, its type, the province, the date, the headline, the title length, the presence of a photograph, or its kind, the news size, and the page number. In the content analysis of the news, we also examined 9 variables: the topic, the message of the headline, the property of the words in the title, the identification of photographs in the news, the age, gender of actors in the news, as well as donor or recipient. In a summary, print media and television channels, failed to show sufficient information about organ donation and transplantation. The percentage of news about organ donation and transplantation was small and mostly negative items in the media. On television channels, sufficient place was not given to organ donation and transplantation. The news in printed media and on television channels was not about motivated or altruistic behavior. The pattern of organ donation and transplantation news is important in terms of perception and comment by the public. Furthermore it directly affects the perception of the news by the reader.


Montage electron tomography of vitrified specimens.
Cryo-electron tomography provides detailed views of macromolecules in situ. However, imaging a large field of view to provide more cellular context requires reducing magnification during data collection, which in turn restricts the resolution. To circumvent this trade-off between field of view and resolution, we have developed a montage data collection scheme that uniformly distributes the dose throughout the specimen. In this approach, sets of slightly overlapping circular tiles are collected at high magnification and stitched to form a composite projection image at each tilt angle. These montage tilt-series are then reconstructed into massive tomograms with a small pixel size but a large field of view. For proof-of-principle, we applied this method to the thin edge of HeLa cells. Thon rings to better than 10 Å were detected in the montaged tilt-series, and diverse cellular features were observed in the resulting tomograms. These results indicate that the additional dose required by this technique is not prohibitive to performing structural analysis to intermediate resolution across a large field of view. We anticipate that montage tomography will prove particularly useful for lamellae, increase the likelihood of imaging rare cellular events, and facilitate visual proteomics.


Improving the de-agglomeration and dissolution of a poorly water soluble drug by decreasing the agglomerate strength of the cohesive powder.
Influence of ternary, poorly water-soluble components on the agglomerate strength of cohesive indomethacin mixtures during dissolution was studied to explore the relationship between agglomerate strength and extent of de-agglomeration and dissolution of indomethacin (Ind). Dissolution profiles of Ind from 20% Ind-lactose binary mixtures, and ternary mixtures containing additional dibasic calcium phosphate (1% or 10%; DCP), calcium sulphate (10%) and talc (10%) were determined. Agglomerate strength distributions were estimated by Monte Carlo simulation of particle size, work of cohesion and packing fraction distributions. The agglomerate strength of Ind decreased from 1.19 MPa for the binary Ind mixture to 0.84 MPa for 1DCP:20Ind mixture and to 0.42 MPa for 1DCP:2Ind mixture. Both extent of de-agglomeration, demonstrated by the concentration of the dispersed indomethacin distribution, and extent of dispersion, demonstrated by the particle size of the dispersed indomethacin, were in descending order of 1DCP:2Ind>1DCP:20Ind>binary Ind. The addition of calcium sulphate dihydrate and talc also reduced the agglomerate strength and improved de-agglomeration and dispersion of indomethacin. While not definitively causal, the improved de-agglomeration and dispersion of a poorly water soluble drug by poorly water soluble components was related to the agglomerate strength of the cohesive matrix during dissolution.


Management of Invasive Fungal Infections in Patients Undergoing Allogeneic Hematopoietic Stem Cell Transplantation: The Turin Experience.
Allogeneic hematopoietic stem cell transplant (allo-HSCT) recipients are exposed to an increased risk of invasive fungal infections (IFIs) due to neutropenia, immunosuppressive treatments, graft-versus-host disease (GvHD) and incomplete immune reconstitution. Although clinical benefit from antifungal prophylaxis has been demonstrated, IFIs remain a leading cause of morbidity and mortality in these patients. In the last decades, attention has also been focused on potential risk factors for IFI to tailor an antifungal prevention strategy based on risk stratification. This retrospective single-center study aimed to assess the epidemiology and the prognostic factors of IFI in a large cohort of allo-HSCT patients. Between January 2004 and December 2020, 563 patients with hematological malignancies received an allo-HSCT at the Stem Cell Transplant Unit in Turin: 191 patients (34%) received grafts from a matched sibling donor, 284 (50.5%) from a matched unrelated donor, and 87 (15.5%) from an haploidentical family member. The graft source was peripheral blood in 81.5% of the patients. Our policy for antifungal prophylaxis included fluconazole in matched related and unrelated donors, while micafungin was administered in patients receiving haploidentical transplant. According to this practice, fluconazole was administered in 441 patients (79.6%) and micafungin in 62 (11.2%), while only 9 patients received mold-active prophylaxis. Galactomannan testing was routinely performed twice a week; patients with persisting fever unresponsive to broad spectrum antibiotics were evaluated with lung high-resolution computed tomography (HRCT) scan. In case of imaging suggestive of IFI, bronchoalveolar lavage (BAL) was performed whenever feasible. Only probable/proven IFI (PP-IFI) occurring during the first 12 months after transplant have been evaluated. IFIs were classified as probable or proven according to the new revised European Organization for Research and Treatment of Cancer (EORTC)/Mycoses Study Group (MSG) consensus criteria. Multivariate competing risk regression, binary logistic, and proportional hazard models were performed to identify risk factors for PP-IFI. A total of 58 PP-IFIs (n = 47 probable; n = 11 proven) occurred in our patients resulting in a cumulative incidence of 4.1%, 8.1%, and 9.6% at 30, 180, and 365 days, respectively. Molds were the predominant agents (n = 50 Aspergillus; n = 1 Mucor), followed by invasive candidemia (n = 5 non-albicans Candida; n = 1 Candida albicans; n = 1 Trichosporon). Lung was the most frequent site involved in patients with mold infections (47/51, 92.2%). Median time from HSCT to IFI was 98.44 days (0-365 days). Only 34.5% of patients with IFI were neutropenic at the time of infection. The presence of IFI had a significant impact on overall survival at 1 year (IFI, 32.8% vs. non-IFI, 54.6%; p < 0.001). IFI-related mortality rate was 20.7% in the overall population, 17% in patients with probable IFI, and 36% in patients with proven IFI. Multivariate competing risk regression revealed that donor type was the factor significantly associated to the risk of IFI [subdistribution hazard ratio (SDHR), 1.91, IC 1.13-3.20; p = 0.015]. BAL was informative in a consistent number of cases (36/57, 63.2%) leading to the identification of fungal (21), bacterial (4), viral (3), and polymicrobial (8) infections. Overall, 79 patients (14%) received a diagnostic-driven treatment, and 63 patients (11.2%) received a fever-driven treatment. Liposomal amphoteric B was the drug used in the majority of patients receiving diagnostic-driven therapy (30/79, 38%), while caspofungin was administered more frequently in patients who received a fever-driven strategy (27/63, 42.9%). According to our experience, a non-mold active prophylaxis in patients undergoing allo-HSCT is feasible when combined with an intensive diagnostic work-up including CT scan and BAL. BAL performed at the onset of the disease may provide informative results in most patients. A diagnostic-driven treatment strategy may contribute to limit the use of costly antifungal therapies.


Hand Fractures-Management and the Complications That Inevitably Occur: Metacarpal Fractures.
Metacarpal fractures are among the most common hand fractures. To properly manage these injuries, surgeons must understand the anatomy, biomechanics, clinical assessment, surgical and nonsurgical treatment options, and potential complications. Metacarpal head fractures often require surgical treatment to restore the joint surface by using a variety of techniques. Metacarpal neck fractures are usually stable injuries that do not require surgical intervention, but surgeons must know when surgical intervention is indicated. Fractures of the metacarpal shaft can be treated surgically and nonsurgically and may be associated with large bone defects or soft-tissue injuries that require careful consideration. Finally, fractures involving the carpometacarpal joints must be promptly managed to avoid long-term complications, potentially requiring salvage procedures.


[The effect of selenium on enzymes metabolizing xenobiotics in rat liver].
Activity of enzymes involved in metabolism of xenobiotics was not altered in liver tissue of rats kept on a ration enriched with selenium at a dose of 2.5 mg/kg. Both organic form of selenium (yeast meal, selenomethionine) and inorganic derivatives (sodium selenite) at a dose of 5 mg/kg of ration caused distinct activation of epoxide hydrolase, UDP-glucuronosyl transferase and glutathione transferase within 6 weeks after the experiment beginning, while content of cytochrome P-450, glutathione-SH and glutathione peroxidase activity were not significantly altered. Within 9 weeks the enzymatic activity remained at the higher rate only in rats kept on the ration with sodium selenite. Relationship between toxic effects of selenium high doses and alterations in activity of enzymes involved in metabolism of xenobiotics is discussed.


Creation of readily accessible analogue of cortistatin A as an antitumor drug lead.
Angiogenesis, the formation of new blood capillaries from preexisting blood vessels, is a vital process in growth of solid tumors and development to malignant stage. This makes angiogenesis a desirable target for cancer treatment. In this work, design and syntheses of structurally simplified analogues of cortistatin A (1), a novel antiangiogenic steroidal alkaloid from Indonesian marine sponge, and their biological activities were investigated. One of the analogues, in which the isoquinoline moiety was appended to the planar tetracyclic core structure, showed potent antiproliferative activity against human umbilical vein endothelial cells (HUVECs) together with high selectivity and also showed in vivo antiangiogenic activity and significant antitumor effect by oral administration.


[Genome instability and neoplasia].
Necessity of oncogene for the neoplastic cell transformation is substantiated. The oncogene may be introduced into the cell by a virus; be present in the cell from its formation but be repressed by products of regulatory genes; be arisen from separated DNA segments. The latter possibility is discussed in view of data on the presence of mobile elements and Sarc sequences in the cell genome.


The Serotonin Transporter Polymorphism (5-HTTLPR) and Alcohol Problems in Heavy Drinkers: Moderation by Depressive Symptoms.
Heavy alcohol use in young adults has been prospectively associated with a host of psychosocial and alcohol-related problems. Recent studies have supported the interaction between serotonin transporter polymorphism and adverse environmental factors, as a predictor of alcohol use and the development of alcohol dependence. The current study examined the role of depressive symptoms in combination with the serotonin transporter polymorphism as a predictor of alcohol use and alcohol-related problems. Results revealed a significant genotype by depressive symptom interaction, such that heavier alcohol use was associated with depressive symptoms in L allele homozygotes but not among S allele carriers. These results remained significant after controlling for ethnicity and gender effects. These findings extend the emerging literature supporting 5-HTTLPR genotype as a risk factor for alcohol-related problems in the context of co-occurring symptoms of depression.


Study protocol for CELLAR (COVID-related Eating Limitations and Latent dietary effects in the Atlantic Region): population-based observational study to monitor dietary intakes and purchasing during COVID-19 in four Atlantic Canadian provinces.
Poor diet is a leading preventable risk for the global burden of non-communicable disease. Robust measurement is needed to determine the effect of COVID-19 on dietary intakes and consumer purchasing, given the widespread changes to consumer food environments and economic precarity. The research objectives are as follows: (1) describe dietary intakes of foods, beverages and nutrients of concern during the COVID-19 pandemic; (2) quantify change in diet during COVID-19 as compared with prepandemic, previously captured in the provincial samples of the population-representative 2015 Canadian Community Health Survey-Nutrition and (3) examine how household purchasing practices predict dietary intakes during COVID-19. Observational study of diet, using a population-based stratified probability sampling strategy allocated via dual-frame (landline and cellphone) calls to random-digit dialled numbers, followed by age-sex group quotas. The base population comprises the four provinces of the Atlantic region of Canada, jurisdictions with an excess burden of pre-existing dietary risk, compared with the rest of Canada. Our aim is n=1000 to obtain reliable estimates at a regional level to describe intakes and compare with prepandemic baseline. Data collection entails 12 weeks participation: (1) enrolment with sociodemographics (key dietary risk predictors such as age, sex, gender, pre-COVID-19 income, employment, household composition, receipt of economic relief, rural residence); (2) two 24hour diet recalls using the online ASA-24 Canada 2018 tool; and (3) online uploads of household food purchase receipts over the 12 weeks enrolled. Participation incentives will be offered. This research protocol received funding from the Canadian Institutes of Health Research (FRN VR5 172691) and ethics review approval from the Dalhousie University Research Ethics Board. Study protocol and instruments and a de-identified dataset will be made publicly available. We will submit the findings to peer-reviewed journals, as well as conferences geared towards scientific and decision-maker audiences.


Evaluating the protective effects of melatonin on di(2-ethylhexyl) phthalate-induced testicular injury in adult mice.
Di (2-ethylhexyl) phthalate (DEHP) is a common phthalate derivative, interfering with normal function of reproductive system. The present study evaluated effects of melatonin on DEHP-induced testicular injury in mice. Thirty-two adult male mice were randomly divided to four groups; group I received normal saline, group II received DEHP, group III received DEHP and melatonin, and group IV was treated with melatonin alone. Body and testes weights, total antioxidant capacity (TAC), glutathione level and superoxide dismutase, glutathione peroxidase and catalase activities were measured. Serum testosterone, luteinizing hormone (LH) and follicle-stimulating hormone (FSH) levels and interleukin 1 beta (IL-1β) and tumor necrosis factor (TNF-α) concentration were evaluated by ELISA assay. Also, malondialdehyde (MDA) and nitric oxide (NO) levels, sperm characteristics and histological changes of testes were analyzed. Body and testes weights were decreased in DEHP group. DEHP also reduced the number of spermatogonia, primary spermatocyte and sertoli cells as well as sperm vitality and progressive motility; these toxic effects were associated with alterations in serum hormone levels. Melatonin remarkably inhibited DEHP-induced reduction of body weight and antioxidant capacity. Melatonin reduced DEHP-induced elevation of NO, MDA, IL-1β and TNF-α levels. Melatonin improved DEHP-induced changes in hormonal levels, number of sertoli cells, spermatogonia, and sperm viability and motility. Melatonin considerably inhibits DEHP-induced gonadotoxicity through reducing oxidative stress and inflammatory responses. These results suggest that melatonin may be considered as a promising agent to reduce toxic effects of endocrine disrupting chemicals such as DEHP on the male reproductive system.


Animal welfare and farm economy-exploring the relationship between dairy animal welfare indicators and economic efficiency.
This paper explores the associations between a set of mainly health and productivity related animal welfare indicators in dairy farming calculated based on herd recordings, and short-term economic efficiency. To explore this association, we merged a dataset with animal welfare indicators and a dataset with farm economic data from 459 Norwegian farms. Gross margin including direct payments together with milk quota, farmland and number of animals and were used to calculate economic efficiency. The analysis was performed in two steps. First, Data Envelopment Analysis (DEA) was used to rank the farms according to efficiency. Second, the machine learning method boosting was applied to analyze the associations between economic efficiency and the different welfare indicators. The findings show that for most welfare indicators, at indicator values below the national average a positive association exists between increasing indicator values and economic efficiency. For some indicators such as for example Fertility, Calves and Youngstock, this positive association continues also beyond the national average. However, for other indicators such as for example Claw and Udder health, a curvilinear relationship exists with a turning point from where the efficiency is negatively associated with increasing indicator values. Possible explanations for the associations between economic efficiency and animal welfare indicators are outlined.


Seamless Lesion Insertion for Data Augmentation in CAD Training.
The performance of a classifier is largely dependent on the size and representativeness of data used for its training. In circumstances where accumulation and/or labeling of training samples is difficult or expensive, such as medical applications, data augmentation can potentially be used to alleviate the limitations of small datasets. We have previously developed an image blending tool that allows users to modify or supplement an existing CT or mammography dataset by seamlessly inserting a lesion extracted from a source image into a target image. This tool also provides the option to apply various types of transformations to different properties of the lesion prior to its insertion into a new location. In this study, we used this tool to create synthetic samples that appear realistic in chest CT. We then augmented different size training sets with these artificial samples, and investigated the effect of the augmentation on training various classifiers for the detection of lung nodules. Our results indicate that the proposed lesion insertion method can improve classifier performance for small training datasets, and thereby help reduce the need to acquire and label actual patient data.


Antagonistic and cooperative actions of the EGFR and Dpp pathways on the iroquois genes regulate Drosophila mesothorax specification and patterning.
In Drosophila, restricted expression of the Iroquois complex (Iro-C) genes in the proximal region of the wing imaginal disc contributes to its territorial subdivision, specifying first the development of the notum versus the wing hinge, and subsequently, that of the lateral versus medial notum. Iro-C expression is under the control of the EGFR and Dpp signalling pathways. To analyze how both pathways cooperate in the regulation of Iro-C, we isolated several wing disc-specific cis-regulatory elements of the complex. One of these (IroRE(2)) integrates competing inputs of the EGFR and Dpp pathways, mediated by the transcription factors Pointed (downstream of EGFR pathway) and Pannier/U-shaped and Mothers against Dpp (Mad), in the case of Dpp. By contrast, a second element (IroRE(1)) mediates activation by both the EGFR and Dpp pathways, thus promoting expression of Iro-C in a region of elevated levels of Dpp signalling, the prospective lateral notum near the anterior-posterior compartment boundary. These results help define the molecular mechanisms of the interplay between the EGFR and Dpp pathways in the specification and patterning of the notum.


[Study of hydrolysis of aminoalcohol ethers, phenol and choline under the action of horse blood serum cholinesterase].
Hydrolysis of ethers of saturated and unsaturated alcohols and ethers, e.g. phenol and choline, under the action of horse blood serum cholinesterase, was studied. The reactivity towards enzymatic hydrolysis is decreased due to a greater length of the chain in the alcohol residue of the benzoic acid aminoethers; at nCH2 = 4 the compound is a poor substrate. An increase in nydrophobicity of the acyl residue of the ether molecule also leads to a decrease in the Vmax and Km values. In case of cholinesterase substrates, an increase in the molecule hydrophobicity results in an increase of its non-productive absorption on the active surface of the enzyme, which decreases its hydrolysis. Aminobutynol benzoates are hydrolyzed by cholinesterase more rapidly as compared to the ethers of corresponding aminobutanols and their homologs.


Effects of Brassica vegetable juice on the induction of apoptosis and aberrant crypt foci in rat colonic mucosal crypts in vivo.
Glucosinolates are sulfur-containing glycosides found in the Brassica vegetables. Their breakdown products include isothiocyanates, which are produced following exposure to the endogenous plant enzyme myrosinase. Isothiocyanates are pungent, biologically active compounds that suppress carcinogenesis in vivo, and induce apoptosis in vitro. We have shown previously that oral administration of the isolated glucosinolate sinigrin induces apoptosis, and suppresses aberrant crypt foci in the colonic mucosa of rats treated previously with 1,2-dimethylhydrazine (DMH). In this study we explored the effects of both raw and thermally processed Brussels sprout tissue on the modulation of crypt cell apoptosis and mitosis, and the frequency of aberrant crypt foci in the colon. Freeze-dried raw and microwave-cooked Brussels sprouts contained high levels of intact glucosinolates, but they were absent from freshly prepared sprout juice. Oral administration of uncooked Brussels sprouts, whether as a juice, or as a freeze-dried powder, was associated with significantly enhanced levels of apoptosis and reduced mitosis in the colonic crypts. However, this effect was confined to rats previously injected (48 h) with DMH, in which levels of apoptosis and mitosis following DNA damage were already high. There was no effect of treatment in control animals. There was also little evidence of these effects when intact glucosinolates were administered in blanched sprout tissue, which lacked active myrosinase. We conclude that glucosinolate breakdown products derived from Brassica vegetables can exert a profound effect on the balance of colorectal cell proliferation and death in an animal model of colorectal neoplasia.


Mesoscopic modelling of Cy3 and Cy5 dyes attached to DNA duplexes.
Cy3 and Cy5 dyes linked to the 5' end of a double stranded DNA molecule are known to attach to both strands in a way that is very similar to an additional base pair and has a stabilizing effect on the oligonucleotide. Here we adapt the Peyrard-Bishop mesoscopic model to incorporate cyanine dyes and use the technique of thermal equivalence to obtain the appropriate parameters from existing melting temperatures. We have found that the stacking parameters are in the same range of ordinary AT and CG base pairs, in particular Cy3-A was found to be most rigidly stacked. While the cyanines stabilize the AT hydrogen bonds quite strongly the CG bonds are mostly unaffected.


Routine magnification cerebral angiography: a practical system designed for community hospital application.
Magnification cerebral angiography is no longer to be considered a tool only of clinical investigation; its routine application is clearly advantageous. A relatively inexpensive system has been developed which can be applied in a special procedure suite where cardiac and abdominal angiography as well as other radiographic procedures must be performed.


Epibenthic predators control mobile macrofauna associated with a foundation species in a subarctic subtidal community.
Foundation species (FS) are strong facilitators providing habitat for numerous dependent organisms. The communities shaped by FS are commonly structured by interplay of facilitation and consumer control. Predators or grazers often indirectly determine community structure eliminating either FS or their principal competitors. Alternatively, they can prey on the dependent taxa directly, which is generally buffered by FS via forming complex habitats with numerous refuges. The latter case has been never investigated at high latitudes, where consumer control is widely considered weak. We manipulated the presence of common epibenthic crustacean predators to assess their effect on mobile macrofauna of the clusters developed by a FS (barnacle Balanus crenatus and its empty tests) in the White Sea shallow subtidal (65° N). While predation pressure on the FS itself here is low, the direct effects of a spider crab Hyas araneus and a shrimp Spirontocaris phippsii on the associated assemblages were unexpectedly strong. Removing the predators did not change species diversity, but tripled total abundance and altered multivariate community structure specifically increasing the numbers of amphipods, isopods (only affected by shrimp), and bivalves. Consumer control in the communities shaped by FS may not strictly follow the latitudinal predation gradient rule.


BDNF/TrkB content and interaction with gastrin-releasing peptide receptor blockade in colorectal cancer.
Neurotrophin and neuropeptide pathways are emerging targets in cancer. Here we show that brain-derived neurotrophic factor (BDNF) and its receptor, TrkB, are present in colorectal cancer and that BDNF levels are increased in tumors compared to nontumor tissue. In addition, we investigate the role of BDNF in influencing the response of colorectal cancer cells to inhibition of gastrin-releasing peptide receptors (GRPR). Fresh-frozen sporadic colorectal adenocarcinoma specimens and adjacent nonneoplastic tissue from 30 patients, as well as paraffin-embedded colorectal cancer samples from 21 patients, were used in this study. Cell proliferation and mRNA and protein levels were examined in HT-29 or SW620 cells treated with a GRPR antagonist, human recombinant BDNF (hrBDNF), a Trk antagonist K252a, or cetuximab. Expression of BDNF and TrkB was detected in tumor samples and cell lines. BDNF levels were higher in tumor samples compared to nonneoplastic tissue. BDNF expression and secretion were increased by GRPR blockade in HT-29 cells through a mechanism dependent on epidermal growth factor receptors. Treatment with hrBDNF prevented the effect of GRPR blockade on cell proliferation, whereas a Trk inhibitor reduced proliferation. BDNF and TrkB are present in colorectal cancer and might contribute to resistance to GRPR antagonists.


Estrogen-induced hypothalamic synaptic plasticity and pituitary sensitization in the control of the estrogen-induced gonadotrophin surge.
Proper gonadal function requires coordinated (feedback) interactions between the gonads, adenohypophysis, and brain: the gonads elaborate sex steroids (progestins, androgens, and estrogens) and proteins (inhibin-activin family) during gamete development. In both sexes, the brain-pituitary gonadotrophin-regulating interaction is coordinated by estradiol through its opposing actions on pituitary gonadotrophs (sensitization of the response to gonadotrophin-releasing hormone [GnRH]) versus hypothalamic neurons (inhibition of GnRH secretion). This dynamic tension between the gonadotrophs and the GnRH cells in the brain regulates the circulating gonadotrophins and is termed reciprocal/negative feedback. In females, reciprocal/negative feedback dominates approximately 90% of the ovarian cycle. In a spectacular exception, the dynamic tension is broken during the surge of circulating estrogen that marks follicle and oocyte(s) maturation. The cause is an estradiol-induced disinhibition of the GnRH neurons that releases GnRH secretion to the highly sensitized pituitary gonadotrophs that in turn release the gonadotrophin surge (the estrogen-induced gonadotrophin surge [EIGS], also known as positive feedback). Studies during the past 4 decades have shown this disinhibition to result from estrogen-induced synaptic plasticity (EISP), including a reversible approximately 50% loss in arcuate nucleus synapses. The disinhibited GnRH secretion occurs during maximal gonadotroph sensitization and results in the EIGS. Specific immunoneutralization of estradiol blocks the EISP and EIGS. The EISP is accompanied by increases in insulinlike growth factor 1, polysialylated neural cell adhesion molecule, and ezrin, 3 proteins that the authors believe are the links between estrogen-induced astroglial extension and the EISP that releases GnRH secretion at the moment of maximal sensitization of the pituitary gonadotrophs. The result is the paradoxical surge of gonadotrophins at the peak of ovarian estrogen secretion and the triggering of ovulation. This enhanced understanding of the mechanics of gonadotrophin control clarifies elements of the involved feedback loops and opens the way to a better understanding of the neurobiology of reproduction.


Microglial phenotypes in Parkinson's disease and animal models of the disease.
Over the last decade the important concept has emerged that microglia, similar to other tissue macrophages, assume different phenotypes and serve several effector functions, generating the theory that activated microglia can be organized by their pro-inflammatory or anti-inflammatory and repairing functions. Importantly, microglia exist in a heterogenous population and their phenotypes are not permanently polarized into two categories; they exist along a continuum where they acquire different profiles based on their local environment. In Parkinson's disease (PD), neuroinflammation and microglia activation are considered neuropathological hallmarks, however their precise role in relation to disease progression is not clear, yet represent a critical challenge in the search of disease-modifying strategies. This review will critically address current knowledge on the activation states of microglia as well as microglial phenotypes found in PD and in animal models of PD, focusing on the expression of surface molecules as well as pro-inflammatory and anti-inflammatory cytokine production during the disease process. While human studies have reported an elevation of both pro- or anti-inflammatory markers in the serum and CSF of PD patients, animal models have provided insights on dynamic changes of microglia phenotypes in relation to disease progression especially prior to the development of motor deficits. We also review recent evidence of malfunction at multiple steps of NFκB signaling that may have a causal interrelationship with pathological microglia activation in animal models of PD. Finally, we discuss the immune-modifying strategies that have been explored regarding mechanisms of chronic microglial activation.


In vivo production of fluorine-18 in a chicken egg tumor model of breast cancer for proton therapy range verification.
Range verification of clinical protontherapy systems via positron-emission tomography (PET) is not a mature technology, suffering from two major issues: insufficient signal from low-energy protons in the Bragg peak area and biological washout of PET emitters. The use of contrast agents including 18O, 68Zn or 63Cu, isotopes with a high cross section for low-energy protons in nuclear reactions producing PET emitters, has been proposed to enhance the PET signal in the last millimeters of the proton path. However, it remains a challenge to achieve sufficient concentrations of these isotopes in the target volume. Here we investigate the possibilities of 18O-enriched water (18-W), a potential contrast agent that could be incorporated in large proportions in live tissues by replacing regular water. We hypothesize that 18-W could also mitigate the problem of biological washout, as PET (18F) isotopes created inside live cells would remain trapped in the form of fluoride anions (F-), allowing its signal to be detected even hours after irradiation. To test our hypothesis, we designed an experiment with two main goals: first, prove that 18-W can incorporate enough 18O into a living organism to produce a detectable signal from 18F after proton irradiation, and second, determine the amount of activity that remains trapped inside the cells. The experiment was performed on a chicken embryo chorioallantoic membrane tumor model of head and neck cancer. Seven eggs with visible tumors were infused with 18-W and irradiated with 8-MeV protons (range in water: 0.74 mm), equivalent to clinical protons at the end of particle range. The activity produced after irradiation was detected and quantified in a small-animal PET-CT scanner, and further studied by placing ex-vivo tumours in a gamma radiation detector. In the acquired images, specific activity of 18F (originating from 18-W) could be detected in the tumour area of the alive chicken embryo up to 9 h after irradiation, which confirms that low-energy protons can indeed produce a detectable PET signal if a suitable contrast agent is employed. Moreover, dynamic PET studies in two of the eggs evidenced a minimal effect of biological washout, with 68% retained specific 18F activity at 8 h after irradiation. Furthermore, ex-vivo analysis of 4 irradiated tumours showed that up to 3% of oxygen atoms in the targets were replaced by 18O from infused 18-W, and evidenced an entrapment of 59% for specific activity of 18F after washing, supporting our hypothesis that F- ions remain trapped within the cells. An infusion of 18-W can incorporate 18O in animal tissues by replacing regular water inside cells, producing a PET signal when irradiated with low-energy protons that could be used for range verification in protontherapy. 18F produced inside cells remains entrapped and suffers from minimal biological washout, allowing for a sharper localization with longer PET acquisitions. Further studies must evaluate the feasibility of this technique in dosimetric conditions closer to clinical practice, in order to define potential protocols for its use in patients.


A comparison of reconstruction algorithms for breast tomosynthesis.
Three algorithms for breast tomosynthesis reconstruction were compared in this paper, including (1) a back-projection (BP) algorithm (equivalent to the shift-and-add algorithm), (2) a Feldkamp filtered back-projection (FBP) algorithm, and (3) an iterative Maximum Likelihood (ML) algorithm. Our breast tomosynthesis system acquires 11 low-dose projections over a 50 degree angular range using an a-Si (CsI:Tl) flat-panel detector. The detector was stationary during the acquisition. Quality metrics such as signal difference to noise ratio (SDNR) and artifact spread function (ASF) were used for quantitative evaluation of tomosynthesis reconstructions. The results of the quantitative evaluation were in good agreement with the results of the qualitative assessment. In patient imaging, the superimposed breast tissues observed in two-dimensional (2D) mammograms were separated in tomosynthesis reconstructions by all three algorithms. It was shown in both phantom imaging and patient imaging that the BP algorithm provided the best SDNR for low-contrast masses but the conspicuity of the feature details was limited by interplane artifacts; the FBP algorithm provided the highest edge sharpness for microcalcifications but the quality of masses was poor; the information of both the masses and the microcalcifications were well restored with balanced quality by the ML algorithm, superior to the results from the other two algorithms.


Towards a transparent, flexible, scalable and disposable image sensor using thin-film luminescent concentrators.
Most image sensors are planar, opaque, and inflexible. We present a novel image sensor that is based on a luminescent concentrator (LC) film which absorbs light from a specific portion of the spectrum. The absorbed light is re-emitted at a lower frequency and transported to the edges of the LC by total internal reflection. The light transport is measured at the border of the film by line scan cameras. With these measurements, images that are focused onto the LC surface can be reconstructed. Thus, our image sensor is fully transparent, flexible, scalable and, due to its low cost, potentially disposable.


Quantification of the Animal Tuberculosis Multi-Host Community Offers Insights for Control.
Animal tuberculosis (TB) is a multi-host zoonotic disease whose prevalence in cattle herds in Europe has been increasing, despite a huge investment in eradication. The composition of the host community is a fundamental driver of pathogen transmission, and yet this has not been formally quantified for animal TB in Europe. We quantified multi-host communities of animal TB, using stochastic models to estimate the number of infected domestic and wild hosts in three regions: officially TB-free Central-Western Europe, and two largely TB-endemic regions, the Iberian Peninsula and Britain and Ireland. We show that the estimated number of infected animals in the three regions was 290,059-1,605,612 and the numbers of infected non-bovine domestic and wild hosts always exceeded those of infected cattle, with ratios ranging from 3.3 (1.3-19.6):1 in Britain and Ireland to 84.3 (20.5-864):1 in the Iberian Peninsula. Our results illustrate for the first time the extent to which animal TB systems in some regions of Europe are dominated by non-bovine domestic and wild species. These findings highlight the need to adapt current strategies for effective future control of the disease.


Beyond repeated-measures analysis of variance: advanced statistical methods for the analysis of longitudinal data in anesthesia research.
Research in the field of anesthesiology relies heavily on longitudinal designs for answering questions about long-term efficacy and safety of various anesthetic and pain regimens. Yet, anesthesiology research is lagging in the use of advanced statistical methods for analyzing longitudinal data. The goal of this article was to increase awareness of the advantages of modern statistical methods and promote their use in anesthesia research. Here we introduce 2 modern and advanced statistical methods for analyzing longitudinal data: the generalized estimating equations (GEE) and mixed-effects models (MEM). These methods were compared with the conventional repeated-measures analysis of variance (RM-ANOVA) through a clinical example with 2 types of end points (continuous and binary). In addition, we compared GEE and MEM to RM-ANOVA through a simulation study with varying sample sizes, varying number of repeated measures, and scenarios with and without missing data. In the clinical study, the 3 methods are found to be similar in terms of statistical estimation, whereas the parameter interpretations are somewhat different. The simulation study shows that the methods of GEE and MEM are more efficient in that they are able to achieve higher power with smaller sample size or lower number of repeated measurements in both complete and missing data scenarios. Based on their advantages over RM-ANOVA, GEE and MEM should be strongly considered for the analysis of longitudinal data. In particular, GEE should be used to explore overall average effects, and MEM should be used when subject-specific effects (in addition to overall average effects) are of primary interest.


The effect of habituating vestibular and optokinetic nystagmus on each other.
Currently, the vestibulo-ocular and optokinetic reflexes are both thought to require a velocity storage mechanism within their neural pathways. To test whether these storage mechanisms are shared by both reflexes, animals were given programs of stimulation known to change the status of the storage mechanism of one of the type of nystagmus. The other type of nystagmus was examined then to ascertain whether the characteristic of its velocity storage system had remained invariant. Horizontal eye movements of three macaque monkeys were recorded during post-rotatory nystagmus and optokinetic afternystagmus (OKAN) before and after 20 habituating exposures to either vestibular or optokinetic stimulation. Repetitive exposures to vestibular stimulation alone markedly reduced the time constant of post-rotatory nystagmus and this effect was accompanied only occasionally by a reduction in the time constant of OKAN. Repetitive exposure to optokinetic stimulation alone reliably reduced the time constant of OKAN but produced no reliable change in the time constant of post-rotatory nystagmus. These results suggest that the vestibulo-ocular and optokinetic reflexes do not share a single common velocity storage mechanism.


Cortical afferents onto the nucleus Reticularis thalami promote plasticity of low-threshold excitability through GluN2C-NMDARs.
Thalamus and cortex represent a highly integrated processing unit that elaborates sensory representations. Interposed between cortex and thalamus, the nucleus Reticularis thalami (nRt) receives strong cortical glutamatergic input and mediates top-down inhibitory feedback to thalamus. Despite growing appreciation that the nRt is integral for thalamocortical functions from sleep to attentional wakefulness, we still face considerable gaps in the synaptic bases for cortico-nRt communication and plastic regulation. Here, we examined modulation of nRt excitability by cortical synaptic drive in Ntsr1-Cre x ChR2tg/+ mice expressing Channelrhodopsin2 in layer 6 corticothalamic cells. We found that cortico-nRt synapses express a major portion of NMDA receptors containing the GluN2C subunit (GluN2C-NMDARs). Upon repetitive photoactivation (10 Hz trains), GluN2C-NMDARs induced a long-term increase in nRt excitability involving a potentiated recruitment of T-type Ca2+ channels. In anaesthetized mice, analogous stimulation of cortical afferents onto nRt produced long-lasting changes in cortical local field potentials (LFPs), with delta oscillations being augmented at the expense of slow oscillations. This shift in LFP spectral composition was sensitive to NMDAR blockade in the nRt. Our data reveal a novel mechanism involving plastic modification of synaptically recruited T-type Ca2+ channels and nRt bursting and indicate a critical role for GluN2C-NMDARs in thalamocortical rhythmogenesis.


[Alpha1 desensitization of the heart during adaptation to stress].
Results of the experiments evidence that a combination of three factors, limiting the Ca2+ concentration increase in myocardial cell, can play a role in the cardioprotective effect of adaptation of rats to short-term immobilization stress (every second day for a month) are presented. Those factors are as follows: desensitization of alpha 1-adrenoreceptors, M-cholinoreceptors up-regulation and reduced number of voltage-dependent Ca2+ channels in the myocardial membranes.


[Morphofunctional immaturity of the urinary tract and vesicoureteral reflux in young children].
On the basis of rich clinical experience--593 patients and 916 renoureteral units (RUU) with vesicoureteral reflux (VUR) in infants the authors showed that early recognition of VUR is necessary, which is ensured by adequate organization of service to infants of the "risk group". Complex examination of children by functional diagnostic methods conducted under general anesthesia makes it possible to undertake some methods of examination for the detection and control of morphofunctional immaturity, while early application of purposeful rehabilitation measures in the stages inpatient treatment--outpatient treatment--treatment in the family produces a "positive dynamics" of the course of the pathological process in most patients (up to 82.1%). All this in complex confirms that morphofunctional immaturity of the urinary tract is the main cause of VUR in infants.


Transcriptional changes in human palate and skin healing.
Most human tissue injuries lead to the formation of a fibrous scar and result in the loss of functional tissue. One adult tissue that exhibits a more regenerative response to injury with minimal scarring is the oral mucosa. We generated a microarray gene expression dataset to examine the response to injury in human palate and skin excisional biopsies spanning the first 7 days after wounding. Differential expression analyses were performed in each tissue to identify genes overexpressed or underexpressed over time when compared to baseline unwounded tissue gene expression levels. To attribute biological processes of interest to these gene expression changes, gene set enrichment analysis was used to identify core gene sets that are enriched over the time-course of the wound healing process with respect to unwounded tissue. This analysis identified gene sets uniquely enriched in either palate or skin wounds and gene sets that are enriched in both tissues in at least one time point after injury. Finally, a cell type enrichment analysis was performed to better understand the cell type distribution in these tissues and how it changes over the time course of wound healing. This work provides a source of human wound gene expression data that includes two tissue types with distinct regenerative and scarring phenotypes.


Changes in nonmelanoma skin cancer incidence between 1977-1978 and 1998-1999 in Northcentral New Mexico.
Nonmelanoma skin cancer (NMSC) is a highly common form of malignant disease in light-skinned populations. In 1977-1978, the National Cancer Institute sponsored a population-based skin cancer survey that found marked geographic variability in the incidence of NMSC within the United States. Some of the highest rates were observed in the southwestern state of New Mexico within its non-Hispanic white population. We recently undertook a follow-up survey of NMSC in New Mexico and report here incidence rate data for non-Hispanic white residents of a three-county area in northcentral New Mexico for two 12-month time periods: June 1, 1977 to May 31, 1978 and July 1, 1998 to June 31, 1999. Our results show that incidence rates of basal cell carcinoma increased by 50% in males and 20% in females, whereas rates of squamous cell carcinoma roughly doubled in both males and females. Temporal analysis of rates according to major anatomical site showed the head and neck was consistently the most frequent site of occurrence, however, the greatest percentage increase in rates over time occurred at the upper and lower limbs. These findings are consistent with those reported for various other populations showing the incidence of NMSC has measurably increased since the 1970s.


Assessment of a Comprehensive Early Childhood Education Program and Cardiovascular Disease Risk in Midlife.
Educational attainment is an underrecognized factor associated with racial disparities in cardiovascular disease (CVD) risk. Early childhood programs providing comprehensive family services can promote educational and socioeconomic success, but few, if any, studies of large-scale programs have assessed their associations with midlife CVD risk and mediating factors. To examine the association between an early childhood program providing multisystemic services from ages 3 to 9 years with midlife CVD risk and whether years of education mediate these associations. Using a matched-group, quasi-experimental design, a cohort of Black and Hispanic children in the Chicago Longitudinal Study enrolled in an established early childhood program from 1983 to 1989 or the usual services were followed up for 30 years after the end of the intervention. A midlife survey on well-being was administered when the participants were aged 37 years. Analyses were conducted from September 1, 2020, to October 15, 2020. The Child-Parent Center (CPC) Education Program provides school-based educational enrichment and comprehensive family services for 6 years, from ages 3 to 9 years (preschool to third grade [P-3]). General and hard Framingham Risk Scores (FRSs) were calculated from self-reported physical health and behavior profiles in the midlife Chicago Longitudinal Study survey. Years of education completed at age 34 years were measured primarily from administrative records. There were 1539 participants in the original sample (1430 Black participants [92.9%]; 108 Hispanic participants [7.0%]; 1 White participant [0.1%]); 1104 of 1401 participants (78.8%) in the tracked sample completed a midlife survey on well-being by age 37 years, and 1060 participants had data available for analysis (mean [SD] age, 34.9 [1.4] years; 565 women [53.3%]), including 523 participants who grew up in high-poverty contexts. After adjusting for 17 baseline attributes and differential attrition via propensity score weighting, CPC preschool was associated with significantly lower general FRS (marginal coefficient, -2.2 percentage points [% hereafter]; 95% CI -0.7% to -3.6%; P = .004) and hard FRS (marginal coefficient, -1.6%; 95% CI -0.5% to -2.6%; P = .004), for a 20% reduction in cardiovascular disease risk. The program group was also less likely to have high-risk FRS status, including being in the top quartile of hard FRS (marginal coefficient, -7.2%; 95% CI, -0.3% to -11.6%; P = .02). Those who participated for 4 to 6 years (CPC P-3) had lower general FRS than those who participated for fewer years, but the difference was not significant (marginal coefficient, -1.2%; 95% CI, -2.5% to 0.2%; P = .09); longer participation was associated with placement in the median or higher risk category for general FRS (marginal coefficient, -7.9%; 95% CI -0.7% to -12.4%; P = .007) and hard FRS (marginal coefficient, -9.0%; 95% CI -0.6% to -11.4%; P = .02). Years of education completed by age 34 years accounted for up to 23% of the observed group differences for FRSs, including CPC preschool (general FRS, from -2.16% to -1.66%; difference = -0.5%) and P-3 (general FRS, from -1.16% to -0.71%; difference = -0.45%). In this quasi-experimental design study, a comprehensive early childhood program for a community cohort at high risk of disparities was associated with lower CVD risk later in life. Corroborating previous research, years of education through college matriculation accounted for a sizable percentage of this association. Early childhood enrichment may contribute to CVD prevention.


The significance of hyperphagia and diet composition on the metabolism in ventromedial hypothalamic lesioned male rats.
The metabolic consequences of ventromedial hypothalamic lesion were studied in a group of aged male rats which were obese and had decreased response to insulin. The effects of hyperphagia and ventromedial hypothalamic lesion per se were separated by comparing experimental animals fed isocalorically with controls and animals fed ad libitum. Ventromedial hypothalamic lesion as such led to increases in the glucose conversion to fatty acid and in lipoprotein lipase activity in adipose tissue. Protein catabolism as reflected by plasma urea levels, was enhanced. The lipoprotein lipase activity in heart tended to be lower after VMH lesion. These metabolic changes were amplified in the VMH lesioned rats fed ad libitum. The liver glycogen content was lowered by VMH lesion, but this effect was abolished by hyperphagia. In parallel experiments the influence of diet composition was studied by feeding similar groups with diet of high fat content. The glucose incorporation in fatty acids was in all groups markedly and similarly inhibited by the high fat diet. The increase in lipoprotein lipase activity in heart and adipose tissue of control rats with high fat intake could not be demonstrated in any of the groups with ventromedial hypothalamic lesion. The plasma urea level in the control group was not affected by the diet, but tended to increase in the ventromedial hypothalamic lesioned groups on high fat intake. These findings demonstrate that the well known metabolic effects of ventromedial hypothalamic lesions are also manifest in obese insulin resistant male rats. Furthermore, the responses to changes in diet composition are different from those of the control rats.


Mutations affecting expression of the rosy locus in Drosophila melanogaster.
The rosy locus in Drosophila melanogaster codes for the enzyme xanthine dehydrogenase (XDH). Previous studies defined a "control element" near the 5' end of the gene, where variant sites affected the amount of rosy mRNA and protein produced. We have determined the DNA sequence of this region from both genomic and cDNA clones, and from the ry+10 underproducer strain. This variant strain had many sequence differences, so that the site of the regulatory change could not be fixed. A mutagenesis was also undertaken to isolate new regulatory mutations. We induced 376 new mutations with 1-ethyl-1-nitrosourea (ENU) and screened them to isolate those that reduced the amount of XDH protein produced, but did not change the properties of the enzyme. Genetic mapping was used to find mutations located near the 5' end of the gene. DNA from each of seven mutants was cloned and sequenced through the 5' region. Mutant base changes were identified in all seven; they appear to affect splicing and translation of the rosy mRNA. In a related study (T. P. Keith et al. 1987), the genomic and cDNA sequences are extended through the 3' end of the gene; the combined sequences define the processing pattern of the rosy transcript and predict the amino acid sequence of XDH.


MyoD Overexpressed Equine Adipose-Derived Stem Cells Enhanced Myogenic Differentiation Potential.
Mesenchymal stem cells could potentially be used in the clinical treatment of muscle disorders and muscle regeneration. Adipose-derived stem cells (ADSCs) can be easily isolated from adipose tissue, as opposed to stem cells of other tissues. We believe that cell therapy using ADSCs could be applied to muscle disorders in horses and other species. We sought to improve the myogenic differentiation potential of equine ADSCs (eqADSCs) using a MyoD lentiviral vector. MyoD lentiviruses were transduced into eqADSCs and selected using puromycin. Cells were cultured in differentiation media containing 5% horse serum, and after 5 days the MyoD-transduced cells differentiated into myogenic cells (MyoD-eqADSCs). Using green fluorescent protein (GFP), MyoD-eqADSCs were purified and transplanted into the tibialis anterior muscles of mice after they were injured with the myotoxin notexin. The mice were sacrificed to examine any regeneration in the tibialis anterior muscle 4 weeks after the MyoD-eqADSCs were injected. The MyoD-eqADSCs cultured in growth media expressed murine and equine MyoD; however, they did not express late differentiation markers such as myogenin (MYOG). When cells were grown in differentiation media, the expression of MYOG was clearly observed. According to our reverse transcription polymerase chain reaction and immunocytochemistry results, MyoD-eqADSCs expressed terminal myogenic phase genes, such as those encoding dystrophin, myosin heavy chain, and troponin I. The MyoD-eqADSCs fused to each other, and the formation of myotube-like cells from myoblasts in differentiation media occurred between days 5 and 14 postplating. In mice, we observed GFP-positive myofibers, which had differentiated from the injected MyoD-eqADSCs. Our approaches improved the myogenic differentiation of eqADSCs through the forced expression of murine MyoD. Our findings suggest that limitations in the treatment of equine muscle disorders could be overcome using ADSCs.


Location, incidence, and malignant potential of duodenal gastrinomas.
Duodenal gastrinomas are increasingly found at surgery, yet information about their location and characteristics is based on the results of either pooled series or retrospective reviews of small numbers of selected cases. To address these issues we have analyzed the location, incidence, and malignant potential of duodenal gastrinomas in 65 consecutive patients who underwent removal of all tumor as part of a 10-year prospective study to resect gastrinomas in patients with sporadic Zollinger-Ellison syndrome. The primary gastrinoma was located in the duodenum in 24 patients (37%). There were 19 men and five women aged 32 to 69 years (mean 49.4 years), with symptoms for 0.6 to 35 years (mean 7.9 years). Preoperative studies included serum gastrin levels of 114 to 35,798 pg/ml (mean 2060 pg/ml), basal acid output of 7 to 95 mEq/hr (mean 37.6 mEq/hr), and a positive secretin test result in 22 patients. Preoperative imaging studies identified tumor in the duodenal area in 11 patients (46%), but most positive imaging findings were metastatic gastrinoma in lymph nodes, and the primary duodenal tumor itself was identified in only two patients. Portal venous sampling had a localizing gastrin gradient in the inferior or superior pancreaticoduodenal vein in 17 of 23 patients (74%). Each of the 24 patients had a single, small duodenal wall tumor of 2.8 to 10.1 mm diameter (mean 6 mm). Each tumor stained positive for gastrin by immunohistochemistry. Seventeen tumors (71%) were located in the first portion of the duodenum, five (21%) in the second, and two (8%) in the third. Each tumor originated in the submucosa, and 13 (54%) were limited to the submucosa, whereas 11 (46%) were locally invasive, four (16%) extending into the muscularis mucosa and seven (29%) into the muscularis propria. Thirteen patients (54%) had spread to regional lymph nodes, whereas two (8%) had liver metastases. Lymph node metastases were seen with larger duodenal tumors (mean 7.1 vs 5.4 mm; p less than 0.01). The data suggest that a single duodenal wall gastrinoma is a common cause of Zollinger-Ellison syndrome (37%). These small (less than 1 cm) tumors are located in the submucosal layer of the proximal duodenum (92%) and are malignant more often than previously thought (54%).


Three novel subunits of Arabidopsis chloroplastic NAD(P)H dehydrogenase identified by bioinformatic and reverse genetic approaches.
Chloroplastic NAD(P)H dehydrogenase (NDH) plays a role in cyclic electron flow around photosystem I to produce ATP, especially in adaptation to environmental changes. Although the NDH complex contains 11 subunits that are homologous to NADH:ubiquinone oxidoreductase (complex I; EC 188.8.131.52), recent genetic and biological studies have indicated that NDH also comprises unique subunits. We describe here an in silico approach based on co-expression analysis and phylogenetic profiling that was used to identify 65 genes as potential candidates for NDH subunits. Characterization of 21 Arabidopsis T-DNA insertion mutants among these ndh gene candidates indicated that three novel ndf (NDH-dependent cyclic electron flow) mutants (ndf1, ndf2 and ndf4) had impaired NDH activity as determined by measurement of chlorophyll fluorescence. The amount of NdhH subunit was greatly decreased in these mutants, suggesting that the loss of NDH activity was caused by a defect in accumulation of the NDH complex. In addition, NDF1, NDF2 and NDF4 proteins co-migrated with the NdhH subunit, as shown by blue native electrophoresis. These results strongly suggest that NDF proteins are novel subunits of the NDH complex. Further analysis revealed that the NDF1 and NDF2 proteins were unstable in the mutants lacking hydrophobic subunits of the NDH complex, but were stable in mutants lacking the hydrophilic subunits, suggesting that NDF1 and NDF2 interact with a hydrophobic sub-complex. NDF4 protein was predicted to possess a redox-active iron-sulfur cluster domain that may be involved in the electron transfer.