Millennials considered: A new generation, new approaches, and implications for nursing education.
A new generation is pursuing degrees in nursing. These students, 18 to 24 years old, aptly called Millennials, are technologically competent and described as optimistic and group oriented. Their propensity for multitasking, reliance on electronics, and need for immediate feedback often perplex and frustrate faculty and challenge customary approaches to teaching and learning. They often express doubt regarding their academic readiness for college. This article explores the authors' experiences with freshmen students of the Millennial generation and shares insights for working successfully with this cohort of learners. The authors assert that by developing strong partnerships with students, faculty may be able to bridge the generational gap to promote mutual understanding, personal growth, and academic success.


Community Interagency Connections for Immigrant Worker Health Interventions, King County, Washington State, 2012-2013.
Cross-sector community partnerships are a potentially powerful strategy to address population health problems, including health disparities. US immigrants - commonly employed in low-wage jobs that pose high risks to their health - experience such disparities because of hazardous exposures in the workplace. Hazardous exposures contribute to chronic health problems and complicate disease management. Moreover, prevention strategies such as worksite wellness programs are not effective for low-wage immigrant groups. The purpose of this article was to describe an innovative application of social network analysis to characterize interagency connections and knowledge needed to design and deliver a comprehensive community-based chronic disease prevention program for immigrant workers. Using iterative sample expansion, we identified 42 agencies representing diverse community sectors (service agencies, faith-based organizations, unions, nonprofits, government agencies) pertinent to the health of Chinese immigrant workers. To capture data on shared information, resources, and services as well as organizational characteristics, we jointly interviewed 2 representatives from each agency. We used social network analysis to describe interagency network structure and the positions of agencies within the networks. Agency interconnections were established primarily for information sharing. In the overall interagency network, a few service-oriented agencies held central or gatekeeper positions. Strong interconnectedness occurred predominately across service, public, and nonprofit sectors. The Chinese and Pan-Asian service sectors showed the strongest interconnectedness. Network analysis yields critical understanding of community structural links and assets needed to inform decisions about actual and potential community collaborations. Alternative intervention strategies may be needed to address health disparities among immigrant workers.


Infection and its effect on the growth of young children: a comparison of The Gambia and Uganda.
Longitudinal studies of 152 children from Keneba, The Gambia, and 45 from Namulonge, Uganda, investigating the relationship between growth and different types of infection in the two areas are reported and their relevance to the patterns of malnutrition seen in Africa are discussed. The relative ineffectiveness of curative medicine in the real health problems of rural Africa and the need for prevention are stressed.


Italian Credit Mobility Students Significantly Increase Their Alcohol Intake, Risky Drinking and Related Consequences During the Study Abroad Experience.
To examine changes in alcohol intake and consequences in Italian students studying abroad. Italian exchange students planning to study abroad were invited to report on their drinking and alcohol-related negative consequences before and after their time abroad. After excluding those who abstained throughout, data on 121 students were analysed and showed that they tended to consume more alcohol and experience more alcohol-related negative consequences compared to their pre-departure levels. The added alcohol risk of study abroad for Italian students merits consideration of possible opportunities for intervention.


The uses of AFLP for detecting DNA polymorphism, genotype identification and genetic diversity between yeasts isolated from Mexican agave-distilled beverages and from grape musts.
The objectives were to determine the variability and to compare the genetic diversity obtained using amplified fragment length polymorphism (AFLP) markers in analyses of wine, tequila, mezcal, sotol and raicilla yeasts. A molecular characterization of yeasts isolated from Mexican agave musts, has been performed by AFLP marker analysis, using reference wine strains from Italian and South African regions. A direct co-relation between genetic profile, origin and fermentation process of strains was found especially in strains isolated from agave must. In addition, unique molecular markers were obtained for all the strains using six combination primers, confirming the discriminatory power of AFLP markers. This is the first report of molecular characterization between yeasts isolated from different Mexican traditional agave-distilled beverages, which shows high genetic differences with respect to wine strains.


Lack of a Clinically Significant Pharmacokinetic Interaction between Etravirine and Raltegravir Using an Original Approach Based on Drug Metabolism, Protein Binding, and Penetration in Seminal Fluid: A Pharmacokinetic Substudy of the ANRS-163 ETRAL Study.
The ANRS163-ETRAL study showed that etravirine 200 mg/raltegravir 400 mg twice-daily dual therapy was highly effective in the treatment of human immunodeficiency virus (HIV)-infected patients older than 45 years, with virologic and therapeutic success rates at week 48 of 99.4% and 94.5%, respectively. The objective of this study was to determine whether a clinically significant pharmacokinetic interaction between etravirine and raltegravir exists by assessing steady-state total and unbound etravirine, raltegravir, and inactive raltegravir-glucuronide concentrations 12 hours after last intake (C12h ) in blood plasma (BP) and seminal plasma (SP). Pharmacokinetic analysis of data from the ANRS163-ETRAL study. One hundred forty-six HIV-1-infected patients (of the 165 patients included in the ANRS-163 ETRAL study) who were receiving etravirine 200 mg and raltegravir 400 mg twice daily. Blood was collected from all 146 patients at weeks 2-4, 12, 24, and 48, and semen was collected from 21 patients at week 48. The extent of BP and SP protein binding was determined by using ultrafiltration assay. Total and unbound etravirine, raltegravir, and raltegravir-glucuronide C12h were determined by ultra high performance liquid chromatography coupled with tandem mass spectrometry and interpreted by using the in vitro calculated protein-bound 95% inhibitory concentration (PBIC95 ) for wild-type (WT) HIV: etravirine (116 ng/ml) and raltegravir (15 ng/ml). Median (interquartile range [IQR]) total BP etravirine C12h (536 ng/ml [376-719]) and raltegravir (278 ng/ml [97-690]) were adequate in 99% and 96% of patients, respectively. Median (IQR) SP:BP C12h ratio and BP unbound fraction were etravirine 0.3 (0.2-0.5) and < 1%, respectively, raltegravir 1.8 (1.3-3.3) and 12%, respectively, and raltegravir-glucuronide 12.0 (6.5-17.7) and > 99%, respectively. The BP raltegravir metabolic ratio (raltegravir glucuronide:raltegravir ratio) was 1.7, suggesting only weak induction of raltegravir glucuronidation by etravirine. Only three patients had etravirine and raltegravir C12h < PBIC95 simultaneously. No clinically significant pharmacokinetic interaction between etravirine and raltegravir was detected. Total etravirine and raltegravir BP concentrations were adequate in most patients, favoring virologic efficacy and confirming good treatment adherence (> 95%), despite twice-daily administration. The long half-life of etravirine and higher unbound fraction SP of raltegravir (57%) ensured adequate concentrations of dual therapy in genital compartments. Our results indicate that etravirine and raltegravir have good, complementary pharmacokinetic profiles, suggesting that they could be used in a dual-treatment strategy.


Oxidation of hydroquinone by both cellular and extracellular grapevine peroxidase fractions.
The oxidation of hydroquinone by two peroxidase (EC 22.214.171.124) fractions obtained from the cells and spent medium of cell cultures of grapevine (Vitis vinifera cv Monastrell) has been studied, and their comparative efficacy (kcat/KM ratio) studied in both the H2O2-consuming and hydroquinone-consuming reactions. While the efficacy in the H2O2-consuming reaction is practically identical for both enzyme fractions, the cellular peroxidase has five-fold more efficacy in the hydroquinone-consuming reaction than the peroxidase located in the spent medium. Screening of cellular peroxidases capable of oxidizing hydroquinone on polyacrylamide gels, by means of a staining reaction based on the nucleophilic attack of 4-aminoantipyrine on p-benzoquinone in acidic media, reveals that all the cellular peroxidase isoenzymes are capable of oxidizing hydroquinone, probably yielding a quinone-diimine as a product of the staining reaction. Since isoperoxidases found in cellular fractions are also present in the spent medium, the values found for the different efficacies in the hydroquinone-consuming reaction must be considered as the results of the different proportions in which each peroxidase isoenzyme was found in the two fractions. The localization of a benzoquinone-generating system of high efficacy inside the plant cell, and probably located in vacuoles, is discussed with respect to the harmful role which the quinone/semiquinone pair might play in cell death, as part of the hypersensitive response expressed within the mechanism of plant disease resistance.


Differentiation between benign and malignant hilar obstructions using laboratory and radiological investigations: a prospective study.
Preoperative determination of the aetiology of bile duct strictures at the hilum is difficult. We evaluated the diagnostic accuracy of laboratory parameters and imaging modalities in differentiating between benign and malignant causes of hilar biliary obstruction. Fifty-eight patients (26 men) with a history of obstructive jaundice and liver function tests (LFTs) and ultrasound suggestive of biliary obstruction at the hilum were studied. They were evaluated by tumour marker assay (CA19-9), CT and MRI/MRCP. A single experienced radiologist, blinded to the results of other tests, evaluated the imaging. The final diagnosis was made either from histology of the resected specimen, operative findings or image-guided biopsy in inoperable patients. A receiver operator characteristic (ROC) curve was constructed for each laboratory parameter to determine optimal diagnostic cut-off to predict malignant biliary stricture (MBS). In all, 34 patients had a benign and 24 had malignant aetiology. The mean age of benign patients was 38 years compared with 54 years for MBS. Forty-seven patients were treated with surgery while 11 had ERCP/PTC and stenting. The ROC curve showed that preoperative bilirubin level >8.4 mg/dl (sensitivity 83.3%, specificity 70%), alkaline phosphatase level >478 IU (sensitivity 63%, specificity 49%) and CA19-9 levels >100 U/L (sensitivity 45.8%, specificity 88.2%) for predicting MBS. The sensitivity, specificity and diagnostic accuracy of MRI/MRCP (87.5%, 85.3%, 82.7%, respectively) was marginally superior to CT (79.2%, 79.4%, 79.3%, respectively). Patients with a bilirubin level of >8.4 mg% and CA19-9 level >100 U/L were more likely to have malignant aetiology. MRI/MRCP was a better imaging modality than CT.


LINC01232 targeting miR-1250-3p/MSH2 axis attenuates mesangial cell proliferation and fibrosis in diabetic nephropathy.
The significance of long non-coding RNA (ncRNAs) in the initiation and progression of diabetic nephropathy (DN) has attracted much interest. The purpose of this work was to ascertain the role of LINC01232 in cell models and animal models of DN. C57BL/6 J mice were administered with streptozotocin (STZ) to develop animal models of DN, and mouse glomerular mesangial cells (MCs) were exposed to high glucose (HG) to establish cell models of DN. Expression levels of LINC01232, miR-1250-3p and MSH2 were identified by quantitative real-time PCR (qPCR) or western blotting. Fibrosis-related proteins were quantified by western blotting. MC proliferative capacity was checked by EdU assay. DN progression and fibrosis level in animal models were assessed by hematoxylin and eosin (HE) and Masson staining. The potential binding sites between miR-1250-3p and LINC01232 or MSH2 were examined by dual-luciferase reporter assay. LINC01232 expression was heightened in kidney tissues of DN patients. Its overexpression in HG-treated MCs alleviated MC proliferation and fibrosis. Overexpression of LINC01232 alleviated the pathological state of glomerular hypertrophy, MC hyperplasia, basement membrane thickening, and fibrosis in the DN models. LINC01232 bound to miR-1250-3p and competed for miR-1250-3p binding sites with MSH2. LINC01232 overexpression decoyed miR-1250-3p to increase MSH2 expression, and MSH2 depletion restored LINC01232 overexpression-inhibited MC proliferation and fibrosis. LINC01232 alleviated the mesangial cell proliferation and fibrosis in the progression of DN by targeting miR-1250-3p/MSH2 pathway.


Changes in subcutaneous adipose tissue microRNA expression in response to exercise training in obese African women.
The mechanisms that underlie exercise-induced adaptations in adipose tissue have not been elucidated, yet, accumulating studies suggest an important role for microRNAs (miRNAs). This study aimed to investigate miRNA expression in gluteal subcutaneous adipose tissue (GSAT) in response to a 12-week exercise intervention in South African women with obesity, and to assess depot-specific differences in miRNA expression in GSAT and abdominal subcutaneous adipose tissue (ASAT). In addition, the association between exercise-induced changes in miRNA expression and metabolic risk was evaluated. Women underwent 12-weeks of supervised aerobic and resistance training (n = 19) or maintained their regular physical activity during this period (n = 12). Exercise-induced miRNAs were identified in GSAT using Illumina sequencing, followed by analysis of differentially expressed miRNAs in GSAT and ASAT using quantitative real-time PCR. Associations between the changes (pre- and post-exercise training) in miRNA expression and metabolic parameters were evaluated using Spearman's correlation tests. Exercise training significantly increased the expression of miR-155-5p (1.5-fold, p = 0.045), miR-329-3p (2.1-fold, p < 0.001) and miR-377-3p (1.7-fold, p = 0.013) in GSAT, but not in ASAT. In addition, a novel miRNA, MYN0617, was identified in GSAT, with low expression in ASAT. The exercise-induced differences in miRNA expression were correlated with each other and associated with changes in high-density lipoprotein concentrations. Exercise training induced adipose-depot specific miRNA expression within subcutaneous adipose tissue depots from South African women with obesity. The significance of the association between exercise-induced miRNAs and metabolic risk warrants further investigation.


A network signal amplification strategy of ultrasensitive photoelectrochemical immunosensing carcinoembryonic antigen based on CdSe/melamine network as label.
Taking advantage of CdSe/melamine network as label and Au-TiO2 as substrate, this work developed a novel kind of signal amplification strategy for fabricating photoelectrochemical (PEC) immunoassay. The melamine, a star-shaped triamino molecule, was firstly used for readily capturing CdSe QDs and forming a CdSe/melamine network, which was formed through strong interactions between the carboxyl groups of TGA-stabilized CdSe QDs and the three amino groups of each melamine molecule. In this strategy, the primary antibody (Ab1) was immobilized onto Au-TiO2 substrate, which made the photoelectric conversion efficiency increase significantly. After the formed Ab2-CdSe/melamine network labels were captured onto the electrode surface via the specific antibody-antigen interaction, the photoelectric activity could be further enhanced via the interaction between the Au-TiO2 substrate and CdSe/melamine network. Due to this amplification of PEC signals and the special structure of the label, the fabricated PEC immunosensor was applied for sensitive and specific detection of cancer biomarker carcinoembryonic antigen (CEA), and displayed a wide linear range (0.005-1000ngmL(-1)) and low detection limit (5pgmL(-1)). In addition, the immunosensor was performed with good stability and reproducibility, and the results to analyze human serum samples were satisfactory.


Degradation of 2,4-dinitrophenol by combining sonolysis and different additives.
Based on the effects of various additives on ultrasonic degradation of 2,4-dinitrophenol (DNP) in aqueous solution, the degradation mechanisms and reaction kinetics of DNP in different processes were proposed. The results showed that some additives, such as CuO, CCl(4), O(3), NaCl and KI, were favorable for DNP sonochemical degradation. On the contrary, DNP degradation efficiency was restrained by Na(2)CO(3), indicating that OH radicals oxidation played an important role in DNP ultrasonic removal. The significant increases in DNP degradation in US/CuO/H(2)O(2), US/CCl(4) and US/O(3) systems were also related to the intermediates formed during the reactions, such as HO(2)/O(2)(-) radicals, chlorine-containing radicals and HClO. In addition, DNP ultrasonic removal was observed to behave as pseudo-first-order kinetics under different experimental conditions tested in the present work.


Nicotine-induced catecholamine synthesis after lesions to the dorsal or ventral noradrenergic bundle.
The role of projections from coerulear (A6) and lateral tegmental (A1-A5) noradrenergic cell groups in the induced catecholamine response to (-)-nicotine was studied following lesions to the dorsal (DNAB) or ventral (VNAB) noradrenergic bundle by 6-hydroxydopamine. The lesions produced large reductions in basal noradrenaline levels in hippocampus (after DNAB lesions) and hypothalamus (after VNAB lesions), while not affecting basal levels of dopamine or 5-hydroxytryptamine. Vehicle and sham operated controls showed a significant increase in DOPA accumulation in response to (-)-nicotine (0.8 mg/kg s.c.) following inhibition of amino acid decarboxylase. In DNAB lesioned rats, the response induced by (-)-nicotine in both the hippocampus and hypothalamus was significantly attenuated, whereas in VNAB lesioned rats the induced response was still evident. The effect of (-)-nicotine was also studied in the nucleus accumbens and was found not to be affected by either lesion. These data suggest that increases in catecholamine synthesis in the hippocampus and hypothalamus reflect increased noradrenaline synthesis, and that this effect occurs specifically in noradrenergic projections originating in the locus coeruleus.


Stimuli-responsive electrospun nanofibers based on PNVCL-PVAc copolymer in biomedical applications.
Poly(N-vinylcaprolactam) (PNVCL) is a suitable alternative for biomedical applications due to its biocompatibility, biodegradability, non-toxicity, and showing phase transition at the human body temperature range. The purpose of this study was to synthesize a high molecular weight PNVCL-PVAc thermo-responsive copolymer with broad mass distribution suitable for electrospun nanofiber fabrication. The chemical structure of the synthesized materials was detected by FTIR and 1HNMR spectroscopies. N-Vinyl caprolactam/vinyl acetate copolymers (159,680 molecular weight (g/mol) and 2.51 PDI) were synthesized by radical polymerization. The phase transition temperature of N-vinyl caprolactam/vinyl acetate copolymer was determined by conducting a contact angle test at various temperatures (25, 26, 28, and 30 [Formula: see text]). The biocompatibility of the nanofibers was also evaluated, and both qualitative and quantitative results showed that the growth and proliferation of 929L mouse fibroblast cells increased to 80% within 48 h. These results revealed that the synthesized nanofibers were biocompatible and not cytotoxic. The results confirmed that the synthesized copolymers have good characteristics for biomedical applications.


The tear-drop sign: a trap door for the unwary?
We describe a patient who presented with features suggestive of an orbital blowout fracture. These were apparently confirmed on imaging and operation was planned. However, further radiological and orthoptic opinions diagnosed both a pre-existing eye condition and benign antral mucosal disease that had been misinterpreted as a single acute problem. The patient was discharged with no need for intervention.


Genome wide in silico SNP-tumor association analysis.
Carcinogenesis occurs, at least in part, due to the accumulation of mutations in critical genes that control the mechanisms of cell proliferation, differentiation and death. Publicly accessible databases contain millions of expressed sequence tag (EST) and single nucleotide polymorphism (SNP) records, which have the potential to assist in the identification of SNPs overrepresented in tumor tissue. An in silico SNP-tumor association study was performed utilizing tissue library and SNP information available in NCBI's dbEST (release 092002) and dbSNP (build 106). A total of 4865 SNPs were identified which were present at higher allele frequencies in tumor compared to normal tissues. A subset of 327 (6.7%) SNPs induce amino acid changes to the protein coding sequences. This approach identified several SNPs which have been previously associated with carcinogenesis, as well as a number of SNPs that now warrant further investigation This novel in silico approach can assist in prioritization of genes and SNPs in the effort to elucidate the genetic mechanisms underlying the development of cancer.


Zero-profile implant (Zero-p) versus plate cage benezech implant (PCB) in the treatment of single-level cervical spondylotic myelopathy.
Anterior cervical discectomy and fusion is the golden standard for anterior surgery treating elderly cervical degenerative disease, but the previous implant has some problems such as looseness, translocation, sinking and dysphagia, So Zero-p implant and PCB implant have been developed to decrease the complications. The clinical data of 57 patients with single level cervical spondylotic myelopathy were retrospectively analyzed. 27 patients adopting Zero-p interbody fusion cage as implant (Zero-p group) and 30 patients adopting integrated plate cage benezech (PCB) as implant (PCB group) from January 2010 to October 2012. Observe whether are differences between the two groups of patients on operation time, intraoperatve blood loss,Japanese Orthopaedic Association (JOA) scores before and after operation, intervertebral height, cervical physiological curvature, fusion rate, Postoperative dysphagia rate and complications. Zero-p group's operation time is 98.2 + 15.2 min and its intraoperatve blood loss is 88.2 + 12.9 ml, both of which are lower than those of PCB group (109.8 + 16.9 min,95.2 + 11.6 ml ), so their differences are statistically significant (P < 0.05). The two groups' JOA scores 3 months after operation and in the last follow-up are significantly higher than those before operation, so the differences are statistically significant (P < 0.05). Coob angle 3 months after operation and in the last follow-up improves obviously compared with before operation, so the difference is statistically significant (P < 0.05). The two groups' operation segments intervertebral height 3 months after operation and in the last follow-up improves obviously compared with before operation, so the difference is statistically significant (P < 0.05) Zero-p group has one patient with dysphagia after operation and PCB group has four patients with dysphagia after operation, so there is no statistical differences between the two groups on dysphagia rate (P > 0.05, P = 0.415). PCB group has two patients with screws backing out and two patients with hoarseness after operation, the two groups' operation segments all saw bony union in the last follow-up. Zero-p group postoperative complications are lower than PCB group (P < 0.05, P = 0.044). Zero-profile implant and PCB implant both achieved good clinical effects on the treatment of cervical spondylotic myelopathy, the two groups both saw bony union in operation segments, but Zero-profile implant has the advantages of easy operation, short operation time, less intraoperatve blood loss and less complications.


Record of the subgenus, Gryllitara Chopard, 1931 (Orthoptera: Gryllidae: Itarinae) and description of a new species from India.
In the present paper, we report the occurrence of the cricket subgenus Gryllitara Chopard, 1931 from India along with the description of a new species, Itara (Gryllitara) pilosa sp. nov., from the state of Kerala, South India. Suitable plates with close-up details have been presented to describe the new species, as well as for Itara (Itara) vietnamensis Gorochov being reported from North East India.


Development and restructuring of a burn centre in an emerging nation, based on the example of Kyrgyzstan.
To implement and review a four-tier plan to develop a burn centre in an emerging nation (Kyrgyzstan). From 2001, the developing burn centre in Bishkek was supported with numerous material donations and, once a year, teams from Germany operatively treated numerous burn victims on site. Training and further education for our Kyrgyzstani colleagues were based on 'helping them to help themselves', with additional consultations on conceptual hygienic and structural improvements. Material donations were delivered personally by the project teams. Education and training were also limited due to physician emigration. However, in the seven trips made by the team from Germany, numerous operations were performed and there was fundamental progress in the areas of bandaging protocols, hygienic concepts and structural improvements. An exact plan patiently tailored to the local situation and culture must be followed, to provide cost-effective support.


[Endocervical form of the vaginal adenosis (author's transl)].
Vaginal adenosis is a rare kind of disease without specific symptoms and does not represent a very serious disorder from the beginning. Concerning the etiology teratogenetic dependencies are known which still are interpreted in different ways in the recent literature. The increased number of cases, including those with clear-cell-carcinoma of the vagina, in juveniles after exposition to DES during the early embryonal development shows in particular (Central index for the stilbestrol-adenosis-carcinoma-syndrome in the USA, since 1971) a wrong pathway in the organogenesis of cervix and vagina. This occurs at the interference zone between the epithelium of the Müllerian ducts and the epithelium of the sinus urogenitalis of the female fetus and in so far constitutes the origin of vaginal adenosis. The basics of the possibility of the development of such irregular differentiation and heterotopic dissemination are proven again by the case report of a vaginal adenosis showing the highly differentiated endocervical type. Not every case of vaginal adenosis does signify a precancerous lesion. The separation of the most frequently endocervical manifestation observed from endometrial and intermediate forms might be difficult. A positive distinction is prognostically essential, however, since clear-cell-carcinomas of the vagina are known in occurence with the endometrial type of vaginal adenosis only. Scanning electron microscopy as a method to follow tissue organization reveals the way of differentiation between the various types via ultrastructural analysis for vaginal adenosis is an example for failures of developmental organization. The same analysis is feasible in cytological smears also. The therapeutic requirements of vaginal adenosis depend on the tissular differentiation and the given local extent of the adenomatous lesions.


When population genetics serves genomics: putting adaptation back in a spatial and historical context.
Recent advances in molecular biology have opened new perspectives for the study of plant adaptation, especially at the intraspecific level. Nowadays, scientists employing -omic results in multiple scientific fields can be optimistic of their chances of revealing mechanisms involved in adaptive population divergence. However, the investment required by integrative studies greatly reduces the number of experiments that can be performed. In this context, a comprehensive choice of accessions under study is crucial. We maintain this choice could be appreciably enlightened by population genetics because it helps putting adaptive population divergence in a spatial and historical context. As an example, we highlight the usefulness of knowledge about population genetic structure in the integrative study of metal tolerance in Arabidopsis halleri.


Inhibition of oestradiol production by epidermal growth factor in human granulosa cells of normal and polycystic ovaries.
Anovulation in women with polycystic ovary syndrome results from a disorder of FSH-mediated follicular maturation which may involve paracrine modulation of FSH action by intra-ovarian factors. Epidermal growth factor (EGF) is a potent inhibitor of FSH-stimulated oestradiol production in the rat and has also been shown to inhibit aromatase activity in human granulosa cells obtained after superovulation. The purpose of this study was to investigate the action of EGF on granulosa cell function in human ovaries which had not been previously exposed to treatment with exogenous gonadotrophins and to compare the responses in tissue obtained from normal and from polycystic ovaries. Granulosa cells were obtained from antral follicles less than 10 mm in diameter after dissection of unstimulated normal or polycystic ovaries (PCO). Cells were pooled, washed, plated and incubated for 48h in the presence of 10(-7) M testosterone and various doses of human FSH. FSH dose responses were obtained with or without the addition of purified EGF (50 pg/ml). Testosterone in the absence of FSH resulted in a fourfold (range 2-7.5) increase in oestradiol accumulation in the medium after incubation of granulosa cells from both normal and polycystic ovaries. This increase was reversed by addition of EGF. FSH treatment stimulated a dose-related increase in oestradiol regardless of the origin of the granulosa cells. The peak E2 response to FSH, obtained at a dose of 1-2.5 ng/ml was a 20-fold increase above testosterone alone (range 4-55) in cells from PCO compared to sixfold (2.5-13) in cells from normal ovaries.(ABSTRACT TRUNCATED AT 250 WORDS)


Guide catheter suction with novel double wire technique for successful management of large thrombus in occluded right coronary artery.
Patients with large intracoronary thrombi represent a difficult management problem for the interventional cardiologist. Deep guide catheter engagement and thrombus aspiration have been described but are associated with serious risk of dissection and vessel closure. We describe here thrombus aspiration using a guide catheter with novel double wire technique to achieve rapid reperfusion of the coronary artery while avoiding possible complications associated with the technique.


Satisfactory clinical outcome of operative and non-operative treatment of avulsion fracture of the hamstring origin with treatment selection based on extent of displacement: a systematic review.
To compare outcome of operative and non-operative treatment of avulsion fractures of the hamstring origin, with minor (< 1.5 cm) and major (≥ 1.5 cm) displacement, and early (≤ 4 weeks) and delayed (> 4 weeks) surgery. A systematic literature search was performed using PubMed, Cochrane, Embase, CINAHL and SPORTDiscus. A quality assessment was performed using the Physiotherapy Evidence Database (PEDro) scale. Eight studies with 90 patients (mean age: 16 years) were included. All studies had low methodological quality (PEDro score ≤ 5). Operative treatment yielded a return to preinjury activity rate (RTPA) of 87% (95% CI: 68-95), return to sports (RTS) rate of 100% (95% CI: 82-100), Harris hip score (HHS) of 99 (range 96-100) and a University of California Los Angeles activity scale (UCLA) score of 100%. Non-operative treatment yielded a RTPA rate of 100% (95% CI:68-100), RTS rate of 86% (95% CI: 69-94), HHS score of 99 (range 96-100), and non-union rate of 18% (95% CI: 9-34). All patients with minor displacement were treated non-operatively (RTPA: 100% [95% CI: 21-100], RTS: 100% [95% CI: 51-100]). For major displacement, operative treatment led to RTPA and RTS rates of 86% (95% CI: 65-95) and 100% (95% CI: 84-100), and 0% (0/1, 95% CI: 0-79) and 100% (95% CI: 51-100) for non-operative treatment. Early surgery yielded RTPA and RTS rates of 100% (95% CI: 34-100 & 57-100) compared to 100 (95% CI: 72-100) and 90% (95% CI: 60-98) for delayed repair. All included studies have high risk of bias. There is only low level of evidence with a limited number of included patients to compare outcome of operative and non-operative treatment. Overall outcome was satisfactory. There is a treatment selection phenomenon based on displacement, with acceptable outcome in both groups. There is insufficient data to draw conclusions regarding timing of surgery. IV.


Greater contribution of N-methyl-D-aspartic acid receptors in ventral compared to dorsal hippocampal slices in the expression and long-term maintenance of epileptiform activity.
Functional segregation along the dorso-ventral axis of the hippocampus is a developing concept. The higher susceptibility of the ventral hippocampus to epileptic activity compared with dorsal hippocampus is one of the main features, which still has obscure mechanisms. Using the model of magnesium-free medium and field recordings, single epileptiform discharges displayed higher incidence (77% vs 57%), rate (41.7+/-3.1 vs 13.5+/-0.7 events/min), duration (173.9+/-17.7 vs 116.8+/-13.6 ms) and intensity (coastline, 25.4+/-2.5 vs 9.5+/-1.8) in ventral compared with dorsal rat hippocampal slices. In addition, the decay phase of the evoked synaptic potentials was 110% slower in ventral slices. The N-methyl-D-aspartate (NMDA) receptor antagonist d-(-)-2-amino-5-phosphonopentanoic acid (50-100 microM) decreased the discharge rate and coastline similarly in ventral and dorsal slices, but it shortened the discharges in ventral slices (by 40%) only. The NMDA receptor antagonist 3-((R)-2-carboxypiperazin-4-yl)-propyl-1-phosphonic acid (10 microM) decreased the rate in both groups and additionally shortened discharges in both kinds of slices, an effect which was greater in ventral ones (31% vs 13%). Furthermore, both drugs shortened the evoked potentials more in ventral (77%) than in dorsal slices (52%). On the other hand, 1 microM of 3-((R)-2-carboxypiperazin-4-yl)-propyl-1-phosphonic acid shortened the discharges and evoked synaptic potentials only in ventral slices, and slowed down the discharge rate only in dorsal slices. Addition of NMDA, in the magnesium-free medium, enhanced activity in both kinds of slices. At 5 and 10 microM of NMDA 51% of the ventral but only 9% of the dorsal slices displayed persistent epileptiform discharges, which were recorded for at least one hour after reintroduction of magnesium in the medium. At 10-20 microM the enhancement of activity was transient, followed by suppression of discharges in 40% and 76% of the ventral and dorsal slices, respectively. Most of the slices having experienced suppression did not develop persistent activity. We propose that the NMDA receptors contribute to the higher susceptibility of the ventral hippocampus to expression and long-term maintenance of epileptiform discharges. This diversification may be related to other aspects of hippocampal dorso-ventral functional segregation.


Systematic mutagenesis of the yeast mating pheromone receptor third intracellular loop.
Signal transduction in the mating pathway of the yeast Saccharomyces cerevisiae is initiated by binding of a peptide pheromone to a G protein-coupled receptor (Ste2). We systematically have mutated the third intracellular loop of the Ste2 receptor to investigate its functional significance. We substituted each of the 13 amino acids in the loop with alanine individually or together with one other residue. In addition, we used a site-directed random replacement mutagenesis technique to replace a region encoding three amino acids in the loop with random sequence. Over 80 such Ste2 mutants have been analyzed by several functional and biochemical criteria in a yeast strain that carries a genomic deletion of the STE2 gene. The mutant phenotypes range from fully functional to severely compromised in signaling. The observation that amino acid substitutions in the third intracellular loop of the Ste2 receptor can affect activation of the yeast mating response implicates the loop in this signal transduction pathway. The types of mutations that compromise the function of the receptor may provide clues to the physical interaction between the receptor and the G protein.


OCP2 exists as a dimer in the organ of Corti.
OCP2 is one of the most abundant proteins in the organ of Corti (OC), comprising approximately 5% of the total protein in the supporting cell population. Although the very close homolog, Skp1p, has been implicated in regulating cell-cycle progression, the function of OCP2 in the terminally differentiated cochlea is presently unknown. We have purified recombinant OCP2 from Escherichia coli and examined the protein by analytical ultracentrifugation. Interestingly, sedimentation equilibrium data collected at 20 degrees C unequivocally indicate that, at the concentrations present in the OC, free OCP2 would exist as a dimeric species. The apparent sedimentation coefficient is independent of concentration at loading concentrations between 10 and 100 microM, indicating the absence of a significant monomer-dimer equilibrium in this concentration range. The functional significance of this finding is discussed.


Termination of sustained ventricular tachycardia by external noninvasive pacing.
Cardiac pacing has proven useful in the termination of sustained ventricular tachycardia (VT). In this study, the effectiveness of external noninvasive temporary pacing was compared with traditional endocardial burst ventricular pacing for the termination of sustained and hemodynamically stable VT. In 14 patients, 16 VT morphologies induced by programmed right ventricular extrastimulation were reproducibly terminated by endocardial burst pacing (3 to 9 complexes). VT cycle lengths averaged 392 +/- 97 ms (standard deviation) and ranged from 300 to 690 ms. The endocardial burst pacing cycle length used to terminate VT averaged 298 +/- 93 ms (range 220 to 600 ms). External burst pacing terminated 14 of 16 VT morphologies (88%). The pacing cycle length used to terminate these 14 VTs averaged 282 +/- 44 ms. The number of ventricular captures ranged from 5 to 20 beats. Failure to terminate 2 VT morphologies probably represented a failure of the device to capture the ventricle. Acceleration of VT occurred in 1 patient with burst external noninvasive pacing. These observations suggest that external burst pacing may be an effective means of terminating sustained VT in some patients.


Simvastatin reduces TGF-β1-induced SMAD2/3-dependent human ventricular fibroblasts differentiation: Role of protein phosphatase activation.
Excessive cardiac fibrosis due to maladaptive remodeling leads to progression of cardiac dysfunction and is modulated by TGF-β1-activated intracellular phospho-SMAD signaling effectors and transcription regulators. SMAD2/3 phosphorylation, regulated by protein-phosphatases, has been studied in different cell types, but its role in human ventricular fibroblasts (hVFs) is not defined as a target to reduce cytokine-mediated excessive fibrotic response and adverse cardiac remodeling. Statins are a class of drugs reported to reduce cardiac fibrosis, although underlying mechanisms are not completely understood. We aimed to assess whether simvastatin-mediated reduction in TGF-β1-augmented profibrotic response involves reduction in phospho-SMAD2/3 owing to activation of protein-phosphatase in hVFs. Cultures of hVFs were used. Effect of simvastatin on TGF-β1-treated hVF proliferation, cytotoxicity, myofibroblast differentiation/activation, profibrotic gene expression and protein-phosphatase activity was assessed. Simvastatin (1 μM) reduced effect of TGF-β1 (5 ng/mL) on hVF proliferation, myofibroblast differentiation (reduced α-smooth muscle actin [α-SMA-expression]) and activation (decreased procollagen-peptide release). Simvastatin also reduced TGF-β1-stimulated time-dependent increases in SMAD2/3 phosphorylation and nuclear translocation, mediated through catalytic activation of protein-phosphatases PPM1A and PP2A, which physically interact with SMAD2/3, thereby promoting their dephosphorylation. Effect of simvastatin on TGF-β1-induced fibroblast activation was annulled by okadaic acid, an inhibitor of protein-phosphatase. This proof-of-concept study using an in vitro experimental cell culture model identifies the protective role of simvastatin against TGF-β1-induced hVF transformation into activated myofibroblasts through activation of protein phosphatase, a novel target that can be therapeutically modulated to curb excessive cardiac fibrosis associated with maladaptive cardiac remodeling.


Imagined risk of suffocation as a trigger for hyperventilation.
Although hyperventilation has been hypothesized to play a role in many pathologies, its critical triggers remain poorly understood. The present experiment aimed to test whether stronger hyperventilation responses occur in response to suggested risk of suffocation compared with other fearful situations in high- and low-trait anxious women. Fractional end-tidal CO2-concentration (FetCO2), respiratory frequency, and inspiratory volume were measured nonintrusively in high- (n = 24) and low- (n = 24) trait anxious women during imagery of 3 fear, 1 tension, 1 depressive, and 3 relaxation scripts. The fear scripts were equal in ratings of unpleasantness and arousal but differed regarding the inclusion of suggested risk of suffocation and entrapment. After each imagery trial, participants rated the emotional dimensions of pleasantness, arousal, and dominance and the vividness of their imagery. Decreases in FetCO2 occurred in all fear scripts. High-trait anxious women showed a stronger reduction in FetCO2 compared with low-trait anxious women during the fear script suggesting risk of suffocation but not during the other fear scripts. This effect was unrelated to any of the self-reported fear ratings. Self-reported fear of entrapment was associated with an overall lower FetCO2 but not with enhanced reactivity to imagined entrapment. High-trait anxiety is associated with stronger respiratory responsivity to imagined risk of suffocation and may constitute a specific vulnerability factor for the development of panic disorder and claustrophobia.


Serum cytokines in second trimester pregnancy and their relationship with spontaneous preterm births in the Ribeirão Preto and São Luiz cohorts.
To evaluate the association between second trimester plasma cytokine levels in asymptomatic pregnant women and preterm births (PTB) in an attempt to identify a possible predictor of preterm birth. The study design was a nested case-control study including women with singleton a gestational age between 20-25(+ 6) weeks from two Brazilian cities. The patients were interviewed, Venous blood samples were collected. The participants were again evaluated at birth. A total of 197 women with PTB comprised the case group. The control group was selected among term births (426 patients). Forty-one cytokines were compared between groups. When only spontaneous PTB were analyzed, GRO, sCD40L and MCP-1 levels were lower in the case group (p < 0.05). Logarithmic transformation was performed for cytokines with discrepant results, which showed increased levels of IL-2 in the group of spontaneous PTB (p < 0.05). In both analyses, the incidence of maternal smoking and of a history of preterm delivery differed significantly between the case and control groups. In multivariate analysis, only serum GRO levels differed between the case and control groups. Lower second trimester serum levels of GRO in asymptomatic women are associated with a larger number of PTB. This finding may reflect a deficient maternal inflammatory response.


Maintenance of B cell anergy requires constant antigen receptor occupancy and signaling.
Immunological tolerance can be mediated by anergy, in which self-reactive B cells persist in the periphery yet remain unresponsive to immunogen. Whether anergy is induced after transient exposure to self antigen and is 'remembered' or requires continuous antigen receptor occupancy and transduction of signals remains unclear. We have explored this using an immunoglobulin-transgenic mouse in which B cells were hapten specific (arsonate) yet cross-reacted with a self antigen that induced anergy in vivo. Many features of anergic cells were rapidly reversed after dissociation of self antigen using hapten competition and these cells regained antigen responsiveness. Our findings indicate that continuous binding of antigen and subsequent receptor signaling are essential for the maintenance of anergy.


Dissociation of cells from sea urchin embryos alters the synthesis of actins and other proteins.
The effects of altered cellular microenvironments on patterns of protein synthesis at various periods during sea urchin development were quantitated by comparing the relative incorporation of [35S]methionine into selected polypeptides of intact embryos and cells dissociated from them. The effects of increasing times of reassociation were also determined. Quantitative, but not qualitative, differences in incorporation were noted. Actins, as well as heterogeneous acidic polypeptides with an Mr of about 80 kDa, showed increased incorporation in dissociated cells labeled at the time control embryos were recently hatched blastulae. Labeling of another acidic group of polypeptides with an Mr of about 100 kDa was decreased. Possible mechanisms regulating these shifts in incorporation were investigated by the use of inhibitors. The dissociation-triggered changes were insensitive to actinomycin D, cordycepin, dibutyryl cAMP, 3-isobutyl-1-methylxanthine, and trifluoperazine; however, the latter two stimulated incorporation into some polypeptides in intact blastulae. Age-dependent shifts in incorporation were also detected in both intact embryos and dissociated/reassociating cells.


(-)-Tetrahydroberberrubine∙acetate accelerates antioxidant potential and inhibits food associated Bacillus cereus in rice.
A protoberberine alkaloid, (-)-tetrahydroberberrubine∙acetate (THBA) was assessed for its antioxidant potential and ability to inhibit the growth of a food hazard bacterium Bacillus cereus in vitro and in situ. THBA displayed significant and dose-dependent cellular antioxidant potential against hydrogen peroxide-induced oxidative stress in NIH 3T3 fibroblast cells and decreased the ROS levels as well as increased the expression levels of SOD1 and SOD2 enzymes. The inhibitory spectrum of THBA confirmed its mechanistic role in the disruption of the membrane integrity of B. cereus as evidenced by the results of time-inactivation, cell membrane integrity, NPN membrane uptake, membrane potential, and electron microscopy analyses. Moreover, THBA inhibited biofilm formation by B. cereus and disrupted pre-established biofilms on a glass surface. Furthermore, THBA was also able to inhibit B. cereus in raw rice with a significant amount of reduction in CFU counts, suggesting its potential role as a natural antioxidant and antimicrobial agent.


Dynamical observation on biological progression of VX2 liver tumors to identify the optimal time for intervention in animal models.
Based on practice guideline of "management of hepatocellular carcinoma (HCC): update" published by American Association for the Study of Liver Diseases (AASLD) and "Barcelona Clinic Liver Cancer staging system (BCLC)," this study investigated how to enroll the optimal VX2 liver tumor model for HCC researches by dynamically observing the biological progression of the tumor. Thirty-two healthy New Zealand white rabbits were implanted VX2 liver tumor by cell suspension method (n=24) and tissue fragment method (n=8). All the rabbits underwent CT scans on day 7, 14, 21 and 28 after implantation to observe the size of the tumors, the time when metastases and ascites occurred and the survival time. Appropriate intervention times were estimated corresponding to different clinical HCC stages by using tumor diameter-time curve. The VX2 liver tumors grew rapidly within 28 days after implantation. And the tumors in the cell suspension group grew faster than those of the tissue fragment group. The appropriate intervention time corresponding to very early stage, early stage and intermediate stage were <11 days, 11-16.9 days and >16.9 days, respectively in the cell suspension group, and <19.9 days, 19.9-25.5 days and >25.5 days, respectively in the tissue fragment group. Preclinical animal research needs to improve on different levels to yield best predictions for human patients. Researchers should seek for an individualized proposal to select optimal VX2 liver tumor models for their experiments. This approach may lead to a more accurate determination of therapeutic outcomes.


[Biological diagnosis of leishmaniasis (author's transl)].
Serochematological, immunological and direct parasitological methods used in the diagnosis of man, dog and wild host reservoirs of Leishmaniasis are described. From their own investigations of more than 5.000 cases, the authors compare the respective technical advantages and the specific indications of each method.


[Coronary artery bypass grafting in cases with poor left ventricular function].
From January 1987 through June 1992, 18 patients with poor left ventricular function (left ventricular ejection fraction [LVEF] less than 0.3) underwent elective isolated primary coronary artery bypass surgery. The mean age was 56.4 years (range, 46 to 72 years), and 15 were males and 3 were females. Mean pre-operative LVEF measured by ventriculography was 0.26 +/- 0.03 (range, 0.19 to 0.30). Sixteen patients (88.9%) had a prior myocardial infarction and 9 (50%) had a history of congestive heart failure. Complete revascularization was the goal for all patients, and the mean number of bypass grafts was 3.0 +/- 0.8 per patient. The left anterior descending coronary artery (LAD) was revascularized in all patients. There were no operative deaths. Post-operative LVEF improved significantly from 0.26 +/- 0.03 to 0.42 +/- 0.11 (p = 0.0002), and the regional left ventricular wall motion improved in the diaphragmatic and posterobasal regions (p < 0.01). The patency of the grafts was 93.9% in all, and 100% for LAD. The mean follow-up period was 77 months, and the overall actuarial survival rate was 88.9% at 10 years. During follow-up periods, two patients died of congestive heart failure (CHF), and two required three rehospitalizations because of CHF. The overall cardiac event free rate was 75.8% at 10 years. In patients with poor left ventricular function, surgical revascularization can be performed safely, but congestive heart failure sometimes occurs during follow-up periods and may be the cause of death. Therefore alternate forms of therapy such as cardiac transplantation and/or TMLR should be considered in selected patients.


Complement activation in thrombotic thrombocytopenic purpura.
Ultra-large von Willebrand factor and deficiency of its cleaving protease are important factors in the events leading to thrombotic microangiopathy; however, the mechanisms involved are only partly understood. Whereas pathological activation of the alternative complement pathway is linked to atypical hemolytic uremic syndrome, the role of complement activation in thrombotic thrombocytopenic purpura (TTP) is unknown. The aim of this study was to investigate whether signs of complement activation are characteristic of TTP. Twenty-three patients with TTP (18 women, median age 38 years) and 17 healthy controls (13 women, median age 38 years) were included. Complement parameters (C3, Factors H, I, B and total alternative pathway activity) together with complement activation fragments (C3a) or complexes (C1rs-INH, C3bBbP, sC5b9) were measured by ELISA or RID. ADAMTS13 activity and anti-ADAMTS13 inhibitory antibodies were measured by the VWF-FRET73 assay. Increased levels of C3a, and SC5b9 were observed in TTP during acute episodes, as compared with healthy controls. Decreased complement C3 levels indicative of complement consumption occurred in 15% of acute TTP patients. Significant decrease of complement activation products C3a and SC5b9 was observed during plasma exchange (PEX). The sustained presence of anti-ADAMTS13 inhibitory antibodies in complete remission was associated with increased complement activation. These data document in an observational study the presence of complement activation in TTP. Further investigation is needed to determine its potential pathogenetic significance.


Structural elucidation of phenidate analogues via the ESI-MS/MS spectra of their sodium adduct ions.
The identification of phenidate new psychoactive substances (NPS) by implementing MS (Mass spectrometry) techniques is a challenging task. Phenidate analogues present information-poor mass spectra, both in GC-EI-MS and LC-ESI-MS/MS of the protonated molecules [M+H]+, with a high abundance fragment/product ion representing the secondary amine-containing residue. This lack of EI-MS and ESI-MS/MS information is attributed to the strong tendency of the amine residue to stabilize the positive charge and leads to unavoidable ambiguity in the identification process. Moreover, thermal decomposition of these compounds occurs in the injection port and/or on the column under standard GC conditions. Herein, we demonstrate how structural information can be attained instantaneously through the LC-ESI-MS/MS fragmentation of the accompanied sodium adducts [M+Na]+. The sodium cation alters the charge distribution during ESI-MS/MS fragmentation, generating a major product ion corresponding to the Na+ adduction of the carbonyl group, providing new structural information of the main core of phenidate derivatives (alkylaryl acetate/acetic acid), enabling their reliable structural elucidation. This quick, simple and easy technique can be implemented to confirm the identity or identify various structurally related phenidate analogues in forensic toxicology and doping analysis without the need for sample handling.


Efficient production of glutathione in Saccharomyces cerevisiae via a synthetic isozyme system.
Glutathione, a tripeptide consisting of cysteine, glutamic acid, and glycine, has multiple beneficial effects on human health. Previous studies have focused on producing glutathione in Saccharomyces cerevisiae by overexpressing γ-glutamylcysteine synthetase (GSH1) and glutathione synthetase (GSH2), which are the rate-limiting enzymes involved in the glutathione biosynthetic pathway. However, the production yield and titer of glutathione remain low due to the feedback inhibition on GSH1. To overcome this limitation, a synthetic isozyme system consisting of a novel bifunctional enzyme (GshF) from Gram-positive bacteria possessing both GSH1 and GSH2 activities, in addition to GSH1/GSH2, was introduced into S. cerevisiae, as GshF is insensitive to feedback inhibition. Given the HSP60 chaperonin system mismatch between bacteria and S. cerevisiae, co-expression of Group-I HSP60 chaperonins (GroEL and GroES) from Escherichia coli was required for functional expression of GshF. Among various strains constructed in this study, the SKSC222 strain capable of synthesizing glutathione with the synthetic isozyme system produced 240 mg L-1 glutathione with glutathione content and yield of 4.3% and 25.6 mgglutathione /gglucose , respectively. These values were 6.6-, 4.9-, and 4.3-fold higher than the corresponding values of the wild-type strain. In a glucose-limited fed-batch fermentation, the SKSC222 strain produced 2.0 g L-1 glutathione in 67 h. Therefore, this study highlights the benefits of the synthetic isozyme system in enhancing the production titer and yield of value-added chemicals by engineered strains of S. cerevisiae.


ECMO for intractable status asthmaticus following atracurium.
Intraoperative allergic reactions are rare but serious events associated with increased morbidity and mortality. We report the salvage of intraoperative anaphylaxis leading to extreme hypercapnic respiratory failure by veno-venous extracorporeal membrane oxygenation (ECMO). A 38-year-old woman undergoing thyroidectomy developed intractable bronchospasm after administration of atracurium, leading to extreme hypercapnic respiratory failure (PaCO2 > 250 mmHg, pH 6.773). After the failure of conventional medical therapy and ventilatory optimization, the patient was connected to a veno-venous ECMO circuit. PaCO2 of 45.6 mmHg and pH of 7.25 were achieved in 1 h, by slowly increasing sweep gas flows up to 3.5 L/min and using continuous end-tidal CO2 monitoring to gauge the procedure. After extubation and disconnection from ECMO, the patient was discharged on the 6th day without sequelae. Rapid reversal of extreme hypercapnic acidosis by ECMO was feasible, without any neurologic sequelae. Veno-venous ECMO support may be a valuable option for the salvage of intraoperative anaphylaxis.


Effects of denervation on the sensitizing effect to noradrenaline induced by morphine in the vas deferens of mice treated chronically with morphine.
The acute administration of morphine to the isolated vas deferens from mice chronically exposed to this analgesic, induced a facilitatory effect on the responses of the muscle to exogenous noradrenaline. It has been suggested that this sensitizing property of morphine might reflect a dependence-like state of the vas deferens. In the present paper, the capability of met- and leu-enkephalin to substitute for morphine was studied, as well as the influence of innervation on the apparent dependence state. The contractile responses to noradrenaline and to acetylcholine were increased after the administration of morphine to the bath containing a denervated vas deferences, prepared from chronically morphinized mice. Morphine administration facilitated noradrenaline- but not acetylcholine-induced contractile effects in vas deferens isolated from mice which had been chronically treated with either morphine or morphine plus guanethidine. The presence of met- or leu-enkephalin in the isolated vas deferens from chronically morphinized mice (either intact, denervated or treated with guanethidine) failed to sensitize contractile responses to noradrenaline or acetylcholine. It is concluded that (a) the sensitizing effect induced by morphine in the vas deferens from mice chronically treated with morphine is specific for the adrenergic neurotransmitter; (b) the effect of morphine is not mimicked by opiate peptides; and (c) denervation of the vas deferens of mice treated chronically with morphine does not suppress the noradrenaline-sensitizing property of morphine.


Arginase: an old enzyme with new tricks.
Arginase has roots in early life-forms. It converts L-arginine to urea and ornithine. The former provides protection against NH3; the latter serves to stimulate cell growth and other physiological functions. Excessive arginase activity in mammals has been associated with cardiovascular and nervous system dysfunction and disease. Two relevant aspects of this elevated activity may be involved in these disease states. First, excessive arginase activity reduces the supply of L-arginine needed by nitric oxide (NO) synthase to produce NO. Second, excessive production of ornithine leads to vascular structural problems and neural toxicity. Recent research has identified inflammatory agents and reactive oxygen species (ROS) as drivers of this pathologic elevation of arginase activity and expression. We review the involvement of arginase in cardiovascular and nervous system dysfunction, and discuss potential therapeutic interventions targeting excess arginase.


Understanding basal cell adenocarcinoma of the head and neck: Population-based study.
Using a population-based database, this study investigates the risk factors, epidemiology, and outcomes of basal cell adenocarcinoma (BCAC) of the head and neck. The Surveillance, Epidemiology, and End Results database was analyzed for all patients with BCAC of the head and neck from 1973 to 2015. Three hundred and twenty-two cases of BCAC of the head and neck were identified. Mean age of diagnosis was 64.1 years. 52.5% were male and 77.3% were white. The most common primary site was the parotid gland (71.7%). Most patients underwent surgery alone (51.9%). Five-year disease-specific survival (5Y-DSS) was 95.6%, and 10Y-DSS was 90.3%. Highest survival was seen with surgery alone followed by combined surgery and radiation (10Y-DSS: 93.9% vs. 88.9%, p = 0.001). Age, primary site, T-classification, grade, and treatment type significantly affected survival. BCAC of the head and neck presents most frequently in the parotid glands. Surgery alone is associated with highest survival.


Poly(ADP-ribose) polymerase-1-induced NAD(+) depletion promotes nuclear factor-κB transcriptional activity by preventing p65 de-acetylation.
NF-κB is a transcription factor that integrates pro-inflammatory and pro-survival responses in diverse cell types. The activity of NF-κB is regulated in part by acetylation of its p65 subunit at lysine 310, which is required for transcription complex formation. De-acetylation at this site is performed by sirtuin 1(SIRT1) and possibly other sirtuins in an NAD(+) dependent manner, such that SIRT1 inhibition promotes NF-κB transcriptional activity. It is unknown, however, whether changes in NAD(+) levels can influence p65 acetylation and cellular inflammatory responses. Poly(ADP-ribose)-1 (PARP-1) is an abundant nuclear enzyme that consumes NAD(+) in the process of forming (ADP-ribose)polymers on target proteins, and extensive PARP-1 activation can reduce intracellular NAD(+) concentrations. Here we tested the idea that PARP-1 activation can regulate NF-κB transcriptional activity by reducing NAD(+) concentrations and thereby inhibiting de-acetylation of p65. Primary astrocyte cultures were treated with the alkylating agent N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) to induce PARP-1 activation. This resulted in sustained acetylation of p65 and increased NF-κB transcriptional activity as monitored by a κB-driven eGFP reporter gene. These effects of MNNG were negated by a PARP-1 inhibitor, in PARP-1(-/-) cells, and in PARP-1(-/-) cells transfected with a catalytically inactive PARP-1 construct, thus confirming that these effects are mediated by PARP-1 catalytic activity. The effects of PARP-1 activation were replicated by a SIRT1 inhibitor, EX-527, and were reversed by exogenous NAD(+). These findings demonstrate that PARP-1-induced changes in NAD(+) levels can modulate NF-κB transcriptional activity through effects on p65 acetylation.


The developmental approach to psychopathology in childhood and adolescence.
The task of developmental psychopathology is to integrate biological processes of maturation with psychological interactions and social influences. Attempts to explain behavioural maladaptation during childhood therefore require multi-level models. Examples are given in which structural lesions are obvious (fetal alcohol syndrome) or less clear (learning disabilities) and in which the expression of psychopathology varies with development. Recent work suggest adult schizophrenia to be related to prenatal neuropathology, thereby opening new perspectives for developmental studies.


Cyclic guanidines: synthesis and antiplatelet activity of 4,6,7,8-tetrahydro-1H-imidazo[1,2-a]pyrazolo[3,4-d]pyrimidin-7-ones and 1,4,6,7,8,9-hexahydropyrazolo [3',4':4,5]pyrimido[2,1-c] [1,2,4]triazin-7-ones.
A series of 4,6,7,8-tetrahydro-1H-imidazo[1,2-a]pyrazolo [3,4-d]pyrimidin-7-ones (1b-n) and 1,4,6,7,8,9-hexahydropyrazolo [3',4':4,5]pyrimido [2,1-c][1,2,4]triazin-7-ones (2a-d) has been synthesized. In view of their potential anti-aggregating activity the compounds were tested in vitro for inhibitory activity towards ADP- and collagen-induced aggregation of human platelets. Among the compounds studied, 8-benzyl-1-(2,5-dichlorophenyl)-4,6,7,8-tetrahydro-1H-imidazo [1,2-a]pyrazolo[3,4-d]pyrimidin-7-one (1n) exhibited the most favorable activity. The 2,5-dichlorophenyl side chain is an important lipophilic and/or steric pharmacophore.


High-Energy Aqueous Magnesium Hybrid Full Batteries Enabled by Carrier-Hosting Potential Compensation.
Underachieved capacity and low voltage plateau is ubiquitous in conventional aqueous magnesium ion full batteries. Such limitations originate from the electrochemistry and the low carrier-hosting ((de)intercalation) potential of electrode materials. Herein, via a strategy of enhancing the electrochemistry through carrier-hosting potential compensation, high-energy Mg2+ /Na+ hybrid batteries are achieved. A Mg1.5 VCr(PO4 )3 (MVCP) cathode is coupled with FeVO4 (FVO) anode in a new aqueous/organic hybrid electrolyte, giving reliable high-voltage operation. This operation enables more sufficient (de)intercalation of hybrid carriers (Mg2+ /Na+ ), thereby enhancing the reversible capacity remarkably (233.4 mA h g-1 at 0.5 A g-1 , 92.7 Wh kg-1 electrode , that is, ≥1.75-fold higher than those in conventional aqueous electrolytes). The relatively high Na+ -hosting potential of the electrodes compensates for the low Mg2+ -hosting potential and widens/elevates the discharge plateau of the full battery up to 1.50 V. Mechanism study further reveals an unusual phase transformation of FVO to Fe2 V3 and the low-lattice-strain pseudocapacitive (de)intercalation chemistry of MVCP.


Self-doped conducting polymers in biomedical engineering: Synthesis, characterization, current applications and perspectives.
Recent studies willingly agree that conducting polymers (CPs) are attractive materials for biomedical engineering purposes, mainly because of their unique physicochemical characteristics combining electrical conductivity and high biocompatibility. Nevertheless, the applicability of CPs is restricted by their limited stability under physiological conditions, associated with a decrease in electrical conductivity upon dedoping. Accordingly, modifying chemical structure of CPs to exhibit a self-doping effect seems to be an appealing approach aimed to enhance their functionality. The aim of this review is to provide a current state-of-the-art in the research concerning self-doped CPs, particularly those with potential biomedical applications. After presenting a library of available structure modifications, we describe their physicochemical characteristics, focusing on achievable conductivities, electrochemical, optical and mechanical behaviour, as well as biological properties. To highlight high applicability of self-doped CPs in biomedical engineering, we elaborate on biomedical areas benefiting most from using this type of conducting materials.


Activity of acid phosphatase in resting and mitogen-stimulated human peripheral blood lymphocytes.
Activity of acid phosphatase was found to be higher in T than in non-T cells of human peripheral blood lymphocytes. The T cell mitogen PHA induced an increase in the magnitude of acid phosphatase activity, while the B cell mitogen LPS was not able to produce any effect on acid phosphatase activity. Acid phosphatase activity in lymphocytes correlated with their spontaneous proliferative activity.


Terminalia catappa: Chemical composition, in vitro and in vivo effects on Haemonchus contortus.
Haemonchus contortus is the most important nematode in small ruminant systems, and has developed tolerance to all commercial anthelmintics in several countries. In vitro (egg hatch assay) and in vivo tests were performed with a multidrug strain of Haemonchus contortus using Terminalia catappa leaf, fruit pulp, and seed extracts (in vitro), or pulp and seed powder in lambs experimentally infected with H. contortus. Crude extracts from leaves, fruit pulp and seeds obtained with 70% acetone were lyophilized until used. In vitro, the extracts had LC50=2.48μg/mL (seeds), LC50=4.62μg/mL (pulp), and LC50=20μg/mL (leaves). In vitro, seed and pulp extracts had LC50 similar to Thiabendazole (LC50=1.31μg/mL). Condensed tannins were more concentrated in pulp extract (183.92g of leucocyanidin/kg dry matter) than in either leaf (4.6g) or seed (35.13g) extracts. Phytochemical tests established that all extracts contained alkaloids, flavonoids, saponins, phenols, and terpenoids. Based on these results, in vivo tests were performed to evaluate the anthelmintic activity of T. catappa whole fruit (pulp+seed) powder. Male Santa Ines lambs were artificially infected with multidrug-resistant H. contortus and divided, according to similar fecal egg count (FEC) and weight, into two groups: Control (infected/untreated) and treated (infected/treated with whole fruit powder). Whole fruit powder was mixed with concentrate and provided at 2g/kg of body weight (BW) for five days. After treatment, parasitological analysis (FEC and egg hatch assay), renal profile (urea and creatinine), liver profile (aspartate aminotransferase) and BW were determined. In vitro (based on LC50), seed/pulp extracts had ovicidal effect similar to Thiabendazole but whole fruit powder had no anthelmintic effect on adult nematodes in the abomasum. We discuss the plausible causes of the lack of in vivo activity.


[Phenotypic heterogeneity and phenotype-genotype correlations in dystrophinopathies: Contribution of genetic and clinical databases].
The objective of this work was to study the natural history of dystrophinopathies and the genotype-phenotype correlations made possible by the development of the clinical part of the French DMD database. The collection of 70,000 clinical data for 600 patients with an average longitudinal follow-up of 12years enabled clarification of the natural history of Duchenne and Becker muscular dystrophies and clinical presentations in symptomatic females. We were able to specify the phenotypic heterogeneity of motor, orthopedic and respiratory involvements (severe, standard and intermediary form), of the cardiac disorder (severe, standard or absent cardiomyopathy, absence of correlation between motor and cardiac involvements), and of brain function (mental deficiency in the patients with Becker muscular dystrophy, psychopathological disorders in dystrophinopathies). Phenotypic variability did not correlate with a specific mutational spectrum. We propose a model of phenotypic analysis based on the presence or not of muscular and cardiac involvements (described by age at onset and rate of progression) and brain involvement (described by the type and the severity of the cognitive impairment and of the psychological disorders). The methodology developed for the DMD gene can be generalized and used for other databases dedicated to genetic diseases. Application of this model of phenotypic analysis for each patient and further development of the database should contribute substantially to clinical research providing useful tools for future clinical trials.


Ablative radioembolization of hepatocellular carcinoma with total arterial supply originating from the superior adrenal artery achieved complete pathologic necrosis.
Exclusively extrahepatic arterial perfusion to previously untreated, early-stage, hepatocellular carcinoma (HCC) is uncommon. We present a case of right superior adrenal artery completely supplying a subcapsular HCC in a 68-year-old male. The extrahepatic arterial territory targeted excluded structures potentially vulnerable to radiation, which allowed safe ablative Y-90 radioembolization that achieved complete pathological necrosis.


Five critical elements to ensure the precision medicine.
The precision medicine as a new emerging area and therapeutic strategy has occurred and was practiced in the individual and brought unexpected successes, and gained high attentions from professional and social aspects as a new path to improve the treatment and prognosis of patients. There will be a number of new components to appear or be discovered, of which clinical bioinformatics integrates clinical phenotypes and informatics with bioinformatics, computational science, mathematics, and systems biology. In addition to those tools, precision medicine calls more accurate and repeatable methodologies for the identification and validation of gene discovery. Precision medicine will bring more new therapeutic strategies, drug discovery and development, and gene-oriented treatment. There is an urgent need to identify and validate disease-specific, mechanism-based, or epigenetics-dependent biomarkers to monitor precision medicine, and develop "precision" regulations to guard the application of precision medicine.


cIAP1/TRAF2 interplay promotes tumor growth through the activation of STAT3.
Cellular inhibitor of apoptosis-1 (cIAP1) is a signaling regulator with oncogenic properties. It is involved in the regulation of signaling pathways controlling inflammation, cell survival, proliferation, differentiation and motility. It is recruited into membrane-receptor-associated signaling complexes thanks to the molecular adaptor TRAF2. However, the cIAP1/TRAF2 complex exists, independently of receptor engagement, in several subcellular compartments. The present work strengthens the importance of TRAF2 in the oncogenic properties of cIAP1. cIAPs-deficient mouse embryonic fibroblasts (MEFs) were transformed using the HRas-V12 oncogene. Re-expression of cIAP1 enhanced tumor growth in a nude mice xenograft model, and promoted lung tumor nodes formation. Deletion or mutation of the TRAF2-binding site completely abolished the oncogenic properties of cIAP1. Further, cIAP1 mediated the clustering of TRAF2, which was sufficient to stimulate tumor growth. Our TRAF2 interactome analysis showed that cIAP1 was critical for TRAF2 to bind to its protein partners. Thus, cIAP1 and TRAF2 would be two essential subunits of a signaling complex promoting a pro-tumoral signal. cIAP1/TRAF2 promoted the activation of the canonical NF-κB and ERK1/2 signaling pathways. NF-κB-dependent production of IL-6 triggered the activation of the JAK/STAT3 axis in an autocrine manner. Inhibition or downregulation of STAT3 specifically compromised the growth of cIAP1-restored MEFs but not that of MEFs expressing a cIAP1-mutant and treating mice with the STAT3 inhibitor niclosamide completely abrogated cIAP1/TRAF2-mediated tumor growth. Altogether, we demonstrate that cIAP1/TRAF2 binding is essential to promote tumor growth via the activation of the JAK/STAT3 signaling pathway.


Bilateral sequential and simultaneous rhegmatogenous retinal detachments: anatomic and functional outcomes.
To assess the anatomic and functional outcomes in sequential and simultaneous bilateral rhegmatogenous retinal detachment (BRRD) repair. Retrospective cohort study. A total of 218 eyes in 109 patients. Patients diagnosed with BRRD between 2014 and 2020 at the University Centre of Ophthalmology-CHU de Québec-Laval University were reviewed. Main outcomes were single-surgery anatomic success (SSAS) and final pinhole visual acuity (PHVA) in logMARs. Of the 109 patients, 86 (79%) were male, and median (Q1, Q3) age at presentation for the first eye was 60 years (range, 54-66 years). Of these patients, 92% (n = 100) had sequential BRRD and 8% (n = 9) simultaneous BRRD. Median (Q1, Q3) duration of symptoms was shorter for second eyes (first, 7 days [range, 3-15 days] vs second, 4 days [range, 2-10 days]; p = 0.028). Second eyes also had less retinal tears (first, 2.94 ± 2.76 tears vs second, 2.38 ± 1.79 tears; p = 0.031) and better median preoperative PHVA (first, 0.46 logMAR [range, 0.14-2.30 logMAR] vs second, 0.24 logMAR [range, 0.06-0.95 logMAR]; p = 0.012). SSAS was achieved in 100 (92%) and 101 (93%) of first and second eyes, respectively (p = 1.00). Final PHVA was better for the first eye (first, 0.14 logMAR [range, 0.04-0.30 logMAR] vs second, 0.20 logMAR [range, 0.04-0.43 logMAR]; p = 0.010) but comparable at 3 months (first, 0.30 logMAR [range, 0.14-0.48 logMAR] vs second, 0.34 logMAR [range, 0.13-0.70 logMAR]; p = 0.36). SSAS was similar for both eyes. The subsequent eye was more likely to be treated earlier with less advanced presentations, but at 3 months, PHVA was not significantly different between eyes. Difference in final PHVA may be attributable to longer follow-up in first eyes.


Reconstructing the portal vein through a posterior pancreatic tunnel: New choice for portal vein thrombosis during liver transplantation.
Thrombectomy and anatomical anastomosis (TAA) has long been considered the optimal approach to portal vein thrombosis (PVT) in liver transplantation (LT). However, TAA and the current approach for non-physiological portal reconstructions are associated with a higher rate of complications and mortality in some cases. To describe a new choice for reconstructing the portal vein through a posterior pancreatic tunnel (RPVPPT) to address cases of unresectable PVT. Between August 2019 and August 2021, 245 adult LTs were performed. Forty-five (18.4%) patients were confirmed to have PVT before surgery, among which seven underwent PV reconstruction via the RPVPPT approach. We retrospectively analyzed the surgical procedure and postoperative complications of these seven recipients that underwent PV reconstruction due to PVT. During the procedure, PVT was found in all the seven cases with significant adhesion to the vascular wall and could not be dissected. The portal vein proximal to the superior mesenteric vein was damaged in one case when attempting thrombolectomy, resulting in massive bleeding. LT was successfully performed in all patients with a mean duration of 585 min (range 491-756 min) and mean intraoperative blood loss of 800 mL (range 500-3000 mL). Postoperative complications consisted of chylous leakage (n = 3), insufficient portal venous flow to the graft (n = 1), intra-abdominal hemorrhage (n = 1), pulmonary infection (n = 1), and perioperative death (n = 1). The remaining six patients survived at 12-17 mo follow-up. The RPVPPT technique might be a safe and effective surgical procedure during LT for complex PVT. However, follow-up studies with large samples are still warranted due to the relatively small number of cases.


Open reduction and internal fixation of three- and four-part proximal humeral fractures by intrafocal distraction: a clinical and radiographic study of thirty-two cases with five to ten years of follow-up.
The disadvantage of ORIF of proximal humerus fracture (PHF) by intrafocal distraction is that once the device is inserted, it cannot be removed. This study was designed to evaluate the tolerance of such a titanium alloy device at five years of minimal follow-up (FU) and secondarily to assess the relevance of the osteosynthesis of these fractures. Thirty-two patients operated for PHF and implanted with a titanium alloy device between 2009 and 2011 were reviewed: fifteen three-part fracture (3-PF) with mean age 65.2 ± seven years and mean FU at 86 months, and seventeen four-part (4-PF) with mean age 62.9 ± 12 years and mean FU at 88 months. There were no signs of clinical or radiological incompatibility to the metal. The lateral cortex of the humeral shaft at the wedge component level of the device was thinner than pre-operatively in half of the cases. For the fifteen 3-PF, the median raw and weighted constant score (CS) were 75 and 100, respectively. Only one case presented avascular necrosis (AVN). For the seventeen 4-PF, the median raw and weighted CS were 64 and 88, respectively. Seven cases presented AVN and none of them seven had initially a dorsomedial metaphyseal extension of the humeral head. Kaplan-Meier survivorship analysis, with 95% confidence interval, was calculated at 89.7% (79.7-100%) survivorship at 7.18 years of follow-up. This study shows no incompatibility of the titanium alloy device, radiological signs of localized stress shielding in half of the cases with no functional impact, excellent clinical and radiological evolution of the 3-PF, and AVN in all 4-PF without dorsomedial metaphyseal extension of the humeral head whereas most of cases without AVN had dorsomedial metaphyseal extension.


Studies on the arrangement of the collagenous fibers in infant epiphyseal plates using polarized light and the scanning electron microscope.
The architecture of the collagenous fibers in epiphyseal plates of infantile long bones was investigated using polarized light and scanning electron microscope. In the zone of columnar cartilage the collagenous fiber bundles are arranged longitudinally in the intercolumnar cartilaginous septa. In addition to this main longitudinal orientation, some of the fibers branch off at an acute angle and flow through the cartilage columns surrounding rhombus-shaped groups of chondrocytes. In the area adjaced to the inert cartilage the collagen fibers become curved and form a network of arcades. In the zone of inert cartilage the collagenous fibers are arranged in a honeycomb-like fashion. The importance of the arrangement of the collagen fibers to the loading strength of the epiphyseal plate is discussed.


Adolescents' adherence to Centers for Disease Control and Prevention guidelines during the COVID-19 pandemic.
The outbreak of the coronavirus (COVID-19) pandemic in the United States resulted in safety guidelines from the Centers for Disease Control and Prevention (CDC) intended to curb the spread of the virus. Adolescents are potentially at risk for disregarding these guidelines due to their reduced psychosocial maturity compared with adults. The current study examined the relationship between adolescents' psychosocial maturity, perceived importance of the CDC guidelines and adherence to the CDC guidelines within some of the highest risk groups for contracting COVID-19 in a county particularly impacted by the pandemic (i.e., Hispanic and low-SES youth in El Paso, Texas). Participants completed a phone interview with a research assistant regarding their thoughts and behaviours in the initial months of the COVID-19 pandemic. Adolescents (N = 68) were 15.38 years old on average (SD = 1.05, range = 13, 17), predominantly male (60.3%) and nearly exclusively Hispanic/Latino (94.1%). Results indicated that although more psychosocially mature adolescents reported greater adherence to the CDC guidelines than less psychosocially mature adolescents, the association between psychosocial maturity and adherence was fully mediated by how important adolescents felt it was to follow the guidelines. Specifically, greater perceived importance was associated with greater adherence to the guidelines. The current study found that more psychosocially mature adolescents adhere to CDC's safety guidelines better than less psychosocially mature adolescents because they are more likely to view the guidelines as important. Information that attempts to increase adolescent adherence to the guidelines should therefore emphasize not only that following the guidelines is important, but why following the guidelines is so important. Less psychosocially mature adolescents may benefit most from interventions efforts and targeted messages regarding the importance of following the CDC's guidelines, as more psychosocially mature adolescents already recognize this importance.


Sphingosine 1-phosphate mediates chemotaxis of human primary fibroblasts via the S1P-receptor subtypes S1P₁ and S1P₃ and Smad-signalling.
The sphingolipid sphingosine 1-phosphate (S1P) induces chemotaxis of primary fibroblasts. Thus, S1P exhibited a chemotactic effect in a concentration-dependent manner from 10⁻⁶ to 10⁻⁹ M; higher concentrations resulted in a loss of migration, and lower amounts were ineffective to evoke movement toward a concentration gradient of S1P. In congruence with the migratory response, S1P caused an extension of lamellipodia at the cell periphery of human fibroblasts and a rearrangement of the cytoskeleton. These effects were visible by phalloidin staining of actin filaments as well as focal adhesion turnover. As the molecular mechanism of S1P-mediated migration of fibroblasts has not been well characterized, we investigated whether S1P-receptors are involved in the chemotactic response. Indeed, inhibition of G(i) signalling markedly reduced motility towards S1P, suggesting an involvement of S1P-receptor subtypes. Moreover, downregulation of S1P₁ and S1P₃ indicated that these S1P-receptor subtypes are responsible for the chemotactic action of the bioactive sphingolipid. After having identified a crosstalk between Smad-proteins and S1P-signalling, we investigated whether Smad-activation is involved in the chemotactic response induced by S1P. Indeed S1P caused a Smad-activation via the S1P receptor subtypes S1P₁ and S1P₃. Moreover, downregulation of Smad3 diminished the ability of S1P to mediate a chemotactic response in fibroblasts, indicating a crosstalk between TGF-β- and S1P-signalling.


Point-activated ESR1Y541S has a dramatic effect on the development of sexually dimorphic organs.
Mutations in the estrogen receptor α (ERα) occur in endocrine-resistant metastatic breast cancer. However, a major gap persists with the lack of genetically tractable immune competent mouse models to study disease. Hence, we developed a Cre-inducible murine model expressing a point-activated ESR1Y541S (ESR1Y537S in humans) driven by its endogenous promoter. Germline expression of mutant ESR1Y541S reveals dramatic developmental defects in the reproductive organs, mammary glands, and bones of the mice. These observations provide critical insights into the tissue-specific roles of ERα during development and highlights the potential use of our model in further developmental and cancer studies.


Experimental intraocular proliferation and neovascularization.
An experimental model of massive periretinal proliferation and intraocular neovascularization, produced in rabbits by the intravitreal injection of 250,00 cultured heterologous fibroblasts, showed no significant difference in the detachment rate (69% to 100%) or neovascularization rate (45% to 88%) between the animals injected with autologous cells and those injected with heterologous cells. Dermal fibroblasts produced a slightly higher detachment rate than conjunctival fibroblasts and were equally effective after reconstitution and subculture from liquid nitrogen storage in 7% dimethyl sulfoxide. Heterologous cells produced no clinical or histologic evidence of rejection when compared with autologous cells in the same animal and had the following advantages: (1) elimination of several biopsies and extended cell culture time; (2) a ready source of cryopreserved cells is available; (3) multiple injections of many animals can be performed within a short time; (4) in vivo and in vitro drug testing can be correlated on the same cell line.


Construction and application of surveillance and response systems for parasitic diseases in China, led by NIPD-CTDR.
Parasitic diseases have been widely epidemic in China with a long history. Great endeavours made in past 70 years led to significant decrease in morbidity and mortablity caused by several major parasitic diseases, while challenges existed to eliminate parasitic diseases. Surveillance-response system has play a crucial role in identifying public health problems, ascertaining the distribution and epidemic dynamics, discovering outbreaks and epidemic anomalies, evaluating the effects of on-site intervention activities and identifying risk factors. In this article, we reviewed the progress of the surveillance system for parasitic diseases, analysed the role of NIPD in the construction and application of surveillance-response system of parasitic diseases through elaborating the surveillance activities and typical surveillance-response events led by NIPD. Suggestion and comments for improve the surveillance-response system were put forward for further control or elimination of parasitic diseases.


Purification and characterization of a fatty acyl-CoA hydrolase from the uropygial glands of Peking ducks (Anas domesticus).
In a previous communication the occurrence of a medium-chain acyl-CoA hydrolase designated thioesterase-B in the uropygial gland of mallard ducks was reported [L. Rogers, P. E. Kolattukudy, and M. J. de Renobales (1982) J. Biol. Chem. 257, 880-886]. In the present study, thioesterase-B was purified from the postmicrosomal supernatant of homogenized uropygial glands from Peking ducks (Anas domesticus). Most of the contaminating thioesterase activities were removed by ammonium sulfate fractionation. The 55% ammonium sulfate supernatant, containing thioesterase-B, was chromatographed on hydroxylapatite followed by gel filtration on Sephadex G-100. The remaining contaminants were removed by chromatofocusing followed by desalting on Sephadex G-75. This procedure gave a 26% yield with a nearly 200-fold purification. Gel filtration of the purified enzyme showed that the molecular weight of the native enzyme was 56,300, whereas sodium dodecyl sulfate-gel electrophoresis of components separated by chromatofocusing showed that the purified enzyme contained enzymatically active proteins of molecular weights 59,400, 58,300, 56,000, and 55,800. The four species differed slightly in pI (4.9, 4.7, 4.45, and 4.40) but they were kinetically and immunologically indistinguishable. All four had the same N-terminal sequence. The purified thioesterase preparation showed a pH optimum of 9.3 with C12-CoA but the pH optimum was dependent on the chain length of the acyl group. At pH 8.0, C10 was the preferred substrate with less activity on C12, C8, and C14. The enzymatic activity was stimulated by bovine serum albumin and was inhibited by p-hydroxymercuribenzoate. Involvement of active serine in catalysis was suggested by inhibition of the enzyme by diethylpyrocarbonate, diisopropylfluorophosphate and phenylmethylsulfonyl fluoride.


Reliability of Graders and Comparison with an Automated Algorithm for Vertical Cup-Disc Ratio Grading in Fundus Photographs.
We aimed to investigate the intergrader and intragrader reliability of human graders and an automated algorithm for vertical cup-disc ratio (CDR) grading in colour fundus photographs. Two-hundred fundus photographs were selected from a database of 3000 photographs of patients screened at a tertiary ophthalmology referral centre. The graders included glaucoma specialists (n = 3), general ophthalmologists (n = 2), optometrists (n = 2), family physicians (n = 2) and a novel automated algorithm (AA). In total, 2 rounds of CDR grading were held for each grader on 2 different dates, with the photographs presented in random order. The CDR values were graded as 0.1-1.0 or ungradable. The grading results of the 2 senior glaucoma specialists were used as the reference benchmarks for comparison. The intraclass correlation coefficient values ranged from 0.37-0.74 and 0.47-0.97 for intergrader and intragrader reliability, respectively. There was no significant correlation between the human graders' level of reliability and their years of experience in grading CDR (P = 0.91). The area under the curve (AUC) value of the AA was 0.847 (comparable to AUC value of 0.876 for the glaucoma specialist). Bland Altman plots demonstrated that the AA's performance was at least comparable to a glaucoma specialist. The results suggest that AA is comparable to and may have more consistent performance than human graders in CDR grading of fundus photographs. This may have potential application as a screening tool to help detect asymptomatic glaucoma-suspect patients in the community.


Characterization of carrot pectin methylesterase.
The most alkaline form of pectin methylesterase was purified from ripe carrot roots and used for structural analysis. Determination of an N-terminal blocking group and of the primary structure allowed comparisons with other forms, and facilitated crystallographic determination of the three-dimensional structure. The mature enzyme has 319 residues and the N-terminal blocking group was shown to be a pyroglutamyl residue derived from a glutaminyl cyclization. Few other methylesterases have been isolated and assigned to exact mature forms, and together with the present enzyme, only two have been analyzed in three-dimensional structure. However, comparison of 39 forms, mainly from GenBank data, reveals clear relationships and identifies subgroups of this enzyme type, deviating in structure but centering around two functionally important and conserved Asp residues at positions 136 and 157 in the carrot enzyme.


Oxidative cleavage of premithramycin B is one of the last steps in the biosynthesis of the antitumor drug mithramycin.
Mithramycin is a member of the clinically important aureolic acid group of antitumor drugs that interact with GC-rich regions of DNA nonintercalatively. These drugs contain a chromophore aglycon that is derived from condensation of ten acetate units (catalyzed by a type II polyketide synthase). The aglycones are glycosylated at two positions with different chain length deoxyoligosaccharides, which are essential for the antitumor activity. During the early stages of mithramycin biosynthesis, tetracyclic intermediates of the tetracycline-type occur, which must be converted at later stages into the tricyclic glycosylated molecule, presumably through oxidative breakage of the fourth ring. Two intermediates in the mithramycin biosynthetic pathway, 4-demethyl-premithramycinone and premithramycin B, were identified in a mutant lacking the mithramycin glycosyltransferase and methyltransferase genes and in the same mutant complemented with the deleted genes, respectively. Premithramycin B contains five deoxysugars moieties (like mithramycin), but contains a tetracyclic aglycon moiety instead of a tricyclic aglycon. We hypothesized that transcription of mtmOIV (encoding an oxygenase) was impaired in this strain, preventing oxidative breakage of the fourth ring of premithramycin B. Inactivating mtmOIV generated a mithramycin nonproducing mutant that accumulated premithramycin B instead of mithramycin. In vitro assays demonstrated that MtmOIV converted premithramycin B into a tricyclic compound. In the late stages of mithramycin biosynthesis by Strepyomyces argillaceus, a fully glycosylated tetracyclic tetracycline-like intermediate (premithramycin B) is converted into a tricyclic compound by the oxygenase MtmOIV. This oxygenase inserts an oxygen (Baeyer-Villiger oxidation) and opens the resulting lactone. The following decarboxylation and ketoreduction steps lead to mithramycin. Opening of the fourth ring represents one of the last steps in mithramycin biosynthesis.


T-cell populations responsive to bovine respiratory syncytial virus in seronegative calves.
Calves lacking detectable serum antibodies against bovine respiratory syncytial virus (BRSV) were screened for virus-specific T-cell memory. Peripheral blood mononuclear cells were cultured in vitro with live BRSV and analyzed by dual-color flow cytometry for surface expression of CD25 on CD4(+), CD8(+), and gammadeltaT-cells. Significant recall responses were detected in some of the seronegative calves. Modified live BRSV vaccine was administered to these and to a group of non-responding calves. Following vaccination, virus-specific IgG, virus neutralizing antibody, and T-cell recall responses were all elevated more rapidly in the group with BRSV-sensitive T-cells than in the T-cell-negative group, which suggested that calves in the first group were previously exposed to BRSV. This demonstrates that exposure to BRSV can induce T and B cell memory in young calves without causing seroconversion. The calves were presumably exposed to BRSV while they had maternal antibody, which inhibited the calves from developing an antibody response.


A problem-oriented analysis of database models.
Which is the most convenient database model considering specific applications? The goal of this paper is to try to answer this question by the use of a chemical example. Examples of requests describe the problems of insertion, deletion, and updating; these requests are analyzed for the hierarchical model and are expressed in a relational language defined by the authors and in Socrate for the network model.


The role of implementation organizations in scaling evidence-based psychosocial interventions.
To bring evidence-based interventions (EBIs) to individuals with behavioral health needs, psychosocial interventions must be delivered at scale. Despite an increasing effort to implement effective treatments in communities, most individuals with mental health and behavioral problems do not receive EBIs. We posit that organizations that commercialize EBIs play an important role in disseminating EBIs, particularly in the USA. The behavioral health and implementation industry is growing, bringing the implementation field to an important inflection point: how to scale interventions to improve access while maintaining EBI effectiveness and minimizing inequities in access to psychosocial intervention. We offer a first-hand examination of five illustrative organizations specializing in EBI implementation: Beck Institute for Cognitive Behavioral Therapy; Incredible Years, Inc.; the PAXIS Institute; PracticeWise, LLC; and Triple P International. We use the Five Stages of Small Business Growth framework to organize themes. We discuss practical structures (e.g., corporate structures, intellectual property agreements, and business models) and considerations that arise when trying to scale EBIs including balancing fidelity and reach of the intervention. Business models consider who will pay for EBI implementation and allow organizations to scale EBIs. We propose research questions to guide scaling: understanding the level of fidelity needed to maintain efficacy, optimizing training outcomes, and researching business models to enable organizations to scale EBIs.