Gender- and Sexuality-Minoritised Adolescents in DBT: A Reflexive Thematic Analysis of Minority-Specific Treatment Targets and Experience.

Gender- and sexuality-minoritised (GSM) adolescents are at increased risk of self-harm and suicidal behaviours compared to their cisgender and heterosexual peers. This increased risk is thought to be explained in part by exposure to stigma and societal oppression. Dialectical Behaviour Therapy (DBT) is an evidence-based intervention for self-harm and suicidal behaviour that may have advantages for supporting GSM young people in distress. No study has yet sought to understand what GSM-associated difficulties may be important to consider in DBT for adolescents, or the experiences of GSM young people in a standard DBT programme. Therefore, this study aimed to understand the experiences of GSM young people in DBT and what difficulties and dilemmas associated with their gender and sexuality diversity were thought by them to be important to target in DBT. Qualitative interviews were conducted with 14 GSM young people in a comprehensive DBT programme and were analysed using Reflexive Thematic Analysis. The analysis was supported by two further GSM young people who had finished DBT. The findings were split into three overarching themes (Identity, Impact of Others, and Space for Sexual and Gender Identity in DBT), each with themes within. The identity-based theme included "identity confusion and acceptance"; the relationship-based themes included "cis-Heterosexism" and "community connectedness"; and the space within DBT themes included "negotiating focus and targeting in DBT" and "creating safety in DBT". Findings are discussed in relation to implications and recommendations for therapists working with GSM young people within and outside of DBT.

Melatonin strongly enhances the Agrobacterium- mediated transformation of carnation in nitrogen-depleted media.

With the rising demand for new cultivars of carnation, efficient transformation protocols are needed to enable the bioengineering of new traits. Here, we established a novel and efficient Agrobacterium-mediated transformation system using callus as the target explant for four commercial carnation cultivars. Leaf-derived calli of all cultivars were inoculated with Agrobacterium tumefaciens strain LBA4404 containing the plasmid pCAMBIA 2301 harboring genes for ß-glucuronidase (uidA) and neomycin phosphotransferase (nptII). Polymerase chain reaction (PCR) and histochemical assays confirmed the presence of uidA and ß-glucuronidase (GUS), respectively in transgenic shoots. The effect on transformation efficiency of medium composition and the presence of antioxidants during inoculation and co-cultivation was investigated. The transformation efficiency was increased in Murashige and Skoog (MS) medium lacking KNO3 and NH4NO3, and also in MS medium lacking macro and micro elements and Fe to 5% and 3.1% respectively, compared to 0.6% in full-strength medium. Transformation efficiency was increased dramatically to 24.4% across all carnation cultivars by the addition of 2 mg/l melatonin to nitrogen-depleted MS medium. Shoot regeneration was also doubled in this treatment. The establishment of this efficient and reliable transformation protocol can advance the development of novel carnation cultivars through molecular breeding approaches.

Hunting the eagle killer: A cyanobacterial neurotoxin causes vacuolar myelinopathy.

Vacuolar myelinopathy is a fatal neurological disease that was initially discovered during a mysterious mass mortality of bald eagles in Arkansas in the United States. The cause of this wildlife disease has eluded scientists for decades while its occurrence has continued to spread throughout freshwater reservoirs in the southeastern United States. Recent studies have demonstrated that vacuolar myelinopathy is induced by consumption of the epiphytic cyanobacterial species Aetokthonos hydrillicola growing on aquatic vegetation, primarily the invasive Hydrilla verticillata Here, we describe the identification, biosynthetic gene cluster, and biological activity of aetokthonotoxin, a pentabrominated biindole alkaloid that is produced by the cyanobacterium A. hydrillicola We identify this cyanobacterial neurotoxin as the causal agent of vacuolar myelinopathy and discuss environmental factors-especially bromide availability-that promote toxin production.

Incidence of cleft-related speech problems in children with an isolated cleft lip.

OBJECTIVES: Clinicians agree that children with isolated cleft lip have fewer cleft-associated problems than children with cleft lip and palate. Unfortunately, for isolated cleft lip children, the risk of cleft-associated problems is unknown and maybe underestimated. Often, these children do not get the required follow-up by a multidisciplinary team and thereby not the known benefits in supporting their development. This study examines the incidence of cleft-related speech problems and ear problems in children with isolated cleft lip. MATERIALS AND METHODS: A prospective study was performed on all children born with an isolated cleft lip and treated at the Wilhelmina Children's Hospital in Utrecht between January 2007 and April 2014. Data were collected for sex, date of birth, genetics, cleft lip type, date of cleft lip repair, type of repair, speech/language problems, and ear problems. RESULTS: This study included 75 patients (59% male). The mean age of the children at the moment of speech examination was 32.5 months (SD 6.1). Eighteen of the 75 children (24%) needed speech and language therapy; however, only one child (1.3%) had a cleft-related speech problem. Sixteen of the 75 patients (21%) reported a history of one or more episodes of acute otitis media (AOM)/otitis media with effusion (OME) during the first 6 years. CONCLUSION/CLINICAL RELEVANCE: This is the first prospective study analyzing the incidence of cleft-related speech problems in children with an isolated cleft lip. These children do not have a higher risk of cleft-related speech problems or AOM/OME when compared to the general population. However, children with an isolated cleft do have a higher incidence of speech therapy.

Intra- and interspecific diversity analyses in the genus Eremurus in Iran using genotyping-by-sequencing reveal geographic population structure.

Eremurus species, better known as 'Foxtail Lily' or 'Desert Candle', are important worldwide in landscaping and the cut-flower industry. One of the centers of highest diversity of the genus Eremurus is Iran, which has seven species. However, little is known about the genetic diversity within the genus Eremurus. With the advent of genotyping-by-sequencing (GBS), it is possible to develop and employ single nucleotide polymorphism (SNP) markers in a cost-efficient manner in any species, regardless of its ploidy level, genome size or availability of a reference genome. Population structure and phylogeographic analyses of the genus Eremurus in Iran using a minimum of 3002 SNP markers identified either at the genus level or at the species level from GBS data showed longitudinal geographic structuring at the country scale for the genus and for the species E. spectabilis and E. luteus, and at the regional scale for E. olgae. Our analyses furthermore showed a close genetic relatedness between E. olgae and E. stenophyllus to the extent that they should be considered subspecies within an E. olgae/stenophyllus species complex. Their close genetic relatedness may explain why crosses between these two (sub)species have been found in the wild and are exploited extensively as ornamentals. Last, current species identification, while robust, relies on flower morphology. A subset of seven SNPs with species-specific (private) alleles were selected that differentiate the seven Eremurus species. The markers will be especially useful for cultivar protection and in hybrid production, where true hybrids could be identified at the seedling stage.

Development of embryogenic dogwood cultures and the regeneration of plants.

Somatic embryogenesis in flowering dogwood (Cornus florida) has been achieved, but not the regeneration of plants with active shoot growth. To improve plant regeneration, eight media treatments were tested for induction of somatic embryogenesis from immature zygotic embryos. Somatic embryogenesis was obtained on three media containing the plant growth regulators (PGRs) 0.1 mg l-1 picloram, 2.0 mg l-1 2,4-dichlorophenoxyacetic acid, or 0.1 mg l-1 indole-3-butyric acid (IBA). Somatic embryogenesis was also induced on Woody Plant Medium without PGRs. Overall, 92% of the somatic embryos examined converted into plants with active root and shoot growth. This is the first report of somatic embryo-derived plants of C. florida that have active shoot growth and that could be transferred to soil. Embryogenic suspensions were established from IBA-treated cultures that could serve as a target for C. florida bioengineering.

Internationally adopted children with cleft lip and/or palate: A retrospective cohort study.

OBJECTIVE: The treatment approach for internationally adopted children with cleft lip and/or palate differs from locally born children with cleft lip and/or palate. They are older at initial presentation, may have had treatment abroad of different quality, and are establishing new and still fragile relationships with their adoptive parents. The aim of this study was to describe the characteristics and initial care and treatment of this group. METHODS: A retrospective cohort study was performed including all internationally adopted children with cleft lip and/or palate presenting to the cleft team outpatient clinic in the Wilhelmina Children's Hospital between January 1994 and December 2014. Medical records of all patients were reviewed; information concerning demographic characteristics, characteristics at initial presentation, and treatment were obtained. RESULTS: A total number of 132 adopted patients were included: 15% had cleft lip, 7% had cleft palate, and 78% had cleft lip and palate. The average age at the time of adoption was 26.5 months. In most cases, China was the country of origin. Seventy-eight percent had surgery in their country of origin, primarily lip repair. Fistulae in need of revision surgery were found in 8% of the patients. Pharyngoplasty was needed in 48% of the patients. No significant differences were found for mean age at adoption, gender, cleft type, and one- or two-stage palatal closure. CONCLUSION: Internationally adopted children with cleft lip and/or palate are a very diverse group of patients with challenging treatment. These children undergo surgery late and frequently need additional surgery.

State of the science and challenges of breeding landscape plants with ecological function.

Exotic plants dominate esthetically-managed landscapes, which cover 30-40 million hectares in the United States alone. Recent ecological studies have found that landscaping with exotic plant species can reduce biodiversity on multiple trophic levels. To support biodiversity in urbanized areas, the increased use of native landscaping plants has been advocated by conservation groups and US federal and state agencies. A major challenge to scaling up the use of native species in landscaping is providing ornamental plants that are both ecologically functional and economically viable. Depending on ecological and economic constraints, accelerated breeding approaches could be applied to ornamental trait development in native plants. This review examines the impact of landscaping choices on biodiversity, the current status of breeding and selection of native ornamental plants, and the interdisciplinary research needed to scale up landscaping plants that can support native biodiversity.

Methemoglobinemia and hemolytic anemia after rasburicase administration in a child with leukemia.

CASE: We report a case of simultaneous methemoglobinemia and hemolytic anemia, probably related to the use of rasburicase, in a child starting treatment for acute lymphoblastic leukemia (ALL). In addition, the patient developed symptoms of pancreatitis. All complications resolved after 3 days of supportive therapy. CONCLUSION: Although usually well tolerated in pediatrics, physicians prescribing rasburicase should be aware of these possible life-threatening adverse reactions.

Mutation scanning of peach floral genes.

BACKGROUND: Mutation scanning technology has been used to develop crop species with improved traits. Modifications that improve screening throughput and sensitivity would facilitate the targeted mutation breeding of crops. Technical innovations for high-resolution melting (HRM) analysis are enabling the clinic-based screening for human disease gene polymorphism. We examined the application of two HRM modifications, COLD-PCR and QMC-PCR, to the mutation scanning of genes in peach, Prunus persica. The targeted genes were the putative floral regulators PpAGAMOUS and PpTERMINAL FLOWER I. RESULTS: HRM analysis of PpAG and PpTFL1 coding regions in 36 peach cultivars found one polymorphic site in each gene. PpTFL1 and PpAG SNPs were used to examine approaches to increase HRM throughput. Cultivars with SNPs could be reliably detected in pools of twelve genotypes. COLD-PCR was found to increase the sensitivity of HRM analysis of pooled samples, but worked best with small amplicons. Examination of QMC-PCR demonstrated that primary PCR products for further analysis could be produced from variable levels of genomic DNA. CONCLUSIONS: Natural SNPs in exons of target peach genes were discovered by HRM analysis of cultivars from a southeastern US breeding program. For detecting natural or induced SNPs in larger populations, HRM efficiency can be improved by increasing sample pooling and template production through approaches such as COLD-PCR and QMC-PCR. Technical advances developed to improve clinical diagnostics can play a role in the targeted mutation breeding of crops.

Methemoglobinemia and hemolytic anemia after rasburicase administration in a child with leukemia.

CASE: We report a case of simultaneous methemoglobinemia and hemolytic anemia, probably related to the use of rasburicase, in a child starting treatment for acute lymphoblastic leukemia (ALL). In addition, the patient developed symptoms of pancreatitis. All complications resolved after 3 days of supportive therapy. CONCLUSION: Although usually well tolerated in pediatrics, physicians prescribing rasburicase should be aware of these possible life-threatening adverse reactions.

Soluble HLA-G is an independent factor for the prediction of pregnancy outcome after ART: a German multi-centre study.

BACKGROUND: Soluble HLA-G (sHLA-G) has been suggested as a non-invasive marker for embryo selection to improve pregnancy rates after assisted reproduction technique (ART). Our study aimed at the identification of parameters influencing the detection of sHLA-G in embryo cultures (ECs) and at the prognostic relevance of sHLA-G in a multi-centre study. METHODS: In total 4212 EC from 2364 cycles were randomly collected from 29 German ART centres and analysed for sHLA-G by Luminex-based technology. RESULTS: Among test and culture conditions, only the cleavage stage of the embryo was identified as an independent factor for sHLA-G detection (P < 0.001). Overall, sHLA-G was significantly associated with pregnancy after ART [P < 0.001; odds ratio: 2.0 (95% CI: 1.7-2.4)], suggesting that sHLA-G testing might improve the pregnancy rate from 30 to 40%. Importantly, the sHLA-G status of embryos could be associated with pregnancy after single embryo transfer [P = 0.002; odds ratio: 3.3 (95% CI: 1.5-6.8)] doubling the probability of pregnancy rate to 26% after sHLA-G testing. The patient's age, number of transferred embryos, morphological grading [EXP(B): 4.3 (95% CI: 2.1-8.9)] of embryos and sHLA-G status [EXP(B): 2.3 (95% CI: 1.8-3.1)] were independent predictors of pregnancy, with the latter two being most powerful. CONCLUSIONS: This study provides significant evidence that the morphological scoring system is still the best strategy for the selection of embryos but that sHLA-G might be considered as a second parameter if a choice has to be made between embryos of morphologically equal quality.

The prognostic significance of soluble NKG2D ligands in B-cell chronic lymphocytic leukemia.

Soluble or membrane-anchored ligands of NKG2D and their receptor have a critical role in the elimination of tumor cells and disease progression. Plasma samples of 98 patients with B-cell chronic lymphocytic leukemia (CLL) were analyzed with specific ELISA systems for soluble major histocompatibility complex class I-related chains (sMICA and sMICB) and UL-16-binding proteins (ULBP1, 2, and 3). The flow cytometric analysis of MICA on CLL cells and natural killer group 2 member D (NKG2D) receptors on NK cells was performed after thawing of frozen peripheral blood lymphocytes of CLL patients (N=51). Levels of sMICA, sMICB, and sULBP2 were significantly increased (P<0.001) compared with 48 controls, whereas sULBP1 3 were not detectable in patients and controls. Levels of sMICA>990 pg/ml (P=0.014), sMICB>200 pg/ml (P=0.0001), and sULBP2>105 pg/ml (P<0.0001) were associated with poor treatment-free survival (TFS). Neither MICA nor NKG2D expression could be related to clinical parameters. In multivariate analysis Binet stage (P=0.002), sULBP2 (P=0.002) and ZAP-70 (P=0.002) were independent predictive factors for TFS. In patients with Binet stage A, sULBP2 levels>105 pg/ml were strongly associated (P=0.0025) with poor TFS. Our data show that soluble but not membrane-anchored NKG2D ligands or receptors are of prognostic significance in CLL. Moreover, sULBP2 seems to be useful to identify early-stage patients with risk of disease progression.

TNF-alpha and IL-10 gene polymorphisms show a weak association with pemphigus vulgaris in the Slovak population.

BACKGROUND: Pemphigus vulgaris is a rare chronic autoimmune disease of skin and mucous membranes, with several cytokines participating in its development. The role of their gene polymorphisms in susceptibility to the disease is, however, not fully understood. OBJECTIVE: The aim of our case-control study was to investigate whether some of 22 single nucleotide polymorphisms (SNPs) in 13 cytokine genes (IL-1alpha, IL-1beta, IL-1RI, IL-1Ra, IL-4Ralpha, IL-12, IFN-gamma, TGF-beta1, TNF-alpha, IL-2, IL-4, IL-6 and IL-10) are associated with pemphigus vulgaris in the Slovak population. METHODS: DNA samples were obtained from 34 pemphigus vulgaris patients and 140 healthy controls of Slovak origin. Cytokine gene SNPs were determined using the polymerase chain reaction with sequence-specific primers (PCR-SSP) method. Results We found a weak association between pemphigus vulgaris and polymorphic variants in TNF-alpha and IL-10 genes only, with haplotypes TNF-alpha-308G/-238G and IL-10 -1082A/-819C/-592C being significantly overrepresented in pemphigus vulgaris patients (TNF-alpha GG: 94.12% vs. 82.86%, P = 0.0216; IL-10 ACC: 44.12% vs. 30.00%, P = 0.0309). CONCLUSIONS: Our preliminary results suggest that certain TNF-alpha and IL-10 gene polymorphisms might contribute to genetic susceptibility to pemphigus vulgaris; however, their overall impact on disease development will be rather limited.

One or three intradermal injections within one week for rabies pre-exposure immunization.

Pre-exposure rabies vaccination is recommended fortravellers to endemic countries and forselected populations in highly endemic regions. It consists of three injections administered over 3-4 weeks. Travellers often ignore or do not have enough time to receive a complete course prior to departure or leave with only one or two injections. In this study, the secondary antibody response in volunteers who received one or three injections within one week is evaluated. Results are compared to the recommended three injection regimen when administered with two booster vaccinations on days 0 and 3 one year later. Long-lasting immune memory in volunteers who received only one time vaccination of 0.1 ml (1 site) intradermally and then received two booster vaccinations of 0.1 ml (1 site) intradermally on days 0 and 3, one or three years later is also reported. All volunteers developed an accelerated and adequate neutralizing antibody response within seven days after they received two boosters. These data suggest that three injections within one week are as immunogenic as three injections within four weeks. One clinic visit with two-site 0.1 ml ID injections or a one-site single 0.1 ml pre-exposure rabies vaccination is enough to prime the host immune memory for at least one to three years.

Ten-day observation of live rabies suspected dogs.

This study aimed at analyzing a ten-day observation period of rabies suspected dogs and cats according to six criteria. Dogs and cats suspected of being rabid were brought for observation when they had either bitten a person or another animal or when abnormal behaviour or unusual illness was observed. Between 1985 and 2005, retrospective and prospective data from 1,222 dogs and 303 cats was collected during the ten-day observation period. If an animal had died, brain examination using fluorescent antibody testing was routinely performed. If an animal had survived for > or =10 days, it was released to its owner or transferred to the municipal dog shelter. A total of 644 dogs and 58 cats found rabid died within 10 days of observation. In addition, for 208 dogs confirmed rabid with laboratory tests between 1997 and 2005, six criteria were analysed from the day of submission. This experience with the implemented 10-day observation period confirms the WHO recommendation on identifying suspected rabid dogs or cats under veterinary supervision following a human exposure.

Prevalence of latent tuberculosis infection in German radiologists.

Healthcare workers have an increased risk of tuberculosis infection compared with the general population. There have been few attempts to quantify the prevalence of latent tuberculosis infection amongst German healthcare workers, due to inadequacy of the current tuberculin skin test (TST). Therefore, it was our aim to investigate the prevalence of latent tuberculosis in this cohort using a tuberculosis-specific ELISpot (T-SPOT.TB) test and to compare the performance of this test to that of the TST. Ninety-five healthy participants working in departments of radiology were examined by ELISpot, lymphocyte transformation test and TST. For cellular in-vitro tests, tuberculosis-specific peptides and purified protein derivate (PPD) were used as antigens. These tests were combined with a questionnaire on prior tuberculosis exposure. Out of 95 healthcare workers, only one (1%) was defined as positive by T-SPOT.TB, 92 (97%) by PPD-ELISpot, 78 (82%) by PPD-lymphocyte transformation test and 32 (34%) by TST. Multivariate analysis showed that the TST was significantly affected (P<0.0001 and P=0.001, respectively) by foreign birth and prior skin testing. The T-SPOT.TB test results were independent of foreign birth, prior skin testing and prior vaccination against tuberculosis. In contrast to the TST, T-SPOT.TB appears to be an accurate and useful tool to track tuberculosis infection in this at-risk group. With only one of 95 participants having acquired latent tuberculosis, these preliminary results argue for a low incidence of latent tuberculosis in German radiologists.

Detection of chromium allergy by cellular in vitro methods.

BACKGROUND: The standard assay for the detection of chromium sensitization, the patch test, does not allow discrimination between patients with and without clinical symptoms of allergy. OBJECTIVE: The aim of this study was to prove whether cellular in vitro tests are predictive of chromium allergy. METHODS: Chromium-sensitized volunteers with and without clinically manifest allergy and non-sensitized healthy controls (n=37, 19, and 26, respectively) were analysed by cellular in vitro methods using tri- and hexavalent chromium (chromium chloride and potassium dichromate) as stimuli. The results were correlated with clinical and anamnestic data. RESULTS: Sensitized individuals with an allergy displayed significantly higher lymphocyte transformation test (LTT) responses than sensitized volunteers without allergy and controls (P<0.05 and P<0.01, respectively). 12.5 microg/mL of chromium chloride and 50 ng/mL of potassium dichromate were found to be optimal to discriminate between sensitized individuals with and without allergy. Combining the results of chromium chloride and potassium dichromate LTT, a positive reaction to at least one of the stimuli was highly predictive of allergy [sensitization with vs. without allergy: Odds ratio (OR)=6.4, P=0.004; sensitization with allergy vs. controls: OR=11.5, P<0.0001]. On the contrary, IFN-gamma, IL-2, IL-4, IL-10, and IL-12 production to the ELISpot, patch test results, sensitization against other metals, and atopy score did not significantly discriminate between sensitization with and without allergy. However, IFN-gamma responses towards chromium chloride were significantly correlated with the strength of patch test reactivity (r=0.49, P=0.002). By IFN-gamma ELISpot, the average precursor cell frequency reactive to trivalent chromium could be defined as 26, 15, and 11 : 10(6) in volunteers with sensitization and allergy, with sensitization without allergy, and controls, respectively. CONCLUSIONS: In contrast to the patch test, the LTT appears to be a method that is predictive of chromium allergy.

Cellular and humoral immune response to a third generation hepatitis B vaccine.

Liver transplantation is often the ultimate option of therapy for chronically hepatitis B virus (HBV) infected patients. Adoptive transfer of HBV immunity with the liver after vaccination of living liver donors (LLD) could be a new approach to prevent reinfection in the recipients. The time to achieve HBV immunity in LLD is usually short (1-2 months). Therefore, we established a short time immunization protocol (four injections in 2 weeks intervals) using Hepimmune, a recombinant vaccine that contains the L, M and S proteins of HBV. We examined cellular and humoral immune responses after immunization with Hepimmune and compared its immunogenicity to that of a standard HBV vaccine containing only the S protein (HBVAXPRO). Cellular immunity was measured by interferon (IFN)-gamma ELISpot and proliferation assay. HBV-specific T cells were detectable in the Hepimmune group after the second and in the standard group after the third vaccination. IFN-gamma production of T cells was significantly higher (P < 0.001) after the third vaccination with Hepimmune. Proliferative responses were also significantly (P < 0.01) higher in the Hepimmune group after the second to fourth vaccination. The humoral immune response could already be detected after the first immunization in nine of 15 Hepimmune vaccinated test persons while it was only observed in one of 15 probands of the later group. Titres differed significantly (P < 0.01) following all four vaccinations. Thus, Hepimmune appears to be a good candidate for short time immunization protocols.