Optimised germination protocol for jackfruit seeds and evaluation of methods for extraction of DNA suitable for genetic analysis

Jackfruit (Artocarpus heterophyllus) is an underutilised plant that is promising in curbing food and nutritional security in sub-Saharan Africa. However, high level of secondary metabolites in its tissues significantly hampers its genetic characterisation for breeding purposes. Primarily, the compounds react with DNA during the extraction process, thus reducing its yield and quality. The utilisation of leaves from jackfruit seedlings is a potentially effective approach of addressing the challenge, however, limited information is available on efficient jackfruit seed germination procedures. Elucidating effective methods of jackfruit seed germination, and optimising protocols for DNA extraction is crucial in promoting its genetic characterisation studies for identification of superior varieties for propagation. The objective of this study was to evaluate methods of jackfruit seed germination, and DNA extraction procedures using jackfruit leaves. Pre-treatment of seeds with 3% hydrogen peroxide was effective in enhancing seed germination within a short time, compared to distilled water and 3% hydrochloric acid. We optimised a DNA extraction technique by combining CTAB-SDS based approach with an enhanced solvent extraction method. The technique yielded high quantity and quality of DNA from jackfruit leaves, which was appropriate for downstream polymerase chain reaction analysis. The sequence-related amplified polymorphism (SRAP) and simple sequence repeat (SSR) amplifications confirmed the effectiveness of the optimised CTAB-SDS based protocol for extraction of high quality DNA. &nbsp

Intranasal administration of olanzapine has beneficial outcome in a rat activity-based anorexia model

The atypical antipsychotic drug olanzapine is prescribed despite clinical studies on olanzapine treatment showing mixed results on treatment efficacy in anorexia nervosa. We investigated the effect of systemic and intranasal administration of olanzapine in the activity-based anorexia (ABA) model. Rats were habituated to a running wheel and exposed to the ABA model while treated with olanzapine. During ABA rats had 1.5 h of daily access to food and ad libitum access to a running wheel for seven consecutive days. Olanzapine was administered via an osmotic minipump (1, 2.75, and 7.5 mg/kg) or intranasally 2 h before dark onset (1 and 2.75 mg/kg). We monitored body weight, food intake, wheel revolutions, body temperature, and adipose tissue. We found 2.75 and 7.5 mg/kg systemic olanzapine decreased wheel revolutions during ABA. Relative adipose tissue mass was increased in the 7.5 mg/kg olanzapine-treated group while body weight, food intake, and body temperature were unaltered by the systemic olanzapine. 1 and 2.75 mg/kg intranasal olanzapine diminished wheel revolutions and body temperature during the first 2 h after administration. The intranasal olanzapine-treated rats had a higher body weight at the end of ABA. We find that olanzapine has beneficial outcomes in the ABA via two administration routes by acting mainly on running wheel activity. Intranasal olanzapine showed a rapid effect in the first hours after administration in reducing locomotor activity. We recommend further exploring intranasal administration of olanzapine in anorectic patients to assist them in coping with restlessness

The effects of polyunsaturated fatty acid (PUFA) administration on the microbiome-gut-brain axis in adolescents with anorexia nervosa (the MiGBAN study): study protocol for a longitudinal, double-blind, randomized, placebo-controlled trial

Background Anorexia nervosa (AN) is a severe psychiatric disease that often takes a chronic course due to insufficient treatment options. Emerging evidence on the gut-brain axis offers the opportunity to find innovative treatments for patients with psychiatric disorders. The gut microbiome of patients with AN shows profound alterations that do not completely disappear after weight rehabilitation. In previous studies, the administration of polyunsaturated fatty acids (PUFA) resulted in effects that might be beneficial in the treatment of AN, affecting the microbiome, body weight and executive functions. Therefore, the MiGBAN study aims to examine the effects of a nutritional supplementation with PUFA on the gut microbiome and body mass index (BMI) in patients with AN. Methods This is a longitudinal, double-blind, randomized, placebo-controlled trial. Within 2 years, 60 adolescent patients aged 12 to 19 years with AN will receive either PUFA or placebo for 6 months additional to treatment as usual. After 1 year, the long-term effect of PUFA on the gut microbiome and consecutively on BMI will be determined. Secondary outcomes include improvement of gastrointestinal symptoms, eating disorder psychopathology, and comorbidities. Additionally, the interaction of the gut microbiome with the brain (microbiome-gut-brain axis) will be studied by conducting MRI measurements to assess functional and morphological changes and neuropsychological assessments to describe cognitive functioning. Anti-inflammatory effects of PUFA in AN will be examined via serum inflammation and gut permeability markers. Our hypothesis is that PUFA administration will have positive effects on the gut microbiota and thus the treatment of AN by leading to a faster weight gain and a reduction of gastrointestinal problems and eating disorder psychopathology. Discussion Due to previously heterogeneous results, a systematic and longitudinal investigation of the microbiome-gut-brain axis in AN is essential. The current trial aims to further analyse this promising research field to identify new, effective therapeutic tools that could help improve the treatment and quality of life of patients. If this trial is successful and PUFA supplementation contributes to beneficial microbiome changes and a better treatment outcome, their administration would be a readily applicable additional component of multimodal AN treatment

The effects of probiotics administration on the gut microbiome in adolescents with anorexia nervosa—A study protocol for a longitudinal, double-blind, randomized, placebo-controlled trial

Objective Knowledge on gut?brain interaction might help to develop new therapies for patients with anorexia nervosa (AN), as severe starvation-induced changes of the microbiome (MI) do not normalise with weight gain. We examine the effects of probiotics supplementation on the gut MI in patients with AN. Method This is a study protocol for a two-centre double-blind randomized-controlled trial comparing the clinical efficacy of multistrain probiotic administration in addition to treatment-as-usual compared to placebo in 60 patients with AN (13?19 years). Moreover, 60 sex- and age-matched healthy controls are included in order to record development-related changes. Assessments are conducted at baseline, discharge, 6 and 12 months after baseline. Assessments include measures of body mass index, psychopathology (including eating-disorder-related psychopathology, depression and anxiety), neuropsychological measures, serum and stool analyses. We hypothesise that probiotic administration will have positive effects on the gut microbiota and the treatment of AN by improvement of weight gain, gastrointestinal complaints and psychopathology, and reduction of inflammatory processes compared to placebo. Conclusions If probiotics could help to normalise the MI composition, reduce inflammation and gastrointestinal discomfort and increase body weight, its administration would be a readily applicable additional component of multi-modal AN treatment

The effects of polyunsaturated fatty acid (PUFA) administration on the microbiome-gut-brain axis in adolescents with anorexia nervosa (the MiGBAN study): study protocol for a longitudinal, double-blind, randomized, placebo-controlled trial

Background: Anorexia nervosa (AN) is a severe psychiatric disease that often takes a chronic course due to insufficient treatment options. Emerging evidence on the gut-brain axis offers the opportunity to find innovative treatments for patients with psychiatric disorders. The gut microbiome of patients with AN shows profound alterations that do not completely disappear after weight rehabilitation. In previous studies, the administration of polyunsaturated fatty acids (PUFA) resulted in effects that might be beneficial in the treatment of AN, affecting the microbiome, body weight and executive functions. Therefore, the MiGBAN study aims to examine the effects of a nutritional supplementation with PUFA on the gut microbiome and body mass index (BMI) in patients with AN. Methods: This is a longitudinal, double-blind, randomized, placebo-controlled trial. Within 2 years, 60 adolescent patients aged 12 to 19 years with AN will receive either PUFA or placebo for 6 months additional to treatment as usual. After 1 year, the long-term effect of PUFA on the gut microbiome and consecutively on BMI will be determined. Secondary outcomes include improvement of gastrointestinal symptoms, eating disorder psychopathology, and comorbidities. Additionally, the interaction of the gut microbiome with the brain (microbiome-gut-brain axis) will be studied by conducting MRI measurements to assess functional and morphological changes and neuropsychological assessments to describe cognitive functioning. Anti-inflammatory effects of PUFA in AN will be examined via serum inflammation and gut permeability markers. Our hypothesis is that PUFA administration will have positive effects on the gut microbiota and thus the treatment of AN by leading to a faster weight gain and a reduction of gastrointestinal problems and eating disorder psychopathology. Discussion: Due to previously heterogeneous results, a systematic and longitudinal investigation of the microbiome-gut-brain axis in AN is essential. The current trial aims to further analyse this promising research field to identify new, effective therapeutic tools that could help improve the treatment and quality of life of patients. If this trial is successful and PUFA supplementation contributes to beneficial microbiome changes and a better treatment outcome, their administration would be a readily applicable additional component of multimodal AN treatment. Trial registration: German Clinical Trials Register DRKS00017130. Registered on 12 November 2019

Investigation of common, low-frequency and rare genome-wide variation in anorexia nervosa

Anorexia nervosa (AN) is a complex neuropsychiatric disorder presenting with dangerously low body weight, and a deep and persistent fear of gaining weight. To date, only one genome-wide significant locus associated with AN has been identified. We performed an exome-chip based genome-wide association studies (GWAS) in 2158 cases from nine populations of European origin and 15 485 ancestrally matched controls. Unlike previous studies, this GWAS also probed association in low-frequency and rare variants. Sixteen independent variants were taken forward for in silico and de novo replication (11 common and 5 rare). No findings reached genome-wide significance. Two notable common variants were identified: rs10791286, an intronic variant in OPCML (P=9.89 × 10 -6), and rs7700147, an intergenic variant (P=2.93 × 10 -5). No low-frequency variant associations were identified at genome-wide significance, although the study was well-powered to detect low-frequency variants with large effect sizes, suggesting that there may be no AN loci in this genomic search space with large effect sizes

Translational genetics: expectations for eating disorders and other psychiatric disorders

Achtergrond Translationeel (genetisch) onderzoek is gericht op de vertaalslag van preklinisch onderzoek naar de praktijk. Hoewel de afgelopen jaren veel genetische studies zijn uitgevoerd, kunnen we de resultaten daarvan niet simpelweg naar de kliniek vertalen.Translationeel (genetisch) onderzoek is gericht op de vertaalslag van preklinisch onderzoek naar de praktijk. Hoewel de afgelopen jaren veel genetische studies zijn uitgevoerd, kunnen we de resultaten daarvan niet simpelweg naar de kliniek vertalen. Doel De stappen zichtbaar maken waarlangs translationeel genetisch onderzoek bijdraagt aan de ontrafeling van biologische achtergronden van psychiatrische aandoeningen, in het bijzonder van de eetstoornissen. Methode Literatuuronderzoek. Resultaten Genetische studies hebben geleid tot ontrafeling van een mechanisme dat ten grondslag ligt aan het honger- en verzadigingssysteem. Er is hoop dat genoomwijde studies naar eetstoornissen leiden tot identificatie van neurale circuits waarin geassocieerde genen clusteren. Nieuwe technieken zoals opto- en chemogenetica geven dan de mogelijkheid de precieze rol van die circuits bij eetstoornissen te onderzoeken. Conclusie De nieuwe technieken in de moleculaire neurowetenschappen maken het mogelijk de complexiteit van de werking van de hersenen te ontrafelen en sommige van die technieken (zoals chemogenetica) worden verder ontwikkeld voor toepassing bij de mens. Toch zal het nog jaren duren voor we dit definitief kunnen vertalen naar de behandeling van psychiatrische aandoeningen

Design heuristic for parallel many server systems

We study a parallel queueing system with multiple types of servers and customers. A bipartite graph describes which pairs of customer-server types are compatible. We consider the service policy that always assigns servers to the first, longest waiting compatible customer, and that always assigns customers to the longest idle compatible server if on arrival multiple compatible servers are available. For a general renewal stream of arriving customers, general service time distributions that depend both on customer and on server types, and general customer patience distributions, the behavior of such systems is very complicated. Key quantities for their performance are the matching rates, the fraction of services for each pair of compatible customer-server. Calculation of these matching rates in general is intractable, it depends on the entire shape of service time distributions. We suggest through a heuristic argument that if the number of servers becomes large, the matching rates are well approximated by matching rates calculated from the tractable bipartite infinite matching model. We present simulation evidence to support this heuristic argument, and show how this can be used to design systems with desired performance requirements