Green NP-HPTLC and green RP-HPTLC methods for the determination of thymoquinone: A contrast of validation parameters and greenness assessment.

INTRODUCTION: Thymoquinone (TQ) is a naturally derived bioactive compound with several therapeutic effects. OBJECTIVE: The highly sensitive, rapid and green normal-phase (NP)/reversed-phase (RP) high-performance thin-layer chromatography (HPTLC) densitometry technique was developed for the determination of TQ in various plant extracts of different geographical regions, commercial capsules, creams and essential oils. METHODOLOGY: The NP densitometry estimation of TQ was performed using a cyclohexane-ethyl acetate (90:10, v/v) green solvent system, while, the RP-densitometry estimation of TQ was performed using an ethanol-water (80:20, v/v) green solvent system. The estimation of TQ was conducted at 259 nm. RESULTS: The NP and RP densitometry techniques were observed linear in the range of 25-1000 and 50-600 ng/band, respectively. All validation parameters such as accuracy, precision, robustness and sensitivity of NP/RP densitometry were observed within the limit of regulatory requirements and hence found to be suitable for the determination of TQ. The TQ contents were found to be highest in the Saudi Arabian extract followed by the Syrian extract, Indian extract, commercial capsules, commercial creams, Jordanian extract, Egyptian extract, Palestinian extract and commercial essential oils using NP densitometry. The TQ contents were found in same order using RP densitometry, but they were much lower than those recorded using NP densitometry. The Analytical GREEnness (AGREE) scores of NP and RP densitometry were found to be 0.82 and 0.84, respectively, suggesting an excellent greenness profile. CONCLUSIONS: Based on these results, NP/RP densitometry was found to be suitable for the pharmaceutical assay of TQ.

Avaliação da saúde óssea em pacientes com anorexia nervosa e bulimia nervosa / Bone health assessment in patients with anorexia nervosa and bulimia nervosa

Introdução: Anorexia nervosa (AN) e bulimia nervosa (BN) são transtornos alimentares (TA), cujos sintomas típicos de restrição alimentar, exercícios físicos excessivos, vômitos auto-induzidos e abuso de laxantes e diuréticos podem resultar em severas complicações clínicas. Mulheres com AN e BN apresentam risco elevado para osteoporose, osteopenia e fraturas, associado com baixo peso, atividade física excessiva, história e duração da doença, amenorreia, perda de força muscular e insuficiência de vitamina D. Objetivo: Avaliar a saúde óssea e a composição corporal de mulheres adultas com diagnóstico de AN e BN atual ou prévio. Métodos: estudo transversal incluindo 53 mulheres adultas (18-49 anos) com diagnóstico atual ou pregresso de AN (n=25) ou BN (n=28). Foi realizada absorciometria com raios-X de dupla energia (DXA) para mensuração da densidade mineral óssea (DMO) (total, coluna lombar e colo do fêmur), massa gorda e massa magra e seus índices ajustados pela estatura; administração de diluição isotópica de deutério (D2O) para determinação de água corporal total e massa magra, e teste de prensão palmar para avaliação da força muscular. A determinação sérica das concentrações de glicose, insulina (cálculo de HOMA-IR), colesterol total e frações (LDL-c, HDL-c e VLDL-C), triglicérides e 25-hydroxyvitamin D (25(OD)D) foram realizadas em amostra de sangue obtida após jejum de 12 horas. Foram aplicados questionários sobre história da doença, uso de suplementos, atividade física habitual e sintomas do TA. Análises estatísticas incluíram testes T Independente, ANOVA, Qui Quadrado, correlação linear de Pearson, modelos de regressão linear univariados e múltiplos e análise de Bland-Altman. Resultados estão apresentados em média, desvio-padrão, frequências e intervalo de confiança de 95%. Revisão sistemática e meta-análise da DMO em indivíduos com TA em comparação a controles saudáveis foi realizada de acordo com o PRISMA guidelines. Resultados: Mulheres com AN apresentaram menor DMO de corpo total [1.09 g/cm2 (0.07) vs BN: 1.18 g/cm2 (0.12); p<0.01] e do colo do fêmur [0.95 g/cm2 (0.13) vs BN: 1.05 g/cm2 (0.16); p<0.01] em comparação com BN. Mulheres com AN apresentaram menor gordura corporal, razão androide/ginoide, tecido adiposo visceral, massa magra e maior percentual de massa magra em comparação àquelas com BN. Comparação entre DXA e D2O mostrou nenhuma diferença significante para gordura corporal (massa, percentual e índice de massa/estatura2) e massa magra (massa e índice de massa/estatura2). Possível viés sistemático do DXA em superestimar gordura corporal e massa magra, exceto para o índice de massa livre de gordura (IMLG), foi relacionado com o diagnóstico de BN, estado de hidratação e atividade física. DMO total foi positivamente associada com gordura corporal (Modelo DXA: ß=0.59; p=0.007; Modelo D2O: ß=0.59; p<0.001) e força muscular (Modelo DXA: ß=0.32; p=0.013). DMO da coluna foi negativamente associada com duração da amenorreia (Modelo DXA: ß=-0.32; p=0.027; Modelo D2O: ß=-0.38; p=0.024). A DMO do colo do fêmur foi associada com gordura corporal (Modelo D2O: ß=0.37; p=0.038) e massa magra (Modelo D2O: ß=0.43; p=0.025). Não houve efeito da duração da doença e atividade física na DMO. Foram observadas diferenças entre os TA nos valores de HOMA-IR [AN: 1.3 (0.8) vs BN: 2.0 (1.2); p<0.05] e HDL-c [AN: 76.6mg/dL (13.2) vs BN: 61.9 mg/dL (16.3); p<0.05]. Inadequação de 25(OH)D (<30ng/mL) foi observada na AN (100%) e na BN (72%). Destacam-se as associações independentes de HOMA-IR e LDL-c com a DMO do colo do fêmur em mulheres com AN (R2=0.45; p<0.00; HOMA ß=0.503; LDL-c: ß=0.398). Na meta-análise (n=42 estudos), a DMO (total, coluna, quadril e fêmur) foi menor na AN e a DMO (total e coluna) foi menor BN (principalmente com história de AN) em comparação aos controles saudáveis. Conclusão: mulheres com AN e BN com história de AN são mais susceptíveis às perdas de massa óssea. É importante que indivíduos com AN ou história de AN tenham sua saúde óssea avaliada, independente do diagnóstico atual. Recomenda-se a utilização do IMLG e o desenvolvimento de protocolos específicos para realização da avaliação da composição corporal pelo DXA nessa população. A avaliação da saúde óssea deve considerar composição corporal, história da doença, função menstrual e estado de hidratação. A inclusão de teste de força e análise de parâmetros bioquímicos pode auxiliar na identificação de indivíduos que apresentam maior necessidade de exames específicos.

Background: Anorexia nervosa (AN) and bulimia nervosa (BN) are eating disorders (ED) whose typical symptoms of food restriction, excessive exercise, self-vomit and misuse of laxatives and diuretics can lead to severe clinical complications. Women with AN and BN have increased risk of developing osteopenia, osteoporosis and fractures, associated with low weight, low physical activity, history and duration of the disease, amenorrhea, lack of muscle strength and vitamin D insufficiency. Aim: to assess bone health and body composition of women with current or previous AN and BN. Methods: cross-sectional study including 53 adult (18-49 years) with current or previous diagnosis of AN (n=25) or BN (n=28). Dual-energy x-ray absorptiometry (DXA) was performed to measure total, lumbar spine and femoral neck BMD, fat mass and lean mass; and administration of deuterium oxide (D2O) solution to determine total body water and lean mass. Serum determinations of glucose, insulin (HOMA-IR calculation), total cholesterol and fractions (LDL-c, HDL-c and VLDL-C), triglycerides, and 25-hydroxyvitamin D (25(OD)D were obtained after a 12-hour fasting blood test. Questionnaires about lifestyle, clinical history of the disease, use of supplements, physical activity and ED symptoms were applied. Statistical analyses included Independent T, ANOVA, chi-Square, Pearson's linear correlation tests, univariate and multivariate linear regressions and Bland-Altman analysis. Results are presented in means, standard deviation, frequencies, and 95% confidence interval. Systematic review and meta-analysis of BMD in individuals with ED in comparison to healthy controls was performed according to PRISMA guidelines. Results: Women with AN had lower total DMO [1.09 g/cm2 (0.07) vs BN: 1.18 g/cm2 (0.12); p<0.01] and femoral neck BMD [0.95 g/cm2 (0.13) vs BN: 1.05 g/cm2 (0.16); p<0.01] than women with BN. Compared to BN, women with AN presented less fat mass, android/gynoid ratio, visceral adipose tissue, lean mass and higher lean mass percentage. DXA and D2O dilution comparison showed absence significant difference for body fat (mass, percentage and index) and lean mass (mass and index). Possible bias of DXA in overestimating fat and lean mass, except for lean mass index (LMI) was related to BN diagnosis, hydration status and physical activity. Total BMD was positively associated with body fat (Model DXA: ß=0.59; p=0.007; Model D2O: ß=0.59; p<0.001) and muscle strength (Model DXA: ß=0.32; p=0.013). Lumbar spine BMD was negatively associated with duration of amenorrhea (Model DXA: ß=-0.32; p=0.027; Model D2O: ß=-0.38; p=0.024). Femoral neck BMD associated with body fat (Model D2O: ß=0.37; p=0.038) and lean mass (Model D2O: ß=0.43; p=0.025). Illness duration and physical activity did not present effect on BMD. Differences were observed for HOMA-IR [AN: 1.3 (0.8) vs BN: 2.0 (1.2); p<0.05] e HDL-c [AN: 76.6mg/dL (13.2) vs BN: 2.0 mg/dL (1.2); p<0.05]. 25(OH)D (<30ng/mL) inadequacy was observed in AN (100%) and BN (72%). HOMA-IR and LDL-c were independently associated with femoral neck BMD in women with AN (R2=0.45; p<0.00; HOMA ß=0.503; LDL-c: ß=0.398). In the meta-analysis (n=42 studies), BMD (total, spine, hip and femur) were lower in AN and BMD (total and spine) lower in BN than in healthy controls. Conclusion: Women with AN and BN and a history of AN are prone to bone loss. It is important that individuals with AN or a history of AN have their bone health assessed, irrespective of the current diagnosis. The use of the LMI and the development of specific protocols to carry out the assessment of body composition by DXA are recommended for this population. Bone health assessment should consider body composition, disease history, menstrual function, and hydration status. The inclusion of grip strength test and analysis of biochemical parameters can help identify individuals who have a greater need for specific tests.

In situ, high-resolution measurement of labile phosphate in sediment porewater using the DET technique coupled with optimized imaging densitometry.

Obtaining two-dimensional distributions of reactive phosphorus in sediment porewater is very important for understanding fine-scale phosphorus mobilization and sequestration processes in sediments. In this study, the diffusive equilibrium in thin films (DET) measurement based on computer imaging densitometry (CID) was studied in detail with optimal conditions described. This study focuses on evaluating the two-dimensional colorimetric DET method coupled with CID (DET-CID method) for porewater labile phosphate measurements. The result shows that the red channel filter is the optimum channel for sensitivity to process the image. Additionally, staining time and temperature have great influence on the method, and 20 min staining time and ≥25 °C staining temperature were recommended. The minimum detection limit of labile phosphate of this method was 0.300 mg P/L, and the maximum detection limit could reach 50.00 mg P/L. The DET-CID technique can be used to measure labile phosphate in a wide range of acidic and alkaline water bodies (pH = 2-10 and water hardness from 0 to 2000 mg/L as CaCO3). The linear regression analysis shows that this technique presents very similar results compared with other two existing methods (R2 = 0.999). Our results would give insights into the precisely measurements of labile phosphate in field applications.

Determination of Cassiarin A Level of Cassia siamea Leaf Obtained from Various Regions in Indonesia Using the TLC-Densitometry Method.

Cassia siamea leaf has been proven in vitro and in vivo to have a strong antimalarial activity with Cassiarin A as its active compound. To obtain a source of C. siamea medicinal plant with high level of active antimalarial compound (Cassiarin A), a valid method for determining Cassiarin A level is needed. For this reason, this research conducts the validation of the Cassiarin A content with determination method using thin-layer chromatography (TLC) densitometry which includes the determination of selectivity (Rs), linearity (r), accuracy, precision, limit of detection (LOD), and limit of quantification (LOQ). Cassiarin A was chromatographed on silica gel 60 F254 TLC plate using chloroform : ethanol (85 : 15 v/v) as a mobile phase. Cassiarin A was quantified by densitometric analysis at 368 nm. The linear regression analysis data for the calibration plots showed good linear relationship with r = 0.9995. The method was validated for precision, recovery, repeatability. The minimum detectable amount was found to be 0.0027 µg/spot, whereas the limit of quantitation was found to be 0.008 µg/spot. The results of this validation are then used to determine the Cassiarin A level of C. siamea leaf from various regions in Indonesia. Based on the results of the study, it can be concluded that the TLC-densitometry method can be used to determine level of the Cassiarin A compound with the advantages of being fast, easy, accurate, and inexpensive. In addition, it showed that C. siamea leaves from Pacitan have the highest level of Cassiarin A compared to other areas studied.

Parâmetros ósseos de codornas de corte suplementadas com diferentes níveis de vitamina A, de 15 a 35 dias de idade / [Parameters of bone of meat-type quail supplemented with different levels of vitamin A from 15 to 35 days of age]

Avaliou-se o efeito da utilização de níveis crescentes de vitamina A sobre os parâmetros ósseos de codornas de corte de 15 a 35 dias de idade. Foram utilizadas 1520 codornas, não sexadas, com peso médio inicial de 85±4,25g, distribuídas em um delineamento inteiramente ao acaso, totalizando oito tratamentos com cinco repetições e 38 codornas por unidade experimental. Os tratamentos consistiram de oito níveis de suplementação de vitamina A (0; 4.500; 6.000; 7.500; 9.000; 10.500; 12.000 e 13.500UI/kg da dieta). O período experimental foi de 15 a 35 dias de idade. Os ossos analisados apresentaram uma redução linear para o índice de Seedor, resistência óssea e teor de cinzas do fêmur e da tíbia de acordo com os níveis de suplementação de vitamina A. A suplementação de vitamina A para codornas de corte de 15 a 35 dias de idade mostrou resultados negativos para a qualidade óssea, com a necessidade de mais estudos sobre seu antagonismo com a vitamina D.(AU)

This study evaluated the effect of using increasing levels of vitamin A on bone parameters in meat-type quails from 15 to 35 days old. A total of 1520 unsexed quail with an average initial weight of 85g distributed in a completely randomized design, totaling eight treatments with five replicates and 38 quails per experimental unit were used. The treatments consisted of eight vitamin supplementation levels A (0; 4,500; 6,000; 7,500; 9,000; 10,500; 12,000 and 13,500IU / kg diet). The experimental period was 15 to 35 days of age. The bones analyzed showed a linear reduction to the Seedor index, bone strength and ash content of the femur and tibia according to vitamin supplementation levels Vitamin A supplementation for cutting quails from 15 to 35 days of age showed negative results for bone quality, with the need for more studies on its antagonism to vitamin D.(AU)

Thin-layer chromatographic quantification of magnolol and honokiol in dietary supplements and selected biological properties of these preparations.

Two isomeric biphenyl neolignans, magnolol and honokiol, are considered as constituents responsible for the healing effect of magnolia bark, a traditional Oriental medicine. To survey the increasing number of dietary supplements that contain magnolia bark or its extract, an affordable quantitative thin-layer chromatography (TLC) - densitometry method was developed. The methanol extracts were analyzed on the silica gel plates after manual sample application using n-hexane - ethyl acetate - ethanol (16:3:1, v/v/v) as a mobile phase. For quantitation, the chromatograms were scanned in the absorbance mode at the wavelength λ = 290 nm. The limits of detection and quantitation were 90 and 280 ng/zone for magnolol and 70 and 200 ng/zone for honokiol, respectively. None of the two targeted neolignans were detected in two of the six analyzed supplements. In the other four samples, the measured amounts were between 0.95-114.69 mg g-1 for magnolol and 4.88-84.86 mg g-1 for honokiol. Moreover, separations of these two neolignans on the TLC and high-performance TLC (HPTLC) layers were compared and HPTLC was combined with antioxidant (DPPH) and antibacterial (Bacillus subtilis and Aliivibrio fischeri) assays and mass spectrometry (MS), using the elution-based interface. Both magnolol and honokiol exhibited effects in all bioactivity assays. The HPTLC-MS tests confirmed purity of neolignan zones in the extracts of dietary supplements and supported tentative identification of the alkaloid piperine and the isoflavone daidzein as additional bioactive components of the investigated dietary supplements. Using the same mobile phase in the orthogonal directions 2D-HPTLC-MS experiments proved degradation, i.e., instability of magnolol and honokiol on the silica gel adsorbent.

Neuroprotective Role of Dietary Supplementation with Omega-3 Fatty Acids in the Presence of Basal Forebrain Cholinergic Neurons Degeneration in Aged Mice.

As major components of neuronal membranes, omega-3 polyunsaturated fatty acids (n-3 PUFA) exhibit a wide range of regulatory functions. Recent human and animal studies indicate that n-3 PUFA may exert beneficial effects on aging processes. Here we analyzed the neuroprotective influence of n-3 PUFA supplementation on behavioral deficits, hippocampal neurogenesis, volume loss, and astrogliosis in aged mice that underwent a selective depletion of basal forebrain cholinergic neurons. Such a lesion represents a valid model to mimic a key component of the cognitive deficits associated with dementia. Aged mice were supplemented with n-3 PUFA or olive oil (as isocaloric control) for 8 weeks and then cholinergically depleted with mu-p75-saporin immunotoxin. Two weeks after lesioning, mice were behaviorally tested to assess anxious, motivational, social, mnesic, and depressive-like behaviors. Subsequently, morphological and biochemical analyses were performed. In lesioned aged mice the n-3 PUFA pre-treatment preserved explorative skills and associative retention memory, enhanced neurogenesis in the dentate gyrus, and reduced volume and VAChT levels loss as well as astrogliosis in hippocampus. The present findings demonstrating that n-3 PUFA supplementation before cholinergic depletion can counteract behavioral deficits and hippocampal neurodegeneration in aged mice advance a low-cost, non-invasive preventive tool to enhance life quality during aging.

The paleoradiology importance in the study of relics: the unique densitometric analysis of a bone relic of Saint Nicholas.

INTRODUCTION: DXA have greatly contributed to the development of paleoradiology, a branch of diagnostic imaging that allows to obtain information about human remains in contexts of archaeological and/or forensic interest. In this manuscript we report the unique experience of DXA performed on the relic of a Saint; in particular we analyzed a skeletal fragment of St. Nicholas, kept in the Basilica of Bari (Italy) since 1087. MATERIALS AND METHODS: The bone to be examined consisted of the posterior arch of the ninth left rib that was 12cm long, 1.2cm maximum width and 1.7cm thick at the body. The data acquired from the densitometric study were performed using the anthropometric measures reported in historical records of St. Nicholas' life: sex (male), age (75 years), weight (70kg), height (167cm), and ethnicity (Caucasian). In addition to the examination of the relic, a comparison assessment was made with the rib of a healthy 60 years old man (height of 170cm, without known skeletal pathologies). This sample had a length of 19cm, maximum width at the head 1cm, and 0.7cm thick at the body. The analysis of bone fragments is different from the analysis of bones in the context of the human body (where soft tissues are placed around the skeleton); for this reason, one of the most critical issues was to create a support that would allow the analysis of bone fragments. We simulated conditions similar to those occurring in patients: a density scale was established, using a specific plexiglass phantom on which the bone fragments to be examined were placed. From the analysis it was calculated the parameter bone mineral density (BMD), express in g/cm2, that indicates the relation between mass of bone mineral content and area of examined bone segment. BMD data was compared to a range normalized by age, sex and ethnicity (BMD-N). RESULTS: The results of the scannnig of St. Nicholas' rib showed a BMD of 0.97g/cm2 with a BMD-N between 0.77 and 1.08g/cm2. Simultaneous measurements of the relic compared with a reference rib showed highlighted BMD of 0.84g/cm2 for the relic and 0.50g/cm2 for the reference rib. The St. Nicholas data are 168% higher than reference bone. All our measurements of the relic indicated a high bone mineral density, most likely due to the presence of a high concentration of calcium salts. A relatively higher mineral density of the relic was seen compared to the healthy subject's rib. From the history of St Nicholas' life, we know of the long imprisonment at the age of 51 in damp and unhealthy environment. The results of this study suggest that a good bone mineral density was maintained by the Saint even in old age. An additional element that can influence bone mineral density is diet, certainly different during the time of St. Nicholas. The good bone densitometry indicates that the Saint maintained a proper diet, with a generally fair state of health. CONCLUSION: For this first DXA analysis of the rib relic of Saint Nicholas was necessary a long and complex experimental work to modify standard technique procedure to particular and unusual sample and Create specific supports and complementary instruments. Perform DXA analysis on relics permit to obtain additional information to living conditions, economical situation, behaviours, diet, diseaes, conservations conditions of remains, change of life style in different age. Our experimental work, the first of its kind, creates the way to analyze precious relics that often include only few bone fragments and data obtained by our work can be useful for a better management and movement of fragile relics. We ourselves are working on a new challenge for the analysis of bone finds from shipwrecks found at the bottom of the sea.

Testosterone propionate activated the Nrf2-ARE pathway in ageing rats and ameliorated the age-related changes in liver.

The present study aimed to evaluate the protective efficacy of testosterone propionate (TP) on age-related liver changes via activation of the nuclear factor erythroid 2-related factor 2-antioxidant response element (Nrf2-ARE) pathway in aged rats. Aged rats received subcutaneous injections of TP (2 mg/kg/d, 84 days). Oxidative stress parameters and the expression levels of signal transducer and activator of transcription 5b (STAT5b), Kelch-like ECH associating protein-1 (Keap1), Nrf2, haem oxygenase-1 (HO-1) and NAD(P)H: quinone oxidoreductase-1 (NQO1) in liver tissues were examined to check whether the Nrf2-ARE pathway was involved in the age-related changes in liver. Our results showed that TP supplementation alleviated liver morphology, liver function and liver fibrosis; improved oxidative stress parameters; and increased the expression of STAT5b, Nrf2, HO-1 and NQO-1 and decreased the expression of Keap1 in the liver tissues of aged rats. These results suggested that TP increased the expression of STAT5b, and then activated the Nrf2-ARE pathway and promoted antioxidant mechanisms in aged rats. These findings may provide new therapeutic uses for TP in patients with age-related liver changes.

Diagnostic accuracy of digital X-ray radiogrammetry on hand bone loss for patients with rheumatoid arthritis.

BACKGROUND: This study will aim to evaluate the diagnostic accuracy of digital X-ray radiogrammetry (DXR) on hand bone loss (HBL) for rheumatoid arthritis (RA). METHODS: In this study, we will search the literature from PubMed, EMBASE, Cochrane Library, PsycINFO, Web of Science, Google Scholar, Cumulative Index to Nursing and Allied Health Literature, Allied and Complementary Medicine Database, Chinese Biomedical Literature Database, China National Knowledge Infrastructure, and WANFANG from the inception to June 1, 2019 without language restrictions. All case-controlled studies on assessing diagnostic accuracy of DXR on HBL for diagnosis of RA will be included. Quality Assessment of Diagnostic Accuracy Studies tool will be used for eligible studies. We will apply RevMan V.5.3 software and Stata V.12.0 software for statistical analysis. RESULTS: We will evaluate diagnostic accuracy of DXR on HBL in patients with RA by assessing the sensitivity, specificity, positive likelihood ratio, negative likelihood ratio, and diagnostic odds ratio. CONCLUSION: This study will detect the diagnostic accuracy of DXR evaluation on HBL in patients with RA. SYSTEMATIC REVIEW REGISTRATION: PROSPERO CRD42019139489.

Determinants of bone mineral density through quantitative ultrasound screening of healthy children visiting ambulatory paediatric clinics.

OBJECTIVES: To investigate the determinants of bone mineral density through screening healthy children using a non-invasive quantitative ultrasound measurement device. METHODS: A descriptive cross-sectional study carried out at King AbdulAziz University Hospital, Jeddah, Kingdom of Saudi Arabia. between May 2018 and January 2019 through interviewing, examining, and screening healthy children visiting general paediatric. Total sample size encompassed 450 children. The inclusion criteria were healthy children between the ages of 2 and 20 years. Exclusion criteria were previous pathological fractures, chronic medical diseases, or long-term medications. Data entry and analysis was conducted using Statistical Package for Social Sciences version 24 (IBM Corp, Armonk, NY, USA). Chi-square tests were used to determine the association between categorical variables, with calculated p<0.05 considered significant. With one-way Anova testing to study the relationship between categorical variables and continuous variables. Results: A significant association with bone mineral density (BMD) was found during first 2 years with height (p=0.015), vitamin D supplementation (p=0.03), and breastfeeding (p=0.025). A directly proportional relationship with BMD was found with pubertal status, physical activity, diet, sun exposure, and calcium supplement intake. CONCLUSION: This is a novel study in the investigation of the dietary, lifestyle and demographic determinants of bone mineral density in the healthy middle-eastern child otherwise unaffected by chronic medical or metabolic disease or exposed to long term medications that could have affected bone metabolism.

Development and Validation of High-Performance Thin-layer Chromatographic Method for Quantification of Berberine in Rhizomes of Coptis teeta Wall, an Endangered Species Collected from Arunachal Pradesh, India.

The high polarity of the protoberberine alkaloids present in Coptis teeta has made it difficult to quantify the alkaloids. This study was designed to develop and validate a thin-layer chromatography (TLC) densitometric-based method using high-performance thin-layer chromatography for quantification of berberine. The separation was achieved in a solvent system consisting of butanol:ethyl acetate:formic acid:water by volume on TLC aluminum plate precoated with silica gel 60 F254. Determination and quantification were performed by densitometric scanning under mercury lamp at a wavelength 351 nm in absorbance mode. The validated method gave compact bands for berberine at an Rf of 0.70. The precision, accuracy and reproducibility of the method were validated by following International Conference on Harmonization guidelines. Graphically, linear results were obtained for berberine with correlation coefficient of 0.997 ± 0.09% (R ± SD) in the concentration range of 90-210 ng/band. The limit of quantification and limit of detection from the analysis were found to be 70 and 30 ng/band, respectively. The berberine concentration in the methanol extract of C. teeta was found to be 30.97 ± 0.55 mg in 100 mg of the crude drug. The method developed here in can be implemented in the analysis and routine quality control of herbal materials and formulations containing C. teeta and berberine.

Impacto de la cirugía bariátrica en el tejido óseo / Impact of bariatric surgery on bone tissue

La importante prevalencia y morbimortalidad de la obesidad ha ocasionado un aumento de pacientes sometidos a cirugía bariátrica. Son numerosos los beneficios reportados de la cirugía de la obesidad en distintas esferas de la salud. Sin embargo, no ocurre lo mismo sobre el hueso, donde tiene un impacto negativo. Los mecanismos fisiopatológicos que subyacen en el deterioro del tejido óseo de estos pacientes son complejos y requieren de un estudio en profundidad. El adecuado conocimiento de estos factores permitirá adoptar las herramientas más oportunas para un adecuado abordaje terapéutico

The important prevalence and morbidity of obesity has generated an increase in bariatric surgery. It has a positive effect in obesity-related comorbidities. However, it's detrimental to bone health. The underline pathophysiological mechanisms are complex and heterogeneous. The knowledge of these factors may lead us to develop an adequate therapeutic intervention

HPTLC Screening of Weight Loss Products for Deliberately Added Undeclared Synthetic Drugs.

Background: The proposed HPTLC method combines features of the existing methods for (1) the detection of sibutramine and (2) for the detection of phosphodiesterase type 5 inhibitors and analogs. Objective: The method permits effective screening for the presence of nine adulterants in finished products, including tablets, capsules, and "instant coffee" powders. Methods: All products were prepared for analysis using the same simple procedure: ultrasound-assisted extraction in methanol for 30 min followed by centrifugation or filtration. Results: The retardation factor (RF) values of individual zones afford preliminary identification of potential adulterants. Scanning densitometry enables comparison of recorded UV spectra with those of known standard compounds and provides further structural information. Conclusions: The method was successfully applied to 12 commercial products. Of those, nine products tested positive for at least one undeclared component.

Extracts from Paeonia lactiflora Pallas, Rehmannia Glutinosa var. Purpurea Makino, Perilla Frutescens var. Acuta Kudo may increase the endometrial receptivity through expression of leukemia inhibitory factor and adhesion molecules.

OBJECTIVE: To find out the combination of the extracts from Paeonia lactiflora Pallas (PL), Reh- mannia Glutinosa var. Purpurea Makino (RG), Perilla Frutescens var. Acuta Kudo (PF) to increase endometrial receptivity. METHODS: Herbal medicines were extracted with boiling water and polysaccharides were removed. We examined the effect of PL, RG, and PF (PRP), a most effective herbal formula deduced from constitutive ingredient herbs of Antai Yin which is composed of PRP, on the leukemia inhibitory factor (LIF) expression and endometrial receptivity. RESULTS: The combination of the extracts from PRP induced the LIF expression in Ishikawa cells and increased the adhesion between Ishikawa and JAr cells. In addition, PRP-induced attachment of JAr cells onto Ishikawa cells and expression of adhesion molecules, ITGAV, ITGB5, CD44s, and L-selectin, are significantly reduced by knock-down of LIF expression. CONCLUSION: Induced by the combination of the PRP extracts, the adhesion between trophoblast and endometrial cells are mediated by expression of LIF and adhesion molecules. Thus, we suggest the combination of the PRP extracts may be a novel therapy for enhancing embryo implantation rate.

Análise da densidade mineral óssea em pacientes com fenilcetonúria e sua correlação com parâmetros nutricionais / Bone mineral density assessment in patients with phenylketonuria and its correlation with nutritional parameters

Introdução: Redução da densidade mineral óssea (DMO) está associada à Fenilcetonúria (PKU), mas a causa desta associação não é completamente entendida. O objetivo desse estudo foi avaliar a ingestão de nutrientes relacionados ao metabolismo ósseo (cálcio, fósforo, magnésio, potássio), e sua associação com a DMO em pacientes com PKU. Métodos: Estudo transversal, observacional. Foram incluídos 15 pacientes (PKU Clássica=8; Leve=7; mediana de idade=16 anos, IQ=15-20), todos em tratamento com dieta restrita em fenilalanina (Phe) e 13 em uso de fórmula metabólica. Foi realizado recordatório alimentar de 24 horas de um dia e demais dados (histórico de fraturas, parâmetros antropométricos, DMO e níveis plasmáticos de Phe, Tyr, cálcio) foram obtidos por revisão de prontuário. Resultados: Nenhum paciente apresentou histórico de fraturas e seis realizavam suplementação de cálcio (alteração prévia da DMO=5; baixa ingestão=1). A mediana dos níveis de Phe foi 11,6 mg/dL (IQ=9,3-13,3). Em relação ao recordatório alimentar, dez indivíduos apresentaram inadequado consumo de carboidratos; 14, de lipídeos; 9, de cálcio; 11, de magnésio; 13, de fósforo; e todos de potássio. A mediana da DMO foi de 0,989 g/cm2 (IQ=0,903-1,069), sendo duas classificadas como reduzidas para idade, ambas de pacientes com PKU Leve que recebiam suplementação de cálcio. Não foi observada correlação entre níveis de Phe, DMO e demais variáveis analisadas. Conclusão: Redução da DMO não foi frequente na amostra, embora ingestão inadequada de cálcio assim o seja. Estudos adicionais são necessários para esclarecer o efeito da Phe e da ingestão dietética sobre o metabolismo ósseo na PKU. (AU)

Introduction: Reduced bone mineral density (BMD) is associated with phenylketonuria (PKU), but this association is not completely understood. This research aimed to evaluate intake of nutrients related to bone metabolism (calcium, phosphorus, magnesium, potassium) and its association with BMD in patients with PKU. Methods: In this cross-sectional, observational study, 15 patients with PKU (Classical=8, Mild=7; median age=16 years, IQ=15-20 years) were included, all of them on phenylalanine (Phe) restricted diet and 13 being supplemented with a metabolic formula. A 24-hour dietary recall was performed and remaining data (history of fractures, anthropometric parameters, BMD and plasma Phe, tyrosine and calcium levels) were obtained through medical chart review. Results: No patient had any fractures and six received calcium supplements, five due to previous change in BMD and one due to inadequate nutritional intake. Median Phe level was 11.6 mg/dL (IQ=9.3-13.3). In relation to dietary recall, all individuals had inadequate intake of some nutrient (carbohydrate=10; lipids=14; calcium=9; magnesium=11; phosphorus=13; potassium=15). The median BMD was 0.989 g/cm2 (IQ=0.903-1.069). Two cases were classified as low BMD for age, both in patients with mild PKU receiving calcium supplements. Conclusion: Reduced BMD was not common in this sample, although inadequate calcium intake was frequently reported. Additional studies are needed to clarify the effect of Phe and dietary intake on bone metabolism in patients with PKU.(AU)

HPTLC fingerprint profile analysis of cocoa proanthocyanidins depending on origin and genotype.

Cocoa beans are rich in bioactive phytochemicals such as alkaloids, anthocyanins, as well as monomeric and oligomeric flavan-3-ols. A high performance thin layer chromatography (HPTLC) method was developed for a fast analysis of a fingerprint of bioactive compounds present in cocoa beans depending on genotype and origin. Evaluation was performed using HPTLC followed by densitometry. The best separation for a fingerprint consisting of eight phenolic compounds as markers was achieved on silica gel 60 F254 HPTLC plates with a solvent mixture of ethyl formate, formic acid, water, toluene 30/4/3/1.5 (v/v/v/v). Staining with Fast Blue Salt B enabled visualization and quantitative evaluation. Compounds of interest were confirmed by eluting zones using a TLC-MS interface. LODs were ≤ 100ng/zone for all compounds. The fingerprints obtained are useful for quality control of cocoa beans allowing their differentiation according to cocoa or chocolate sorts, varieties, and origins.

Fractal dimension: A complementary diagnostic indicator of osteoporosis to bone mineral density.

Morbidity of osteoporosis is increasing as the world population grows and ages, but bone mineral density (BMD) as a common-used diagnostic indicator is not omnipotent in predicting the bone fragility. According to the definition of osteoporosis by World Health Organization (WHO), the present hypothesis proposes an additional fractal-dimension indicator less than 3 to measure the structural change of bones, and further to diagnose osteoporotic patients with complications, to which BMD is insensitive. Literature reports support that the fractal dimension is more sensitive than BMD in specific cases. The hypothesis shows a promise not only in improving the accuracy of screening osteoporotic patients as a complementary indicator to BMD, but also in evaluating mechanical properties of osteoporotic bone and bone-repair effect of bone tissue engineering.

Phase Separation of FUS Is Suppressed by Its Nuclear Import Receptor and Arginine Methylation.

Cytoplasmic FUS aggregates are a pathological hallmark in a subset of patients with frontotemporal dementia (FTD) or amyotrophic lateral sclerosis (ALS). A key step that is disrupted in these patients is nuclear import of FUS mediated by the import receptor Transportin/Karyopherin-ß2. In ALS-FUS patients, this is caused by mutations in the nuclear localization signal (NLS) of FUS that weaken Transportin binding. In FTD-FUS patients, Transportin is aggregated, and post-translational arginine methylation, which regulates the FUS-Transportin interaction, is lost. Here, we show that Transportin and arginine methylation have a crucial function beyond nuclear import-namely to suppress RGG/RG-driven phase separation and stress granule association of FUS. ALS-associated FUS-NLS mutations weaken the chaperone activity of Transportin and loss of FUS arginine methylation, as seen in FTD-FUS, promote phase separation, and stress granule partitioning of FUS. Our findings reveal two regulatory mechanisms of liquid-phase homeostasis that are disrupted in FUS-associated neurodegeneration.

TLC determination of some flavanones in the buds of different genus Populus species and hybrids.

Flavonoids in the buds of eight Populus species and hybrids were detected and compared with the aid of an optimized TLC method. Separation of 17 flavonoid aglycones belonging to different groups, namely, flavones, flavonols, flavanones and flavanonols, previously described as constituents of poplar buds, was performed on silica gel plates using a hexane/ethyl acetate/formic acid (60:40:1.3, V/V/V) mixture as the mobile phase. Pinocembrin and pinostrobin were found in the majority of analyzed poplar buds. For quantitative analysis of both compounds, two TLC evaluation modes, densitometric and videodensitometric, were compared and the established methods were validated. Concentrations of flavanones in some extracts differed slightly or significantly due to the analyzed plant matrix complexity and the TLC evaluation mode applied. Poplar buds rich in flavanones originated from P. × canadensis 'Robusta' (1.82 and 2.23 g per 100 g, resp.) and P. balsamifera (1.17 and 2.24 g per 100 g, resp.).