Combined Hormone Research Articles

Combined pituitary hormone deficiency harboring CHD7 gene missense mutation without CHARGE syndrome: a case report

BackgroundHeterozygous loss-of-function mutations in the chromodomain helicase DNA-binding protein 7 (CHD7) gene cause CHARGE syndrome characterized by various congenital anomalies. A majority of patients with CHARGE syndrome present with congenital hypogonadotropic hypogonadism (HH), and combined pituitary hormone deficiency (CPHD) can also be present. Whereas CHD7 mutations have been identified in some patients with isolated HH without a diagnosis of CHARGE syndrome, it remains unclear whether CHD7 mutations can be identified in patients with CPHD who do not fulfill the criteria for CHARGE syndrome.Case presentationA 33-year-old woman was admitted to our hospital. She had primary amenorrhea and was at Tanner stage 2 for both pubic hair and breast development. She was diagnosed with CPHD (HH, growth hormone deficiency, and central hypothyroidism), and a heterozygous rare missense mutation (c.6745G > A, p.Asp2249Asn) in the CHD7 gene was identified. Our conservation analysis and numerous in silico analyses suggested that this mutation had pathogenic potential. She had mild intellectual disability, a minor feature of CHARGE syndrome, but did not fulfill the criteria for CHARGE syndrome.ConclusionsWe report a rare case of CPHD harboring CHD7 mutation without CHARGE syndrome. This case provides valuable insights into phenotypes caused by CHD7 mutations. CHD7 mutations can have a continuous phenotypic spectrum depending on the severity of hypopituitarism and CHARGE features. Therefore, we would like to propose a novel concept of CHD7-associated syndrome.

Regenerative plantlets with improved agronomic characteristics caused by anther culture of tetraploid potato (Solanum tuberosum L.).

As the primary means of plant-induced haploid, anther culture is of great significance in quickly obtaining pure lines and significantly shortening the potato breeding cycle. Nevertheless, the methods of anther culture of tetraploid potato were still not well established. In this study, 16 potato cultivars (lines) were used for anther culture in vitro. The corresponding relation between the different development stages of microspores and the external morphology of buds was investigated. A highly-efficient anther culture system of tetraploid potatoes was established. It was shown in the results that the combined use of 0.5 mg/L 1-Naphthylacetic acid (NAA), 1.0 mg/L 2,4-Dichlorophenoxyacetic acid (2,4-D), and 1.0 mg/L Kinetin (KT) was the ideal choice of hormone pairing for anther callus. Ten of the 16 potato cultivars examined could be induced callus with their respective anthers, and the induction rate ranged from 4.44% to 22.67% using this hormone combination. According to the outcome from the orthogonal design experiments of four kinds of appendages, we found that the medium with sucrose (40 g/L), AgNO3 (30 mg/L), activated carbon (3 g/L), potato extract (200 g/L) had a promotive induction effect on the anther callus. In contrast, adding 1 mg/L Zeatin (ZT) effectively facilitated callus differentiation. Finally, 201 anther culture plantlets were differentiated from 10 potato cultivars. Among these, Qingshu 168 and Ningshu 15 had higher efficiency than anther culture. After identification by flow cytometry and fluorescence in situ hybridization, 10 haploid plantlets (5%), 177 tetraploids (88%), and 14 octoploids (7%) were obtained. Some premium anther-cultured plantlets were further selected by morphological and agronomic comparison. Our findings provide important guidance for potato ploidy breeding.

Effect of 12-months testosterone replacement therapy on bone mineral density and markers of bone turnover in testicular cancer survivors – results from a randomized double-blind trial

Background Testicular cancer survivors (TCS) are at risk of Leydig cell insufficiency, which is a condition characterized by elevated luteinising hormone (LH) in combination with low levels of testosterone. It has been suggested that this condition is associated with impaired metabolic profile and low bone mineral density (BMD). The primary aim of the randomized double-blind trial NCT02991209 was to evaluate metabolic profile after 12-months testosterone replacement therapy (TRT) in TCS with mild Leydig cell insufficiency. Here we present the secondary outcomes of changes in BMD and markers of bone turnover. Methodology In total, 69 TCS with mild Leydig cell insufficiency were randomized 1:1 to 12 months TRT (n = 35) (Tostran, gel, 2%, applied transdermally, with a maximum daily dose of 40 mg) or placebo (n = 34). BMD and markers of bone turnover were evaluated at baseline, after 6- and 12-months TRT, and 3-months post-treatment. Linear mixed effects models were used to analyse changes in BMD, N-terminal propeptide of type 1 procollagen (P1NP) and C-terminal telopeptide of type I collagen (CTX). Results After 12 months treatment, TRT was not associated with a statistically significant difference in BMD compared to placebo; total body BMD: 0.01 g/cm2 (95% confidence interval (CI): −0.01 − 0.02), BMD of the lumbar spine: 0.01 g/cm2, (95% CI: −0.01–0.03), BMD of the left femoral neck: 0.00, (95% CI: −0.01–0.02). TRT was associated with a small but statistically significant increase in P1NP: 11.65 µg/L (95% CI: 3.96, 19.35), while there was no difference in CTX. Conclusion 12 months of TRT did not change BMD, while there was as small and clinically irrelevant increase in P1NP compared to placebo in TCS with mild Leydig cell insufficiency. The findings need validation in a larger cohort.

Combined levothyroxine and testosterone treatment for restoring erectile dysfunction in propylthiouracil-induced hypothyroid rats.

Sexual dysfunction may indicate severe endocrine diseases. Recent research has suggested a link between hypothyroidism, low testosterone (T) levels, and erectile dysfunction (ED); however, the exact cause is unknown. We sought to investigate possible beneficial effects of levothyroxine and T alone or in combination on ED in propylthiouracil (PTU)-induced hypothyroid rats. Adult Wistar rats (n = 35) were divided into 5 groups: control, PTU-induced hypothyroidism, PTU + levothyroxine, PTU + Sustanon (a mixture of 4 types of T: propionate, phenylpropionate, isocaproate, and decanoate) and PTU + levothyroxine + Sustanon. PTU was given in drinking water for 6 weeks. Four weeks after PTU administration, levothyroxine (20μg microgram kg/day, oral) and Sustanon (10mg/kg/week, intramuscular) were given for 2weeks. Serum levels of total T, triiodothyronine (T3), and thyroxine (T4) were determined. In vivo erectile response and in vitro relaxant responses were measured. Localization of neuronal nitric oxide synthase (nNOS), endothelial NOS (eNOS), and phosphodiesterase type 5 (PDE5) were determined using immunohistochemical analysis. The relative area of smooth muscle to collagen was measured using Masson trichrome staining. Outcome variables included in vivo erectile function, in vitro relaxant and contractile responses of corpus cavernosum (CC) strips; protein localization of eNOS, nNOS, and PDE5; and smooth muscle content in penile tissue. The rat model of hypothyroidism showed a significant decline in serum levels of total T, T3, and T4. Levothyroxine increased T3 and T4 levels, whereas Sustanon normalized only total T levels. Combined treatment enhanced all hormone levels. Rats with hypothyroidism displayed the lowest erectile response (P < 0.001 vs controls). Combined treatment returned reduced responses, while partial amelioration was observed after levothyroxine and Sustanon treatment alone. Acetylcholine (P < 0.01 vs controls), electrical field stimulation (P < 0.001 vs controls), and sildenafil-induced relaxant responses (P < 0.05 vs controls) were decreased in the CC strips from hypothyroid rats. The combined treatment increased the reduction in relaxation responses. Levothyroxine and Sustanon restored decreases in eNOS and nNOS expression in the hypothyroid group. There was no significant difference in PDE5 expression among groups. Monotreatment partially enhanced reduced smooth muscle mass, while combined therapy completely recovered. The combination of thyroid hormones and T is likely to be a therapeutic approach for treatment of hypothyroidism-induced ED in men. Beneficial effects of levothyroxine and Sustanon treatment were shown in vitro and in vivo in PTU-induced hypothyroid rats. The main limitation of the study was the lack of measurement of androgen-sensitive organ weights and luteinizing hormone, follicle-stimulating hormone, and prolactin levels. These findings demonstrate that neurogenic and endothelium-dependent relaxation responses are reduced by hypothyroidism, which is detrimental to T levels and erectile responses. Levothyroxine and Sustanon combination medication was able to counteract this effect.

Efficacy and Safety of Neoadjuvant Luteinizing Hormone-Releasing Hormone Antagonist and Tegafur-Uracil Chemohormonal Therapy for High-Risk Prostate Cancer.

This retrospective single-center cohort study evaluated the efficacy and safety of a combination of neoadjuvant luteinizing hormone-releasing hormone (LHRH) antagonist and tegafur-uracil (UFT) therapy (NCHT) and investigated the medical records of patients with high-risk PCa who underwent robot-assisted radical prostatectomy (RARP). The therapy was followed by RARP for high-risk PCa. The enrolled patients were divided into two groups: low-intermediate-risk PCa patients who underwent RARP without neoadjuvant therapy (non-high-risk) and those who underwent NCHT followed by RARP (high-risk group). This study enrolled 227 patients (126: non-high-risk and 101: high-risk group). Patients in the high-risk-group had high-grade cancer compared to those in the non-high-risk-group. At the median follow-up period of 12.0 months, there were no PCa deaths; two patients (0.9%) died of other causes. Twenty patients developed biochemical recurrence (BCR); the median time until BCR was 9.9 months after surgery. The 2-year biochemical recurrence-free survival rates were 94.2% and 91.1% in the non-high-risk and high-risk-group, respectively (p = 0.465). Grade ≥3 NCHT-related adverse events developed in nine patients (8.9%). This study indicates that combining neoadjuvant LHRH antagonists and UFT followed by RARP may improve oncological outcomes in patients with high-risk PCa.

Autoimmune glial fibrillary acidic protein astrocytopathy: clinical analysis and review of 15 cases

BackgroundTo review clinical characteristics, auxiliary examination results, treatment effects, and outcomes of patients with autoimmune glial fibrillary acidic protein astrocytopathy (GFAP-A).MethodsWe collated and retrospectively analyzed clinical data of 15 patients admitted with clinical characteristics of an autoimmune GFAP-A acute encephalitis or meningitis phenotype.ResultsAll patients were diagnosed with acute-onset meningoencephalitis and meningoencephalomyelitis. Initial presentations included pyrexia and headache at onset; dual symptoms of prominent tremor with urinary and bowel dysfunction; ataxia, psychiatric and behavioral abnormalities, and impaired consciousness; neck resistance; reduced extremity muscle strength; blurred vision; epileptic seizures; and reduced basic blood pressure. Cerebrospinal fluid (CSF) examination showed that the degree of protein elevation was significantly higher than the degree of increase in white blood cells. Moreover, in the absence of obvious low chloride and glucose levels, CSF chloride levels decreased in 13 patients, accompanied by a CSF glucose level decrease in four. Brain abnormalities were found in magnetic resonance imaging of ten patients, with a linear radial perivascular enhancement present in the lateral ventricles of two patients and symmetric abnormalities in the splenium of the corpus callosum in three patients.ConclusionsAutoimmune GFAP-A may be a spectrum disorder, with acute- or subacute-onset meningitis, encephalitis, and myelitis being the main phenotypes. When used for acute stage treatment, combined hormone and immunoglobulin therapy was superior to hormone pulse therapy or immunoglobulin pulse therapy alone. However, hormone pulse therapy alone without immunoglobulin pulse therapy was associated with a greater number of remaining neurological deficits.

Evaluations of memory, anxiety, and the growth factor IGF-1R after post-surgical menopause treatment with a highly selective progestin

Progestogens are a key component of menopausal hormone therapies. While some progestogens can be detrimental to cognition, there is preclinical evidence that progestogens with a strong progesterone-receptor affinity benefit some molecular mechanisms believed to underlie cognitive function. Thus, a progestin that maximizes progesterone-receptor affinity and minimizes affinities to other receptors may be cognitively beneficial. We evaluated segesterone-acetate (SGA), a 19-norprogesterone derivative with a strong progesterone-receptor affinity and no androgenic or estrogenic-receptor activity, hypothesizing that it would enhance cognition. Middle-aged rats underwent Sham or Ovariectomy (Ovx) surgery followed by administration of medroxyprogesterone-acetate (MPA; used as a positive control as we have previously shown MPA-induced cognitive deficits), SGA (low or high dose), or vehicle (one Sham and one Ovx group). Spatial working and reference memory, delayed retention, and anxiety-like behavior were assessed, as were memory- and hormone- related protein assays within the frontal cortex, dorsal hippocampus, and entorhinal cortex. Low-dose SGA impaired spatial working memory, while high-dose SGA had a more extensive detrimental impact, negatively affecting spatial reference memory and delayed retention. Replicating previous findings, MPA impaired spatial reference memory and delayed retention. SGA, but not MPA, alleviated Ovx-induced anxiety-like behaviors. On two working memory measures, IGF-1R expression correlated with better working memory only in rats without hormone manipulation; any hormone manipulation or combination of hormone manipulations used herein altered this relationship. These findings suggest that SGA impairs spatial cognition after surgical menopause, and that surgical menopause with or without progestin administration disrupts relationships between a growth factor critical to neuroplasticity.

Transcription Factor Immunohistochemistry in the Classification of Pituitary Neuroendocrine Tumor/Adenoma: Proposal in a Limited-Resource Setting.

Pituitary neuroendocrine tumors/adenomas are common intracranial tumors that require accurate subtyping because each tumor differs in its biologic behavior and response to treatment. Pituitary-specific transcription factors allow for improved lineage identification and diagnosis of newly introduced variants. To assess the usefulness of transcription factors and design a limited panel of immunostains for classification of pituitary neuroendocrine tumors/adenoma. A total of 356 tumors were classified as per expression of pituitary hormones and transcription factors T-box family member TBX19 (TPIT), pituitary-specific POU-class homeodomain (PIT1), and steroidogenic factor-1 (SF-1). The resultant classification was correlated with patients' clinical and biochemical features. The performance and relevance of individual immunostains were analyzed. Reclassification of 34.8% (124 of 356) of pituitary neuroendocrine tumors/adenoma was done after application of transcription factors. The highest agreement with final diagnosis was seen using a combination of hormone and transcription factors. SF-1 had higher sensitivity, specificity, and predictive value compared with follicle-stimulating hormone and luteinizing hormone. On the other hand, TPIT and PIT1 had similar performance and Allred scores compared with their respective hormones. SF-1 and PIT1 should be included in the routine panel for guiding the classification. PIT1 positivity needs to be followed by hormone immunohistochemistry, especially in nonfunctional cases. TPIT and adrenocorticotropin can be used interchangeably as per availability of the lab.

Effect of leuprorelin plus growth hormone on serum luteinizing hormone, follicle stimulating hormone, and estradiol levels in children with idiopathic central precocious puberty

Purpose: To evaluate the effects of leuprorelin plus growth hormone on serum luteinizing hormone (LH), follicle-stimulating hormone (FSH), and estradiol (E2) levels in children with idiopathic central precocious puberty. Methods: A total of 102 girls with idiopathic central precocious puberty who met the inclusion criteria between April 2020 and September 2021 were enrolled in this study, and assigned randomly to either a control group given leuprorelin only, or a combined treatment group given leuprorelin plus growth hormone. There were 51 cases in each group. The parameters/indices evaluated include sex hormone levels (serum LH, FSH, and E2), height standard deviation scores (HtSDS), predicted adult height (PAH), and insulin-like growth factors-1 (IGH-1). Results: Patients administered combined therapy showed lower levels of LH, FSH, and E2 than those on mono-therapy (p &lt; 0.05). Leuprorelin plus growth hormone resulted in significantly higher HtSDS and PAH, and lower IGF-1 levels compared with those who received leuprorelin (p &lt; 0.05). Conclusion: The combination of leuprorelin and growth hormone effectively reduces the serum levels of LH, FSH, and E2 in children with idiopathic central precocious puberty, and improves their linear growth rate and adult height. However, economic benefits and patient acceptance should be considered in future studies to assess its overall applicability.

Estetrol Increases Progesterone Genetic Response without Triggering Common Estrogenic Effects in Endometriotic Cell Lines and Primary Cultures

Estetrol (E4), a natural estrogen produced by the human fetal liver, is actively studied for menopause and breast cancer treatment. It has low side effects and preferential estrogen receptor alpha (ERα) affinity. There are no data about its effects on endometriosis, a common gynecological disease affecting 6-10% of cycling women, generating painful pelvic lesions and infertility. Current combined hormone treatment (progestins and estrogens) is safe and efficient; nevertheless, one-third of patients develop progesterone (P4) resistance and recurrence by reducing P4 receptors (PRs) levels. We aimed to compare E4 and 17β-estradiol (E2) effects using two human endometriotic cell lines (epithelial 11Z and stromal Hs832 cells) and primary cultures from endometriotic patients. We evaluated cell growth (MTS), migration (wound assay), hormone receptors levels (Western blot), and P4 response by PCR array. Compared to E2, E4 did not affect cell growth or migration but increased estrogen receptor alpha (ERα) and PRs, and reduced ERβ. Finally, the incubation with E4 improved the P4 gene response. In conclusion, E4 increased PRs levels and genetic response without inducing cell growth or migration. These results suggest that E4 might be useful for endometriosis treatment avoiding P4 resistance; however, evaluating its response in more complex models is required.

EFEKTIFITAS KOMBINASI FORMULA OIL CRAB DAN EKSTRAK DAUN KARAMUNTING (Melastoma malabathricum) PADA PROSES PEMATANGAN OVARI INDUK KEPITING BAKAU (Scylla sp.)

Mud crab are one of the fisheries commodities that have important economic value. Today the demand for mud crab on the market is increasing, so that the capture of mud crab is also increasing. To overcome this, various efforts have been made to meet market demand, namely by conducting seeding, fattening and enlargement of mud crab in the pond. This study aims to determine the maturity level of mud crab ovaries (Scylla sp.) by administering a combination of karamunting (M. malabathricum) leaf extract and thyroxine hormone. This study used a complete randomized design with 4 treatments and 3 replications. The treatments in this study were treatment A (control), treatment B (M. Malabathricum 0.25 mg/g and thyroxine hormone 0.1 µg/g), treatment C (M. Malabathricum 0.5 mg/g and thyroxine hormone 0.1 µg/g), treatment D (M. Malabathricum 1 mg/g and thyroxine hormone 0.1 µg/g). The results showed that the administration of a combination of karamunting leaf extract (M. malabathricum) and the thyroxine hormone in treatments B and C can accelerate the process of egg removal (berried), which is within 20 days. The combination of karamunting leaf extract (M. malabathricum) and the thyroxine hormone can accelerate the ripening ovaries of mud crab from control treatment.

HYPOGLYCEMIA AND JAUNDICE IN A NEWBORN REVEALING PITUITARY STALK INTERRUPTION SYNDROME

Pituitary stalk interruption syndrome (PSIS) is a rare congenital abnormality of the pituitary gland. It is associated with either isolated growth hormone deficiency or combined pituitary hormone deficiency (CPHD). Neonatal PSIS is extremely rare and difficult to diagnose and can be life threatening if diagnosed late. In this manuscript, we descripted a case of neonatal PSIS who is treated at the department ofPediatric Endocrinology atHassan II University Hospital Center of Fez.Sustained hypoglycemia and jaundice in newborns, indicate the presentation of PSIS. Early recognition is of great importance to avoid a life-threatening crisis.

Individual and combined effects of arbuscular mycorrhizal fungi and phytohormones on the growth and physiobiochemical characteristics of tea cutting seedlings.

Both arbuscular mycorrhizal fungi (AMF) and phytohormones collectively regulate plant growth and root development, but their individual and combined effects on tea [Camellia sinensis (L.) O. Kuntze] cutting seedings remain unclear. This study examined the individual and combined effects of two species of AMF (Rhizophagus intraradices, RI and Funneliformis mosseae, FM) and two types of palnt hormones (strigolactones, SLs; polyamines, PAs) on tea cutting seedings, by evaluating the growth and physiobiochemical characteristics of plants treated with the AMFs and/or hormones. The results showed that inoculation with either AMF individually or hormones treatment alone could significantly enhanced mycorrhizal colonization, growth target and physiobiochemical characteristics of tea cutting seedlings. Interestingly, the addition of a combination of AMFs and hormones showed superior effects, while SL and RI exhibited the most improvements to the colonization rate, plant growth, root-morphological traits, root DHA activity, photosynthesis, chlorophyll content, soluble sugar content in leaves, and the activities of antioxidant enzymes (SOD, POD, and CAT), compared to other treatment combinations (SL + FM, PA + RI, and PA + FM). Correlation analyses revealed a significantly (p < 0.05) positive correlation of root AMF colonization with root-related traits (e.g., DHA, root total length, surface area, and volume) and leaf-related traits (e.g., leaf area, shoot biomass, total chlorophyll, and antioxidant enzyme activities). This study demonstrated that while the apllication of individual AMF or plant hormones had a certain good effects on most growth and physiobiochemical characteristics parameters of tea cutting seedings, the additive effect was from specific combined of AMF and plant hormones. These results highlight the possibility for combined of AMF and plant hormones to improve the asexual reproduction of tea plants via cuttings.

Index based selection of treatments and genotypes for the in-vitro shoot regeneration in banana

The in vitro shoot regeneration studies for four banana cultivars (Poovan, Karpuravalli, Rasthali andMonthan) was conducted via shoot tip culture using three plant bioregulators (PBRs) viz., 6-Benzylaminopurine(BAP), Kinetin (KIN) and 1-naphthaleneacetic acid (NAA) in different combinations and concentrations. Shootregeneration revealed the significant effects of various PBRs combinations. This study utilized index basedselection, for the selection of best treatments and cultivars for shoot regeneration in bananas. Based on theSmith’s selection index score, the PBRs combination T6 (BAP@5.0mg/l+NAA@0.5mg/l) which ranked first (0.24)was selected as the best PBRs combination for shoot regeneration. Among the cultivars, ‘Rasthali’ (-0.76) whichranked first was selected as the best genotype for shoot regeneration in bananas. However, the combinedeffect of hormonal combination (T6) and cultivar ‘Poovan’ (2.61) ranked first. These results also showed that theresponse of ABB type (Karpuravalli and Monthan) was poorer than that of the AAB group (Poovan and Rasthali)indicating that, the in vitro regeneration was much influenced by the genotype of plant species and the type ofexplants material as well as the type of PBR combinations. Overall, the cultivar-specific regeneration protocolproved essential for better in vitro regeneration of banana.

Low-intensity pulsed ultrasound and parathyroid hormone improve muscle atrophy in estrogen deficiency mice

Due to aging and long-term estrogen deficiency, postmenopausal women suffer muscle atrophy (MA), which is characterized by decreased muscle mass and muscle quality. Low-intensity pulsed ultrasound (LIPUS) is an acoustic wave inducing biological effects mainly by the mechanical stimulation and used as a non-invasive physical therapy for muscle repair. Parathyroid hormone (PTH) is an 84-amino-acid polypeptide, and its bioactive fragment [PTH (1–34)] has potential application in the treatment of MA. We speculate that the combination of physical therapy (i.e., the LIPUS) and regulatory hormone (i.e., the PTH) would be more effective in the treatment of MA. The objective of this study was to evaluate the individual and combined effects of LIPUS and PTH therapy on MA in estrogen deficiency mice. Seventy 8-week-old female C57BL/6J mice were used in this study and the MA model was induced by an intraperitoneal injection of 4-vinylcyclohexene diepoxide (VCD) for 20 consecutive days. The VCD-induced MA mice were randomly divided into MA, LIPUS, PTH and LIPUS + PTH (Combined) groups (n = 10/group). In the LIPUS group, the mice were treated by LIPUS in bilateral quadriceps muscles for 20 min, five times a week for 6 weeks. In the PTH group, the mice received subcutaneous injection of PTH (1–34) (80 ug/kg/d) five times a week, for 6 weeks. In the Combined group, the PTH was administrated 30 min before each LIPUS session. Hematoxylin-eosin (H&E) staining, serum biochemical analysis and quantitative real-time polymerase chain reaction (qRT-PCR) were applied to evaluate the therapeutic effects of related treatments. The results showed that the MA mice had a disordered estrus cycle, significantly decreased muscle mass and myofibers cross-sectional area (CSA). After treatments, LIPUS, PTH and Combined groups had a significantly increased CSA, compared with the MA mice without treatment. In addition, Combined group had a significantly increased mRNA expression of Pax7, MyoD and MyoG, compared with LIPUS and PTH monotherapy groups. Our findings indicated that the combination of LIPUS and PTH treatment improves muscle regeneration ability, which might have potential for treating MA in postmenopausal women.

Influence of plant growth regulators on in vitro biomass production and biosynthesis of cytotoxic Amaryllidaceae alkaloids in Caliphuria tenera Baker

Plants of the Amaryllidoideae subfamily are characterized by the presence of isoquinoline alkaloids called Amaryllidaceae alkaloids. These compounds have exhibited a broad spectrum of pharmacological effects, including anti-Alzheimer and anti-cancer. In this study, Caliphruria tenera Baker, Amaryllidaceae, an endangered and endemic species, was selected to evaluate the effect of growth regulators on plant micropropagation, biomass increase, and production of cytotoxic alkaloids. Different growth regulators were used individually and mixed, and the Amaryllidaceae alkaloids were identified and quantified by GC-MS. The maximum number of in vitro shoots (4.8) was recorded with 6-benzylaminopurine (BAP) (1 mg/L) + Kinetin (KIN) (1 mg/L) + Naphthaleneacetic acid (NAA) (0.2 mg/L). In addition, nineteen alkaloids were identified as crinine, galantamine, lycorine, narciclasine, and tazzetine-type alkaloids in cultivated and micropropagated plants. In general, the biomass was maximized with the growth hormone combination BAP + KIN + NAA, but the biosynthesis of alkaloids was minimized. However, the combination BAP + NAA showed the greatest productivity of trisphaeridine (632.01 μg/g). The growth regulators showed also a positive effect on the production of lycorine-type alkaloids with a content of 396.33 μg/g for KIN, 142.38 μg/g for BAP, and 41.67 μg/g for NAA. All alkaloid fractions were evaluated for their antiproliferative effects against human lung carcinoma (A549) and human keratinocytes (HaCat) cells. The alkaloids produced in the BAP + NAA treatment present a potent and selective cytotoxic against the A549 cell line at 30 μg/mL. This study shows that micropropagation is a promising approach for the production of cytotoxic Amaryllidaceae alkaloids in C. tenera without affecting natural populations.

Optimization of bud disinfection technique and influence of growth regulators on micropropagation in ginger (Zingiber officinale Rosc.)

Ginger (Zingiber officinale Rosc.) is a spice, considered as a food medicine because of its numerous beneficial actions on health. However, its production faces several constraints, including the lack of efficient planting material. This is a limiting factor for industrial ginger production. The present study therefore aimed at developing an effective in vitro regeneration protocol for ginger. First, three disinfecting agents (sodium hypochlorite, calcium hypochlorite and mercury chloride) were tested. Then, different combinations of naphthalene acetic acid (0.5 mg/L NAA) and/or benzyl amino purine (1, 3 and 5 mg/L BAP) were evaluated on in vitro shoots regeneration. The results revealed that 3.6% sodium hypochlorite, used for 20 min, induced the best disinfection rates (100%) and healthy buds (84.66%). Furthermore, this study showed that the tested hormonal combinations significantly influenced shoots proliferation in ginger. However, the Proliferation Medium 4 (PM4) [Murashige and Skoog medium including vitamin B5 (MSB) + 0.5 mg/L of Naphthalene Acetic Acid (ANA) + 5 mg/L of Benzyl Amino Purine (BAP)] was the most effective. It induced the highest average number of shoots (22.83 shoots) with an induction rate of 80.50%. As a result of this study, 3.6% sodium hypochlorite used for 20 min and MP4 medium (MSB + 0.5 mg/L ANA + 5 mg/L BAP) were selected for in vitro ginger regeneration.

FEATURES OF ENDOCRINE PROFILE IN HYPERTROPHIC AND KELOID SCARS OF SKIN

Background. Given the role of hormones in the regulation of scar formation and remodeling processes, it is advisable to study the features of the hormonal profile in conditions of pathological scars. This will improve the efficiency of diagnosing of scar type and form a pathogenetic basis for the treatment of hypertrophic and keloid scars. Aim. To identify the features of the hormonal profile in conditions of hypertrophic and keloid scars. Materials and methods. The clinical state of the scars and the hormonal profile (10 hormones) were studied in 90 women with hypertrophic, keloid and normotrophic scars (control). Blood was taken before treatment on the 5th–7th day of the menstrual cycle. The results were processed by non-parametric statistics (Me (Q25-Q75), U-test at p&lt;0.05). Correlations between hormones were determined by the method of multiple regression at the coefficient of determination R2&gt;0.7. Results. All patients with pathological scars showed a decrease in cortisol levels. Hypertrophic scars are characterized by an increase in the concentration of follicle-stimulating hormone, a decrease in the concentration of estradiol and testosterone. In patients with keloids, the concentration of growth hormone is increased, the levels of prolactin, luteinizing hormone are reduced. Regression analysis showed that the interdependence of hormones is maximally expressed in conditions of normotrophic scars, while hormones had 1-2 correlations in conditions of pathological scars. At the same time, the qualitative composition and direction of interrelations have changed. Conclusion. Pathological scarring is characterized by a decrease in the level of cortisol in the blood, which is expressed in patients with keloid scars in combination with reduced prolactin. A weakening of hormonal interactions and an imbalance in the system of sex hormones were revealed in conditions of hypertrophic scars. and the increased production of growth hormone in combination with a stable interaction between steroidogenesis and a weakened adaptive ability took place in conditions of keloid scars.

Biochemical identification of prepubertal boys with Klinefelter syndrome by combined reproductive hormone profiling using machine learning.

Klinefelter syndrome (KS) is the most common sex chromosome disorder and genetic cause of infertility in males. A highly variable phenotype contributes to the fact that a large proportion of cases are never diagnosed. Typical hallmarks in adults include small testes and azoospermia which may prompt biochemical evaluation that typically shows extremely high FSH and low/undetectable inhibin B serum concentrations. However, in prepubertal KS individual biochemical parameters are largely overlapping those of prepubertal controls. We aimed to characterize clinical profiles of prepubertal boys with KS in relation to controls, and to develop a novel biochemical classification model to identify KS before puberty. Retrospective, longitudinal data from 15 prepubertal boys with KS and data from 1475 controls were used to calculate age- and sex-adjusted standard deviation scores (SDS) for height and serum concentrations of reproductive hormones and used to infer a decision tree classification model for KS. Individual reproductive hormones were low but within reference ranges and did not discriminate KS from controls. Clinical and biochemical profiles including age- and sex-adjusted SDS from multiple reference curves provided input data to train a 'random forest' machine learning (ML) model for detection of KS. Applied to unseen data, the ML model achieved a classification accuracy of 78% (95% CI, 61% - 94%). Supervised ML applied to clinically relevant variables enabled computational classification of control and KS profiles. The application of age- and sex-adjusted SDS provided robust predictions irrespective of age. Specialized ML models applied to combined reproductive hormone concentrations may be useful diagnostic tools to improve the identification of prepubertal boys with KS.

Hormone exposure and venous thromboembolism in commercially insured women aged 50 to 64 years

BackgroundMenopausal hormone therapy (MHT) can elevate venous thromboembolism (VTE) risk, but less is known about formulations and routes of exposures. ObjectiveTo estimate hormone-associated VTE risk by route and formulation in exposed and unexposed women aged 50 to 64 years in the US. MethodsIn a nested case-control study of US commercially insured women aged 50 to 64 years (2007-2019), cases were defined as incident VTE diagnoses and matched to 10 controls by date of VTE and age, excluding prior VTE, inferior vena cava filter placement, or anticoagulants. Filled prescriptions in the prior year defined hormone exposures. International Classification of Diseases and Current Procedural Terminology codes identified risk factors and comorbidities. ResultsOdds ratios (ORs) were estimated with conditional logistic regression controlling for differences between cases (n = 20,359) and controls (n = 203,590) in comorbidities and VTE risk factors. For exposures within 60 days, oral MHT risk was almost twice as high as transdermal MHT (OR = 1.92; 95% CI, 1.43-2.60); transdermal MHT did not elevate risk compared with no exposure (unopposed OR = 0.70; 95% CI, 0.59-0.83; combined OR = 0.73; 95% CI, 0.56-0.96). Risk was highest for MHT combinations with ethinyl estradiol, followed by conjugated equine estrogen (CEE) (ethinyl estradiol-CEE: OR = 1.55; 95% CI, 1.07-2.25), and lowest for estradiol (CEE-estradiol: OR = 1.33; 95% CI, 1.02-1.72). Combined hormonal contraceptives elevated risk 5 times higher than no exposure (OR = 5.22; 95% CI, 4.67-5.84) and 3 times higher than oral MHT (OR = 3.65; 95% CI, 3.09-4.31). ConclusionThe risk of VTE is much lower with MHT than combined hormone contraceptives and varies by hormone formulation and route of exposure. Transdermal MHT did not elevate risk. Oral MHT combinations with estradiol were lower risk than other forms of estrogen. Oral combined hormone contraceptives had much higher risk than oral combined hormonal MHT.