Weight gain in menopause: systematic review of adverse events in women treated with black cohosh.

Weight gain is a frequent problem in perimenopausal and postmenopausal women. Cimicifuga racemosa (CR) is a popular treatment option for menopausal symptoms. The aim of this review was to investigate whether there is scientific evidence that CR causes weight gain. We searched our database for medically confirmed, spontaneous adverse events regarding weight gain, literature for case reports and randomized controlled trials. Thirty cases in total were spontaneously reported in 15 years. The causality was not considered certain/likely in any of the cases. A nurse (consumer) assessed the causality as possible. Only one case was published in the literature. However, no change in body fat composition was reported, and the causality seems unlikely. Of the 31 identified studies, 17 were double-blind placebo-controlled, five were double-blind reference-controlled and nine were open reference-controlled. In total, 1839 women were treated with CR for up to 12 months. Two studies reported weight gain as an adverse event; however, no significant differences in weight changes were observed between the groups. One case of weight gain (about 2 kg) was reported, but the authors did not specify in which treatment group. In conclusion, this study provides no scientific evidence that the use of Cimicifuga racemosa causes weight gain in menopausal women.

Review & meta-analysis: isopropanolic black cohosh extract iCR for menopausal symptoms - an update on the evidence.

A systematic literature search revealed 35 clinical studies and one meta-analysis comprising 43,759 women, of which 13,096 were treated with isopropanolic Cimicifuga racemosa extract (iCR). Compared to placebo, iCR was significantly superior for treating neurovegetative and psychological menopausal symptoms, with a standardized mean difference of -0.694 in favor of iCR (p < 0.0001). Effect sizes were larger when higher dosages of iCR as monotherapy or in combination with St. John's wort (Hypericum perforatum [HP]) were given (-1.020 and -0.999, respectively), suggesting a dose-dependency. For psychological symptoms, the iCR+HP combination was superior to iCR monotherapy. Efficacy of iCR was comparable to low-dose transdermal estradiol or tibolone. Yet, due to its better tolerability, iCR had a significantly better benefit-risk profile than tibolone. Treatment with iCR/iCR+HP was well tolerated with few minor adverse events, with a frequency comparable to placebo. The clinical data did not reveal any evidence of hepatotoxicity. Hormone levels remained unchanged and estrogen-sensitive tissues (e.g. breast, endometrium) were unaffected by iCR treatment. As benefits clearly outweigh risks, iCR/iCR+HP should be recommended as an evidence-based treatment option for natural climacteric symptoms. With its good safety profile in general and at estrogen-sensitive organs, iCR as a non-hormonal herbal therapy can also be used in patients with hormone-dependent diseases who suffer from iatrogenic climacteric symptoms.

DIAGNOSIS OF ENDOCRINE DISEASE: Drug-induced endocrinopathies and diabetes: a combo-endocrinology overview.

In the currently overwhelming era of polypharmacy, the balance of the dynamic and delicate endocrine system can easily be disturbed by interfering pharmaceutical agents like medications. Drugs can cause endocrine abnormalities via different mechanisms, including direct alteration of hormone production, changes in the regulation of the feedback axis, on hormonal transport, binding and signaling, as well as similar changes to counter-regulatory hormone systems. Furthermore, drugs can interfere with the hormonal assays, leading to erroneous laboratory results that disorientate clinicians from the right diagnosis. The purpose of this review is to cover a contemporary topic, the drug-induced endocrinopathies, which was presented in the monothematic annual Combo Endo Course 2018. This challenging part of endocrinology is constantly expanding particularly during the last decade, with the new oncological therapeutic agents, targeting novel molecular pathways in the process of malignancies. In this new context of drug-induced endocrine disease, clinicians should be aware that drugs can cause endocrine abnormalities via different mechanisms and mimic a variety of clinical scenarios. Therefore, it is extremely important for clinicians not only to promptly recognize drug-induced hormonal and metabolic abnormalities, but also to address the therapeutic issues for timely intervention.

Genes involved in hormone metabolism and cellular response in human ovarian granulosa cells.

Endocrinal interactions are one of the most crucial regulatory mechanisms that maintain the state of homeostasis in humans. Processes such as oogenesis, folliculogenesis, menstruation and pregnancy remain under hormonal control. A key role in folliculogenesis is played by granulosa cells. Moreover, granulosa cells take part in corpus luteum formation after ovulation. Because of that, it is important to understand the ways in which the granulosa cells, associated with those processes, respond to hormonal stimulus. In the present study, a transcriptomic analysis of human granulosa cells (GCs) was carried out with the use of expression microarrays. The results were validated by RT-qPCR. The total RNA was isolated after 1st, 7th, 15th and 30th days of long-term primary cultures. The main focus of this work was placed on the genes belonging to "Response to estradiol", "Response to follicle-stimulating-hormone", "Cellular response to hormone stimulus", "Cellular hormone metabolic process" and "Hormone biosynthetic process" gene ontology groups. These groups of genes have been associated with GC hormone metabolism and cellular response to hormones. Eighty genes belonging to these groups were identified. Those that were members of more than one of the analyzed gene ontology groups, or exhibited unique expression patterns, were selected for further analysis. All of the selected genes were described, with their expression patterns detailed. In this manuscript, two gene expression patterns have been described. The first one showed large downregulation of genes in the later stages of culture, with the second one presenting upregulation of expression after day 1 of IVC. The present research was focused on six genes found to be the most important for steroidogenesis: STAR, POR, CYP11A1, ADM, GCLC, IL1B, as well as three genes of higher expression at the later stages of long-term in vitro culture: NR2F2, BMP4, COL1A1. The main goal of the presented study was to select genes involved in response to hormonal stimulus and hormone metabolism in GC long-term in vitro culture.

Transcriptomic profile of cell cycle progression genes in human ovarian granulosa cells.

The ovarian granulosa cells (GCs) that form the structure of follicle undergo substantial modification during the various stages of human folliculogenesis. These modifications include morphological changes, accompanied by differential expression of genes, encoding proteins which are mainly involved in cell growth, proliferation and differentiation. Recent data bring a new insight into the aspects of GCs' stem-like specificity and plasticity, enabling their prolonged proliferation and differentiation into other cell types. This manuscript focuses attention on emerging alterations during GC cell cycle - a series of biochemical and biophysical changes within the cell. Human GCs were collected from follicles of women set to undergo intracytoplasmic sperm injection procedure, as a part of remnant follicular fluid. The cells were primarily cultured for 30 days. Throughout this time, we observed the prominent change in cell morphology from epithelial-like to fibroblast-like, suggesting differentiation to other cell types. Additionally, at days 1, 7, 15 and 30, the RNA was isolated for molecular assays. Using Affymetrix® Human Genome U219 Array, we found 2579 human transcripts that were differentially expressed in GCs. From these genes, we extracted 582 Gene Ontology Biological Process (GO BP) Terms and 45 KEGG pathways, among which we investigated transcripts belonging to four GO BPs associated with cell proliferation: "cell cycle phase transition", "G1/S phase transition", G2/M phase transition" and "cell cycle checkpoint". Microarray results were validated by RT-qPCR. Increased expression of all the genes studied indicated that increase in GC proliferation during long-term in vitro culture is orchestrated by the up-regulation of genes related to cell cycle control. Furthermore, observed changes in cell morphology may be regulated by a presented set of genes, leading to the induction of pathways specific for stemness plasticity and transdifferentiation in vitro.

Effects of metformin or an oral contraceptive containing cyproterone acetate on serum c-reactive protein, interleukin-6 and soluble vascular cell adhesion molecule-1 concentrations in women with polycystic ovary syndrome.

The aim of the present study was to evaluate the effects of commonly used treatment regimens such as metformin (MET) or an oral contraceptive pill (OC) containing ethynyloestradiol and cyproterone acetate (EE-CPA) on surrogate serum CVD risk factors and markers of endothelial dysfunction (CRP, IL-6, sVCAM) in women with PCOS.This study was conducted in a crossover design in order to compare the effects of 2 different treatment regimens in the same subject and has been registered under the number NCT01798875 in the ClinicalTrials.gov registry.42 women with PCOS (age range 18-36 years, median BMI 24.9) were randomly assigned to treatment with MET (850 mg bid) or EE-CPA containing OC for 4 months. After 2 months washout period, treatments were crossed over.Treatment with and OC increased significantly serum CRP concentrations (from 0.77 mg/l [95% CI: 0.70; 2.18] to 1.70 mg/l [95% CI: 1.65; 3.69], P<0.001). Treatment with MET slightly reduced serum CRP levels, but this difference did not reach statistical significance (P=0.08). 4 months treatment with MET or EE-CPA had no effect on serum IL-6 and sVCAM-1 concentrations (P>0.05).Treatment with EE-CPA containing OC for 4 months in women with PCOS significantly raises serum CRP. Since this rise was not accompanied by the increase in serum concentrations of IL-6, which is the most potent and effective stimulant of hepatic CRP production, we can speculate that this effect is caused by the liver first-pass effect of oral oestrogen administration. If this in turn can confer, cardiovascular risk among these women warrants further -studies.

Differential diagnosis of hyperandrogenism in women with polycystic ovary syndrome.

According to the Androgen Excess and Polycystic Ovary Syndrome Society (AE&PCOS), the main feature of PCOS is clinical hyperandrogenism or laboratory hyperandrogenaemia. Therefore, in diagnosing PCOS one must always exclude other causes of androgen excess. In a woman with hyperandrogenism, the diagnosis of PCOS can usually be made according to the patient's history and careful clinical examination. Signs of mild hyperandrogenaemia usually start after the menarche and cycles continue to be anovulatory in adult life. Non-classical congenital adrenal hyperplasia (NCCAH) can be another cause of hyperandrogenism with oligomenorrhea. This can be diagnosed in a patient with elevated basal or ACTH stimulated serum 17OH-progesterone (17-OHP) levels or in a case of a significant decrease in serum testosterone (TST) and dehydroepiandrosterone sulphate (DHEA-S) in a two day dexamethasone suppression test. Cushing's disease (ACTH producing pituitary adenoma) is a rare cause of hyperandrogenaemia in women with recent onset of hyperandrogenism. However, it must always be taken into the consideration in a patient with accompanying signs of hypercortisolism. It can usually be excluded by performing an overnight dexamethasone suppression test or the measurement of 24 h urinary free cortisol levels. Severe signs of hyperandrogenism which lead to virilization should always lead to the exclusion of androgen secreting tumors of ovarian or adrenal origin. These are very rare but should be always taken into the account in a patient with recent onset of severe signs of androgen excess and very high serum androgen levels. Mild signs of hyperandrogenaemia in a woman with recent oligomenorrhea should always lead to the exclusion of hyperprolactinaemia.

Effect of exercise training on insulin sensitivity, mitochondria and computed tomography muscle attenuation in overweight women with and without polycystic ovary syndrome.

AIMS/HYPOTHESIS: Polycystic ovary syndrome (PCOS) is an insulin resistant (IR) state. Increased skeletal muscle lipid content and impaired mitochondrial biogenesis have been implicated in the pathogenesis of IR. We investigated whether differences in these variables explain the IR of women affected by PCOS and whether improvements in IR with exercise are reflected by changes in these variables. METHODS: Sixteen PCOS and 13 non-PCOS overweight women were assessed, and eight PCOS and seven non-PCOS women were reassessed after 12 weeks of moderate and vigorous exercise training. Outcomes included insulin sensitivity (glucose infusion rate [GIR]), skeletal muscle gene expression and protein abundance, enzyme activity of selected mitochondrial components, and computed tomography (CT) attenuation-estimated muscle lipid. RESULTS: GIR was lower in women with PCOS versus those without (p = 0.01) and increased with exercise in both groups. Baseline CT muscle attenuation suggested a trend to less muscle lipid in PCOS, which increased with exercise training, with a difference in the change in muscle lipid (p = 0.01, age-corrected), compared with non-PCOS women. GIR correlated with PGC1A gene expression across the whole group; skeletal muscle expression of mitochondrial biogenesis markers was not different between groups at baseline, or after training. Neither lipid changes nor mitochondrial changes correlated with changes in GIR. CONCLUSIONS/INTERPRETATION: Differences in IR in women with and without PCOS were not explained by differences in skeletal muscle lipid or mitochondrial parameters. Improvements in IR with exercise were dissociated from mitochondrial parameters. CT muscle attenuation suggested a differential capacity of PCOS muscle to store lipid compared with non-PCOS. TRIAL REGISTRATION: Clinicaltrials.gov ISRCTN84763265. FUNDING: National Health & Medical Research Council (Grant number 606553), Monash University and The Jean Hailes Foundation.

Postmenopausal hormone therapy and the risk of venous thromboembolism.

Venous thromboembolism (VTE) is an important determinant of the benefit-to-risk profile of postmenopausal hormone replacement therapy (HRT). Women's health practitioners who prescribe HRT to their patients are often more concerned about the thromboembolic complications than the risk of breast malignancy. This is in contrast to their patients who are eligible and considering commencing hormonal treatment where breast cancer is often the primary concern. This review summarizes the data on the actual HRT-related VTE risk and factors influencing it. It also provides practical guidelines which should support the health professional in ensuring informed choice on HRT for women.

Uterotropic effects of dietary equol administration in ovariectomized Sprague-Dawley rats.

AIM: The aim of the present study was to evaluate the uterotropic effects of the administration of dietary equol, a metabolite of soy-derived daidzein or formononetin present in red clover, in an ovariectomized rat model of menopause. METHOD: Two doses of racemic equol were used (50 mg/kg of chow and 400 mg/kg of chow) and the results were compared with two doses of estradiol-3 benzoate (E2B) (4.3 mg/kg of chow and 17.3 mg/kg of chow). After 3 months, animals were sacrificed and the uteri were removed, weighed and paraffin-embedded for morphometrical and immunohistochemical evaluation. The expression of selected uterine estrogen-responsive genes was also measured using real-time reverse transcription-polymerase chain reaction. RESULTS: Compared to controls, uterine weights in animals treated with high-dose equol were significantly higher, presented histologic features of mild estrogenic stimulation and had greater epithelial height and thickness of the uterine stroma and myometrium. Staining for the presence of the proliferating cell nuclear antigen (PCNA) also showed a greater prevalence of the PCNA-positive cells in the uterine stroma in animals treated with high-dose equol. Conversely, the percentage of PCNA-positive cells in the uterine epithelium was lower compared to the controls. Dietary high-dose equol treatment also increased significantly levels of uterine insulin-like growth factor 1, progesterone receptor and complement protein 3 mRNA. Although statistically significant, all these effects were, however, lower in magnitude compared to the effects of low- and high-dose E2B treatment. Low-dose equol did not have any effects on the above-studied parameters. CONCLUSION: Long-term high-dose dietary equol administration to ovariectomized rats exerts uterotropic effects at the cellular and molecular level which question the safety of uncontrolled and unlimited consumption of soy or red clover supplements by postmenopausal women with intact uteri.

Effects of dietary equol on the pituitary of the ovariectomized rats.

The aim of our study was to evaluate the effects of dietary equol, metabolite of a phytoestrogen daidzein, on the secretion of prolactin (PRL) and lutenizing hormone (LH), as well as the expression of estrogen receptors (ERalpha, ERbeta and truncated estrogen receptor-1 (TERP-1) in the pituitary gland of ovariectomized (ovx) female Sprague-Dawley rats. Two doses of equol (50 mg/kg of chow and 400 mg/kg of chow) were used and the results were compared with the effects of estradiol 3-benzoate (E2B), also given at two doses (4.3 mg/kg of chow and 17.3 mg/kg of chow). Treatment period was 3 months. Dietary equol administration at the high dose increased significantly serum PRL levels. This effect was also observed in the E2B group but this difference did not reach statistical significance. Surprisingly, high dose dietary equol treatment also significantly increased serum LH levels, which was in contrast to E2B treatment where serum LH levels were significantly decreased at both doses. Serum LH levels in the equol low group were unaffected. Equol treatment had no effects on pituitary ERalpha or ERbeta gene expression. In contrast, high dose E2B treatment increased significantly pituitary ERalpha mRNA levels but decreased those of ERbeta. Both doses of E2B also increased significantly pituitary TERP-1 mRNA levels. This effect was also observed in the equol high group but at a much smaller magnitude. In conclusion, high dose dietary equol administration to ovx rats exerts estrogenic like effects on the lactotropes and anti-estrogenic on the gonadotropes.

In vitro effects of genistein and resveratrol on the production of interferon-gamma (IFNgamma) and interleukin-10 (IL-10) by stimulated murine splenocytes.

Phytoestrogens are a group of plant-derived biologically active substances with a chemical structure that resembles that of 17beta-estradiol (E2). As the presence of estrogen receptors (ER) has been identified in several immune cells, phytoestrogens may also have a great impact on the immune system. The aim of our study was to determine the in vitro effects of genistein and resveratrol on the production of interferon-gamma (IFNgamma) and interleukin-10 (IL-10) by stimulated murine splenocytes and compare them with the effects of natural E2. Phorbol 12-miristate 13-acetate (PMA) together with ionomycin was used to stimulate the cells. E2 and genistein did not show any significant effects on the stimulated production of IFNgamma. Resveratrol had a mild inhibitory effect on IFNgamma production at the concentration of 10(-7)M; however, this difference did not reach statistical significance (p>0.05). IL-10 levels in the splenocytes culture supernatants were found to be increased in the presence of E2, genistein and resveratrol; however, these effects were also not statistically significant. To determine whether the exposure to our studied phytoestrogens induced a shift in the T-helper 1/T-helper 2 (Th1/Th2) balance, we calculated the ratio between the production of IFNgamma, the prototypic Th1 cytokine, and the production of IL-10, the prototypic Th2 cytokine, at different concentrations of our tested substances. Genistein at the concentrations of 10(-6) and 10(-7)M and resveratrol at the concentrations of 10(-6)M decreased significantly the IFNgamma/IL-10 ratio. This decrease was comparable to that of E2 at the concentrations of 10(-7)M. From our in vitro experiments we conclude that genistein and resveratrol, similarly to E2, by decreasing the IFNgamma/IL10 ratio may shift the Th1/Th2 balance towards the Th2 response.

High levels of circulating interleukin-10 in diabetic nephropathy patients.

AIMS: The aim of our study was to analyse the level of circulating interleukin-10 (IL-10) and relate it to the grade of albuminuria in patients with diabetic nephropathy (DN) due to type 1 diabetes mellitus (DM). Since IL-10 has met the criteria for an anti-inflammatory and an immunosuppressive cytokine, its activity may be important for clinical outcome of DN. METHODS: The IL-10 level was measured by ELISA in serum samples from thirty patients with DN due to type 1 DM, and compared with thirty patients with type 1 DM without DN and a control group of thirty, healthy, age- and sex-matched people. RESULTS: We observed a greatly elevated concentration of circulating IL-10 in 30/30 DM patients with DN (mean 140 pg/mL +/- 102), compared to DM patients without DN in whom IL-10 was detectable in only 11/30 patients (0.79 pg/mL +/- 1.24), and the group of healthy people in whom IL-10 was detectable in only 3/30 donors (0.92 pg/mL +/- 0.17). IL-10 appeared to be the strongest independent predictor of albuminuria, followed by HbA1c, diastolic blood pressure and DN duration. There was a positive correlation between the values of IL-10 and albuminuria in DM patients with DN. The patients in the fourth quartile of albuminuria had a distinctly higher concentration of IL-10 than those in the lower quartiles. CONCLUSIONS: The increased concentration of IL-10 in the serum samples from DM patients with DN seems to depend on the severity of the nephropathy. The excessive IL-10 production may indirectly contribute towards DN progression. On the other hand, it may explain the relatively long course of diabetic nephropathy.

Natural killer cell activity unaffected by ozonated autohemotherapy in patients with end-stage renal disease on maintenance renal replacement therapy.

Ozonotherapy is a complementary medical approach in the treatment of resistant infections, immune deficiency syndromes, orthopedic pathologies and vascular diseases. The criticism of this method is associated with potentially harmful effects of ozone on cells. The aim of this study was to investigate the influence of ozonated autohemotherapy (O3-AHT) on the cellular response of the immunologic system represented by cytotoxic activity of natural killer cells. 12 hemodialyzed patients (8 M, 4 F) aged 64.8 +/- 7.6 years with peripheral arterial disease as the main reason for the treatment with O3-AHT were examined in a prospective, placebo controlled, single blind study. They received 9 sessions of autohemotherapy without ozone exposure as a placebo-control and subsequent 9 sessions of O3-AHT. The procedures were performed 3 times a week, just before hemodialysis session. Ozone-oxygen gas mixture with ozone concentration of 50 microg/ml produced by ozone generator (ATO3, KrioMetrum, Poland) was used during O3-AHT Natural killer cell activity was measured using lactate dehydrogenase release assay There was no statistical difference between natural killer cell activity (%) at the baseline (16.78 +/- 8.07), after nine sessions of control autohemotherapy (15.98 +/- 6.67), and after nine sessions of O3-AHT (18.26 +/- 8.82). In conclusion, our findings showed that O3-AHT in a dose of 50 mg/mL does not have any significant influence on natural killer cell function in hemodialyzed patients.

Serum interleukin-6 levels and bone mineral density at the femoral neck in post-menopausal women with Type 1 diabetes.

BACKGROUND: Although osteopenia is often reported in Type 1 diabetes mellitus, the pathogenic mechanisms are not fully understood. Oestrogen deficiency also leads to decreased bone mineral density (BMD). Enhanced interleukin-6 (IL-6) production among Type 1 diabetic patients could be involved in the pathogenesis of diabetic bone loss since it is a potent bone resorbing cytokine. AIMS: To evaluate the relationship between serum bioactive IL-6 levels and BMD at the femoral neck of post-menopausal women with Type 1 diabetes. METHODS: We studied BMD, urine excretion of deoxypirydynoline crosslinks, serum bioactive IL-6 and soluble IL-6 receptor (sIL-6R) levels in 20 post-menopausal women with Type 1 diabetes mellitus, and compared these results with 20 matched healthy post-menopausal controls. RESULTS: Post-menopausal women with Type 1 diabetes had significantly lower BMD at the femoral neck and increased serum bioactive IL-6 levels compared with the control group, but no relationship was observed between these variables in a multiple regression analysis. Using BMD at the femoral neck of diabetic women as the dependent variable in the multiple step regression analysis model, we found that independent variables that were strongly associated with bone mass at the femoral neck in this group were: time since menopause and duration of diabetes. CONCLUSIONS: Although our study had a small sample size, we found that post-menopausal women with Type 1 diabetes mellitus present lower bone mass and higher serum bioactive IL-6 levels than matched healthy controls, but we were unable to find a correlation between these two parameters.

Effects of oestrogen deprivation on interleukin-6 production by peripheral blood mononuclear cells of postmenopausal women.

Various hormones can influence the expression of interleukin-6 (IL-6) and oestrogens are the most extensively studied. There is, however, controversy about the nature of the IL-6 secreted by human cells and its regulation by 17beta-oestradiol. The aim of this work was to clarify whether oestrogen deprivation after menopause may contribute to an enhanced IL-6 production by peripheral blood mononuclear cells (PBMC) in postmenopausal women. Twenty-two healthy postmenopausal women, age range 45-63 years, with clinical symptoms of oestrogen deficiency were enrolled in the study. The control group consisted of 16 healthy young women, age range 22-31 years, with regular menses and who were not taking oral contraceptives. Levels of IL-6 in the sera and PBMC culture supernatants were measured by the biological B9 cell-proliferation assay and expression of the IL-6 gene in non-stimulated PBMC was detected by RT-PCR. The effect of 17beta-oestradiol on spontaneous IL-6 production by the PBMC of postmenopausal women was also studied in vitro and in vivo. Seventeen out of the twenty-two postmenopausal women were given hormonal replacement therapy of 50 microg 17beta-oestradiol/day transdermally and the spontaneous production of IL-6 by the PBMC was analysed after 6 and 12 months of treatment. The postmenopausal women had significantly higher serum levels of IL-6 than the young controls. The spontaneous production of IL-6 by non-stimulated PBMC into the culture supernatants was also significantly higher in the postmenopausal women compared with the young. We also found that IL-6 gene expression was present in the non-stimulated PBMC isolated directly from the venous blood of the majority of the postmenopausal women. Women with IL-6 gene expression in the non-stimulated PBMC had significantly lower serum levels of 17beta-oestradiol compared with those where the IL-6 gene was not expressed in the PBMC. Our in vitro experiments showed that 17beta-oestradiol at concentrations of 10(-9) M and 10(-10) M decreased spontaneous IL-6 production by the PBMC of postmenopausal women. In vivo treatment with 17beta-oestradiol transdermally also significantly decreased spontaneous IL-6 production by the PBMC of postmenopausal women after 12 months of the therapy. Our results indicate that oestrogen deprivation after menopause may enhance IL-6 production by the PBMC of postmenopausal women. We suspect that the late complications of oestrogen deficiency, such as osteoporosis, coronary heart disease and Alzheimer's disease, may be mediated by an exaggerated production of IL-6 - a cytokine which seems to play a pivotal role in the pathogenesis of these age-related diseases.

Natural killer activity and thyroid hormone levels in young and elderly persons.

BACKGROUND: On the basis that (1) multiple interactions exist between the hormonal and immune systems, and (2) aging is accompanied by changes in thyroid hormone metabolism and responsiveness, we postulate that thyroid hormones may be involved in the observed decrease in natural killer (NK) activity in a population of apparently healthy elderly subjects. The purpose of the study is to compare NK cytotoxic activity and serum concentrations of TSH and thyroid hormones in healthy old and young people, and to assess in vitro the effects of triiodothyronine (T(3)) on NK activity. MATERIALS AND METHODS: Sixteen of the 47 healthy old people (mean age 64 +/- 5.2) were classified as optimally healthy, and the remainder as 'almost healthy' (according to the criteria of the Senieur protocol) [Ligthart et al., Mech Ageing Dev 1984;28:47-55]; the mean age of the healthy young people was 23.3 +/- 2.3 years. NK cytotoxic activity of peripheral blood mononuclear cells was assessed using (51)Cr release from K562 target cells. The cutoff level for defining low and high NK responses was set at a value of 20%. Serum concentrations of TSH, total thyroxine (T(4)) and total triiodothyronine (T(3)) were measured by radioimmunoassay. RESULTS: NK activity in the 'optimally healthy' elderly was high (mean 41 +/- 12%, SE), whereas 'almost healthy' subjects showed low NK activity (mean 6 +/- 5%). Serum T(4) and TSH levels, but not T(3) concentrations were similar in both the young and old. We observed a significant correlation (r = 0.53, n = 21, p < 0.05) between the serum total T(3) level and the NK activity in the elderly individuals. Under in vitro conditions exogenous T(3) significantly increased NK activity in the elderly subjects who had serum T(3) values at the lower end of the reference range. However, no effect of T(3) on NK activity was observed in peripheral blood mononuclear cells obtained from either old or young individuals who had serum T(3) levels at the midpoint of the range. CONCLUSION: Decreased serum concentrations of total T(3) may contribute to low NK activity in the 'almost healthy' subgroup of the elderly.

Recombinant human erythropoietin stimulates production of interleukin 2 by whole blood cell cultures of hemodialysis patients.

The impairment of function of human T lymphocytes, leading to an inappropriate cytokine production, is partially responsible for defective immunological response in hemodialysis patients (HD). Recent data suggest that recombinant human erythropoietin (rhEPO) may exert immunological effects. The aim of this study was to find out whether rhEPO treatment of the HD patients may have an effect on the interleukin 2 (IL-2) production by their whole blood cell cultures. The study was carried out in 10 HD patients receiving rhEPO for 6 months. Compared with the levels seen before the treatment, the concentration of IL-2 increased in the phytohemagglutinin-stimulated whole blood cell cultures of 7 of 10 patients under study. Addition of rhEPO in vitro to the whole blood cell cultures of the HD patients before implementation of erythropoietin confirmed that rhEPO is able to directly stimulate IL-2 production. Our studies show that the therapy with rhEPO affects IL-2 secretion.

The influence of recombinant human erythropoietin on tumor necrosis factor alpha and interleukin-10 production by whole blood cell cultures in hemodialysis patients.

Impaired immunological response in hemodialysis (HD) patients, which leads to inappropriate cytokine production, is partially caused by the hyperstimulation of both T lymphocytes and monocytes/macrophages. Recent data suggest that human recombinant erythropoietin (rhEPO) may have an immunological action. The goal of our study was to estimate the influence of rhEPO treatment on the production of the inflammatory cytokine tumor necrosis factor alpha (TNFalpha) and antiinflammatory cytokin interleukin-10 (IL-10) in 10 HD patients receiving rhEPO for 6 months. The levels of cytokines were measured in the in vitro cultures of whole blood. The level of IL-10 increased in all treated patients during the therapy, and it was accompanied by a transitory decrease of TNFalpha. The results of our studies suggest that rhEPO may reduce the inflammatory process by decreasing production of TNFalpha and increasing production of IL-10.