Quantitative analyses of cytokine profiles reveal hormone-mediated modulation of cytokine profiles in recurrent spontaneous miscarriage.

PURPOSE: Cytokines play important roles in pregnancy complications. Some hormones such as estrogen, progesterone, and dydrogesterone have been shown to alter cytokine profiles. Understanding how cytokine profiles are affected by these hormones is therefore an important step towards immunomodulatory therapies for pregnancy complications. We analyse previously published data on the effects of estrogen, progesterone, and dydrogesterone on cytokine balances in women having recurrent spontaneous miscarriages. MATERIALS AND METHODS: Levels of eight cytokines (IFN-γ, IL-2, IL-6, IL-10, IL-13, IL-17, IL-23, TNF-α) from n = 22 women presenting unexplained recurrent spontaneous miscarriages were studied. Cytokine values were recorded after in vitro exposure of peripheral blood cells to estrogen, progesterone, and dydrogesterone. We expand on earlier analysis of the dataset by employing different statistical techniques including effect sizes for individual cytokine values, a more powerful statistical test, and adjusting p-values for multiple comparisons. We employ multivariate analysis methods, including to determine the relative magnitude of the effects of the hormone therapies on cytokines. A new statistical method is introduced based on pairwise distances able to accommodate complex relations in cytokine profiles. RESULTS: We report several statistically significant differences in individual cytokine values between the control group and each hormone treated group, with estrogen affecting the fewest cytokines, and progesterone and dydrogesterone both affecting seven out of eight cytokines. Exposure to estrogen produces no large effects sizes however, while IFN-γ and IL-17 show large effect sizes for both progesterone and dydrogesterone, among other cytokines. Our new method for identifying which collections (i.e. subsets) of cytokines best distinguish contrasting groups identifies IFN-γ, IL-10 and IL-23 as especially noteworthy for both progesterone and dydrogesterone treatments. CONCLUSIONS: While some statistically significant differences in cytokine levels after exposure to estrogen are found, these have small effect sizes and are unlikely to be clinically relevant. Progesterone and dydrogesterone both induce statistically significant and large effect-size differences in cytokine levels, hence therapy with these two progestogens is more likely to be clinically relevant. Univariate and multivariate methods for identifying cytokine importances provide insight into which groups of cytokines are most affected and in what ways by therapies.

CytokineExplore: An Online Tool for Statistical Analysis of Cytokine Concentration Datasets.

PURPOSE: Cytokine data sets are increasing both in the number of different cytokines measured and the number of samples assayed. Further, typically data from different groups may be contrasted, eg, normal vs complication subjects. Many univariate and multivariate statistical techniques exist to study such cytokine datasets, but the ability to implement these techniques may be lacking for some practitioners, or may not be available quickly and conveniently. Here, we introduce CytokineExplore, an online tool for multi-cytokine and multi-group data analysis of user-provided Microsoft Excel data files. MATERIALS AND METHODS: In order to illustrate the tool features, we use data from intrauterine growth retardation (IUGR), a pregnancy complication, and normal healthy subjects as a control. The dataset contains levels for 10 cytokines, namely: IL-4, IL-6, IL-8, IL-10, IL-12, IL-13, IL-18, IL-23, interferon-gamma (IFN-γ) and tumour necrosis-alpha (TNF-α), obtained from 34 women with IUGR (further divided into 17 symmetric and 17 asymmetric cases) and 24 gestationally age-matched normal controls. RESULTS: The online tool automatically generates box-plots, histograms, PCA and PLSDA plots, t-tests and Mann-Whitney statistical tests, cytokine importance values for separating two groups, heatmaps for comparing multiple groups, and other functionalities. Figures generated can be directly downloaded for use in presentations or journal articles. CONCLUSION: The tool facilitates quick and easy numerical exploration and multivariate analysis of cytokine datasets, to aid basic understanding and hypothesis generation.

Effects of Progesterone, Dydrogesterone and Estrogen on the Production of Th1/Th2/Th17 Cytokines by Lymphocytes from Women with Recurrent Spontaneous Miscarriage.

Anti-inflammatory Th2 cytokines have been shown to be associated with healthy, successful pregnancy while pro-inflammatory Th1 and Th17 cytokines are associated with pregnancy loss due to recurrent spontaneous miscarriage. This nexus between unexplained recurrent spontaneous miscarriage (uRSM) and maternal inflammatory has led to the possibility of using pregnancy-related hormones to modify the maternal cytokine bias in a manner that is conducive to successful pregnancy. We investigated the ability of progesterone, dydrogesterone and estrogen to modulate cytokine production by peripheral blood lymphocytes from women undergoing uRSM. Peripheral blood mononuclear cells (PBMC) from females with uRSM were stimulated in vitro with phytohemagglutinin (PHA) in the presence and absence of progesterone or dydrogesterone or 17ß-estradiol. Culture supernatants were assayed for IFN-α, TNF-γ, IL-2, IL-6, IL-10, IL-13, IL-17A, and IL-23 by ELISA. Progesterone and dydrogesterone significantly down-regulated the secretion of the Th1 cytokines IFN-α and TNF-γ, and the Th17 cytokine IL-17A, and IL-23. Additionally, the secretion of the Th2 cytokine IL-6 was up-regulated. Estrogen, on the other hand, decreased the production of IFN-α and IL-2, increased the production of IL-6 but did not affect IL-17A and IL-23 secretion. Progestogens and estrogen can decrease the production of some Th1/Th17 inflammatory cytokines secreted by lymphocytes from uRSM and upregulate the production of anti-inflammatory cytokines. These data support the notion that progestogens can be used for altering maternal cytokine profiles to manage pregnancy complications.

Multivariate Comparison of Cytokine Profiles for Normal- and Low-Bone-Density Subjects.

Osteoporosis is a serious worldwide public health concern. The role of the immune system in the onset of osteoporosis in postmenopausal women is an area of current research. Here we study data from a panel of 10 cytokines obtained from postmenopausal women, with both normal and low bone mineral density (BMD). Normal- and low-BMD groups are compared and contrasted, and further low-BMD participants are sub-classified into osteopenic and osteoporotic based on BMD levels, and compared to each other. Via the use of multivariate statistical tools, we examine contrasting groups in relation to: (a) the presence of subgroups/clusters; (b) whether groups have statistically different multivariate distributions; (c) how strongly groups differ (if at all), which relates to the practical/clinical significant of any differences; and (d) which cytokines contribute most to any differences between groups. We find that the normal- vs. low-BMD groups are markedly different (p-value = 0.00013), with IL-23, IL-12, TNF-α, IL-4 and IL-6 being the most important differentiating cytokines. No significant difference between the osteopenic and osteoporotic groups is found (p-value = 0.81). These findings may aid the development of cytokine therapies for osteoporosis, and suggest the use of certain cytokine profiles as biomarkers for osteoporosis risk factors, and ways to quantify the progress of treatment therapies.

Circulatory Levels of RANKL, OPG, and Oxidative Stress Markers in Postmenopausal Women With Normal or Low Bone Mineral Density.

INTRODUCTION: Receptor activator of nuclear factor κB ligand (RANKL), osteoprotegerin (OPG), and oxidative stress markers are suggested to contribute to bone loss in osteoporosis that occurs in menopause. However, the association between these markers and bone mineral density (BMD) is controversial. The aim of this study was to measure circulatory levels of these parameters in postmenopausal women with normal or low BMD. METHODS: The study population included 71 postmenopausal women, of whom 25 had normal BMD, 31 had osteopenia, and 15 had osteoporosis. Serum levels of RANKL, OPG, and 5 oxidative stress markers (catalase, peroxiredoxin 2 [PRX2], superoxide dismutase 1 [SOD1], superoxide dismutase 2 [SOD2], and thioredoxin [TRx1]) were measured using the Multiplex system. RESULTS: As compared with subjects having normal BMD, subjects with low BMD had significantly lower median serum levels of OPG, catalase, SOD2, and PRX2 (P = .004, .031, .044, and .041 respectively). Although levels of RANKL were not different between the 2 groups, the RANKL/OPG ratio was higher in women with low BMD (P = .027). CONCLUSIONS: These data provide insights into the possible roles of OPG, RANKL, and oxidative stress in the pathogenesis of postmenopausal osteoporosis. However, the lack of association between these markers and BMD indicates that osteoporosis is complex and multivariate.

In vitro spleen cell cytokine responses of adult mice immunized with a recombinant PorA (major outer membrane protein [MOMP]) from Campylobacter jejuni.

There is no information on cytokine profiles for use as markers of protection in Campylobacter jejuni infection. To study this, we used outer membrane protein (MOMP [PorA]) as the vaccine for protection and spleen cell cytokines as markers of protection. We cloned and expressed porA from C. jejuni111 and immunized mice by the intraperitoneal route. Subsequently, mice were orally challenged with live C. jejuni 111. The vaccine induced protection as evidenced by reduced fecal excretion of C. jejuni111. Cytokines were measured in vitro after stimulation of spleen cells with MOMP. The levels of pro-inflammatory cytokines, IL-12, TNF-α, IL-17A and IL-17F were similar in control and test mice. The levels of pro-inflammatory cytokines, IL-2 and IFN-γ were higher in control mice than in test mice, and the levels of pro-inflammatory cytokines, IL-8 and IL-1ß were higher in test mice than in control mice. Among the two anti-inflammatory cytokines, the levels were similar for IL-10 but higher for IL-4 in test mice than in control mice. Ratios of pro-inflammatory to anti-inflammatory cytokines showed a bias towards an anti-inflammatory response in favor of antibody production reflecting the role of antibodies in immunity. Cytokine production patterns by spleen cells may be used as markers of protection in the mouse model.

Circulatory pattern of cytokines, adipokines and bone markers in postmenopausal women with low BMD.

Objective: In addition to some well-characterized bone turnover markers (BTMs), cytokines and adipokines have also been suggested to be linked to osteoporosis seen in menopause. However, there is much controversy on the possible association between these markers and bone mineral density (BMD). This study was aimed at measuring circulatory levels of selected cytokines, adipokines and BTMs in postmenopausal women with normal and low BMD. Methods: The study population included 71 post-menopausal women, of whom 25 had normal BMD, 31 had osteopenia and 13 had osteoporosis. Circulatory levels of selected pro-resorptive (TNF-α, IL-1ß, IL-6, IL-8, IL-12, IL-17), anti-resorptive (IFN-γ, IL-4, IL-10, IL-13, TGF-ß) and five adipokine markers (adiponectin, adipsin, lipocalin-2/NGAL, PAI-1 and resistin) were measured using the Multiplex system and read on the Magpix ELISA platform. Further, two bone turnover markers (PINP, CTX) as well as estradiol levels were assayed from the same samples. Results: While circulatory levels of cytokines were comparable between groups, women with low BMD had statistically significantly higher median circulatory levels of adipokines as compared to those with normal BMD. Further, while levels of CTX were not different between the two groups; PINP, PINP/CTX ratio and estradiol levels were significantly lower in women with low BMD. Levels of adiponectin, PINP, PINP/CTX ratio and estradiol correlated significantly with BMD of the hip and spine. Conclusion: The associations between various markers and BMD are complex and multivariate. Our data provide insights into the possible use of circulatory levels of cytokines, adipokines and bone turnover markers on the pathogenesis of postmenopausal osteoporosis because of the well-documented effects of these molecules on bone tissue and their relevance to osteoporosis.

Multivariate analysis of cytokine profiles in pregnancy complications.

PROBLEM: The immunoregulation to tolerate the semiallogeneic fetus during pregnancy includes a harmonious dynamic balance between anti- and pro-inflammatory cytokines. Several earlier studies reported significantly different levels and/or ratios of several cytokines in complicated pregnancy as compared to normal pregnancy. However, as cytokines operate in networks with potentially complex interactions, it is also interesting to compare groups with multi-cytokine data sets, with multivariate analysis. Such analysis will further examine how great the differences are, and which cytokines are more different than others. METHODS: Various multivariate statistical tools, such as Cramer test, classification and regression trees, partial least squares regression figures, 2-dimensional Kolmogorov-Smirmov test, principal component analysis and gap statistic, were used to compare cytokine data of normal vs anomalous groups of different pregnancy complications. RESULTS: Multivariate analysis assisted in examining if the groups were different, how strongly they differed, in what ways they differed and further reported evidence for subgroups in 1 group (pregnancy-induced hypertension), possibly indicating multiple causes for the complication. CONCLUSION: This work contributes to a better understanding of cytokines interaction and may have important implications on targeting cytokine balance modulation or design of future medications or interventions that best direct management or prevention from an immunological approach.

Patterns of circulatory and peripheral blood mononuclear cytokines in rheumatoid arthritis.

An imbalance in pro- and anti-inflammatory cytokines is suggested to contribute to tissue damage in rheumatoid arthritis (RA). This study was aimed at investigating profiles of cytokines in circulation and cytokines produced by mitogen-stimulated peripheral blood mononuclear cells (PBMC) in RA patients and healthy controls, and to explore correlations of cytokines with disease activity. Our aim was to identify patterns of cytokine expression as possible indicators of disease activity. Levels of plasma cytokines and PBMC-secreted cytokines were estimated in 26 female RA patients and 28 controls. Five pro-inflammatory cytokines (IFN-γ, TNF-α, IL-6, IL-17, IL-12) and three anti-inflammatory cytokines (IL-4, IL-10, IL-13) were assayed in a multiplex ELISA. RA patients had significantly higher plasma levels of TNF-α, IL-12, and IL-4 compared to healthy controls. On the other hand, mitogen-activated PBMC secreted significantly higher levels of the pro-inflammatory cytokines TNF-α, IFN-γ, IL-17, and IL-12, but lower levels of the anti-inflammatory cytokine IL-10 in RA compared to healthy subjects. The ratios TNF-α/IL-10, IFN-γ/IL-10, IL-17/IL-10, IL-12/IL-10, and IFN-γ/IL-13 were significantly higher in RA patients compared to healthy controls. The range and expression of cytokines were higher in PBMC than in the plasma in all the groups studied. Multivariate pattern analysis of eight cytokines revealed a prediction accuracy of 69% in differentiating RA patients from healthy controls, and of 73% in classifying patients as in remission or active RA. Our data suggest that it is worthwhile to explore ratios of pro- to anti-inflammatory cytokines produced by mitogen-stimulated PBMC in RA, and the use of multivariate cytokine pattern and algorithms for better delineation of this condition.

Cytokine profiles in osteoporosis suggest a proresorptive bias.

OBJECTIVE: As the immune system is suggested to contribute to the pathophysiology of osteoporosis in menopause, we compared the levels of proresorptive and antiresorptive cytokines produced by peripheral blood mononuclear cells (PBMCs) from postmenopausal women with normal and low bone mineral density (BMD). METHODS: Seventy-one postmenopausal women were studied; 25 had normal BMD and 46 had low BMD. Participants were categorized as normal (n = 25), osteopenic (n = 31), and osteoporotic (n = 15) based on T-scores. Levels of 10 cytokines produced by mitogen-stimulated PBMCs were measured by Multiplex ELISA. RESULTS: PBMCs from women with low BMD produced higher levels of the proresorptive cytokines tumor necrosis factor-alpha, interleukin (IL)-6, IL-12, and IL-17 (P = 0.014, 0.012, 0.011, and 0.049), and lower levels of the antiresorptive cytokines IL-4, IL-10, and IL-23 (P = 0.003, 0.018, and 0.025) compared with women with normal BMD. Proresorptive cytokines were similar in osteopenic and osteoporotic women, but both had higher levels than women with normal BMD. Osteoporotic women produced lower levels of the antiresorptive cytokines IL-4, IL-10, IL-13, and IL-23 compared with the normal BMD group (P = 0.001, 0.05, 0.05, and 0.026), and lower levels of IL-4 as compared with osteopenic women (P = 0.05). Osteopenic women produced lower levels of IL-4 and IL-10 compared with the normal BMD group (P = 0.025 and 0.038). Ratios of proresorptive to antiresorptive cytokines suggest a stronger proresorptive cytokine bias in women with low BMD. Most of the ratios are lowest in the normal BMD group, modest in osteopenic women, and highest in the osteoporotic group. CONCLUSIONS: Women with low BMD have a proresorptive cytokine bias.

Fractures in Kuwait: incidence and distribution.

The absence of descriptive epidemiological data on the frequency and distribution of fractures in a population is serious and may underestimate the actual importance of this public health problem. In this study, we report the crude and standardized incidence rates of fractures in the population of Kuwait during the period 2009-2012. Using the Ministry of Public Health's national registry, demographic data of all fracture cases between 2009 and 2012 were retrieved. These were further categorized into fractures at the hip or any other sites. Average annual incidence rates were calculated and standardized using the world's population in 2010 as a reference. Thus, 18,119 fractures among males and 6,362 among females were recorded. The overall estimated annual incidence rates of fractures per 100,000 person-years were 207 for males and 111.8 for females. Moreover, 13.3% of all fractures were in the hip, with incidence rates of 24.8 for males and 18.9 for females; while 86.7% were in other sites, with corresponding incidence rates of 182.2 and 92.8, respectively. The age-specific fracture incidence rates in females remained below the corresponding rates of males until ≥50 years of age, after which the female age groups showed successively higher rates. The age-standardized incidence rates for all fractures (hip and other sites) were 247.4 for males, 175.4 for females, and 216.2 for both sexes. The burden of this major public health challenge is set to rise, and such population-based incidence data call for an urgent need for action to reduce the projected human impact and socioeconomic costs of fracture.

Association of self-reported symptoms with serum levels of vitamin D and multivariate cytokine profile in healthy women.

BACKGROUND: Although a large number of studies have investigated possible relationships among serum levels of vitamin D or cytokines with disease progress and prognosis, similar studies on self-reported symptoms are still controversial. The overall objective of this study was to look into the association between serum levels of vitamin D or cytokines with self-reported symptoms related to musculoskeletal pain, sleep disorders, and premenstrual syndrome (PMS) in healthy adult women. SUBJECTS AND METHODS: Venous blood samples were collected from 117 healthy adult women, and serum levels of vitamin D, pro-inflammatory cytokines (IL-1ß, IL-6, IL-8, IL-17, IFN-γ, and TNF-α) and anti-inflammatory cytokines (IL-4, IL-10, and IL-13) were measured. Groups were tested for differences in single parameters, pro-:anti-inflammatory cytokine ratios, and differences in multivariate patterns. RESULTS: There were no significant associations between serum levels of vitamin D and any of the self-reported symptoms studied. However, serum levels of certain pro-inflammatory cytokines were significantly higher in subjects with musculoskeletal pain (IL-8, P=0.008), sleep disorders (IFN-γ, P=0.02), and PMS (IL-8 and TNF-α, P=0.009 and 0.002, respectively) compared to subjects who reported no symptoms. The pro-:anti-inflammatory cytokine ratios showed pro-inflammatory cytokine dominance in subjects with self-reported symptoms, particularly in the groups with deficient levels of vitamin D. However, the multivariate cytokine-pattern analysis was significantly different between PMS groups only. CONCLUSION: These data point to a possible role of pro-inflammatory cytokines as a contributing factor in self-reported symptoms related to musculoskeletal pain, sleep disorders, and PMS.

Association between levels of vitamin D and inflammatory markers in healthy women.

BACKGROUND: No one can deny that the biological importance of vitamin D is much beyond its classical role in bone metabolism. Several recent publications have highlighted its potential role in the functioning of the immune system. The overall objective of this study was to look into possible correlations between levels of vitamin D and inflammatory markers in sera of healthy adult women. These markers included proinflammatory cytokines (interleukin [IL]-1ß, IL-6, IL-8, IL-17, interferon [IFN]-γ, and tumor necrosis factor [TNF]-α), anti-inflammatory cytokines (IL-4, IL-10, and IL-13), as well as C-reactive protein (CRP) as a general indicator of inflammation. METHODS: Venous blood samples were collected from 118 healthy adult women and serum levels of vitamin D, CRP, proinflammatory cytokines (IL-1ß, IL-6, IL-8, IL-17, IFN-γ, and TNF-α), and anti-inflammatory cytokines (IL-4, IL-10, and IL-13) were measured. RESULTS: There were no significant direct correlations between serum levels of vitamin D and any of the inflammatory markers measured. However, subjects with deficient levels of vitamin D and high CRP produced significantly higher levels of the proinflammatory cytokines (TNF-α and IL-8) as compared to subjects with low CRP levels with nondeficient and deficient levels of vitamin D. Further, the anti-inflammatory/proinflammatory ratios suggest a role of vitamin D in maintaining an anti-inflammatory environment at low levels of CRP, an association that is weaker at high CRP levels in subjects with subclinical inflammatory situations. CONCLUSION: These data point to a possible role of vitamin D as a contributing factor in balancing cytokines toward an anti-inflammatory role in inflammatory situations.

Incidence of hip fracture in Kuwait: a national registry-based study.

UNLABELLED: The crude and age-standardized rates of hip fractures in elderly Kuwaiti subpopulation were determined for the years 2009-2012. Both rates have increased and are further expected to rise substantially in the coming decades. INTRODUCTION: It is projected that rates of hip fractures will increase in most of the Middle East countries. There are only few population-based studies investigating the incidence of hip fractures in the Arabian Gulf region. The objective of this study is to report the crude and age-standardized incidence rates of hip fracture in the Kuwaiti elderly population. METHODS: Using the Ministry of Public Health's registry data, this population-based study evaluated the incidence of hip fractures in Kuwaiti subpopulation aged 50 and above for the years 2009, 2010, 2011, and 2012. Using the world population in 2010 as a reference, these incidence rates were age-standardized and compared to data from several regions. RESULTS: Hip fracture crude incidence rates varied between 113.7 and 147.4/100,000 for males and between 135.3 and 148.1/100,000 for females, with a female/male ratio of 1.3-1.5. The combined (men and women) incidence rate of hip fractures increased by 17.1 % over the 4-year period of study (125.9/100,000 in 2009 to 147.8/100,000 in 2012). Using the world population in 2010 as a reference, the age-standardized rates were 129.5, 131.5, 154.6, and 169.8 for males and 189.6, 192.9, 197.2, and 214.4 for females, for the years 2009, 2010, 2011, and 2012, respectively. CONCLUSIONS: The hip fracture age-standardized incidence rates in the Kuwaiti subpopulation aged 50 years and above are rising and expected to increase substantially in the coming decades.

Tumor necrosis factor-α and pregnancy complications: a prospective study.

OBJECTIVE: The aim of this study was to compare the levels of tumor necrosis factor-α (TNF-α) produced by peripheral blood mononuclear cells in normal pregnancies and pregnancies with complications. MATERIALS AND METHODS: Maternal peripheral blood mononuclear cells from women with a recurrent spontaneous miscarriage (n = 35), premature rupture of fetal membranes (n = 30), preeclampsia (n = 27) and intrauterine fetal growth retardation (IUGR; n = 36) were stimulated with mitogen or antigen, and the levels of TNF-α produced were compared to those produced by peripheral blood mononuclear cells from a normal pregnancy (n = 35). RESULTS: The median levels of mitogen-induced TNF-α at the 1st, 2nd and 3rd trimester, and at normal delivery were 1,176.4, 4,320.9, 7,307.4 and 2,463.0 pg/ml, respectively, while those produced in the recurrent spontaneous miscarriage, premature rupture of membranes and preeclampsia cases were 4,159.8, 3,489.5 and 4,149.2 pg/ml, respectively. The differences were statistically significantly higher in these pregnancy complications (p = 0.04, 0.024 and 0.014) as compared to the levels in normal pregnancy. Furthermore, antigen-induced TNF-α levels were produced at statistically significantly higher levels by women with IUGR (120.4 pg/ml) compared to women with normal pregnancies (17.9 pg/ml; p = 0.041). CONCLUSION: Higher levels of TNF-α seem to play a role in these pregnancy complications, suggesting its pathogenesis in such conditions.

Evaluation of bone mineral density in patients with chronic low back pain.

STUDY DESIGN: This was designed as a retrospective study. PURPOSE: We investigated the relationship between bone mineral density (BMD) and chronic lower back pain (LBP). OVERVIEW OF LITERATURE: In spite of a large number of epidemiological surveys on the prevalence of LBP and BMD measurements completed separately in the general population, the relationship between the two has not been well documented. METHODS: The study included 171 patients with chronic LBP who underwent the BMD study. The control group was selected from our database regarding BMD without LBP. RESULTS: A total of 678 subjects, aged 18 to 100 years (mean, 49.9±12.9 years) were included in the study, 25% (n=171) of the subjects had LBP. Compared to those patients without LBP, patients exhibiting LBP had statistically significant lower mean weight, hip and spine BMD and T-score. Lower BMD and T-scores were significant regardless of the age group, gender, menopausal status, and obesity classification. CONCLUSIONS: Chronic LBP has a negative correlation with hip and spine bone mineral density.

Relationship between leptin levels and suppressed CD4 counts in HIV patients.

OBJECTIVE: To examine the relationship between serum leptin levels and suppression of CD4 count in HIV-infected individuals with highly active antiretroviral therapy (HAART). SUBJECTS AND METHODS: Thirty seropositive HIV male patients selected from the Infectious Disease Hospital were classified into two groups according to their immunological and virological response to HAART. The first group included 15 male patients with low viral load and low CD4 counts; the second included 15 male patients with low viral load and high CD4 counts. Morning serum leptin and tumor necrosis factor-α levels of HIV patients were measured and correlated with fasting serum insulin, Homeostasis Model Assessment for Insulin Resistance (HOMA-IR), HIV viral load and CD4 count. RESULTS: Serum leptin levels were significantly higher in patients with high CD4 counts than in patients with low CD4 counts (mean serum leptin level 47.3 vs. 10.9 ng/ml, respectively; p < 0.0001). A positive correlation was observed between serum leptin levels and CD4 counts (r = 0.697; p < 0.0001); positive correlations were also seen between leptin levels and fasting serum insulin and HOMA-IR (r = 0.633, p < 0.0001, and r = 0.537, p < 0.003, respectively). CONCLUSION: Serum leptin level was higher in HIV patients with high CD4 count and correlated with fasting serum insulin and HOMA-IR, thereby indicating that HAART treatment could lead to decreased levels of leptin in HIV patients, which might lead to impaired immunological recovery.

Intrauterine growth restriction: cytokine profiles of trophoblast antigen-stimulated maternal lymphocytes.

Intrauterine growth restriction (IUGR) is an important perinatal syndrome that poses several serious short- and long-term effects. We studied cytokine production by maternal peripheral blood lymphocytes stimulated by trophoblast antigens. 36 women with a diagnosis of IUGR and 22 healthy women with normal fetal growth were inducted. Peripheral blood mononuclear cells were stimulated with trophoblast antigens and levels of the proinflammatory cytokines IL-6, IL-8, IL-12, IL-23, IFNγ, and TNFα and the anti-inflammatory cytokines IL-4, IL-10, and IL-13 were measured in culture supernatants by ELISA. IL-8 was produced at higher levels by blood cells of the IUGR group than normal pregnant women, while IL-13 was produced at lower levels. IL-8, IFNγ, and TNFα were higher in IUGR with placental insufficiency than in normal pregnancy. IL-12 levels were higher and IL-10 levels were lower in IUGR with placental insufficiency than in IUGR without placental insufficiency. We suggest that a stronger pro-inflammatory bias exists in IUGR as compared to normal pregnancy and in IUGR with placental insufficiency when compared to IUGR without placental insufficiency. Several ratios of proinflammatory to anti-inflammatory cytokines also support the existence of an inflammatory bias in IUGR.

Evaluation of bone mineral density in postmenopausal women in Kuwait.

Menopause is the major risk factor for the loss of bone mineral density (BMD) and bone mineral content (BMC) in women. In this study, we determined the prevalence of osteoporosis in postmenopausal women in Kuwait and compared it with that of other Middle East and west countries. Two thousand two hundred ninety-six postmenopausal women ranging in age from 40 to 87yr were included in the study and divided into 4 age groups by decade. We measured body weight, height, body mass index (BMI), BMD, and BMC. The mean age, height, and weight were 59.1+7.9yr, 154.7+6.5cm, and 77.3+14.9kg, respectively. The mean BMI and BMC were 32.4+6.6kg/m(2) and 0.9+0.14g/cm(2), respectively. The average T-scores for the hip and lumbar spine were -0.280+1.2 and -1.297+1.33, respectively. BMC significantly decreased with age from 0.95 to 0.81g/cm(2). Four hundred forty-four (19.3%) were found to have osteoporosis. The incidence of osteoporosis significantly increased from 4.3% to 39.9% with age, which is lower than that reported for Saudi (40%) and Moroccan women (39.6%) and higher than that for US/European (31%) and Lebanese women (11%).

Redirection of cytokine production by lymphocytes from women with pre-term delivery by dydrogesterone.

PROBLEM: To study the ability of dydrogesterone to modulate the production of pro-inflammatory and anti-inflammatory cytokines by lymphocytes from women undergoing pre-term delivery (PTD). METHOD OF STUDY: Peripheral blood mononuclear cells (PBMC) from 18 subjects undergoing PTD were stimulated with the mitogen phytohemagglutinin in the presence and absence of progesterone and dydrogesterone. The levels of interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha, interleukin (IL)-4, and IL-10 in culture supernatants were then estimated by enzyme-linked immunoabsorbant assay. Cytokine production in the presence and absence of progesterone and dydrogesterone were compared. RESULTS: The exposure of PBMC to dydrogesterone resulted in a significant inhibition in the production of the pro-inflammatory cytokines IFN-gamma and TNF-alpha and a significant increase in the levels of the anti-inflammatory cytokine IL-4, resulting in a substantial shift in the ratio of Th1/Th2 cytokines. CONCLUSION: Dydrogesterone induces a shift in cytokine bias, by inhibiting pro-inflammatory cytokine production and increasing anti-inflammatory cytokine production.