Lymphocyte Expression of Intracellular Cytokines and Heat Shock Proteins in Peripheral Blood of Patients with Atopic Dermatitis.

INTRODUCTION: The present study evaluates expression by activated CD4+ T helper1 (Th1) and T helper 2 (Th2) T lymphocytes of pro-inflammatory cytokines and cytoprotective heat shock proteins (HSPs) in peripheral blood of atopic dermatitis (AD) patients. METHODS: This research represents preliminary work by the authors to identify correlates between critical immune parameters with the potential to serve as guidelines for the development of pharmacological strategies for altering these factors to promote the restoration of healthy immune profiles in persons afflicted with major atopic diseases. The major experimental strategy used in this research assessed immune activation by peripheral blood mononuclear cells (PBMC) from 21 AD patients and 12 age- and gender-matched healthy control subjects cultured with phorbol myristate acetate (PMA) and ionomycin (PMA/I), which are mutagenic immune activators, to induce expression of pro-inflammatory biomarkers in CD4+ T cells differentiated to express Th1 or Th2 cytokines and heme oxygenase-1 (HO-1) intracellularly (i). Evaluations were performed using an FC500 Beckman-Coulter flow cytometer. Elevated CD4+ T cell expression of cytokines, interleukin-4 (iIL-4), interleukin- 5 (iIL-5), interleukin-10 (iIL-10), interferon-gamma (iIFN-g), tumor necrosis factor-alpha (iTNF-α), were observed. RESULTS: Additionally, the heat shock proteins (HSP) iHO-1 and iHSP-70 were evaluated in cells from the blood of AD patients versus the control subjects. The present study demonstrated an elevated expression of both Th1 and Th2-associated cytokines in CD4+ T cells of AD patients, with a significant direct correlation between Th1 and Th2 cell populations, thus yielding insight into the immune features of the AD-associated systemic inflammatory profile. CONCLUSION: Finally, the observed increased iHO-1 and iHSP-70 expressions likely represent adaptive physiologic countermeasures to AD-associated inflammatory tissue damage, suggesting that HSP inducers are promising candidates for the management of atopic disorders.

Parkinson's Disease: Alpha Synuclein, Heme Oxygenase and Biotherapeutic Countermeasures.

Neurodegenerative disorders have been and remain persistent sources of enormous suffering throughout human history. The tragedy of their impact on human relationships, physical vitality, and fundamental dignity cannot be understated. Parkinson's disease (PD), one of the most common of these terrible illnesses, has a global incidence of approximately two-to-four percent of the human population, along with devastating social and economic impact. The present review analyzes aspects of PD pathophysiology that offer particularly attractive strategies for the development of improved prevention and therapy. The occurrence, symptoms, pathogenesis, and etiology of PD are considered, with focus on how the Alpha synuclein protein, which normally regulates neurotransmitter release, is aggregated by oxidative stressors into toxic inclusions, prominently including Lewy bodies and insoluble fibrils that disrupt the organization of brain areas responsible for motor control. The contribution to a progressively prooxidant tissue environment resulting from interaction between advanced glycation end products (AGEs) and their cognate receptors (RAGEs) is examined here as a significant driver of PD. This review also explores strategies currently being developed by a U.S.-Russian team that may reduce the risk and severity of PD by use of recombinant atoxic derivatives (ad) of botulinum neurotoxins (BoNT/A ad), that traffic inducers of the cytoprotective enzyme heme oxygenase to selected midbrain neurons, at which Alpha synuclein aggregation occurs. Considered together, the topic material presented here provides both researchers and clinicians with a short but concise overview of the current understanding of PD pathology and approaches to biotherapeutic (precision) countermeasures to its onset and progression.

Correlation between heat shock proteins, adiponectin, and T lymphocyte cytokine expression in type 2 diabetics.

Type 2 diabetes mellitus (T2DM) features insulin resistance, hyperglycemia, dyslipidemia, overproduction of inflammatory cytokines, and systemic oxidative stress. Here, heat shock proteins Hsp70 and Hsp 90, adiponectin, and heme oxygenase-1 (HO-1, Hsp32) are profiled in peripheral blood mononuclear cells (PBMC) and serum from 25 T2DM patients and 25 healthy control subjects. Cells cultured with phorbol 12-myristate 13-acetate/ionomycin were evaluated by three-color flow cytometry for immunophenotypic biomarkers. Plasma HO-1, Hsp, and adiponectin levels were assayed by enzyme-linked immunosorbent assay (ELISA). Relative to healthy controls, T2DM patients exhibited significantly elevated plasma Hsp70, and representation of T helper immunophenotypes activated to express inflammatory cytokines, including CD4+ IFN-γ+, CD4+ TNF-α+, CD4+ IL-6+, CD4+ IL-1ß+ T cells, significantly lower representation of CD4+ IL-10+ T cells, plasma adiponectin and cell-associated HO-1 expression-with no significant differences in plasma Hsp90 between T2DM and healthy controls. Plasma HO-1 and adiponectin in T2DM patients inversely correlated with TNF-α and showed inverse correlation between serum LDL and plasma HO-1. Moreover, TNF-α and Hsp90 in T2DM patients correlated positively with fasting blood glucose (FBG). These results demonstrate correlation between potentially pathogenic T cells, HO-1, and adiponectin, additionally revealing a T helper (Th)1-related character of T2DM immunopathogenesis, suggesting potential for novel T cell-related management strategies for T2DM and related co-morbidities.

Activity of Paraoxonase/Arylesterase and Butyrylcholinesterase in Peripheral Blood of Gulf War Era Veterans With Neurologic Symptom Complexes or Post-Traumatic Stress Disorder.

OBJECTIVE: Two groups of Gulf War era veterans, one exhibiting blurred vision, balance problems/dizziness, tremors/shaking, and speech difficulty and a second group with post-traumatic stress disorder (PTSD), but not the neurologic syndrome, were assessed for organophosphate-detoxifying enzyme paraoxonase/arylesterase (PON1) and its Q/R isoforms, butyrylcholinesterase (BuChE) and its U/A isoforms and cytokines. METHODS: Defibrinated peripheral blood was evaluated for enzymes and cytokines. RESULTS: Trends toward elevation of Th2 cytokines interleukin-4 (IL-4) and IL-13 were observed in subjects with neurologic syndrome. Neither the activities nor isoforms of the enzyme, the neurologic symptoms, nor PTSD had any relationship to wartime deployment to the theater of combat. CONCLUSION: The negative outcomes described above suggest that exposure to organophosphates or other agents normally detoxified by PON1 and BuChE may not have contributed significantly to neurologic components of Gulf War Illness.

Effect of Diabetea tea ™ consumption on inflammatory cytokines and metabolic biomarkers in type 2 diabetes patients.

ETHNOPHARMACOLOGICAL RELEVANCE: Diabetea tea™ (DT) is an anti-diabetic alternative medicine in some Asian countries. The main constituent of DT is black tea originating from Camellia sinensis that is supplemented by 12 other medicinal plants. Black tea contains a large amount of the flavonoids catechins especially epigallocatechin gallate (EGCG) which has anti-inflammatory and antioxidative capacity. This study was undertaken to evaluate the possible effects of DT intake on inflammatory cytokines, regulatory T cells (Tregs) and metabolic biomarkers in T2DM. MATERIALS AND METHODS: The study included 50 patients with T2DM. The patients had received 3 cups (600ml) of DT extract or placebo (PL) extract per day during a period of 12 weeks. Intracellular cytokine expression in peripheral blood mononuclear cell (PBMC) as well as the glycemic and lipid profiles were measured at baseline and after the treatment period. The active constituents of the medicinal plants included in DT were investigated by gas chromatography-mass spectrometry (GC/MS). RESULTS: Ingestion of DT suppressed CD4+ T cell expression of IL-1ß and Il-8 (p<0.05) and up-regulated the expression of IL-10 and the Treg/IL-17 ratio (p<0.05) which was not shown in PL. A significant decrease in HbA1c and LDL was observed at the end of the study period (p<0.05) in DT. The GC/MS analyses of DT indicated the presence of lupeol, ß-Amyrin and ß-sitosterol. Also analyses of individual herbs showed the presence of higher levels of lupeol and ß-Amyrin in Nuga Ficus bengalensis and ß-sitosterol in the Attikka Ficus racemosa, indicating that the active ingredients of DT are concentrated in these two herbs. CONCLUSION: The present study provides evidence that DT has hypoglycemic and antihyperlipidemic properties. Interestingly, DT has anti-inflammatory effects. These properties are attributed to the flavonoids, triterpenes and phytosterol contents of the tea. We suggest that DT protects against diabetes complications in the long term.

Effect of Black Tea Consumption on Intracellular Cytokines, Regulatory T Cells and Metabolic Biomarkers in Type 2 Diabetes Patients.

The present study was undertaken to evaluate the effects of black tea intake on inflammatory cytokines and metabolic biomarkers in Type 2 diabetes mellitus (T2DM). Thirty patients with T2DM were randomly assigned either to a High Intake (HI) group, consuming three cups (600 mL) of black tea per day; and a Low Intake (LI) group, administered 1 cup (200 mL) per day, each during a 12-week period. Intracellular cytokine expression, regulatory T cells (Treg), glycemic and lipid profiles were measured at baseline and following the tea intake period. Tea consumption correlated with major effects measured in peripheral blood of subjects that included significantly reduced glycated hemoglobin (HbA1c) levels, along with increased regulatory T cells CD3+ CD4+ CD25+ FOXP3, CD3+ CD4+ IL-10+ cells (an immunosuppressive phenotype), reduced (pro-inflammatory) CD3+ CD4+ IL-17+ cells and reduced Th1-associated CD3+ CD4+ IFN-Υ+ cells. Tea consumption was also observed to abolish the significance of an inverse correlation between total serum cholesterol and representation of CD4+ IL-4+ T cells, which may reflect protection against atopy-related oxidative stress. Outcomes of this study describe both advantages and limitations to consumption of black tea as an aid to sustained health maintenance by persons at-risk for TD2M and related obesity-associated metabolic syndromes.

Amelioration of human osteoarthritis symptoms with topical 'biotherapeutics': a phase I human trial.

Osteoarthritis (OA) treatments presently rely on analgesics, which manage pain but fail to restore imbalances between catabolic and anabolic processes that underlie OA pathogenesis. Recently, biologic (biotherapeutic) drugs, which alter the activity of catabolic agents such as nitric oxide and inflammatory cytokines in ways, allowing tissue regeneration, were evaluated for efficacy in OA treatment. These studies failed to demonstrate dramatic abatement of OA symptoms by these drugs, but suggested strategies by which biologic agents might be used to treat OA. The present review summarizes current understanding of OA pathogenesis and evolving treatments. Preliminary evaluations of a novel biotherapeutic strategy are presented here. Twenty OA patients receiving sour topical cherry seed extract (SCE), an inducer of heme oxygenase-1 (HO-1), a major physiological protectant against oxidative stress exhibited significantly decreased joint pain and activation of CD4+ T cells expressing inflammatory cytokines (p < 0.05), significantly decreased peripheral blood C-reactive protein (CRP), and increased leukocyte HO-1 (p < 0.05) in comparison with ten placebo-treated patients. SCE inhibits joint-damaging inflammatory mediator production. This agent therefore meets the main criterion for classification as a "biotherapeutic," or "biologic" agent. The negligible toxicity and low cost of such materials make them promising contributors to OA treatment, sustainable within resource limitations of a wide range of patients.

Immune responses following McKenzie lumbar spine exercise in individuals with acute low back pain: a preliminary study.

OBJECTIVE: This study explores the immune responses following 4 weeks of McKenzie lumbar spine exercise in individuals with acute low back pain (ALBP). PATIENTS AND METHODS: Fifteen patients with ALBP and 15 healthy individuals volunteered in this study. Ten ml of peripheral blood were obtained from each patient before and after exercise sessions, and from healthy individuals at the beginning of the study. Flow cytometric analysis was used to evaluate the frequencies of CD4+ T lymphocyte sub-populations and the intracellular cytokine expression within this cell population. Pain perceptions were obtained at baseline and following each week of exercise sessions. RESULTS: In comparison with healthy subjects there was an elevated frequency of memory (CD4+CD45RO+) T cells, helper inducer (CD4+CD29+) T cells, CD3+CD16+CD56+ T cells and a lower frequency of naïve/suppressor (CD4+CD45RA+) T cells at base line in back pain patients (p<0.05). After 4 weeks of McKenzie exercise sessions, pain intensity significantly decreased (p<0.05); however, there was no significant difference in the frequency of memory (CD4+CD45RO+) T cells, helper inducer (CD4+CD29+) T cells, CD3+CD16+CD56+ T cells and naïve/ suppressor (CD4+CD45RA+) T cells at base line relative to these cell populations after exercise sessions. The percentage of Pan (CD3+) T cells expressing IL-8 and TNF-α and the CD3+ T cells expressing the anti-inflammatory cytokine IL-4 increased significantly (p<0.05) following exercise sessions in comparison with baseline and healthy references. The reduction in pain scores did not correlate with elevated anti-inflammatory cytokines. CONCLUSION: McKenzie exercise sessions induced an immune activation state and simultaneously up regulated anti-inflammatory IL-4 cytokines that boost pain relief.

Sour cherry (Prunus cerasus) seed extract increases heme oxygenase-1 expression and decreases proinflammatory signaling in peripheral blood human leukocytes from rheumatoid arthritis patients.

Sour cherry seed extract (SCE) was evaluated for its capacity to inhibit lipopolysaccharide-treated human peripheral blood T cells expressing tumor necrosis factor-alpha, and the chemokine interleukin-8. Both proteins are diagnostic biomarkers for inflammatory pathologies. Peripheral blood leukocytes from 11 rheumatoid arthritis (RA) patients and 8 healthy control subjects were co-cultured for 24h in lipopolysaccharide and the extract, then evaluated by flow cytometry for T cell activation and by enzyme-linked immunoassay for lymphocyte-associated heme oxygenase-1 (HO-1) expression. There was a dose-dependent decrease in expression of the immunophenotypes: CD3+TNF-α+, and CD3+IL8+ in cultures from RA patients to a greater extent than in cells from healthy participants. These results suggest that the extract may have a modulatory roll in RA and other inflammatory disorders via the induction of HO-1, thus abating oxidative stress and strengthening regulation of pro-inflammatory signaling pathways.

Inflammatory cytokines and the risk of cardiovascular complications in type 2 diabetes.

This study evaluates peripheral blood T lymphocyte expression of inflammatory and proinflammatory cytokines as well as T regulatory (Treg) (FOXP3+CD25+CD4+) cells in type 2 diabetes (T2DM). Participants included 40 T2DM and 30 healthy control subjects. Twenty-four patients had no complications while 16 were afflicted with coronary heart disease (CHD). Relative to healthy subjects, all T2DM patients showed a significant increase in expression of CD4+IFN- Y +, CD4+TNF- α +, and CD4+IL-8+ T cells (P < 0.001) as well as CD4+IL-6+, CD4+IL-1 ß +, and IL-17+ T cells (P < 0.05) while the ratios of Treg/Th1(CD4+IFN- Y +) and Treg/Th-17(CD4+IL-17+) cells were significantly decreased (P < 0.05 and P < 0.01). T2DM patients with CHD showed a significant increase in CD4+IFN- Y +, CD4+TNF- α +, and CD4+IL-17+ T cells and a significant decrease in Treg/Th1 and Treg/IL-17 cells compared to T2DM patients without CHD (P < 0.05). In CHD-afflicted T2DM, HbA1c correlated positively with CD4+IFN- Y + T cells (P < 0.01), HDL correlated negatively with each of CD4+IL-8+ T cells and CD4+IL-17+ T cells (P < 0.05), and LDL correlated positively with CD4+IL-1 ß + T cells (P < 0.05). Conclusion. This study shows that hyperglycemia and dyslipidemia correlate with increased inflammatory cytokine expression and suggests the involvement of T cells in the development of diabetes and its complications.

Sour cherry seed kernel extract increases heme oxygenase-1 expression and decreases representation of CD3+ TNF-α+ and CD3+IL-8+ subpopulations in peripheral blood leukocyte cultures from type 2 diabetes patients.

The present study evaluates a hypothesis that sour cherry (Prunus cerasus) seed extracts (SCE) modulate CD3+ T lymphocyte activity in ways predictive of potential for uses of SCE in management of inflammatory diseases. Peripheral blood mononuclear cells (PBMC) from 12 type 2 diabetes (T2DM) patients and eight healthy control subjects were cultured 24 h with 100 ng/ml lipopolysaccharide (LPS) to increase inflammatory signaling and co-incubated with 0.5-100 µg/ml SCE. Cultures were evaluated by two-color flow cytometry for percent representation of CD3+ IL8+ and CD3+TNF-α cells which express interleukin-8 (IL-8), and tumor necrosis factor-α, (TNF-α+) respectively, and by enzyme-linked immunoassay for lymphocyte-associated heme oxygenase-1 (HO-1, known to be induced by SCE). SCE dosage ranges of 0.5-100 µg/ml in cell cultures significantly suppressed LPS-increased CD3+TNF-α+ and CD3+IL8+ representation from all participants (p < 0.05), with greater pharmacological effect noted in suppression of CD3+TNF-α+ noted in cells from T2DM patients versus healthy control subjects. These effects correlated with increased HO-1 expression in SCE-treated PBMC from all subjects (p < 0.05). Since TNF-α and IL-8 are diagnostic/prognostic biomarkers for many inflammatory syndromes, the capacity of SCE to down-regulate representation of cells that express them suggests potential for therapeutic use of SCE in T2DM and other diseases.

In vitro suppression of lymphocyte activation in patients with seasonal allergic rhinitis and pollen-related asthma by cetirizine or azelastine in combination with ginkgolide B or astaxanthin.

Novel strategies are evaluated for management of allergic rhinitis and asthma in patients co-afflicted with both disorders. It is hypothesized that the platelet activating factor receptor antagonist ginkgolide B (GB) and the carotenoid antioxidant astaxanthin (ASX) interact with antihistamines cetirizine dihydrochloride (CTZ) and azelastine (AZE) to potentiate their ability to downregulate potentially pathological immune activation. Peripheral blood mononuclear cells from asthmatics and healthy subjects, cultured 24 hours with 50 µg/ml phytohemaglutinin (PHA) or PHA plus each drug are analyzed by flow cytometry for expression of CD25+ or HLA-DR+ by CD3+ (T cells). Results are reported as stimulation indices for CD3+CD25+ (SICD3+CD25+) and CD3+HLA-DR+ (SICD3+HLADR+) cells in cultures treated with PHA alone, versus cultures treated with both PHA and drugs. Optimal suppression of activated cells was observed in cultures stimulated with ASX 10-6 M + CTZ 10-6 M (SICD3+CD25+, p = 0.016; SICD3+HLADR, p = 0.012); ASX 10-6 M + AZE 10-6 M (SICD3+CD25+, p = 0.012; SICD3+HLADR, p = 0.015); GB 10-6 M + CTZ 10-6 M (SICD3+CD25+, p = 0.024, SICD3+HLADR+, p = 0.019). Results demonstrate improved activity of antihistamines by 2 phytochemicals, suggesting dosing strategies for animal trials of ASX- or GB-augmented formulations for seasonal allergic rhinitis and asthma.

Trace elements and cell-mediated immunity in gestational and pre-gestational diabetes mellitus at third trimester of pregnancy.

THE AIM OF THE STUDY: To evaluate the correlations between Zn2+, Cu2+, Mg2+, Se2+ and Cr3+ and alteration in T cell subsets during diabetic and normal pregnancy. METHODS: The study involved 63 women with gestational diabetes mellitus (GD) and 16 pregnant women with Type 2 diabetes and 48 healthy, non-pregnant women were included as controls. Ten ml of whole venous blood from each participant was analyzed for electrolytes by atomic absorption; total antioxidant activity, individual enzymatic antioxidants by spectrophotometry; and lymphocyte sub-populations by flow cytometry. RESULTS: There were significant changes in lymphocyte sub-populations: Naïve T cells were decreased and memory T-cells and activated T cells (CD4+HLA-DR+, CD4+CD29+) were increased in diabetes in pregnancy. Zn2+ and Cr3+ deficiency were observed in Type 2 diabetics with an increase in Cu2+ in all pregnant cohorts. In healthy pregnant subjects, CD4+-HLA-DR+ was increased in direct proportion to serum Mg2+ (p<0.05) and Se2+ (p<0.01). In insulin-treated GD patients, CD4+CD29+ cells were increased proportionally to serum Zn2+ (p<0.05) while in diet controlled GD cohort CD45RO+/ CD45RA+ T cells correlated directly with serum Mg (p<0.05) and Zn2+ (p<0.01) while it correlated inversely with serum Cu2+ (p<0.01). CONCLUSIONS: The results of the present study show a correlation between trace element deficiency and increased lipid peroxidation in diabetes in pregnancy and lymphocyte activation. Dietary manipulation may, therefore, point to improvement in existing approaches to management of diabetes mellitus in pregnancy.

Summative interaction between astaxanthin, Ginkgo biloba extract (EGb761) and vitamin C in suppression of respiratory inflammation: a comparison with ibuprofen.

In this study, combinations of Ginkgo biloba leaf extract (EGb761) plus the carotenoid antioxidant astaxanthin (ASX) and vitamin C were evaluated for a summative dose effect in the inhibition of asthma-associated inflammation in asthmatic guinea-pigs. Ovalbumin-sensitized Hartley guinea-pigs challenged with ovalbumin aerosol to induce asthma, were administered EGb761, ASX, vitamin C or ibuprofen. Following killing, bronchoalveolar lavage (BAL) fluid was evaluated for inflammatory cell infiltrates and lung tissue cyclic nucleotide content. Each parameter measured was significantly altered to a greater degree by drug combinations, than by each component acting independently. An optimal combination was identified that included astaxanthin (10 mg/kg), vitamin C (200 mg/kg) and EGb761 (10 mg/kg), resulting in counts of eosinophils and neutrophils each 1.6-fold lower; macrophages 1.8-fold lower, cAMP 1.4-fold higher; and cGMP 2.04-fold higher than levels in untreated, asthmatic animals (p < 0.05). In conclusion, EGb761, ASX and vitamin C are shown here to interact summatively to suppress inflammation with efficacy equal to or better than ibuprofen, a widely used non-steroidal antiinflammatory drug (NSAID). Such combinations of non-toxic phytochemicals constitute powerful tools for the prevention of onset of acute and chronic inflammatory disease if consumed regularly by healthy individuals; and may also augment the effectiveness of therapy for those with established illness.

T lymphocyte activation profiles in peripheral blood of long- versus short-term residents of Kuwait: comparison with asthmatics.

INTRODUCTION: During the Arabian Gulf Wars of 1991 and 2003, the resident population of Kuwait sustained heavy exposure to environmental toxicants introduced by military activities. No comprehensive studies have been conducted to assess how exposure to the wartime and postwar environment may have altered the fundamental patterns of immune reactivity among Kuwaitis in ways that affect pathogenesis of disease. This present study addresses this issue by characterising immunological features of asthma and allergies in a Kuwaiti population that is unique and possibly correlates with toxicant exposures. MATERIALS AND METHODS: Twenty-fi ve long-term residents of Kuwait afflicted with bronchial asthma concurrent with rhinitis; and 2 healthy control groups: 18 long-term residents and 10 newcomers to Kuwait were evaluated by 2- and 3-colour fl ow cytometry for peripheral blood T cell subpopulation frequencies. RESULTS: Relative to healthy, long-term residents, significantly elevated frequencies of all activated cell phenotypes were observed in the blood of the asthmatic group (P <0.05 to P <0.001), except for CD8+HLA-DR+ cells and a presumed T-regulatory (Treg) subpopulation: CD4+CD25(high). The asthmatic group was also observed to have larger populations of CD3+ (pan-T cells), CD4+ (T helper cells) and CD8+ (cytotoxic T cells), CD3+CD56 (NKT-like cells) and CD56+CD16+ (NK cells) compared to healthy long-term residents. Compared to healthy recent immigrants, the blood of long-term residents contained elevated levels of CD3+CD56+ (NK-like), CD4+CD45RA+/ CD45RO+ (Naive-to-Memory Transitional), but lower CD4+CD25+(high) (Treg) (P <0.05). CONCLUSIONS: Elevated representation of natural killer (NKT)-like and memory phenotypes may predispose long-term residents towards enhanced susceptibility for airway disease; while at the same time, reducing representation of Treg cells which are protective against airway disease, and this may increase vulnerability to these syndromes among the residents of Kuwait. These results may provide insight into the features of immunopathogenesis of asthma and allergies in Kuwait that arise as a result of the special environment of the country.

Arrhythmia in the neonatal intensive care unit.

A random sample of 457 neonates was prospectively studied in order to identify the incidence, common types, and risk factors for arrhythmias in the neonatal intensive care unit (NICU). A 12-lead EKG was studied in all neonates (n = 457). A total of 139 Holter studies was done in every fourth baby with a normal EKG (n = 100) and in all babies with an abnormal EKG (n = 39). Of the 100 infants who were thought to be arrhythmia-free by EKG, nine infants demonstrated an arrhythmia on Holter studies. When we correlated screening results with maternal, obstetrical, and neonatal risk factors; arrhythmias were significantly associated with male gender, more mature gestational age, lower glucose levels, maternal smoking, high umbilical artery lines, and the use of the nebulized beta-2 adrenergic treatment, whereas umbilical venous lines and dopamine infusion did not relate to arrhythmia. We conclude that arrhythmias are more common in the NICU than in the general neonatal population. Compared to Holter monitoring, the sensitivity of the EKG was only 89%.

Levocetirizine modulates lymphocyte activation in patients with allergic rhinitis.

Levocetirizine, a second generation non-sedating antihistamine that blocks the H(1) histamine receptor, may exhibit immunoregulatory properties that augment its primary pharmacological mechanism. To investigate this possibility, 13 Kuwaiti seasonal allergic rhinitis (SAR) patients were treated with levocetirizine for four weeks in comparison with a 7-member placebo-treated control group, followed by clinical evaluation and flow cytometric analysis of peripheral venous blood for inflammatory cell and lymphocyte subpopulation profiles. Relative to the controls, levocetirizine-treated patients exhibited an expected reduction in early phase allergic symptoms, including sneezing (P<0.001), nasal itching (P<0.01), nasal congestion, and running nose (P<0.001); reduced percentages of eosinophils (P<0.05); and three subpopulations of activated T lymphocytes: CD4+CD29+, CD4+CD212+, and CD4+CD54+ (P<0.05). Levocetirizine treatment also correlated with a significant increase in the percentage of CD4+CD25+ T cells (P<0.001). The ability of levocetirizine to reduce percentage representation of cell phenotypes known to contribute to inflammatory tissue damage (eosinophils, CD4+CD29+, CD4+CD212+, and CD4+CD54+) and expand percentages of CD4+CD25+, which may include protective immunoregulatory (Treg) cells, indicates that the drug has pharmacological potential beyond the immediate effects of H(1) histamine-receptor inhibition. Although the present data does not define a therapeutic mechanism, the results reported here establish important trends that may be used to guide future mechanistic examination of immunoregulatory capacity of H(1) inhibitors.

Butyrylcholinesterase activity in gestational diabetes: correlation with lymphocyte subpopulations in peripheral blood.

PROBLEM: Inefficient clearance of pregnancy-threatening toxins may contribute to gestational diabetes (GD) and Type II diabetes mellitus (DM) through mechanisms involving immune dysregulation. METHOD OF STUDY: Peripheral venous blood from pregnant Kuwaiti women in third trimester, including 15 GD and 17 DM patients, 14 healthy pregnant (HP) and eight non-pregnant subjects, was analyzed by two-color flow cytometery for number and percentage representation of T lymphocytes. Buterylcholinesterase (BuChE) activity was measured using buterylthiocholine iodide and spectrophotometry. RESULTS: Relative to HP, GD patients exhibited higher ratios of activated and memory phenotypes, including CD4+ CD25+ (P < 0.01), CD4+ HLA-DR (P < 0.05) and CD4+ CD45RO+ (P < 0.05) cells. Serum BuChE activity exhibited positive correlation within the HP cohort with CD4+ CD25+ (P < 0.05), but not in GD and DM cohorts. CONCLUSIONS: Positive correlation between BuChE and a (presumptive) 'regulatory' T-cell phenotype in HP, but not GD or DM may indicate existence of protective detoxification mechanisms against oxidative stress in normal pregnancies.

Lymphocyte sub-populations in gestational diabetes.

PROBLEM: We hypothesize that the normal immunologic responses by the maternal immune system during pregnancy are not as well-regulated in gestational diabetes (GD) patients as in healthy pregnant women. METHOD OF STUDY: Using two-color flow cytometry we evaluated frequencies of peripheral blood lymphocytes in 20 GD patients being treated with insulin; 43 GD patients treated with dietary therapy but no insulin; 44 women experiencing normal pregnancies; and 48 non-pregnant women. RESULTS: When compared with healthy pregnant women, both GD cohorts showed higher percentages CD4(+)CD25(+) (P < 0.05), CD4(+)CD45RO(+) (P < 0.05) and CD4(+)CD29(+) (P < 0.01) but lower percentages of CD4(+)CD45RA(+) (P < 0.05). Higher percentages of the activated phenotypes CD8(+)CD25(+) and CD8(+)HLA-DR(+) cells in the diet-treated cohort and CD4(+)HLA-DR(+) cells in insulin-treated GB cohort, were observed compared with healthy pregnant subjects (P < 0.05). CONCLUSIONS: Expanded populations of activated peripheral blood T cells are associated with GD, suggesting that normal maternal immunosuppression is less effective in GD-afflicted women.

Major lymphocyte populations and T-cell expression of ICAM-1 and L-selectin adhesion molecules in Kuwaitis with asthma and rhinitis.

The authors evaluate major immunologic features of asthma and allergies in a Kuwaiti population. They analyzed peripheral venous blood from 17 asthmatic and 17 healthy long-term residents of Kuwait by using two-color flow cytometry for major lymphocyte subpopulations; they also evaluated 10 healthy individuals who had recently arrived in Kuwait. Relative to healthy subjects, asthmatics exhibited increased percentages of T+ NK cells (p < .01), T-helper cells (p < .05), T-cytotoxic and NK cells for both total numbers (p < .01-.001) and percentages (p < .05-.01), and increased percentages of T cells expressing CD54 (ICAM-1; p < .001) and CD62 (L-selectin; p < .01). However, B cells were present at significantly lower levels in asthmatics, both in total numbers (p < .05) and percentages (p < .01). In comparison with healthy individuals who had recently arrived in Kuwait, healthy long-term residents exhibited elevated numbers of pan-T cells (p < .01) and T-helper cells (p < .05). These results help establish immunological parameters for asthma and allergies in Kuwaiti populations.