Normal Healthy Adult Volunteers Research Articles

Differential Expression of Human Peripheral Mononuclear Cells Phenotype Markers in Type 2 Diabetic Patients and Type 2 Diabetic Patients on Metformin.

Background: Although peripheral blood mononuclear cells (PBMC) have been demonstrated to be in a pro-inflammatory state in obesity and type 2 Diabetes Mellitus (T2DM), characterization of circulating PBMC phenotypes in the obese and T2DM and the effect of Metformin on these phenotypes in humans is still ill-defined and remains to be determined.Methods: Thirty normal healthy adult volunteers of normal weight, 30 obese subjects, 20 obese newly diagnosed diabetics and 30 obese diabetics on Metformin were recruited for the study. Fasting blood samples were collected and PBMC were isolated from whole blood. Polarization markers (CD86, IL-6, TNFα, iNOS, CD36, CD11c, CD169, CD206, CD163, CD68, CD11b, CD16, and CD14) were measured by RT-qPCR. Gene expression fold changes were calculated using the 2−ΔΔCT method for RT-qPCR.Results: Obesity and T2DM are associated an increased CD68 marker in PBMC. mRNA expression of CD11b, CD11c, CD169, and CD163 were significantly reduced in PBMC from T2DM subjects whereas CD11c was significantly inhibited in PBMC from obese subjects. On the other hand, macrophage M1-like phenotype was observed in T2DM circulation as demonstrated by increased mRNA expression of CD16, IL-6, iNOS, TNFα, and CD36. There were no significant changes in CD14 and CD86 in the obese and T2DM when compared to the lean subjects. Metformin treatment in T2DM reverted CD11c, CD169, IL-6, iNOS, TNFα, and CD36 to levels comparable to lean subjects. CD206 mRNA expression was significantly upregulated in PBMC of T2DM while Metformin treatment inhibited CD206 expression levels.Conclusions: These data support the notion that PBMC in circulation in T2DM express different pattern of phenotypic markers than the patterns typically present in M1 and M2 like cells. These phenotypic markers could be representative of metabolically activated macrophages (MMe)-like cells. Metformin, on the other hand, reduces MMe-like cells in circulation.

Sonographic Measurements of the Ulnar Nerve at the Elbow With Different Degrees of Elbow Flexion

To determine whether there were differences in the cross-sectional area (CSA) and the flattening ratio of the normative ulnar nerve as it passes between the medial epicondyle and the olecranon at 30° of elbow flexion versus 90° of elbow flexion. Bilateral upper extremities of normal healthy adult volunteers were evaluated with ultrasound. The CSA and the flattening ratio of the ulnar nerve at the elbow as it passes between the medial epicondyle and the olecranon were measured, with the elbow flexed at 30° and at 90°, by 2 operators with varying ultrasound scanning experience by using ellipse and direct tracing methods. The results from the 2 different angles of elbow flexion were compared for each individual operator. Finally, intraclass correlations for absolute agreement and consistency between the 2 raters were calculated. An outpatient clinic room at a regional rehabilitation center. Twenty-five normal healthy adult volunteers. The mean CSA and the mean flattening ratio of the ulnar nerve at 30° of elbow flexion and at 90° of elbow flexion. First, for the ellipse method, the mean CSA of the ulnar nerve at 90° (9.93mm(2)) was slightly larger than at 30° (9.77mm(2)) for rater 1. However, for rater 2, the mean CSA of the ulnar nerve at 90° (6.80mm(2)) was slightly smaller than at 30° (7.08mm(2)). This was found to be statistically insignificant when using a matched pairs t test and the Wilcoxon signed-rank test, with a significance level of .05. Similarly, the difference between the right side and the left side was not statistically significant. The intraclass correlations for absolute agreement between the 2 raters were not very high due to different measurement locations, but the intraclass correlations for consistency were high. Second, for the direct tracing method, the mean CSA at 90° (7.26mm(2)) was slightly lower than at 30° (7.48mm(2)). This was found to be statistically nonsignificant when using the matched pairs t test and the Wilcoxon signed-rank test with a significance level of .05. There was no significant difference in the average flattening ratio between the 2 angles for the left arm (0.54 at 30° vs 0.56 at 90°; P= .619 for the matched pairs t test and .274 for the Wilcoxon signed-rank test). However, for the right arm, the flattening ratio at 90° was significantly higher than that at 30° (0.58 at 90° vs 0.50 at 30°; P= .007 for both the matched pairs t test and the Wilcoxon signed-rank test). The mean CSA of the ulnar nerve at the elbow at 30° was not significantly different than at 90°. However, the average flattening ratio at 90° was found to be significantly higher than at 30° for the right arm.

Assessing the accuracy and reliability of direct height measurement for use in adult neurological patients with contractures: a comparison with height from ulna length

It is not possible to measure the height of a patient with contractures in the normal way. On occasions, health professionals need to measure the height directly of patients with contractures because: (i) the patient is unable to report his/her height as a result of a lack of mental capacity and (ii) there is no-one available to report the height or the reported height is obviously inaccurate. Height derived from ulna length is an option but, in some individuals, it can be conspicuously inaccurate. Therefore, we have attempted to develop and validate a simple bedside method using a tape measure. Twenty-four normal healthy adult volunteers were recruited into the study. They were positioned on a hospital bed in three different standardised positions simulating patients with typical contractures. They were measured in each position by four measurers using blanked out tape measures. The participants were then measured standing giving a 'true' height. Ulna length was also measured. The mean of the tape-measured heights was 2.3cm taller than the 'true' height (95% confidence interval=4.6-9.3). The measurement in all three positions gave an over-estimate of the height, whereas the ulna-derived height gave an under-estimate (2.2cm). The estimated reliability between measurers (intraclass correlation coefficient) was 0.94 with a 95% confidence interval (0.91-0.96). This new method is giving an acceptable estimate of the height of contracted patients with a good inter-rater reliability. It comprises a useful additional tool for measuring the height of people with contractures.

Macular Choroidal Thickness in Normal Pediatric Population Measured by Swept-Source Optical Coherence Tomography

To evaluate choroidal thickness in healthy pediatric population by swept-source longer-wavelength optical coherence tomography (SS-OCT). This was a cross-sectional comparative, noninterventional study. The macular area of 83 eyes from 43 pediatric patients (<18 years) was studied with an SS-OCT prototype system. Macular choroidal thickness was manually determined at 750-μm intervals by measuring the perpendicular distance from the posterior edge of the RPE to the choroid/sclera junction, along a horizontal 4500-μm line centered in the fovea. Three observers independently determined choroidal thickness. Pediatric choroidal thickness was compared with choroidal thickness from 75 eyes from 50 normal healthy adult volunteers (18 years or older). Mean age was 10 ± 3 years (3-17) in the pediatric population versus 53 ± 16 (25-85) in the adult population (P < 0.001). Mean spherical equivalent was not different (P = 0.06) between both groups. Mean subfoveal choroidal thickness was 312.9 ± 65.3 μm in the pediatric versus 305.6 ± 102.6 μm in the adult population (P = 0.19). Mean macular choroidal thickness was 285.2 ± 56.7 μm in the pediatric versus 275.2 ± 92.7 μm in the adult population (P = 0.08). The distribution of choroidal thickness along the horizontal line was different for both populations; the temporal choroid was thicker in the pediatric population (320, 322, and 324 μm; P = 0.002, 0.001, and 0.06, respectively), followed by the subfoveal (312 μm) and nasal choroid (281, 239, and 195 μm). Macular choroidal thickness in the pediatric population is not significantly thicker than that of healthy adults. Differences are more remarkable in the temporal side of the fovea.

Increased Monocyte Expression of Sialoadhesin During Acute Cellular Rejection and Other Enteritides After Intestine Transplantation in Children

Sialoadhesin (CD169) facilitates T-cell priming when overexpressed on inflammatory monocytes. Monocyte-derived macrophages prime acute cellular rejection after intestine transplantation (ITx).The purpose of this study was to evaluate whether CD169-expressing activated monocytes associate with or predict ITx rejection. After informed consent (ClinicalTrials.gov NCT No. 01163578), activated CD169+CD14+monocytes were measured by flow cytometry in five normal healthy adult volunteers (group A), and 56 children with ITx sampled cross-sectionally (group B, 26), longitudinally (group C, 18), or during infection/inflammation without rejection (group D: acute enteritis, 9; Helicobacter pylori, 1; Streptococcal pharyngitis 1; and posttransplant lymphoma, 1). Activated monocytes were tested for correlations with donor-specific alloreactivity in simultaneous mixed lymphocyte co-cultures. Median age was 3 years (range 0.5-21 yr), and distribution of ITx-alone:combined liver-ITx was 25:31. Higher frequencies (%) of activated monocytes were seen during rejection in group B and infection/inflammation without rejection in group D (58 ± 28 and 73 ± 26), compared with nonrejectors or normal controls (10.6 ± 7.9 or 10.7 ± 6.5, P=0.001). In longitudinal monitoring, rejectors also showed higher activated monocyte frequencies (%) before ITx (64 ± 26 vs. 13.4 ± 8.6, P=0.0007) and during acute cellular rejection (55 ± 28 vs. 22.4 ± 15, P=0.006) when compared with nonrejectors. Activated monocytes correlated significantly with allospecific CD154+T-cytotoxic memory cells (Spearman r=0.688, P=7.1E-05) and CD154+B cells (r=0.518, P=0.005) in ITx recipients without inflammation/infection but not in group D. Monocytes overexpress sialoadhesin nonspecifically during ITx rejection and systemic or enteritic inflammatory states. When combined with allospecific T and B cells, this information may differentiate between rejection and other enteritides.

Isolation of Human Bone Marrow Mesenchymal Stem Cells by a Novel Monoclonal Antibody, ZUC3.

Objective: Due to absence of a single definitive marker of mesenchymal stem cells (MSCs) and low incidence in human bone marrow, the primary culture of MSCs, conventionally isolated with its characteristic of adherent, were considered to be heterogeneous containing of several subpopulations, which had currently limited our understanding of their biology and therapeutic applications. In our previous study, a novel murine monoclonal antibody (McAb) ZUC3 was produced by hybridoma technology, which was specifically reactive with human MSCs, while showed negative cross-reactivity when screened against a variety of human tissues. Now, ZUC3 antigen positive MSCs population would be further identified by magnetic-activated cell sorting (MACS).Methods: Bone marrow were taken from the iliac crest of normal healthy adult volunteers, and mononuclear cells were separated by density gradient centrifugation, then separated into positively- and negatively-labelled fractions with McAb ZUC3 by immunomagnetic activated cell sorting. The purity of positive cells was analyzed by flow cytometry, then ZUC3 antigen positive and negative cells were plated respectively in human MSCs medium consisting of 10% FBS, LG-DMEM. Characteristics of ZUC3 antigen positive cells phenotype was analyzed by flow cytometry, and proliferation and multiple differentiation potential of the cells was observed in vitro.Results: Flow cytometric analysis showed that ZUC3 antigen expression by cultured MSCs and mononuclear cells derived from bone marrow were 91.31±2.92%, 0.96±0.28% respectively, and western blotting showed the molecular mass of antigen was about 33KD. The purity of the recovered fractions for ZUC3 by MACS was 76.82±6.32%. The positive cells have adhered to culture flask in vitro, and the quantity of adhered cells that had fibroblast-like morphology increased and proliferated during primary expansion period, while the negative cells were observed as round shape cells without any proliferation. It was demonstrated that ZUC3 antigen positive cells continued growth with spindle-shape, extending beyond 30 population doublings in long-term culture. Analyzed by flow cytometry, the culture-expanded positive cells were uniformly positive for CD29, CD44, CD105, CD106, and lack typical hematopoietic antigens such as CD14, CD34, CD45, HLA-DR, which demonstrated that ZUC3 postive cells sorted from bone marrow mononuclear cells by McAb were MSCs. With proper medium, the ZUC3 antigen positive cells could be successfully induced to differentiate into adipocytes, osteoblasts, and neuro-like cells which were positive of neuron markers such as nestin, NSE and NF-M.Conclusion: ZUC3 McAb was a specific surface marker against human MSCs for cell sorting. The ZUC3 antigen positive cells separated from bone marrow mononuclear cells had potential capacity of high proliferation and multiple differentiation.

Comparative Immunogenicity of a Genetically Derived and a Plasma Pooled Hepatitis B Vaccine in Normal Adult Volunteers

ObjectiveThis study compares the seroconversion rate after vaccination with genetically derived and plasma pooled vaccine in normal healthy adult volunteers. Population and methodsForty volunteers of either sex were randomly divided into two groups comprising equal numbers of subjects. The mean (± SD) age and weight of the subjects in group A were 27.52±3.48 years and 65.60±8.06kg, respectively, while the mean (± SD) values for the individuals in group B were 30.75±3.78 years and 68.55±6.59kg. Recombinant and plasma pooled vaccines were administered as single 20μg and 3μg intramuscular injections to the respective groups of volunteers on days 1, 30, and 180. An automated system (AxSYM, based on microparticle enzyme immunoassay) was used to measure the seroconversion rate in serum harvested from an 8mL blood sample collected within 1 month of the third dose (i.e., on day 210), from each volunteer. ResultsBoth of the vaccines were well tolerated by all individuals and no adverse reaction was reported or observed during the study. The subject seroconversion rate corrected with reference to the cut-off rate after vaccination with DNA-derived vaccine was significantly greater (p<0.05) than that observed after the administration of plasma pooled-derived vaccine (51.64±17.00 versus 22.96±12.04). ConclusionThe vaccine response, as indicated by the seroconversion rate (concentration of anti-HBs in mIU/mL) in the group vaccinated with genetically derived vaccine was observed to be better (p<0.01) when matched with the International Antibody Standard.

The Influences of Halo-Vest Fixation and Cervical Hyperextension on Swallowing in Healthy Volunteers

Radiographic and electromyographic evaluation of swallowing functions was performed for different positions with a Halo-vest brace. The aim of this study was to clarify the mechanism of dysphagia of cervical hyperextension with a Halo-vest brace in neurologically normal adult volunteers. Garfin et al reported that 3 of 179 patients had dysphagia attributable to the Halo-vest brace after cervical spinal cord injury. Readjustment of the position of the head in the Halo-vest brace was required in those cases. They concluded that the head-extended position with a Halo-vest brace made it difficult to swallow. However, the details of this dysphagia were not known. Six healthy adults volunteers between the ages of 24 and 33 participated in this study. Subjects were radiographically and electromyographically observed swallowing thin liquids at the neutral position without a Halo-vest brace (N-HV), the neutral position with a Halo-vest brace (N+HV) and at hyperextension with a Halo-vest brace (E+HV). In the durational measurements, there were significant changes between the N-HV and E+HV in pharyngeal transit time. The motion measurements showed that the initial hyoid position placed lower from the mandibular plane, and vertical hyoid movement was prolonged in the E+HV. In the electromyographic measurements, greater activity was observed from the suprahyoid muscles in the E+HV. One subject had laryngeal penetration already in the N-HV, and the same subject exhibited aspiration in the E+HV. Another two subjects exhibited penetration in the E+HV. The result of this study demonstrated that cervical hyperextension with the Halo-vest brace caused mechanical changes in the swallowing of normal healthy adult volunteers.

Differential effects of glucose and alcohol on reactive oxygen species generation and intranuclear nuclear factor-κB in mononuclear cells

It has previously been shown that oral intake of 300 calories of glucose (75 g), lipid, or protein increases reactive oxygen species (ROS) generation by polymorphonuclear cells (PMNL) and mononuclear cells (MNCs). We investigated the effects of 75 g glucose on proinflammatory transcription factor, nuclear factor-κB (NFκB), in mononuclear cells. To further investigate whether the effects of macronutrient-induced oxidative stress are due to consumption of calories or are nutrient specific, we investigated the effects of acute oral challenge of equicaloric amounts of alcohol (300 calories) on ROS generation and NF-κB activation in MNCs and PMNL and compared them with those of glucose and water (control). Sixteen normal healthy adult volunteers were given either vodka (10 subjects), glucose solution (10 subjects), or 300 mL water (7 subjects). Vodka and glucose drinks were equivalent to 300 calories. We measured ROS generation and intranuclear NF-κB activation by PMNL cells and MNCs at 1 hour, 2 hours, and 3 hours following ingestion. ROS generation by both MNC and PMNL increased significantly ( P < .05 for MNC and P < .01 for PMNL) following intake of glucose solution, but did not change significantly following alcohol or water. NF-κB binding activity in MNC nuclear extracts also increased ( P < .001) following ingestion of glucose solution, but did not change after the administration of alcohol or water. We conclude that (1) 75 g oral glucose increases NF-κB binding activity in MNCs. (2) While 75 g glucose (300 calories) induces an increase in ROS generation and intranuclear NF-κB, equicaloric amounts of alcohol did not produce these effects.

Interleukin 10-induced thrombocytopenia in normal healthy adult volunteers: evidence for decreased platelet production

Recombinant human interleukin 10 (rhuIL-10) inhibits the production of proinflammatory cytokines and has shown promise in the treatment of inflammatory bowel disease. Clinical trials have been accompanied by a reversible decline in platelet counts. We conducted a randomized, double-blinded, placebo-controlled, parallel group trial in 12 healthy volunteers to investigate the aetiology of rhuIL-10-induced thrombocytopenia. Eight volunteers received 8 microg/kg/d of rhuIL-10 subcutaneously, while four subjects received a placebo alone for 10 d. A reversible decline in the platelet counts from a mean of 275 x 10(9)/l to 164 x 10(9)/l was observed in the IL-10-treated cohort (P = 0.012). A fall in the haemoglobin mean levels was also observed in the IL-10-treated cohort from 13.7 to 11.7 g/dl (P = 0.011). No significant change was observed in the bone marrow cellularity or myeloid/erythroid ratio or in the number of megakaryocytes per high-powered field (HPF). A fall was observed in the number of megakaryocyte colony-forming units (CFU-MKs) after the administration of IL-10 compared with those receiving the placebo (P = 0.068). No difference in the change in granulocyte-macrophage CFUs (CFU-GMs), mixed lineage CFUs (CFU-GEMMs) or erythroid burst-forming units (BFU-Es) was observed when comparing the IL-10- vs. placebo-treated groups (P > 0.465). Serum cytokine levels of thrombopoietin (TPO). IL-6 and granulocyte-macrophage colony stimulating factor (GM-CSF) were not decreased following IL-10 administration. In fact, both TPO and GM-CSF appeared to be slightly increased in the serum. All subjects underwent In111-labelled platelet survival studies with liver/spleen scans to assess splenic sequestration prior to and then on day 7 of treatment. A significant reduction in splenic sequestration of platelets (P =0.012) was observed in the IL-10-treated group, but not in the placebo-treated subjects.

Interleukin 10‐induced thrombocytopenia in normal healthy adult volunteers: evidence for decreased platelet production

Recombinant human interleukin 10 (rhuIL‐10) inhibits the production of proinflammatory cytokines and has shown promise in the treatment of inflammatory bowel disease. Clinical trials have been accompanied by a reversible decline in platelet counts. We conducted a randomized, double‐blinded, placebo‐controlled, parallel group trial in 12 healthy volunteers to investigate the aetiology of rhuIL‐10‐induced thrombocytopenia. Eight volunteers received 8 μg/kg/d of rhuIL‐10 subcutaneously, while four subjects received a placebo alone for 10 d. A reversible decline in the platelet counts from a mean of 275 × 109/l to 164 × 109/l was observed in the IL‐10‐treated cohort (P = 0·012). A fall in the haemoglobin mean levels was also observed in the IL‐10‐treated cohort from 13·7 to 11·7 g/dl (P = 0·011). No significant change was observed in the bone marrow cellularity or myeloid/erythroid ratio or in the number of megakaryocytes per high‐powered field (HPF). A fall was observed in the number of megakaryocyte colony‐forming units (CFU‐MKs) after the administration of IL‐10 compared with those receiving the placebo (P = 0·068). No difference in the change in granulocyte–macrophage CFUs (CFU‐GMs), mixed lineage CFUs (CFU‐GEMMs) or erythroid burst‐forming units (BFU‐Es) was observed when comparing the IL‐10‐ vs. placebo‐treated groups (P &gt; 0·465). Serum cytokine levels of thrombopoietin (TPO), IL‐6 and granulocyte–macrophage colony stimulating factor (GM‐CSF) were not decreased following IL‐10 administration. In fact, both TPO and GM‐CSF appeared to be slightly increased in the serum. All subjects underwent In111‐labelled platelet survival studies with liver/spleen scans to assess splenic sequestration prior to and then on day 7 of treatment. A significant reduction in splenic sequestration of platelets (P = 0·012) was observed in the IL‐10‐treated group, but not in the placebo‐treated subjects.

The effects of practice on measures of performance

AbstractThe effects of practice on a range of performance measures often used in studies of psychoactive drugs have been assessed in 50 normal healthy adult volunteers, during 10 practice sessions over an 8‐day period. There was no dramatic effect of practice on any of the parameters measured, only one of the mean values at the tenth session exceeded one standard deviation of the mean of the first session (manipulative motor task). Practice did not materially affect performance on elapsing time estimation, tapping rate, body sway measured by a ultrasonic method or rapid visual information processing task variables. There tended to be a non‐significant, exponential learning curve for choice reaction response and movement times to visual stimuli, flash fusion threshold and the manipulative motor task, with a statistically significant practice effect on all 16 visual analogue scales used; a plateau was apparently achieved by the fifth practice session for all these parameters. There was a statistically significant straight‐line improvement in digit span across the 10 sessions. It is concluded that for most simple measures of performance used in psychopharmacology including visual analogue scales, four pre‐study training sessions are necessary, and that direct methods of recording body sway are preferred to indirect methods such as the balance platform.

In vitro effect of Panax ginseng on phytohaemagglutinin-induced lymphocyte transformation.

The effect of hydrocortisone or Panax ginseng, and/or a combination of hydrocortisone and Panax ginseng on phytohaemagglutinin (PHA-P)-induced transformation of peripheral blood lymphocytes were studied in 4 normal healthy adult volunteers. Increasing concentrations of Panax ginseng 0.16-1.60 micrograms X ml-1 caused a dose-related inhibition of PHA-P transformation of lymphocytes. A combination of 500 micrograms X ml-1 hydrocortisone and 0.80 micrograms X ml-1 Panax ginseng produced a greater suppression of PHA-P stimulation than either drug used alone. This suggests that Panax ginseng has a steroid-like effect in vitro, and may have a potentiating effect with hydrocortisone on T-cell-mediated immunity.

Cardiovascular Effects of Topical Glaucoma Therapies in Normal Subjects

AbstractTopical glaucoma therapies may induce adverse systemic side effects. This study was designed to determine the potential for drug-induced cardiovascular side effects of various glaucoma therapies applied topically to the eye. Data from three short-term double-blind studies in a total of 19 normal healthy adult volunteers were used to discriminate potential cardiovascular side effects. Baseline pulse rates and blood pressures were measured before and after exercise. Subsequent measurements were made 3 hr after topical ocular treatment with two drops of each glaucoma medication. Each individual was used to test each glaucoma therapy after washout of a 1 week period between treatments. The therapies included an alpha agonist (ALO 2145), three beta-blockers (betaxolol, bunolol, and timolol), and a placebo. No drug related pre- and post-exercise mean arterial pressure changes were observed. However, the two nonselective beta-blockers, timolol (0.5%) and bunolol (1%), consistently decreased the resting a...