Enrichment of IL-17A+ IFN-γ+ and IL-22+ IFN-γ+ T cell subsets is associated with reduction of NKp44+ ILC3s in the terminal ileum of Crohn's disease patients.

Crohn's disease (CD) is a chronic inflammatory condition of the human gastrointestinal tract whose aetiology remains largely unknown. Dysregulated adaptive immune responses and defective innate immunity both contribute to this process. In this study, we demonstrated that the interleukin (IL)-17A+ interferon (IFN)-γ+ and IL-22+ IFN-γ+ T cell subsets accumulated specifically in the inflamed terminal ileum of CD patients. These cells had higher expression of Ki-67 and were active cytokine producers. In addition, their proportions within both the IL-17A-producer and IL-22-producer populations were increased significantly. These data suggest that IL-17A+ IFN-γ+ and IL-22+ IFN-γ+ T cell subsets might represent the pathogenic T helper type 17 (Th17) population in the context of intestinal inflammation for CD patients. In the innate immunity compartment we detected a dramatic alteration of both phenotype and function of the intestinal innate lymphoid cells (ILCs), that play an important role in the maintenance of mucosal homeostasis. In the inflamed gut the frequency of the NKp44- CD117- ILC1s subset was increased significantly, while the frequency of NKp44+ ILC3s was reduced. Furthermore, the frequency of human leucocyte antigen D-related (HLA-DR)-expressing-NKp44+ ILC3s was also reduced significantly. Interestingly, the decrease in the NKp44+ ILC3s population was associated with an increase of pathogenic IL-17A+ IFN-γ+ and IL-22+ IFN-γ+ T cell subsets in the adaptive compartment. This might suggest a potential link between NKp44+ ILC3s and the IL-17A+ IFN-γ+ and IL-22+ IFN-γ+ T cell subsets in the terminal ileum of CD patients.

Central nervous system delivery of recombinant ciliary neurotrophic factor by polymer encapsulated differentiated C2C12 myoblasts.

Neurotrophic factors hold promise for the treatment of neurodegenerative diseases. Intrathecal transplantation of polymer encapsulated cell lines genetically engineered to release neurotrophic factors provides a means to deliver them continuously behind the blood-brain barrier. Long-term delivery, however, may benefit from the use of conditionally mitotic cells to avoid the overgrowth observed with continuously dividing cell lines. Myoblast lines have all the advantages of dividing cell lines, i.e., unlimited availability, possibility for in vitro screening for the presence of pathogens, suitability for stable gene transfer and clonal selection. Furthermore they can be differentiated into a nonmitotic stage upon exposure to low-serum-containing medium. In this study, mouse C2C12 myoblasts were transfected with a pNUT expression vector containing the human ciliary neurotrophic factor (CNTF) gene. hCNTF expression and bioactivity were demonstrated by Northern blot, ELISA assay, and measurement of choline acetyltransferase (ChAT) activity in embryonic spinal cord motor neuron cultures. One C2C12 clone was found to secrete 200 ng of CNTF/10(6) cells per day. The rate of secretion of hCNTF was not altered upon differentiation of C2C12 myoblasts. A bromodeoxyuridine (BrdU) proliferation assay indicated that approximately 12% of the myoblasts continue to divide after 4 days in low-serum-containing medium. The presence of the herpes simplex thymidine kinase gene (HSV-tk) in the expression vector, however, provides a way to eliminate these dividing myoblasts upon exposure to ganciclovir, therefore increasing the safety of the encapsulation technology using established cell lines. Encapsulated hCNTF-C2C12 cells can partially rescue motor neurons from axotomy-induced cell death. In adult rats, intrathecal implantation of encapsulated hCNTF-C2C12 cells or control C2C12 confirmed the long-term survival of these cells and their potential use as a source of neurotophic factors for the treatment of neurodegenerative diseases.

Reduced thyroid function after thyrotropin stimulation.

The course of serum T4 and T3 return to baseline after TSH stimulation was studied in two groups at six normal subjects over 28 days after im bovine TSH (b TSH; 0.15 U/kg). In the first group of six subjects, serum bTSH rose from undetectable levels to a mean peak of 5.6 +/- 0.5 ng/ml (mean +/- SE) at 2 h, and fell below detectable levels by 24 h with a t1/2 of 7 +/- 1 h. T4 rose to a peak 59 +/- 10% above basal levels within 24 h, returned to basal levels on day 7, then dropped below basal levels on days 9-24, with a nadir of -16 +/- 4% on day 14. Free T4 paralleled T4 levels. T3 rose to a peak 104 +/- 28% above basal at 24 h, then fell faster than T4, reaching basal levels by day 4. During the period of low T4, T3 was at or below basal levels. Human TSH (h TSH) concentration dropped when T4 and T3 rose, but did not rise above basal levels when T4 and T3 fell below basal levels. Neither a T3 elevation nor an increased percentage of free T4 was present during the time of reduced T4 levels. The same pattern of thyroidal response was seen in the second group of six subjects. In this second group, hTSH response to repeated TRH challenge was studied. During the period of reduced T4 and T3, hTSH response to TRH was diminished. On day 28, T4, T3, hTSH, and hTSH response to TRH returned to basal levels. We conclude that the brief elevation of T4 and T3 after bTSH stimulation exerts a suppressive effect on the pituitary which extends beyond the period of elevated thyroid hormone levels, and that delay in pituitary recovery is the mechanism of the decreased thyroid function after acute bTSH stimulation.

Elevated plasma glucagon in amyotrophic lateral sclerosis.

This study shown an abnormality in glucagon levels that may explain the glucose intolerance, abnormal insulin reactions, and abnormal plasma amino acid levels seen in amyotrophic lateral sclerosis (ALS). We randomly administered two test meals, differing only in protein source (soy versus casein) at least 1 week apart and measured fasting and postprandial bloods for glucagon, insulin, and glucose levels in 11 ALS patients. With the soy test meal, glucagon levels were elevated in all ALS patients compared with controls: at fasting (237 +/- 111 versus 108 +/- 46 pg/ml, p less than 0.01) and 1/2 hour (389 +/- 94 versus 133 +/- 68 pg/ml, p less than 0.001), and 2 hours postprandial (379 +/- 75 versus 108 +/- 53 pg/ml, p less than 0.001). Glucagon levels after the casein test meal were also significantly elevated. Insulin was elevated by both test meals. Casein produced significant glucose intolerance.

Indapamide regresses, but transdermal clonidine does not regress, left ventricular hypertrophy in hypertensive diabetic patients.

This report describes the effects of indapamide versus transdermal clonidine on left ventricular hypertrophy (LVH) in hypertensive diabetic patients. A sample of 24 hypertensive diabetic men, aged 40-68 years, with echocardiographically proven LVH was equally divided in to 2 groups. Group 1 was treated with indapamide 2.5 mg/day, and group C with transdermal clonidine weekly. Left ventricular mass and posterior wall and septal thickness were measured by standard echocardiograms done at baseline and every 6 months. At 24 months, treatment crossover was done. Normotension was maintained throughout the study. With indapamide, LVH regression was measurable at 6 months, and left ventricular mass had returned to normal after 18 months. Transdermal clonidine did not regress LVH, but when the patients were switched to indapamide, LVH did regress. Clonidine maintained normal ventricular dimensions after regression had been induced by indapamide.

Glial cell line-derived neurotrophic factor (GDNF), a new neurotrophic factor for motoneurones.

Glial cell line-derived neurotrophic factor (GDNF) has been postulated to be a specific dopaminergic neurotrophic factor since it selectively enhances the survival of dopaminergic neurones in vitro. We report here that GDNF can also act as a neurotrophic factor for motoneurones. GDNF released by GDNF-transfected BHK cells increases the activity of choline acetyltransferase (ChAT) in cultures from embryonic rat ventral mesencephalon containing cholinergic neurones from cranial motor nuclei and in cultured spinal motoneurones. Furthermore, local application of polymer-encapsulated BHK cells releasing GDNF to transected facial nerve in newborn rats diminishes the death of motoneurones normally occurring after axotomy in the neonatal period. The present results indicate that GDNF may have a therapeutic potential in human motoneurone diseases such as amyotrophic lateral sclerosis.

Rescue of motoneurons from axotomy-induced cell death by polymer encapsulated cells genetically engineered to release CNTF.

The neurodegenerative disease amyotrophic lateral sclerosis (ALS) results from the progressive loss of motoneurons, leading to death in a few years. Ciliary neurotrophic factor (CNTF), which decreases naturally occurring and axotomy-induced cell death, may result in slowing of motoneuron loss and has been evaluated as a treatment for ALS. Effective administration of this protein to motoneurons may be hampered by the exceedingly short half-life of CNTF, and the inability to deliver effective concentration into the central nervous system after systemic administration in vivo. The constitutive release of CNTF from genetically engineered cells may represent a solution to this delivery problem. In this work, baby hamster kidney (BHK) cells stably tranfected with a chimeric plasmid construct containing the gene for human or mouse CNTF were encapsulated in polymer fibers, which prevents immune rejection and allow long-term survival of the transplanted cells. In vitro bioassays show that the encapsulated transfected cells release bioactive CNTF. In vivo, systemic delivery of human and mouse CNTF from encapsulated cells was observed to rescue 26 and 27% more facial motoneurons, respectively, as compared to capsules containing parent BHK cells 1 wk postaxotomy in neonatal rats. With local application of CNTF on the nerve stump and by systemic delivery through repeated subcutaneous injections, 15 and 13% more rescue effects were observed. These data illustrate the potential of using encapsulated genetically engineered cells to continuously release CNTF to slow down motoneuron degeneration following axotomy and suggest that encapsulated cell delivery of neurotrophic factors may provide a general method for effective administration of therapeutic proteins for the treatment of neurodegenerative diseases.

The effect of long endurance running on natural killer cells in marathoners.

Ten experienced marathoners were exercised 3 h in the laboratory. Blood samples were collected at 0 h baseline, 1 h exercise, and 5 min, 1.5 h, 6 h, and 21 h recovery and were analyzed for total number of lymphocytes expressing membrane antigens found on natural killer (NK) cells. NK activity was also measured. Four of the seven subpopulations of lymphocytes studied, Leu-11+19+, Leu-11+19-, Leu-11+7-, and Leu-19+11-, showed significant within-subject effects over time, using repeated measures ANOVA. Simple contrasts with baseline values showed that, at 1.5 h and 21 h recovery, total number of lymphocytes bearing three different combinations of NK markers, Leu-11+19+, Leu-11+19-, and Leu-11+7-, were significantly decreased when compared with baseline values. At 1.5 h recovery, NK activity was significantly decreased below baseline levels for four of the six effector NK cell/target K562 myelogenous leukemia cell (E:T) ratios tested. At 6 h recovery, NK activity was still decreased significantly with the 12.5:1 and 3:1 E:T ratios. By 21 h recovery, NK activity did not differ significantly from baseline levels. Cortisol levels at 5 min post-exercise were negatively correlated with NK activity at 1.5 h recovery (r = -0.62, P = 0.05, 50:1 E:T ratio; r = -0.66, P = 0.04, 25:1 E:T ratio). Further research is needed to elucidate the effect these changes have on host immunosurveillance and immunoresponsiveness in vivo.

Quantitative modeling of limitations caused by diffusion.

Transport of nutrients and metabolites in many bioartificial tissue constructs relies exclusively on diffusion, i.e., on the presence of a concentration gradient between the inside of the construct and the surrounding milieu. A quantitative evaluation of the rate of diffusional processes is thus essential for properly designing three-dimensional cell-polymer systems, and for assessing the chemical environment at various locales within the construct.

Neuroendocrine and stress hormone changes during mirthful laughter.

Positive emotional activities have been suggested as modifiers of neuroendocrine hormones involved in the classical stress response. To detect changes in these components during a mirthful laughter experience, the authors studied 10 healthy male subjects. Five experimental subjects viewed a 60 minute humor video and five control subjects did not. Serial blood samples were measured for corticotropin (ACTH), cortisol, beta-endorphin, 3,4-dihydrophenylacetic acid (dopac)--the major serum neuronal catabolite of dopamine, epinephrine, norepinephrine, growth hormone, and prolactin. Repeated measures analysis of variance showed that cortisol and dopac in the experimental group decreased more rapidly from baseline than the control group (p = 0.011, p = 0.025, respectively). Epinephrine levels in the experimental group were significantly lower than the control at all time points (p = 0.017). Growth hormone levels in the experimental group significantly increased during baseline (p = 0.027) and then decreased with laughter intervention (p less than 0.0005), whereas, the controls did not change over time (p = 0.787). ACTH, beta-endorphin, prolactin, and norepinephrine levels did not significantly increase. The mirthful laughter experience appears to reduce serum levels of cortisol, dopac, epinephrine, and growth hormone. These biochemical changes have implications for the reversal of the neuroendocrine and classical stress hormone response.

Modulation of neuroimmune parameters during the eustress of humor-associated mirthful laughter.

CONTEXT: Humor therapy and the related mirthful laughter are suggested to have preventive and healing effects. Although these effects may be mediated by neuroendocrine/neuroimmune modulation, specific neuroimmune parameters have not been fully investigated. OBJECTIVE: To determine the efficacy of mirthful laughter to modulate neuroimmune parameters in normal subjects. DESIGN: A series of 5 separate studies based on a multivariate repeated measures design, with post hoc simple contrast analysis. SETTING: The schools of medicine and public health at Loma Linda University, Loma Linda, Calif. SUBJECTS: 52 healthy men. INTERVENTION: Viewing of a humor video for 1 hour. Blood samples were taken 10 minutes before, 30 minutes into, and 30 minutes and 12 hours after the intervention. MAIN OUTCOME MEASURES: Natural killer cell activity; plasma immunoglobulins; functional phenotypic markers for leukocytes including activated T cells, nonactivated T cells, B cells, natural killer cells, T cells with helper and suppressor markers, and assessment of plasma volume and compartmental shifts; plasma cytokine--interferon-gamma; and total leukocytes with subpopulations of lymphocytes, granulocytes, and monocytes. RESULTS: Increases were found in natural killer cell activity (P < .01); immunoglobulins G (P < .02), A (P < .01), and M (P < .09), with several immunoglobulin effects lasting 12 hours into recovery from initiation of the humor intervention; functional phenotypic markers for leukocyte subsets such as activated T cells (P < .01), active cytotoxic T cells (P < .01), natural killer cells (P = .09), B cells (P < .01), helper T cells (P < .02), uncommitted T cells with helper and suppressor markers (P < .02), helper/suppressor ratio (P = .10) with several leukocyte subset increase effects lasting 12 hours after the humor experience; the cytokine interferon-gamma (P = .02), with increases lasting 12 hours; total leukocytes (P < .05), with specific subpopulation lymphocytes during the intervention (P < .01) and 90 minutes into recovery (P < .05); and granulocytes during the intervention (P < .05) and 90 minutes following the intervention (P < .01). CONCLUSION: Modulation of neuroimmune parameters during and following the humor-associated eustress of laughter may provide beneficial health effects for wellness and a complementary adjunct to whole-person integrative medicine therapies.

Placental chorioangioma: a case report and review.

Placental Chorioangioma occurs in 1% of pregnancies and are generally asymptomatic. However, they are known to cause a number of complications which are detrimental to the mother, fetus or the neonate. A typical case where acute polyhydramnios precipitated premature labour and delivery of the fetus as a result of a large placental chorioangioma is presented. A brief review of the other possible complications are also included.

Birth defects--the state of awareness amongst mothers.

Birth defects have in recent years become the major cause of perinatal morbidity and mortality. The incidence of birth defects is between 2% to 6% of all live births. However, from a survey carried out in Singapore General Hospital (SGH), the majority of mothers enter pregnancy without realising the risks that they or their offspring may face. Only 20% to 25% of mothers were aware of the actual incidence of birth defects, and only 2.5%-10% knew that the risk of Down Syndrome (DS) increases with maternal age. However, the use of ultrasound is now well accepted and is considered a necessary investigation by 90% of the mothers surveyed.

Birth defects--the state of awareness.

As doctors and nurses are the primary sources of medical information, a simple survey was conducted in 1992 among doctors and nurses to determine their level of awareness of the incidence of and risk of having an abnormal as well as their knowledge of the use of ultrasound in pregnancy. Only 10% of doctors and 23% of nurses were aware that the general risk of having an abnormal baby is greater than 1%. Only 37% and 17% of doctors and nurses respectively were aware that the best time to screen for structural abnormalities was indeed at approximately 20 weeks gestation.

Synthesis of nano-sized hydroxyapatite.

In this work, nanometer HA crystals have been synthesized via wet chemical precipitation and characterized. This research studies how key synthesis parameters affect the size and phase purity of the produced HA. Characterization work was carried out using X-ray powder diffraction method and scanning electron microscopy for phase identification and particle sizing, respectively.

Hyperiodotyrosinemia-induced hyperprolactinemia and hyperaldosteronism.

A 21-year-old goitrous hypothyroid Chinese woman had elevated serum iodotyrosines with a monoiodotyrosine level of 85.9 nmol/l (normal 0.49-0.89 nmol/l) and a diiodotyrosine level of 25.3 nmol/l (normal 0.023-0.53 nmol/l). She was amenorrheic with low luteinizing hormone and follicle-stimulating hormone levels at 5.8 and 2.8 U/l, respectively. The hypogonadotropic hypogonadism was due to an elevated prolactin level of 8.8 nmol/l. She also had a low potassium level of 3.2 mmol/l, and a high urinary aldosterone level of 158 nmol/day. The hyperprolactinemia, hypogonadotropic hypogonadism, hyperaldosteronism and hypokalemia subsided with the administration of bromocriptine 5 mg/day. However, bromocriptine accentuated the hyperiodotyrosinemia, and the patient remained hypothyroid. Levothyroxine therapy lowered the monoiodotyrosine and diiodotyrosine levels, ameliorated all her endocrinopathies, started her periods, and shrank the goiter. She probably had a deiodinase defect which permitted the discharge of accumulated iodotyrosines from the thyroid gland. Since iodotyrosines are tyrosine hydroxylase inhibitors, the hyperiodotyrosinemia causes dopamine synthesis inhibition, and induces the hyperprolactinemia and hyperaldosteronism.

Distribution of ciliatine (2-aminoethylphosphonic acid) and phosphonoalanine (2-amino-3-phosphonopropionic acid) in human tissues.

Ciliatine (2-aminoethylphosphonic acid) was detected in the human brain, heart, kidney, liver, intestine, spleen, adrenal glands, and aorta. Phosphonoalanine (2-amino-3-phosphonopropionic acid) was found in the human liver, intestine and spleen. Tissue homogenates were extracted with trichloroacetic acid and a chloroform-methanol mixture. After hydrolysis, each fraction was subfractionated by ion-exchange chromatography and examined by paper chromatography and electrophoresis using a specific ninhydrin-molybdate staining procedure to detect the phosphonic acids. The acids were found bound either to lipid or to protein; no free phosphonic acid was detected.

The problems of delivering neuroactive molecules to the CNS.

At present, the aetiologies of many neurological and neurodegenerative diseases are unknown. However, emergence of a better understanding of these diseases, at both cellular and molecular levels, opens up the possibility of replacement therapies. The presence of the blood-brain barrier complicates the delivery of molecules to the central nervous system. Numerous attempts have been made to bypass this barrier either by delivering the drugs directly into the brain or by transplanting cells to produce the missing molecules in situ. This review explores several methods for delivering bioactive molecules into the CNS, including the use of permeabilizers, osmotic pumps, slow polymer release systems and transplantation of cells with or without the use of the encapsulation technology.