Phenotypic and Genotypic Correction of WASP Gene Mutation in Wiskott-Aldrich Syndrome by Unrelated Cord Blood Stem Cell Transplantation

We present two cases of Wiskott-Aldrich syndrome (WAS), in which nonsense mutations in the WASP gene were corrected phenotypically as well as genotypically by unrelated cord blood stem cell transplantation (CBSCT). Two male patients were diagnosed with WAS at the age of 5-month and 3-month and each received unrelated CBSCT at 16-month and 20-month of age, respectively. The infused cord blood (CB) units had 4/6 and 5/6 HLA matches and the infusion doses of total nucleated cells (TNC) and CD34+ cells were 6.24×10⁷/kg and 5.08×10⁷/kg for TNC and 1.33×10⁵/kg and 4.8×10⁵/kg for CD34+ cells, for UPN1 and UPN2, respectively. Complete donor cell chimerism was documented by variable number tandem repeat (VNTR) with neutrophil engraftment on days 31 and 13 and platelets on days 58 and 50, respectively. Immunologic reconstitution demonstrated that CBSCT resulted in consistent and stable T-, B-, and NK-cell development. Flow cytometric analysis for immunologic markers and sequence analysis of the WASP gene mutation revealed a normal pattern after CBSCT. These cases demonstrate that CBs can be an important source of stem cells for the phenotypical and genotypical correction of genetic diseases such as WAS.     

Hematologic Recovery after Tandem High-Dose Chemotherapy and Autologous Stem Cell Transplantation in Children with High-Risk Solid Tumors

Although the number of studies using tandem high-dose chemotherapy and autologous stem cell transplantation (HDCT/autoSCT) for the treatment of high-risk pediatric solid tumors has been increasing, documentation of hematologic recovery after tandem HDCT/autoSCT is very limited. For this reason, we retrospectively analyzed the hematologic recovery of 236 children with high-risk solid tumors who underwent tandem HDCT/autoSCT. The median numbers of CD34⁺ cells transplanted during the first and second HDCT/autoSCT were 4.3 × 10⁶/kg (range 0.6-220.2) and 4.1 × 10⁶/kg (range 0.9-157.6), respectively (P = 0.664). While there was no difference in neutrophil recovery between the first and second HDCT/autoSCT, platelet and RBC recoveries were significantly delayed in the second HDCT/autoSCT (P < 0.001 and P < 0.001, respectively). Delayed recovery in the second HDCT/autoSCT was more prominent when the number of transplanted CD34⁺ cells was lower, especially if it was < 2 × 10⁶/kg. A lower CD34⁺ cell count was also associated with increased RBC transfusion requirements and a higher serum ferritin level after tandem HDCT/autoSCT. More CD34⁺ cells need to be transplanted during the second HDCT/autoSCT in order to achieve the same hematologic recovery as the first HDCT/autoSCT.     

Gynura procumbens Merr. decreases blood pressure in rats by vasodilatation via inhibition of calcium channels

INTRODUCTION:

Gynura procumbens has been shown to decrease blood pressure via inhibition of the angiotensin‐converting enzyme. However, other mechanisms that may contribute to the hypotensive effect have not been studied.

OBJECTIVES:

To investigate the cardiovascular effects of a butanolic fraction of Gynura procumbens in rats.

METHODS:

Anaesthetized rats were given intravenous bolus injections of butanolic fraction at doses of 2.5–20 mg/kg in vivo. The effect of butanolic fraction on vascular reactivity was recorded in isolated rat aortic rings in vitro.

RESULTS:

Intravenous administrations of butanolic fraction elicited significant (p<0.001) and dose‐dependent decreases in the mean arterial pressure. However, a significant (p<0.05) decrease in the heart rate was observed only at the higher doses (10 and 20 mg/kg). In isolated preparations of rat aortic rings, phenylephrine (1×10^‐6 M)‐ or potassium chloride (8×10^‐2 M)‐precontracted endothelium‐intact and ‐denuded tissue; butanolic fraction (1×10^‐6–1×10^‐1 g/ml) induced similar concentration‐dependent relaxation of the vessels. In the presence of 2.5×10^‐3 and 5.0×10^‐3 g/ml butanolic fraction, the contractions induced by phenylephrine (1×10^‐9–3×10^‐5 M) and potassium chloride (1×10^‐2–8×10^‐2 M) were significantly antagonized. The calcium‐induced vasocontractions (1×10^‐4–1×10^‐2 M) were antagonized by butanolic fraction concentration‐dependently in calcium‐free and high potassium (6×10^‐2 M) medium, as well as in calcium‐ and potassium‐free medium containing 1×10^‐6 M phenylephrine. However, the contractions induced by noradrenaline (1×10^‐6 M) and caffeine (4.5×10^‐2 M) were not affected by butanolic fraction.

CONCLUSION:

Butanolic fraction contains putative hypotensive compounds that appear to inhibit calcium influx via receptor‐operated and/or voltage‐dependent calcium channels to cause vasodilation and a consequent fall in blood pressure.     

Sweat Rates, Sweat Sodium Concentrations, and Sodium Losses in 3 Groups of Professional Football Players

Context:

Sweat sodium losses have never been reported in a large cohort of American football players.

Objective:

To compare sweat rates (SwtRs), sweat sodium concentrations (SwtNa⁺), and sodium losses in 3 groups of players (backs and receivers [BK], linebackers and quarterbacks [LB/QB], and linemen [LM]) to determine if positional differences and, therefore, size differences exist.

Design:

Observational study.

Setting:

Data were collected during practices in the second week of 2 consecutive training camps. The wet bulb globe temperature was 78.5°F ± 3.5°F (25.9°C ± 1.9°C).

Patients or Other Participants:

Eighteen BK, 12 LB/QB, and 14 LM volunteered.

Intervention(s):

Sterile sweat patches were applied to the right forearm after the skin was appropriately cleaned. The patches were removed during practice, placed in sterile tubes, centrifuged, frozen, and later analyzed by flame photometry.

Main Outcome Measure(s):

Sweat rate, SwtNa⁺, and sodium loss. We calculated SwtR by change in mass adjusted for urine produced and fluids consumed divided by practice time in hours.

Results:

Other than age, physical characteristics were different among groups (P < .001). The SwtR was different among groups (F_2,41  =  7.3, P  =  .002). It was lower in BK (1.42 ± 0.45 L/h) than in LB/QB (1.98 ± 0.49 L/h) (P < .05) and LM (2.16 ± 0.75 L/h) (P < .01), but we found no differences between SwtRs for LB/QB and LM. The SwtNa⁺ was not different among groups (BK  =  50 ± 16 mEq/L, LB/QB  =  48.2 ± 23 mEq/L, and LM  =  52.8 ± 25 mEq/L) and ranged from 15 to 99 mEq/L. Sweat sodium losses ranged from 642 mg/h to 6.7 g/h, and findings for group comparisons approached significance (P  =  .06). On days when players practiced 4.5 hours, calculated sodium losses ranged from 2.3 to 30 g/d.

Conclusions:

The BK sweated at lower rates than did the midsized LB/QB and large LM, but LB/QB sweated similarly to LM. Sweat sodium concentration and daily sodium losses ranged considerably. Heavy, salty sweaters require increased dietary consumption of sodium during preseason.     

Bacterial indicators of pollution of the Douala lagoon, Cameroon: Public health implications

Background

Indiscriminate disposal of untreated wastes which are often heavily laden with sewage microorganisms some of which are pathogenic to humans into aquatic environments near cities could serve as potential dangers to human health.

Objective

A prospective study was undertaken to investigate the scope of potential bacterial pathogens and to assess the extent of pollution of the Douala lagoon.

Methods

A total of eighty water samples were collected fortnightly from the lagoon at five stations from March to October 2005 and analysed for heterotrophic bacterial densities, coliform counts, faecal coliform and faecal streptococcal counts. Bacteria were isolated and identified using standard microbiology and biochemical techniques.

Results

High heterotrophic bacterial counts (33 × 10⁵ − 161 × 10⁵ CFU/ mL), total coliform counts (1.8 × 10² − 2.4 × 10² CFU/100 mL), faecal coliform counts (2.2 × 10² − 2.4 × 10² CFU/ 100 mL) and faecal streptococcal counts (2.1 × 102 − 2.3 x 10² CFU/100mL were observed in all sampling stations. Eleven species of bacteria: Bacteroides fragilis, Proteus vulgaris, Klebsiella pneumoniae, E. coli, Enterococcus faecalis, Enterobacter aerogenes, Citrobacter freundii, Aeromonas hydrophila, Pseudomonas aeruginosa, Bacillus mycoides and Serratia marcesens, were frequently isolated.

Conclusion

The presence of potential bacterial agents such as Bacteroides fragilis, Pseudomonas aeruginosa, Aeromonas hydrophila, Klebsiella pneumoniae and E. coli in the lagoon may pose a serious threat to the health and well being of users of the Lagoon and calls for urgent intervention.     

Pre-Clinical Efficacy and Safety Evaluation of Human Amniotic Fluid-Derived Stem Cell Injection in a Mouse Model of Urinary Incontinence

Purpose

Stem cell-based therapies represent new promises for the treatment of urinary incontinence. This study was performed to assess optimized cell passage number, cell dose, therapeutic efficacy, feasibility, toxicity, and cell trafficking for the first step of the pre-clinical evaluation of human amniotic fluid stem cell (hAFSC) therapy in a urinary incontinence animal model.

Materials and Methods

The proper cell passage number was analyzed with hAFSCs at passages 4, 6, and 8 at week 2. The cell dose optimization included 1×10⁴, 1×10⁵, and 1×10⁶ cells at week 2. The in vivo cell toxicity was performed with 0.25×10⁶, 0.5×10⁶, and 1×10⁶ cells at weeks 2 and 4. Cell tracking was performed with 1×10⁶ cells at weeks 2 and 4.

Results

The selected optimal cell passage number was smaller than 6, and the optimal cell dose was 1×10⁶ for the mouse model. In our pre-clinical study, hAFSC-injected animals showed normal values for several parameters. Moreover, the injected cells were found to be non-toxic and non-tumorigenic. Furthermore, the injected hAFSCs were rarely identified by in vivo cell trafficking in the target organs at week 2.

Conclusion

This study demonstrates for the first time the pre-clinical efficacy and safety of hAFSC injection in the urinary incontinence animal model and provides a basis for future clinical applications.     

Multi‐parametric flow cytometric and genetic investigation of the peripheral B cell compartment in human type 1 diabetes

The appearance of circulating islet-specific autoantibodies before disease diagnosis is a hallmark of human type 1 diabetes (T1D), and suggests a role for B cells in the pathogenesis of the disease. Alterations in the peripheral B cell compartment have been reported in T1D patients; however, to date, such studies have produced conflicting results and have been limited by sample size. In this study, we have performed a detailed characterization of the B cell compartment in T1D patients (n = 45) and healthy controls (n = 46), and assessed the secretion of the anti-inflammatory cytokine interleukin (IL)-10 in purified B cells from the same donors. Overall, we found no evidence for a profound alteration of the B cell compartment or in the production of IL-10 in peripheral blood of T1D patients. We also investigated age-related changes in peripheral B cell subsets and confirmed the sharp decrease with age of transitional CD19⁺CD27⁻CD24^hiCD38^hi B cells, a subset that has recently been ascribed a putative regulatory function. Genetic analysis of the B cell compartment revealed evidence for association of the IL2–IL21 T1D locus with IL-10 production by both memory B cells (P = 6·4 × 10⁻⁴) and islet-specific CD4⁺ T cells (P = 2·9 × 10⁻³). In contrast to previous reports, we found no evidence for an alteration of the B cell compartment in healthy individuals homozygous for the non-synonymous PTPN22 Trp⁶²⁰ T1D risk allele (rs2476601; Arg⁶²⁰Trp). The IL2–IL21 association we have identified, if confirmed, suggests a novel role for B cells in T1D pathogenesis through the production of IL-10, and reinforces the importance of IL-10 production by autoreactive CD4⁺ T cells.     

Inhibitory Effect of Fentanyl on Phenylephrine-Induced Contraction of the Rat Aorta

Purpose

Fentanyl was reported to inhibit the α₁-adrenoceptor agonist-induced contraction. The goal of this in vitro study was to identify the α₁-adrenoceptor subtype primarily involved in the fentanyl-induced attenuation of phenylephrine-induced contraction in isolated endothelium-denuded rat aorta.

Materials and Methods

Aortic rings were suspended in order to record isometric tension. Concentration-response curves for phenylephrine (10^-9 to 10^-5 M) were generated in the presence or absence of one of the following drugs: fentanyl (3×10^-7, 10^-6, 3×10^-6 M), 5-methylurapidil (3×10^-8, 10^-7, 3×10^-7 M), chloroethylclonidine (10^-5 M) and BMY 7378 (3×10^-9, 10^-8, 3×10^-8 M). Phenylephrine concentration-response curves were generated in the presence or absence of fentanyl in rings pretreated with either 3×10^-9 M prazosin, 10^-9 M 5-methylurapidil or 3×10^-9 M BMY 7378.

Results

Fentanyl (10^-6, 3×10^-6 M) attenuated phenylephrine-induced contraction in the rat aorta. 5-Methylurapidil and BMY 7378 produced a parallel rightward shift in the phenylephrine concentration-response curve. The pA₂ values for 5-methylurapidil and BMY 7378 were estimated to be 7.71 ± 0.15 and 8.99 ± 0.24, respectively. Fentanyl (10^-6 M) attenuated phenylephrine-induced contraction in rings pretreated with 10^-9 M 5-methylurapidil, but did not alter the rings when pretreated with 3×10^-9 M BMY 7378. Pretreatment of the rings with chloroethylclonidine showed a 72.9 ± 2.3% reduction in phenylephrine-induced maximal contraction.

Conclusion

The results suggest that fentanyl attenuates phenylephrine-induced contraction by inhibiting the pathway involved in the α_1D-adrenoceptor-mediated contraction of the rat aorta.     

Cultivation of Virulent Treponema pallidum in Tissue Culture

In a series of seven experiments, the virulent Nichols strain of Treponema pallidum was shown to attach and replicate on the surface of tissue culture cells of cottontail rabbit epithelium (Sf1Ep) growing in conventional monolayer cultures under an atmosphere of 1.5% oxygen. Five days after inoculation of 10⁶T. pallidum, the number of treponemes had increased to between 8 × 10⁶ and 2.59 × 10⁷. The viability of harvested organisms ranged from 86 to 97%. The number of T. pallidum continued to increase, generally reaching a plateau between days 9 and 12 of incubation, with increases ranging up to 100-fold and averaging 49-fold. There appeared to be a ceiling of multiplication of about 2 × 10⁸ irrespective of the inoculum, which ranged from 10⁶ to 10⁸T. pallidum. Concurrent deoxyribonucleic acid assays were performed on the cultures containing T. pallidum to obtain further evidence of replication. Significant increases in treponemal deoxyribonucleic acid were observed when the inocula ranged from 10⁶ to 10⁷, with the greatest increases, as might be expected, being in the former group. There was also excellent correlation in the amount of deoxyribonucleic acid per treponeme; the averages for the 10⁶, 2.5 × 10⁶, and 10⁷ groups were 3.46 × 10⁻¹⁴, 3.28 × 10⁻¹⁴, and 2.79 × 10⁻¹⁴ g per treponeme, respectively (3.14 ± 0.72 × 10⁻¹⁴ g per treponeme). In each experiment, organisms were harvested from the group inoculated with 10⁶T. pallidum after 7 days of incubation to test for virulence. In all instances, the organisms were virulent; erythematous, indurated, treponeme-containing lesions were produced from an average of six to seven organisms. Scanning electron microscopy revealed that during the course of replication many microcolonies of treponemes formed on the surface of the cells.     

An in vivo rat model for early development of colorectal cancer metastasis to liver

At diagnosis of colorectal cancer, approximately 25% of the patients have established colorectal liver metastasis. Optimal management of disseminated disease requires therapies targeting multiple stages in hepatic colorectal cancer metastasis development. To facilitate this, biologically accurate in vivo models are required. Early colonic cancer liver metastases development was studied using BDIX and Sprague–Dawley rat strains with human HT29 and rat DHDK12 colonic cancer cell lines. Different cancer cell–host combinations were used. Rat DHDK12 was previously chemically induced in the BDIX rat. Real‐time intra‐vital microscopy was employed to analyse the early development of liver metastases in four groups (n = 6 per group) (HT29–BDIX, DHDK12–BDIX, HT29–SD and DHDK12–SD). Data were compared using one‐way anova with Bonferroni’s multiple comparison test. The total number of tumour cells visualized, adherent cells within the hepatic sinusoids, extravasated tumour cells and migration rates were significantly higher in the DHDK12–BDIX combination. Maximum number of visualized cells and maximum migration rate were also significantly higher in this group. No significant differences were observed in these experimental parameters among the other three groups or in the haemodynamic parameters among all groups. In conclusion, cancer cell line–host selection has a significant effect on early colonic cancer liver metastasis development.     

Cytotoxicity in patients with different clinical forms of Chagas' disease

There are few studies on cell-mediated cytotoxicity in human Chagas' disease. In the present study, we evaluated peripheral blood mononuclear cell (PBMC) cytotoxicity activity from chagasic patients with different clinical forms of disease. To verify the cytotoxic response, we performed cell lysis assays using ⁵¹Cr-labelled K562 cells as targets. Results are reported as lytic units (LU=number of cells required for 30% lysis) per million PBMC. Exposure of patients’ PBMC to Trypanosoma cruzi antigen led to an increase in cytotoxic activity compared with unstimulated patient cells against K562. Asymptomatic cardiomyopathy patients had higher responses (37.8±5.0 LU/10⁶ PBMC; mean±s.d.) than indeterminate (11.5±3.6 LU/10⁶) and symptomatic cardiomyopathy (7.8±2.5 LU/10⁶). Normal control PBMC stimulated with T. cruzi antigen had 4.36±1.31 LU/10⁶ PBMC against K562. Addition of recombinant interferon-gamma (IFN-γ) did not lead to significant increase in cytotoxicity in any group of patients. On the other hand, recombinant human IL-12 significantly increased cytotoxic responses from symptomatic cardiomyopathy patients and normal controls who presented low levels of cytotoxicity induced by T. cruzi antigen. The combined use of IL-12 and a neutralizing anti-IFN-γ antibody did not change IL-12-induced cytotoxic responses, showing the direct role of this cytokine on natural killer (NK) cells. NK cells were the main cells responsible for the lysis of K562 target cells as evidenced by testing cell subsets following magnetic cell sorting. These data demonstrate that chagasic patients with different clinical forms of disease have PBMC which respond to T. cruzi antigen with a cytotoxic response, and this response is up-regulated by IL-12.     

Genome-wide association study of coronary artery disease in the Japanese

A new understanding of the genetic basis of coronary artery disease (CAD) has recently emerged from genome-wide association (GWA) studies of common single-nucleotide polymorphisms (SNPs), thus far performed mostly in European-descent populations. To identify novel susceptibility gene variants for CAD and confirm those previously identified mostly in populations of European descent, a multistage GWA study was performed in the Japanese. In the discovery phase, we first genotyped 806 cases and 1337 controls with 451 382 SNP markers and subsequently assessed 34 selected SNPs with direct genotyping (541 additional cases) and in silico comparison (964 healthy controls). In the replication phase, involving 3052 cases and 6335 controls, 12 SNPs were tested; CAD association was replicated and/or verified for 4 (of 12) SNPs from 3 loci: near BRAP and ALDH2 on 12q24 (P=1.6 × 10⁻³⁴), HLA-DQB1 on 6p21 (P=4.7 × 10⁻⁷), and CDKN2A/B on 9p21 (P=6.1 × 10⁻¹⁶). On 12q24, we identified the strongest association signal with the strength of association substantially pronounced for a subgroup of myocardial infarction cases (P=1.4 × 10⁻⁴⁰). On 6p21, an HLA allele, DQB1^*0604, could show one of the most prominent association signals in an ∼8-Mb interval that encompasses the LTA gene, where an association with myocardial infarction had been reported in another Japanese study. CAD association was also identified at CDKN2A/B, as previously reported in different populations of European descent and Asians. Thus, three loci confirmed in the Japanese GWA study highlight the likely presence of risk alleles with two types of genetic effects – population specific and common – on susceptibility to CAD.     

The postnatal development of cerebellar Purkinje cells in the Göttingen minipig estimated with a new stereological sampling technique--the vertical bar fractionator

The postnatal development of total number and perikaryon volume of cerebellar Purkinje cells was estimated in the Göttingen minipig cerebellar cortex using a new stereological approach, the vertical bar fractionator. Data were obtained from the brains of five neonate and five adult female Göttingen minipigs. The total number of Purkinje cells ranged from 1.83 × 10⁶ in the neonate to 2.82 × 10⁶ in the adult Göttingen minipig. The number-weighted mean perikaryon volume of Purkinje cells increased concurrently from around 6800 µm³ in the neonate to 17 600 µm³ in the adult. The study demonstrates that a pronounced postnatal neurogenesis in Purkinje cell number and perikaryon volume is part of the growth and development of the cerebellum in the Göttingen minipig. The Purkinje cells of the Göttingen minipig were found to be substantially large compared with human and represents the largest cells described hitherto from mammalian cerebella. The vertical fractionator is a new sampling technique, which allows the combination of a fractionator design on vertical bar sections excluding exhaustive sampling and bias from artificial edges. By design, the sections are perfect stereological vertical sections and provide the basis for unbiased estimates of total number of structural entities in the brain, including surface area, fibre length and particle volume.     

The effects of Cu-chlorophyllin on the active site formation of each component of guinea-pig complement

The effects of Cu-chlorophyllin on the formation of intermediate complexes (site formation) by each component of guinea-pig complement on sensitized sheep erythrocytes were examined. When the Z value of immune haemolysis varied between 0·30 and 2·66, the 50 per cent inhibition doses of Cu-chlorophyllin on C′1, C′2, C′5, C′6 and C′7 were 2·3–3·2×10^-5 M, 0·31–0·37×10^-5 M, 1·4–1·6×10^-5 M, 1·0–1·2×10^-5 M and 2·4–2·8×10^-5 M, respectively. Site generation of C′3 and C′9 was enhanced whereas no effect was observed on C′4 and C′8 site formation.     

Effects of catecholamines on the neuromuscular junction in the rat diaphragm

1. The effects of noradrenaline, adrenaline and isoprenaline on neuromuscular transmission in the rat diaphragm and the influence of adrenergic blocking agents on these actions were investigated.

2. The resting membrane potential of the muscle fibre was increased by adrenaline (5 × 10^-6-10^-5 g/ml.) and isoprenaline (5 × 10^-6 g/ml.) up to 3-4 mV, but noradrenaline (5 × 10^-6-10^-5 g/ml.) had little effect.

3. The amplitude and the half-decay time of the end-plate potential (e.p.p.) were increased by noradrenaline (1 × 10^-6 g/ml.), adrenaline (1 × 10^-7-10^-5 g/ml.) and isoprenaline (1-5 × 10^-6 g/ml.). The potentiation of the amplitude of the e.p.p. was greater with noradrenaline than with adrenaline and isoprenaline.

4. Noradrenaline (5 × 10^-6 g/ml.) increased the frequency of miniature end-plate potentials (m.e.p.p.), but not their amplitude. However, isoprenaline (5 × 10^-6 g/ml.) increased the amplitude of m.e.p.p.s without change in frequency. Adrenaline (5 × 10^-6 g/ml.) increased both frequency and amplitude of m.e.p.p.s.

5. Adrenaline (5 × 10^-6 g/ml.) and isoprenaline (5 × 10^-6 g/ml.) increased the input resistance of the muscle membrane. The effect was blocked by the β-blocker, pronethalol (2 × 10^-6 g/ml.), but not by the α-blocker, phentolamine (2 × 10^-6 g/ml.). Noradrenaline did not change the input resistance of the muscle fibre.

6. Noradrenaline (5 × 10^-6 g/ml.) and adrenaline (5 × 10^-6 g/ml.) augmented the extracellularly recorded end-plate current (e.p.c.), but they had no effect on the half duration, nor on the action current (a.c.) of the nerve terminal, nor on the synaptic delay. Isoprenaline (5 × 10^-6 g/ml.) had no effect on any of these parameters. The actions of noradrenaline and adrenaline on e.p.c. were abolished by phentolamine (2 × 10^-6 g/ml.), but not by pronethalol (2 × 10^-6 g/ml.).

7. Adrenaline (5 × 10^-6 g/ml.) and isoprenaline (5 × 10^-6 g/ml.) enhanced the amplitude of the acetylcholine potential elicited by iontophoretic application of acetylcholine. No such effect was produced by noradrenaline (5 × 10^-6 g/ml.).

8. It was concluded that noradrenaline acts on the nerve ending increasing the release of transmitter, and that isoprenaline acts on the post-synaptic membrane enhancing the input resistance, while adrenaline has both presynaptic and post-synaptic actions. The effect on the nerve ending is concerned with the α-action, whereas that on post-synaptic membrane with β-action of the catecholamines.

     

Treatment with 8‐OH‐modified adenine (TLR7 ligand)‐allergen conjugates decreases T helper type 2‐oriented murine airway inflammation

A strategy to improve allergen-specific immunotherapy is to employ new adjuvants stably linked to allergens. The study is addressed to evaluate the in vivo and in vitro effects of allergens [natural Dermatophagoides pteronyssinus 2 (nDer p 2) and ovalbumin (OVA)] chemically bound to an 8-OH-modified adenine. Humoral and cellular responses were analysed in allergen-sensitized and challenged mice by using conjugates (Conj) in a therapeutic setting. The in vitro activity of the conjugates on cytokine production induced by bone marrow dendritic cells and the co-culture system was also investigated. The nDer p 2-Conj treatment in nDer p 2-primed and challenged BALB/c mice reduced the numbers of eosinophils in bronchoalveolar lavage fluid and lung, airway allergen-driven interleukin-13 (IL-13) production in lung mononuclear cells and IgE, in comparison with nDer p 2-treated mice. The increase of IgG2a paralleled that of interferon-γ (IFN-γ) and IL-10 in allergen-stimulated spleen cells. Similar effects were elicited by treatment with OVA-Conj in an OVA-driven BALB/c model. The nDer p 2-Conj or OVA-Conj redirected memory T helper type 2 cells towards the production of IL-10 and IFN-γ also in C57BL/6 mice and when subcutaneously administered. Interleukin-10, IL-12 and IL-27 were produced in vitro by Conj-stimulated bone marrow dendritic cells, whereas IL-10 and IFN-γ were up-regulated in co-cultures of CD11c⁺ and CD4⁺ T cells from Conj-treated mice stimulated with allergen. Cytofluorometric analysis indicated that the Conj expanded IFN-γ- and IL-10- producing memory T cells. The Conj effects on IL-10^−/− and IL-12^−/− mice confirmed the role of IL-10 and IFN-γ in inducing a protective and balanced redirection the T helper type 2-mediated airway inflammation.     

Phagocytic responses of peritoneal macrophages and neutrophils are different in rats following prolonged exercise

OBJECTIVE:

To analyze the effects of exhausting long‐duration physical exercise (swimming) sessions of different durations and intensities on the number and phagocytic capacity of macrophages and neutrophils in sedentary rats.

INTRODUCTION:

Exercise intensity, duration and frequency are important factors in determining immune response to physical effort. Thus, the effects of exhausting long‐duration exercise are unclear.

METHODS:

Wistar rats were divided into two groups: an untreated group (macrophage study) and oyster glycogen‐treated rats (neutrophil study). In each group, the animals were subdivided into five groups (10 rats per group): unexercised controls, an unadapted low‐intensity exercise group, an unadapted moderate‐intensity exercise group, a preadapted low‐intensity exercise group and a preadapted moderate‐intensity exercise group. All exercises were performed to exhaustion, and preadaptation consisted of 5, 15, 30 and 45 min sessions.

RESULTS:

Macrophage study: the number of peritoneal macrophages significantly decreased (9.22 ± 1.78 × 10⁶) after unadapted exercise but increased (21.50 ± 0.63 × 10⁶) after preadapted low‐intensity exercise, with no changes in the moderate‐intensity exercise group. Phagocytic capacity, however, increased by more than 80% in all exercise groups (low/moderate, unadapted/preadapted). Neutrophil study: the number of peritoneal neutrophils significantly decreased after unadapted (29.20 ± 3.34 × 10⁶) and preadapted (50.00 ± 3.53 × 10⁶) low‐intensity exercise but increased after unadapted (127.60 ± 5.14 × 10⁶) and preadapted (221.80 ± 14.85 × 10⁶) moderate exercise. Neutrophil phagocytic capacity decreased by 63% after unadapted moderate exercise but increased by 90% after corresponding preadapted sessions, with no changes in the low‐intensity exercise groups.

CONCLUSION:

Neutrophils and macrophages of sedentary rats respond differently to exercise‐induced stress. Adaptation sessions reduce exercise‐induced stress on the immune system.     

Inhibition of in vitro lymphocyte blastogenesis by inhibitor produced by cultured human lymphoblasts

Normal human lymphoblastoid cell lines, growing in continuous suspension culture, produce inhibitors of in vitro lymphocyte blastogenesis. The inhibitor reduces human lymphocyte blastogenic responses to phytohaemagglutinin, streptolysin-O and the mixed lymphocyte culture 90–99%, is non-cytotoxic and can inhibit both newly initiated and on-going responses. The inhibitor is heat-stable at 80°C but labile at 100°C, non-dialysable and degraded by pronase but not DNase or RNase. It is species- and tissue-specific and does not inhibit the proliferation of mouse lymphocytes, human melanoma cells or human bone marrow in vitro colony-forming cells. Inhibitor was produced only under very specific conditions of crowding. Thus, maximal inhibitor production occurred at 5 × 10⁶ lymphocytes per cm² culture surface area while only 0–5% of the maximal amount was produced at 10⁶ or 5 × 10⁷ lymphoblasts per cm². This data is relevant to the nature of feedback control of immunological reactions and may guide the development of new classes of immunosuppressants.