Haemostasis and analgesia with autologous platelet-rich plasma in tonsillectomy.

OBJECTIVE: A single-centre, single-blinded prospective experimental study was conducted to determine the effectiveness of autologous platelet-rich plasma applied to the tonsillar bed post-operatively in reducing post-operative pain and haemorrhage. METHODS: Platelet-rich plasma, prepared prior to surgery, was applied with calcium gluconate to one randomly chosen tonsillar fossa. Pain and haemorrhage were analysed, using a visual analogue scale and a pre-defined grading scale respectively, four times on the day of surgery at 2-hourly intervals, and thrice on the following day. RESULTS: The pain score and haemorrhage grade on the test side were lower than on the control side. These findings were statistically significant. CONCLUSION: This pilot study, conducted in India, revealed valid positive results for a promising new technology. The manual preparation of platelet-rich plasma could be automated in the future to allow a larger sample size.

IgE-associated food allergy alters the presentation of paediatric eosinophilic esophagitis.

BACKGROUND: Links between food allergens and eosinophilic esophagitis (EoE) have been established, but the interplay between EoE- and IgE-associated immediate hypersensitivity to foods remains unclear. OBJECTIVE: We sought to determine the prevalence of IgE-associated food allergy at the time of diagnosis of EoE in children and to determine whether differences existed in presentation and disease compared to subjects with EoE alone. METHODS: Eosinophilic esophagitis patients were stratified based on the diagnosis of IgE-associated immediate hypersensitivity (EoE + IH vs. EoE-IH). Clinical, histologic, pathologic, and endoscopic differences were investigated using a retrospective database. RESULTS: We found that 29% of the 198 EoE patients in our cohort had EoE + IH. These subjects presented at a younger age than those without IH (6.05 vs. 8.09 years, P = 0.013) and were more likely to have comorbid allergic disease. Surprisingly, the EoE + IH group presented with significantly different clinical symptoms, with increased dysphagia, gagging, cough, and poor appetite compared to their counterparts in the EoE-IH group. Male gender, allergic rhinitis, the presence of dysphagia, and younger age were independently associated with having EoE + IH. Specific IgE levels to common EoE-associated foods were higher in EoE + IH, regardless of eliciting immediate hypersensitivity symptoms. In contrast, IgE levels for specific foods triggering EoE were relatively lower in both the groups than IgE levels for immediate reactions. CONCLUSIONS AND CLINICAL RELEVANCE: Immediate hypersensitivity is common in children with EoE and identifies a population of EoE patients with distinct clinical characteristics. Our study describes a subtype of EoE in which IgE-mediated food allergy may impact the presentation of paediatric EoE.

Naturally occurring tolerance acquisition to foods in previously allergic children is characterized by antigen specificity and associated with increased subsets of regulatory T cells.

BACKGROUND: Food allergy affects approximately 6-8% of children, and increasing in prevalence. Some children naturally outgrow their food allergy without intervention, but the mechanisms by which this occurs remain poorly understood. We sought to investigate the role of regulatory T cells in the development of naturally acquired tolerance. METHODS: Fifty-eight children (1-18 years) with either egg or peanut allergy, recent acquisition of natural tolerance to egg or peanut, or no food allergy were studied. Peripheral blood mononuclear cells (PBMC) from these groups were stimulated with relevant antigen for 48 h and flow cytometry performed to characterize both surface (CD3, CD4, CD25, CD14, CD19, and CD127) and intracellular markers (IL-10, Foxp3, and IL-5). RESULTS: Resting PBMC from naturally tolerant patients had significantly increased CD3+CD4+CD25+CD127loFoxp3+ cells, when compared to allergic or control patients (mean 6.36 vs. 2.37 vs. 2.62%, respectively, P < 0.05). Upon stimulation with relevant antigen, naturally tolerant patients also had increased IL-10-expressing CD25+CD127lo cells (6.33 vs. 1.65 vs. 0.7, P < 0.01), Foxp3+ cells (mean 12.6 vs. 5.42 vs. 3%, P < 0.01), and CD4+ cells (mean 4.48 vs. 1.59 vs. 0.87%, P < 0.01); the increase was not observed in PBMCs from allergic or control patients. Additionally, this upregulation was only seen with relevant antigen stimulation and not upon stimulation with unrelated antigen. CONCLUSION: The increased CD3+CD4+CD25+CD127lo cells at baseline and upon stimulation and increased induction of IL-10-producing cells of several types, including Tr1 cells, from naturally tolerant patients suggests an important role for regulatory T cell subsets in the acquisition of natural tolerance.

Recurrent superantigen exposure in vivo leads to highly suppressive CD4+CD25+ and CD4+CD25- T cells with anergic and suppressive genetic signatures.

Staphylococcal enterotoxin B (SEB) activates T cells via non-canonical signalling through the T cell receptor and is an established model for T cell unresponsiveness in vivo. In this study, we sought to characterize the suppressive qualities of SEB-exposed CD4(+) T cells and correlate this with genetic signatures of anergy and suppression. SEB-exposed CD25(+) and CD25(-)Vbeta8(+)CD4(+) T cells expressed forkhead box P3 (FoxP3) at levels comparable to naive CD25(+) T regulatory cells and were enriched after exposure in vivo. Gene related to anergy in lymphocytes (GRAIL), an anergy-related E3 ubiquitin ligase, was up-regulated in the SEB-exposed CD25(+) and CD25(-)FoxP3(+)Vbeta8(+)CD4(+) T cells and FoxP3(-)CD25(-)Vbeta8(+)CD4(+) T cells, suggesting that GRAIL may be important for dominant and recessive tolerance. The SEB-exposed FoxP3(+)GRAIL(+) T cells were highly suppressive and non-proliferative independent of CD25 expression level and via a glucocorticoid-induced tumour necrosis factor R-related protein-independent mechanism, whereas naive T regulatory cells were non-suppressive and partially proliferative with SEB activation in vitro. Lastly, adoptive transfer of conventional T cells revealed that induction of FoxP3(+) regulatory cells is not operational in this model system. These data provide a novel paradigm for chronic non-canonical T cell receptor engagement leading to highly suppressive FoxP3(+)GRAIL(+)CD4(+) T cells.

Asthma exacerbations. 2: aetiology.

The natural history of asthma involves relatively stable periods that are often punctuated by significant exacerbations of symptoms. There are many aetiologies that may lead to an increase in asthma severity including respiratory infection (bacterial/viral), allergens, irritants, and occupational exposures. Each trigger probably acts through different mechanisms, but a final common pathway of multicellular inflammation, enhanced bronchial responsiveness, and airflow obstruction is a likely consequence. This review discusses the most common causes of asthma exacerbations with a focus on their microbiology and immunopathogenesis. Through an understanding of underlying causes of asthma exacerbations, treatments with increased effectiveness may be developed, and it is these future developments that may directly influence the morbidity and mortality of the disease.

Fine needle aspiration cytology of liver: a study of 202 cases.

From January 1990 to December 2000, 202 patients with clinical evidence of liver disease underwent fine needle aspiration cytology of the liver. Of these, 102 patients were diagnosed as non-neoplastic lesions. These include diffuse parenchymal disease of liver, liver abscess, hepatitis, and granulomas. There were 100 patients with malignancies of the liver. Out of the above, 64 were due to metastatic carcinoma, 31 were primary hepatocellular carcinoma, 1 hepatoblastoma and in 4 patients the diagnosis of non-Hodgkin's lymphoma was made. By comparing with clinical and biochemical parameters, the diagnostic accuracy of the fine needle aspiration cytology, in this study, was found to be more accurate in malignant nodules of the liver as compared to other pathological lesion. The findings of fine needle aspiration cytology of the liver reported by other authors are discussed and it is concluded that this diagnostic method is a safe, useful and economic procedure with minimum complication and can be routinely done for assisting diagnosis of liver diseases in our clinical set up.

Influence of calcium, magnesium, or potassium ions on the formation and stability of emulsions prepared using highly hydrolyzed whey proteins.

Oil-in-water emulsions (4 wt % soy oil) containing 4 wt % whey protein hydrolysate (WPH) (27% degree of hydrolysis) and different levels of calcium, magnesium, or potassium chloride were prepared in a two-stage homogenizer. Other emulsions containing 4 wt % WPH but including 0.35 wt % hydroxylated lecithin and different levels of the above minerals were similarly prepared. The formation and stability of these emulsions were determined by measuring oil droplet size distributions using laser light scattering and by confocal scanning laser microscopy and a gravity creaming test. Both lecithin-free and lecithin-containing emulsions showed no change in droplet size distributions with increasing concentration of potassium in the range 0-37.5 mM. In contrast, the diameter of emulsion droplets increased with increasing calcium or magnesium concentration >12.5 mM. Emulsions containing hydroxylated lecithin were more sensitive to the addition of calcium or magnesium than the lecithin-free emulsions. Storage of emulsions at 20 degrees C for 24 h further increased the diameter of droplets and resulted in extensive creaming in emulsions containing >25 mM calcium or magnesium. It appears that both flocculation and coalescence processes were involved in the destabilization of emulsions induced by the addition of divalent cations.

A simple microassay for computing the hemolytic potency of drugs.

A simple microassay and computer program are described for determining the erythrocyte hemolytic potency of drugs in vitro. This microassay is sensitive for both micro as well as macro ranges of hemoglobin concentration. An ELISA reader has been adapted to read erythrocyte lysis (hemolysis), which reduces the number and culture of replicates. A computer program was developed that calculates parameters such as C50 (concentration of drug causing 50% hemolysis), C100 (concentration of drug causing 100% hemolysis) and beta (slope of the curve) and graphically expresses the hemolytic patterns of various drugs simultaneously. The program can obtain optical densities directly from a 96-well plate ELISA reader by interfacing the microplate reader to the computer or by using a keyboard. This method is useful for screening a large number of hemolytic drugs and requires lower amounts of test compounds. It may also be applicable to quantitative functional assays, such as complement-mediated hemolysis and enumeration of antibody-secreting cells. The program can be obtained from the authors on request.

Use of specific polyclonal antibodies for site specific drug targeting to malaria infected erythrocytes in vivo.

The possibility of using specific polyclonal antibodies for effective site specific drug targeting to malaria infected erythrocytes has been examined. For this purpose, rabbit polyclonal antiserum was raised against Plasmodium berghei infected mouse erythrocytes (IRBC) and extensively absorbed with normal erythrocytes (NRBC). Absorbed antiserum specifically recognized IRBC. F(ab')2-fragments of these antibodies were coupled to chloroquine (chq) laden liposomes. These immunoliposomes when tested in vivo significantly suppressed malarial infection in mice.

Anterior mandibulotomy for oral and oropharyngeal tumours.

In the last six years, anterior mandibulotomy was used to approach tumours of the oropharynx and oral cavity in 39 cases. Twenty-six of these had primary lesions in the anterior two-thirds of the tongue. Eight cases had lesions in the base tongue, three in the cheek, and two in the tonsil. Twenty-six cases had T3 tumours, nine had T2 lesions, and four had T4 tumours. Twenty-five patients received post-operative radiotherapy. In 16 cases the mandibulotomy was combined with a marginal mandibulectomy. In 23 cases reconstruction was carried out using a pectoralis major myocutaneous flap. Adequate margins on histopathology obtained in all but eight patients. Bone-related complications occurred in only three patients, all of whom were previously irradiated. Thus the anterior mandibulotomy provides excellent exposure for oral and oropharyngeal tumours, with low complication rate, and avoidance of segmental mandibulectomy.

The impact of migration on women and the family: research, policy and programme issues in developing countries.

"In the context of the pervasive invisibility of migrant women in the standard data collection, policy-making and planning processes, this paper seeks to review some of the major trends and patterns of female migration in developing countries; identify some important research priorities, and to delineate some of the areas in which policies and programmes might be framed so as to assist or change the impact of migration on women and the family. In the discussion of the relevance of these issues to national services and community organisation are emphasised as priority action programmes."