Use of platelet-rich plasma in the healing of chronic ulcers of the lower extremity.

INTRODUCTION AND OBJECTIVES: Platelet-rich plasma (PRP) is used as an adjuvant in the treatment of chronic ulcers of the lower extremity and has shown particularly promising results in the case of neuropathic ulcers. There has been less research, however, into its use in venous and hypertensive ulcers. Our aim was to assess the safety and feasibility of using PRP in the treatment of chronic ulcers of the lower extremity and to evaluate its potential benefits in directed healing. MATERIAL AND METHODS: We prospectively selected 11 patients with nonischemic ulcers of the lower extremity that had been present for at least 6 weeks. PRP was injected subcutaneously into the perilesional tissue and applied topically in 4 sessions held at 1-week intervals. We assessed quality of life (SF-12 questionnaire), pain (visual analog scale), and the circumference of the ulcer before and after treatment. RESULTS: There was a predominance of women (8/11, 73%), and venous ulcers (7/11, 64%) were more common than hypertensive ulcers (4/11, 36%). The median age of the patients was 79 years and the median time since onset of the ulcer was 17 months (range, 6-108 months). We observed a significant reduction in pain (P<.05) and a significant improvement in the physical and mental components of the SF-12 (P<.05). The mean reduction in ulcer size was 60%, and complete healing was achieved in 5 cases. No adverse effects were observed. CONCLUSION: The local application of PRP is a valuable and practical procedure that promotes the healing of chronic ulcers of the lower extremity; it can improve patient quality of life and is particularly effective in local pain relief.

[Ophthalmological manifestations of arterial hypertension]. / Manifestaciones oftalmológicas de la hipertensión arterial.

Systemic Arterial hypertension (AHT) is one of the most frequent diseases in the industrialised countries, with an incidence reaching 30%, a figure that is rising due to the greater life expectancy of the population. This rise in arterial tension causes, or accelerates, changes in the vascular wall of the target organs such as the kidney, brain, heart and eye. At the ocular level, AHT produces lesions in the retina, the choroids and optic nerve head; this can include a wide range of lesions, from slight vascular narrowing to severe visual loss due to ischaemic optical neuropathy. The primary response of the retinal arteries to systemic arterial hypertension is vascular narrowing and the manifestations that appear at the back of the eye in hypertensive retinopathy are diffuse or focal vasoconstriction, extravasation due to increased vascular permeability and arteriosclerosis with swelling of the wall of the vessels. These three entities are responsible for the appearance of different lesions that characterise the stages of the retinal disease, which are: arteriovenous crossings, hard and cotton-like exudates, thrombosis, embolisms, haemorrhages in the retinal parenchyma, vitreous detachment from the retina, papilla edema and ischaemic optical neuropathy in the more severe cases, such as in case malign arterial hypertension.

Evaluation of alternatives to RFLP for the analysis of clustered cases of tuberculosis.

BACKGROUND: Mycobacterial interspersed repetitive unit-variable number tandem repeat (MIRU-VNTR) analysis is a recently developed method which could be suitable as a 'real-time' genotyping tool for Mycobacterium tuberculosis. METHODS: One hundred and thirty-four M. tuberculosis isolates were analysed using the reference method, IS6110-restriction fragment length polymorphism (RFLP), and by MIRU, alone and together with spoligotyping. RESULTS: MIRU reduced the genotyping turnaround time by 21 days. The discriminatory power (HGDI) for MIRU and RFLP was 0.978 and 0.989, respectively. RFLP clustered 41.8% of the isolates (17 clusters; 2-9 representatives), whereas MIRU increased the number and size of the clusters (57.5% of the isolates in 20 clusters; 2-14 representatives). With respect to the RFLP clusters, MIRU data showed full correlation in only 7/ 17 (41%) clusters and low correlation in 8/17 (47%) clusters. When MIRU and spoligotyping were considered together, the analysis fitted better with RFLP data: 1) 42.5% of the isolates were grouped in 20 clusters of 2-6 representatives, and 2) the number of clusters with full correlation with RFLP data increased to 11/17 and those with low correlation decreased to 2/17. CONCLUSION: MIRU-VNTR analysis showed low correlation with RFLP. The addition of spoligotyping to MIRU analysis fitted much better with RFLP analysis, although full correlation was still not achieved.

[Eyelid contact allergic eczema caused by topical ketorolac tromethamine 0.5%]. / Eccema alérgico de contacto palpebral por ketorolaco trometamol 0,5% tópico.

CLINICAL CASE: A 65-year-old man had been treated with eye-drops containing NSAIDs (Ketorolac). He developed conjunctival injection, edematous swelling of the eyelids and periorbital dermatitis due to a contact allergy. Allergy to NSAIDs is uncommon. DISCUSSION: This is an unusual case because topical application of ketorolac is safe in the vast majority of ophthalmologic patients. However adverse events associated with Ketorolac are similar to that of other NSAIDs.

[Intraocular lenses with blue light filter]. / Lentes intraoculares con filtros para luz azul.

PURPOSE: To evaluate if a SN60AT <> IOL (Alcon) with IMPRUV filter (Alcon) offers any advantage in patients having cataract surgery, as compared with Acrysof SA60 IOL (Alcon). METHODS: This was a prospective analytical study. We studied sixty-four eyes from 32 patients with cataracts. After a random assignment, phacoemulsification with the implant was performed using the Acrysof SN60AT tinted lens in one eye and Acrysof SA60 in the other. A variety of variables was subsequently assessed: contrast sensitivity with a CSV 1000E test, colour perception through a Farnsworth 25 test and subjective measures by a patient survey. Differences between the groups were assessed statistically. RESULTS: The contrast sensitivity results were obtained for 3, 6, 12 and 18 cycles per degree (cpd) frequencies under scotopic conditions, with better results obtained following the use of SN60AT: 0.75 (CSV 1000E test units) for 3 cpd, 0.86 for 6 cpd, 0.54 for 12 cpd and 0.35 for 18 cpd. The differences were statistically significant between the two groups for 3 cpd and 6 cpd. A total of 62.5% of patients using a tinted lens and 50% with SA60 achieved good results in the Farnsworth test; 12.5% and 21% had fair results and 25% and 28.1% respectively had poor results. CONCLUSIONS: We obtained an improvement in favour of the tinted lens for all frequencies, with this being statistically significant at lower frequencies (3 and 6 cpd). No significant differences in the Farnsworth test performance or in the patient survey were found. The tinting provided by the IMPRUV filter brings an increase in contrast sensitivity without affecting colour perception.

Evidence that thrombin present in lungs of patients with pulmonary metastasis may contribute to the development of the disease.

Early cellular events in the lung which may lead to the development of pulmonary metastases (PM) are still poorly understood. Thrombin, a key component of the coagulation cascade, may be involved in the development of PM as it has been shown to be an enhancer of platelet-tumor interaction in vitro and metastasis in vivo, and because it has been found in high levels in lungs from patients with PM. In this study, we assessed the potential role of thrombin in promoting PM by inducing an enhancement of tumor cell adhesion to platelets and tumor cell chemoinvasion and proliferation. We used bronchoalveolar lavage fluid (BALF) from 20 patients with PM. Results were compared with those from healthy controls. We found an enhancement of adhesion of PM-BALF-treated tumor cells to untreated platelets. BALF from patients with PM significantly increased chemoinvasion and proliferation in three human tumor cell lines. These activities were attenuated significantly by a thrombin inhibitor: hirudin. These results indicate that the thrombin present in the lungs of patients with PM is, at least in part, responsible for their adhesive, invasive and mitogenic activity on three different tumor cell lines. They also suggest that thrombin may be involved in the development of PM.

Thrombin is present in the lungs of patients with primary extremity osteosarcoma and pulmonary metastases.

Osteosarcoma is a rare cancer, which metastasizes to the lung in up to 80% of cases. Thrombin is involved in metastasis and is present in the lungs of patients with pulmonary metastases (PM). To identify its role in PM and osteosarcoma, we measured thrombin levels in the bronchoalveolar lavage fluid (BALF) of 15 patients. BALF was collected at different stages of the disease and correlated with the diagnosis of PM. We also assessed fibrinogen overexpression in the tumors. We found that 11/15 (73%) patients with high thrombin levels in the lungs developed PM within the first 12 months from primary surgery. The median thrombin concentration in the BALF of these patients increased up to 8x10(-9) M (range, 3x10(-9)M-15x10(-9)M), which represents a more than 100-fold increase compared to patients without PM (p<0.0001). Eight of 15 (53%) primary and 11/15 (73%) metastatic samples showed fibrinogen overexpression. A significant difference between high thrombin levels, fibrinogen overexpression and PM was found compared to patients without PM (p=0.00073 and p=0.025). These results show that thrombin levels are increased in the lungs of patients with primary osteosarcoma and a high risk of developing PM. They suggest that thrombin may be involved in the development of PM.

Antibacterial IgE antibodies in children.

The presence of specific IgE antibodies directed against common microorganisms was analyzed in a population of children. These antibodies included Escherichia coli, Proteus spp., Pseudomonas spp., Klebsiella spp., Streptococcus spp. and Staphylococcus spp. The determination of these antibodies was done by radioimmunoassay. Comparisons were made according to the parameters of age and total IgE levels. These antibodies were found in a high proportion of the cases. Statistical analysis demonstrated that the different specific IgE antibodies were present in all children, none showing special relevance. All of the antibodies studied were found to be present in a higher number of cases in the groups of older children, and in the group of children with increased total IgE levels, a significantly higher number of cases with antibacterial specific IgE antibodies was observed. The presence and the variety of the antibodies might be conditioned by individual as well as environmental factors. The role of antibodies is not well known. Although they are frequently found in allergic children, there have not been enough cases studied to establish their involvement in allergy. Understanding the role of antibodies might be of interest in the study of allergic processes exacerbated by bacterial infections.

Assessment of the doses received by the Cuban population from 40K contained in the body: modelling based on a neural network.

Potassium-40 constitutes the main natural source of potassium present in the body, which influences the effective dose received by people. With the aim of assessing the contribution of this component to the doses received by the Cuban population, a study intended to assess the doses was developed. For this purpose, a representative sample of the Cuban population was selected according to age and sex. The measurements were made using the whole-body counter (WBC) of the Center for Radiation Protection and Hygiene (CPHR). For dose estimations, a uniform distribution of potassium for the whole body was assumed. The methodology used was the one recommended by the ICRP. The values of annual effective dose range between 93 and 209 microSv for females and between 123 and 212 microSv for males. The annual average effective dose for members of the public was estimated as 150 +/- 40 microSv, taking into account the experimental data and the specific features of the Cuban population. With the dose values obtained, it was possible to model dose estimates by means of a neural network, which was trained with the results obtained and using as starting data the sex, age, height and corporal weight of people studied. The modelobtained allows consideration of the anatomical features of each person in the estimation of doses.

Novel use of the Elizabethan collar as a "hoop-skirt" for protecting wounds and catheters in nonhuman primates.

This article describes the novel use of an Elizabethan collar, which is attached to a primate jacket to create a tamper-proof "hoop-skirt" for protecting wounds and catheters. We successfully have used this hoop-skirt to manage juvenile male rhesus monkeys with Foley catheters for 10 days post-prostatectomy. In addition, our hoop-skirt has been used to manage wounds on the hindlimbs of both macaques and baboons.

Antinociceptive activity of and clinical experience with buprenorphine in swine.

We performed antinociceptive testing on swine receiving buprenorphine. Intravenous access was achieved, and animals were allowed to recover for 24 h. Baseline skin-twitch latency to a focused light source was determined for each animal. Animals received intravenous (i.v.) buprenorphine at 0.08 (n =1), 0.16 (n = 1), 0.005 (n = 5), 0.01 (n = 5), or 0.02 mg/kg (n = 6). Skin-twitch latency was determined 15, 30, 60, 120, 180, 240, 300, 360, 420, 480, 540, and 600 min after buprenorphine administration. Analgesic activity as measured by a significant increase in latency time over baseline values occurred at all time points except 480 min in animals that received 0.02 mg/kg buprenorphine i.v. Analgesic activity to 420 min was demonstrated in animals that received 0.01 mg/kg buprenorphine i.v. Analgesic activity was not demonstrated at any time point in animals that received 0.005 mg/kg buprenorphine i.v. A retrospective analysis of postoperative care records was performed to determine whether 0.01 mg/kg buprenorphine i.v. or intramuscularly (i.m.) postoperatively to swine provided clinically relevant analgesia. Records of swine receiving buprenorphine from 1997 to 2000 were reviewed for indications of treatment failure, such as pain or a change in analgesic regimen from that used routinely. Treatment failure occurred in 18 of 416 (4.3%) cases treated with buprenorphine. This failure occurred in 17% of cases with problems categorized as inflammatory in nature and in 15.5% of those with systemic problems or organ failure. We concluded that antinociceptive testing predicted that buprenorphine administered at 0.01 mg/kg i.v. in swine likely would provide analgesic efficacy for 6 h and when administered at 0.02 mg/kg i.v. likely would provide 10 h analgesia. Clinical signs of pain in animals recovering from surgery were not observed in the majority of cases when buprenorphine was administered twice or thrice daily at 0.01 mg/kg i.m. or i.v. However, buprenorphine was less effective at treating signs of pain associated with inflammation, organ failure, or systemic disease than at ameliorating pain associated with surgical incisions and orthopedic, dental, and ophthalmic procedures.

Oropharyngeal administration of colostrum to extremely low birth weight infants: theoretical perspectives.

Studies in adults have shown that the oropharyngeal route can be used to effectively and safely administer interferon-alpha, an immune cell-derived cytokine, to patients who are unable to tolerate its parenteral administration. The mechanism for this appears to be the stimulatory effects of the cytokine, on the oropharyngeal-associated lymphoid tissue system. Own mother's colostrum (OMC) is rich in cytokines and other immune agents that provide bacteriostatic, bacteriocidal, antiviral, anti-inflammatory and immunomodulatory protection against infection. OMC may be especially protective for the extremely low birth weight (ELBW) infant in the first days of life; however clinical instability typically precludes enteral feedings during this period. Oropharyngeal administration is a potential alternative method of providing OMC. Oropharyngeal administration of OMC may have immunomodulatory effects on the recipient infant, and would be especially beneficial to the ELBW infant who would otherwise remain nil per os during the first days of life.

Extramedullary plasmacytoma of the salivary gland in two Syrian hamsters (Mesocricetus auratus).

Two Syrian hamsters developed marked swelling of the ventral neck. Histologic examination of both masses revealed that the submaxillary salivary glands were effaced by large numbers of neoplastic plasma cells. In one hamster, neoplastic cells had infiltrated the adjacent lymph node. The neoplastic cells expressed CD79a antigen and were negative for CD3, lambda, and kappa light chains. Ultrastructural features of neoplastic cells in the salivary gland of one hamster included abundant cytoplasmic rough endoplasmic reticulum profiles, and peripherally displaced nuclei that contained marginated heterochromatin, consistent with plasma cells. Salivary gland plasmacytomas are extremely rare in humans and have not previously been reported in nonhuman species. The occurrence of such neoplasms in two hamsters suggests that this species may be predisposed to developing tumors of this type.

Clinical and histopathological evaluation of 13 cases of adenocarcinoma in aged rhesus macaques (Macaca mulatta).

In recent years, the emphasis on aging research, has led to an increase in the number of aged macaques being maintained in some research facilities with a subsequent increase in the occurrence of age-related diseases. One of the most commonly reported age related diseases is intestinal adenocarcinoma. At the University of Illinois at Chicago (UIC), which maintains a colony of approximately 55 aged rhesus macaques 13 cases of intestinal adenocarcinoma were diagnosed within a 25-month period. This report provides a comprehensive description of the clinical findings for intestinal adenocarcinoma in aged rhesus macaques, including results from physical examinations, laboratory tests, radiographic evaluations, gross and histopathologic findings as well as a comparison with the disease condition in humans. The use of carcinoembryonic antigen as a potential tumor marker was evaluated by immunohistochemical analysis of tissue specimens in 10 cases. Intestinal adenocarcinoma is a disease condition that should be of concern to individuals responsible for the care of aged rhesus macaques.

Ki-67: a proliferative marker that may predict pulmonary metastases and mortality of primary osteosarcoma.

Alterations in Ki-67 activity have been associated with tumor progression and poor outcome in cancer patients. This study was undertaken to identify the potential of this proliferative marker as a predictor of pulmonary metastases (PM) and mortality in osteosarcoma patients. In 38 patients with tissue available for immunohistochemical analysis, overexpression of Ki-67 was assessed. Chi-square and log rank tests were used to determine differences between proportions of the marker with PM and mortality and survival distributions respectively. P values equal or less than .05 were considered statistically significant. The median follow up of this case series was 28 months. Eighteen (47.4%) of 38 patients developed PM, and 17 (44%) overexpressed Ki-67. We found a high frequency of PM (15 of 17) among those cases that overexpressed Ki-67. This relationship was significant (P = .000006) when compared to the rest of the group. We also found a statistically significant correlation between patients with positive and negative Ki-67 scores and higher and lower mortality (P = .000962). These findings suggest that Ki-67 overexpression could be used as a prognostic molecular marker for the development of PM in osteosarcoma patients.

Indomethacin suppresses the anti-proliferative effects of transforming growth factor-beta isoforms on fibroblast cell cultures.

The transforming growth factor-beta (TGFbeta) family of mediators consists of five closely related isoforms, of which three are present in mammals. TGFbeta1 has been shown to exert a biphasic effect on the proliferation of several cell types, including fibroblasts, with stimulation at low concentrations and inhibition at higher concentrations. The stimulatory effects are well characterized, but the mechanisms by which TGFbeta1 inhibits cell proliferation are incompletely understood. In the present study we have compared the effects of all three mammalian TGFbeta isoforms on human lung fibroblast proliferation, and have elucidated the role of the TGFbeta-induced synthesis of prostaglandin E2 (PGE2) in mediating their actions. All three isoforms stimulated fibroblast proliferation with maximal effects at 5 pg/ml (0.2 pM) and an order of potency of TGFbeta3 > TGFbeta2 > TGFbeta1. At higher concentrations, proliferation declined, and at 40 pg/ml and above all isoforms inhibited fibroblast proliferation. Again TGFbeta3 was the most potent, but there were no significant differences between the inhibitory effects of TGFbeta1 and TGFbeta2. Addition of indomethacin, an inhibitor of PGE2 synthesis, did not alter the proliferative activity of any of the TGFbeta isoforms, but completely overcame their inhibitory effects, restoring the stimulatory actions observed at lower TGFbeta concentrations. All TGFbeta isoforms stimulated PGE2 synthesis; TGFbeta3 was approximately twice as potent as TGFbeta1 and TGFbeta2, each of which had similar effects. These data suggest that the inhibition of fibroblast proliferation at higher concentrations of TGFbeta isoforms may be mediated by autocrine stimulation of PGE2 synthesis.

Lung cell toxicity experimentally induced by a mixed dust from Mexicali, Baja California, Mexico.

Lung disease caused by nonoccupational exposures to inorganic particles from the soil has been reported in several areas of the world. We tested the toxic potential of dust samples from a Mexican city (Mexicali) that is frequently affected by dust storms and is geographically related to the area of San Diego, CA, where constituents of the soil have been reported to be fibrogenic. We found that samples of Mexicali dust are a mixture of approximately 75% potassium aluminum silicates (illite) and approximately 20% silica. Respirable size particles were highly hemolytic and induced lactic dehydrogenase release from alveolar macrophages exposed in vitro. Animals instilled intratracheally with the dust developed a multifocal interstitial lung disease associated with deposits of the aluminum silicates, which were identified by X-ray microanalysis. Inhalation studies in rats demonstrated that the majority of particles were deposited preferentially at the first alveolar duct bifurcations. Twenty-four hours later, numerous particles had been ingested by alveolar macrophages that had migrated to those sites of deposition. It is proposed that alveolar macrophages are attracted to the deposited particles by complement fragments since Mexicali dust is capable of activating complement proteins from both serum and bronchoalveolar lavage. Activation resulted in alveolar macrophage chemotaxis. Mexicali dust induced biological activities and lung changes similar to those of asbestos and silica, suggesting that this material could be an etiologic agent of pulmonary fibrosis in exposed individuals.

Diverse cellular TGF-beta 1 and TGF-beta 3 gene expression in normal human and murine lung.

A role for transforming growth factor-beta 1, (TGF-beta 1) has been proposed in lung development and in the pathogenesis of pulmonary disease. However, previous studies have not delineated the cells expressing TGF-beta 1 in normal adult lung, nor compared its gene expression with that of other TGF-beta isoforms. We used digoxigenin-labelled riboprobes to localize TGF-beta 1 and TGF-beta 3 gene expression in normal adult human and mouse lung. This procedure was technically simple, providing excellent resolution. TGF-beta 1 and TGF-beta 3 messenger ribonucleic acid (mRNA) transcripts were detected in a wide variety of cells. In human lung, mRNA for both isoforms was localized to bronchiolar epithelium and alveolar macrophages. TGF-beta 1, but not TGF-beta 3 mRNA was detected in mesenchymal and endothelial cells. In murine tissue, TGF-beta 1, mRNA was localized to bronchiolar epithelium, Clara cells, mesenchymal cells, pulmonary endothelium and alveolar cells, including macrophages. TGF-beta 3 mRNA was similarly distributed but not detected in endothelium. In summary, using a nonisotopic technique in lung tissue, we have detailed the cells expressing the transforming growth factor-beta 1 and beta 3 genes in human and murine lung. There was widespread expression of these cytokines in normal lung consistent with autocrine or paracrine roles in regulating cellular turnover, immune defence and matrix protein metabolism.

Platelet-derived growth factor (PDGF)-AA, -AB, and -BB induce differential chemotaxis of early-passage rat lung fibroblasts in vitro.

Platelet-derived growth factor (PDGF) isoforms are chemoattractants and mitogens for cells of mesenchymal origin that could be important mediators of pulmonary fibrogenesis. We have previously reported that particle-activated alveolar macrophages secrete homologues of PDGF that are composed of all three PDGF isoforms (PDGF-AA, -AB, and -BB). This mixture of macrophage-derived PDGF, once dissociated from the PDGF-alpha-macroglobulin complex, induces chemotaxis of rat lung fibroblasts (RLF) in the nanomolar range. In addition, we have reported that PDGF isoforms induce differential proliferation of RLF (PDGF-BB > PDGF-AB > PDGF-AA). In the present study, we sought to determine the relative chemotactic potency of the three PDGF isoforms and correlate these responses to the relative abundance of the two types of PDGF cell-surface receptors: PDGF-alpha receptor (PDGF-R alpha) and PDGF-beta receptor (PDGF-R beta). We also investigated the chemotactic activity of combinations of two PDGF isoforms simultaneously. Isolates of early-passage RLF were assayed for chemotaxis in 48-microwell chambers. Swiss mouse 3T3 cells were assayed in parallel as a positive control cell line for PDGF-R alpha and PDGF-R beta expression. RLF responded differentially to the PDGF isoforms: PDGF-AB and PDGF-BB were potent chemoattractants and stimulated maximal chemotactic responses between 4 and 8 ng/ml PDGF, whereas PDGF-AA elicited a weak chemotactic response that was maximally 15% of that obtained with either B-chain isoform. PDGF-AB and PDGF-BB were also the most potent chemoattractants for Swiss 3T3 cells, and their response to these B-chain isoforms was approximately 40% greater than that obtained for RLF.(ABSTRACT TRUNCATED AT 250 WORDS)