Clinical variables associated with leprosy reactions and persistence of physical impairment

Introduction Leprosy is a chronic disease that affects skin and peripheral nerves. Disease complications include reactional episodes and physical impairment. One World Health Organization (WHO) goal of leprosy programs is to decrease the number of grade 2 impairment diagnoses by 2015. This study aims to evaluate clinical factors associated with the occurrence of leprosy reactions and physical impairment in leprosy patients. Methods We conducted a retrospective study of data from medical records of patients followed in two important centers for the treatment of leprosy in Aracaju, Sergipe, Brazil, from 2005 to 2011. We used the chi-square test to analyze associations between the following categorical variables: gender, age, operational classification, clinical forms, leprosy reactions, corticosteroid treatment, and physical impairment at the diagnosis and after cure. Clinical variables associated with multibacillary leprosy and/or reactional episodes and the presence of any grade of physical impairment after cure were evaluated using the logistic regression model. Results We found that men were more affected by multibacillary forms, reactional episodes, and grade 2 physical impairment at diagnosis. Leprosy reactions were detected in a total of 40% of patients and all were treated with corticosteroids. However, physical impairment was observed in 29.8% of the patients analyzed at the end of the treatment and our multivariate analysis associated a low dose and short period of corticosteroid treatment with persistence of physical impairments. Conclusions Physical impairment should receive an increased attention before and after treatment, and adequate treatment should be emphasized.

Analysis of the chemical components of hydatid fluid from Echinococcus granulosus

Introduction The aim of this study was to explore the environment of Echinococcus granulosus (E. granulosus) protoscolices and their relationship with their host. Methods Proteins from the hydatid-cyst fluid (HCF) from E. granulosus were identified by proteomics. An inductively coupled plasma atomic emission spectrometer (ICP-AES) was used to determine the elements, an automatic biochemical analyzer was used to detect the types and levels of biochemical indices, and an automatic amino acid analyzer was used to detect the types and levels of amino acids in the E. granulosus HCF. Results I) Approximately 30 protein spots and 21 peptide mass fingerprints (PMF) were acquired in the two-dimensional gel electrophoresis (2-DE) pattern of hydatid fluid; II) We detected 10 chemical elements in the cyst fluid, including sodium, potassium, calcium, magnesium, copper, and zinc; III) We measured 19 biochemical metabolites in the cyst fluid, and the amount of most of these metabolites was lower than that in normal human serum; IV) We detected 17 free amino acids and measured some of these, including alanine, glycine, and valine. Conclusions We identified and measured many chemical components of the cyst fluid, providing a theoretical basis for developing new drugs to prevent and treat hydatid disease by inhibiting or blocking nutrition, metabolism, and other functions of the pathogen.

Management of post-transplant Epstein-Barr virus-related lymphoproliferative disease in solid organ and hematopoietic stem cell recipients

Epstein-Barr virus (EBV)-related post-transplant lymphoproliferative disease (PTLD) is one of the most serious complications associated with solid organ and hematopoietic stem cell transplantation. PTLD is most frequently seen with primary EBV infection post-transplant, a common scenario for pediatric solid organ recipients. Risk factors for infection or reactivation of EBV following solid organ transplant are stronger immunosuppressive therapy regimens, and being seronegative for receptor. For hematopoietic stem cell transplantation, the risk factors relate to the type of transplant, human leukocyte antigen disparity, the use of stronger immunosuppressants, T-cell depletion, and severe graft-versus-host disease. Mortality is high, and most frequent in patients who develop PTLD in the first six months post-transplant. The primary goal of this article is to provide an overview of the clinical manifestations, diagnosis, accepted therapies, and management of EBV infection in transplant recipients, and to suggest that the adoption of monitoring protocols could contribute to a reduction in related complications.

Changes in body fluid and energy compartments during prolonged hunger strike

Prolonged total food deprivation in non-obese adults is rare, and few studies have documented body composition changes in this setting. In a group of eight hunger strikers who refused alimentation for 43 days, water and energy compartments were estimated, aiming to assess the impact of progressive starvation. Measurements included body mass index (BMI), triceps skinfold (TSF), arm muscle circumference (AMC), and bioimpedance (BIA) determinations of water, fat, lean body mass (LBM), and total resistance. Indirect calorimetry was also performed in one occasion. The age of the group was 43.3±6.2 years (seven males, one female). Only water, intermittent vitamins and electrolytes were ingested, and average weight loss reached 17.9%. On the last two days of the fast (43rd-44th day) rapid intravenous fluid, electrolyte, and vitamin replenishment were provided before proceeding with realimentation. Body fat decreased approximately 60% (BIA and TSF), whereas BMI reduced only 18%. Initial fat was estimated by BIA as 52.2±5.4% of body weight, and even on the 43rd day it was still measured as 19.7±3.8% of weight. TSF findings were much lower and commensurate with other anthropometric results. Water was comparatively low with high total resistance, and these findings rapidly reversed upon the intravenous rapid hydration. At the end of the starvation period, BMI (21.5±2.6 kg/m²) and most anthropometric determinations were still acceptable, suggesting efficient energy and muscle conservation. Conclusions: 1) All compartments diminished during fasting, but body fat was by far the most affected; 2) Total water was low and total body resistance comparatively elevated, but these findings rapidly reversed upon rehydration; 3) Exaggerated fat percentage estimates from BIA tests and simultaneous increase in lean body mass estimates suggested that this method was inappropriate for assessing energy compartments in the studied population; 4) Patients were not morphologically malnourished after 43 days of fasting; however, the prognostic impact of other impairments was not considered in this analysis.

Effect of clarithromycin on the cell profile of bronchoalveolar lavage fluid in mice with neutrophil-predominant lung disease

OBJECTIVE: Macrolide antibiotics have anti-inflammatory properties in lung diseases. The aim of this study was to investigate the effect of clarithromycin in pulmonary cellular inflammatory response in mice. METHOD: Eight adult Swiss mice were studied. All animals received an intranasal challenge (80 µL) with dead Pseudomonas aeruginosa (1.0 x 10(12) CFU/mL). Bronchoalveolar lavage was performed 2 days later, with total cell count and differential cell analysis. The study group (n = 4) received clarithromycin treatment (50 mg/kg/day, intraperitoneal) for 5 days. Treatment was initiated 2 days before intranasal challenge. RESULTS: There was no significant difference in total cell count between the groups (mean: 2.0 x 10(6) and 1.3 x 10(6), respectively). In both groups, there was a predominance of neutrophils. However, the study group had a higher percentage of lymphocytes in the bronchoalveolar lavage than the control group (median of 19% vs 2.5%, P = .029). CONCLUSION: Clarithromycin alters the cytological pattern of bronchoalveolar lavage of Swiss mice with neutrophil pulmonary inflammation, significantly increasing the percentage of lymphocytes.

Sôbre u'a modificação do meio de Monteverde

It is well known that the culture media used in the presumptive diagnosis of suspiciuous colonies from plates inoculated with stools for isolation of enteric organisms do not always correctly indicate the major groups of enterobacteria. In an effort to obtain a medium affording more exact indications, several media (1-9) have been tested. Modifications of some of these media have also been tested with the result that a satisfactory modification of Monteverde's medium was finaly selected. This proved to be most satisfactory, affording, as a result of only one inoculation, a complete series of basic indications. The modification involves changes in the formula, in the method of preparation and in the manner of storage. The formulae are: A. Thymol blue indicator: NaOH 0.1/N .............. 34.4 ml; Thymol blue .............. 1.6 g; Water .................... 65.6 ml. B. Andrade's indicator. C. Urea and sugar solution: Urea ..................... 20 g; Lactose ................... 30 g; Sucrose ................... 30 g; Water .................... 100 ml. The mixture (C.) should be warmed slightly in order to dissolve the ingredients rapidly. Sterilise by filtration (Seitz). Keep stock in refrigeratior. The modification of Monteverde's medium is prepared in two parts. Semi-solid part - Peptone (Difco) 2.0 g; NaCl 0.5 g; Agar 0.5 g; Water 100.0 ml. Boil to dissolve the ingredients. Adjust pH with NaOH to 7.3-7.4. Boil again for precipitation. Filter through cotton. Ad indicators "A" 0.3 ml and "B" 1.0 ml. Sterilise in autoclave 115ºC, 15 minutes in amounts not higher than 200 ml. Just before using, add solution "C" asseptically in amounts of 10 ml to 200 ml of the melted semi-solid medium, maintained at 48-50ºC. Solid part - Peptone (Difco) 1.5 g; Trypticase (BBL) 0.5 g; Agar 2.0 g; Water 100,00 ml. Boil to dissolve the ingredients. Adjust pH with NaOH to 7.3-7.4. Boils again. Filter through cotton. Add indicators "A" 0.3 ml and "B" 1.0 ml; ferrous ammonium sulfate 0.02 g; sodiun thiosulfate 0.02 g. Sterilise in autoclave 115ºC, 15 minutes in amounts not higher than 200 ml. Just before using, add solution "C" asseptically in amounts of 10 ml to 200 ml of the melted solid medium, maintained at 48-50ºC. Final medium - The semi-solid part is dispensed first (tubes about 12 x 120 mm) in 2.5 ml amounts and left to harden at room temperature, in vertical position. The solid part is dispensed over the hardened semi-solid one in amounts from 2.0 ml to 2.5 ml and left to harden in slant position, affording a butt of 12 to 15 mm. The tubes of medium should be subjected to a sterility test in the incubator, overnight. Tubes showing spontaneous gas bubbles (air) should then be discarded. The medium should be stored in the incubator (37ºC), for not more than 2 to 4 days. Storage of the tubes in the ice-box produces the absorption of air which is released as bubbles when the tubes are incubated at 37ºC after inoculation. This fact confirmed the observation of ARCHAMBAULT & McCRADY (10) who worked with liquid media and the aplication of their observation was found to be essential to the proper working conditions of this double-layer medium. Inoculation - The inoculation is made by means of a long straight needle, as is usually done on the triple sugar, but the needel should penetrate only to about half of the height of the semi-solid column. Indol detection - After inoculation, a strip of sterelized filter papaer previously moistened with Ehrlich's reagent, is suspended above the surface of the medium, being held between the cotton plug and the tube. Indications given - In addition to providing a mass of organisms on the slant for serological invetigations, the medium gives the following indications: 1. Acid from lactose and/or sucrose (red, of yellowsh with strains which reduce the indicators). 2. Gas from lactose and/or sucrose (bubbles). 3. H[2]S production, observed on the solid part (black). 4. Motility observed on the semi-solid part (tubidity). 5. Urease production, observed on solid and semi-solid parts (blue). 6. Indol production, observed on the strip of filter paper (red or purplish). Indol production is not observed with indol positive strains which rapidly acidify the surface o the slant, and the use of oxalic acid has proved to give less sensitive reaction (11). Reading of results - In most cases overnight incubation is enough; sometimes the reactions appear within only a few hours of incubation, affording a definitive orientation of the diagnosis. With some cultures it is necessary to observe the medium during 48 hours of incubation. A description showing typical differential reaction follows: Salmonella: Color of the medium unchanged, with blackening of the solid part when H[2]S is positive. The slant tends to alkalinity (greenish of bluish). Gas always absent. Indol negative. Motility positive or negative. Shigella: Color of the medium unchanged at the beginning of incubation period, but acquiring a red color when the strain is late lactose/sucrose positive. Slant tending to alkalinity (greenish or purplish). Indol positive or negative. Motility, gas and H[2]S always negative. Proteus: Color of the medium generally changes entirely to blue or sometimes to green (urease positive delayed), with blackening of solid part when H[2]S is positive. Motility positive of negative. Indol positive. Gas positive or negative. The strains which attack rapidly sucrose may give a yellow-greenish color to the medium. Sometimes the intense blue color of the medium renders difficult the reading of the H[2]S production. Escherichiae and Klebsiellae: Color of the medium red or yellow (acid) with great and rapid production of gas. Motility positive or negative. Indol generally impossible to observe. Paracoli: Those lactose of sucrose positive give the same reaction as Esherichia. Those lactose or sucrose negatives give the same reactions as Salmonellae. Sometimes indol positive and H[2]S negative. Pseudomonas: Color of the medium unchanged. The slant tends to alkalinity. It is impossible to observe motility because there is no growth in the bottom. Alkaligenes: Color of the medium unchanged. The slant tends to alkalinity. The medium does not alter the antigenic properties of the strains and with the mass of organisms on the slant we can make the serologic diagnosis. It is admitted that this medium is somewhat more laborious to prepare than others used for similar purposes. Nevertheless it can give informations generally obtained by two or three other media. Its use represents much saving in time, labor and material, and we suggest it for routine laboratory work in which a quick presumptive preliminary grouping of enteric organisms is needed.

Mitsuda's reactions: induced by BCG in the normal Rhesus ("Macacca mulatta")

The reversals of Mitsuda's reactions induced by BCG have been objected to based on the possiblem interference of other determination causes of the phenomenon: tuberculous primo-infections, communicants of unsuspected leprosy, revearsals due to other causes, such as anti-diphteric and anti-tetanic vaccination, etc. In order to study the problem, we have used Rhesus monkeys (Macaca mulatta), which were reared in isolation, in an attempt to avoid the referred to interferences. Prior to the experiments, all animals were tested and found negative to radiograph, tuberculin and lepromin tests and were then submitted to the application of BCG vaccine (from 1 to 3 days old), in different doses and by different via. At different times, after the application of BCG, they were again submitted to the radiographic, tuberculin and lepromin tests. In the tables I to IV the experiences were summarised. From the experiments, the following conclusions were reached: 1 - From 12 Rhesus that received BCG 11 showed reversals of the Mitsuda reaction (91.7%). 2 - These reverseals took place both in tests effected shortly after BCG (from 6 days to 2 months), and tests effected much later (from 7 to 12 months after BCG). 3 - Some differences were found in the results, according to the dosis and the application via of the BCG. a) - The testicular and peritonela via (0,02g) were the only that determined strong positive Mitsuda's reactions (+++). b) - By oral via, animals that received high dosis (0.6g and 1.2 g), there resulted uniform and regular reversals, even though of low intensity (+); but from those who got small doses (0.2 g.) one showed no reversals in all tests, and the other presented reversals in the 2nd and 3rd tests only, also with low positivity (+). 4) In the 2nd and 3rd Mitsuda's reactions in the same animals, positivity was always precocious (generally within 48 hours), one getting the impression that there occurs a sensibilization of the animal body by the antigen with the repetition of the tests, even though the intensity of the reaction always remains the same. This precocious reaction (Fernandez type) occurs both shortly and long time after the application of the BCG. Its precocity depends not of the antigen only because the first Mitsuda's reaction after the BCG application occurs after some time and seems not influenced by the control lepromin test effected on the Rhesus before the BCG. 5) On the control group, the animals which received a.a.f. bacilli suspensions (Mycobacterium sp.; M. avium, and M. smegmatis), did not show reverseals of the Mitsuda's reaction. Two Rhesus, however, which received dead BCG (120ºC autoclave 1 hour), one intradermically (0.006 g) and the other orally (1.2 g), did both present reversals of the Mitsuda's reaction, with weak positivity (+). In all animals of the control-group, the allergic reactions were found negative. 6) Strong local inflammatory reactions were observed in the Rhesus that had received living BCG by intradermal via, and in the one submitted to multipunctures, there occurred the formation of a large caseous abcess. 7) The allergic tuberculinic and infratuberculinic reactions appeared dissociated from the Mitsuda's reactions: sometimes they are more precocious, occurring before of the lepromin test; on other occasions they disappear, when the Mitsuda's reactions still persist; and finally, they may be absent, when the latter occur, especially after the oral application of the BCG. 8) In Rhesus which received BCG by testicular and peritonela via, in the infratuberculinic test (0.1 ml of total BCG extract), besides the classic answer, which occurs between 48 and 96 hours, one could observe a delayed answer (15 to 20 days), represented by a non-erythematous nodule, which persists for 11-14 days.

Cutaneous allergic reactions to Triatoma infestans after xenodiagnosis

We determined the frequency of cutaneous allergic reactions to bites of Triatoma infestans during xenodiagnosis in a rural community where Panstrongylus megistrus is the only domestic vector of Trypanosoma cruzi. Localized urticaria or more intense cutaneous allergic reactions at 48 and 72 hours were observed in 86.7% and 82.1% respectively of the individuals in our study. Urticaria was more severe in children and older adults and in women than in men. The high frequency of reactions suggests either cutaneous reactivity to T. infestans without prior sensitization or cross reactivity between P. megistus and T. infestans. A single application of topical corticosteroid or antihistamine medication did not reduce the cutaneous reactions.

Milky spots reactions to schistosomal mansoni infection

Milky spots (MS), considered by the authors as a Coelomatic Lympho-myelopoietic Organ (CLMO), present a strong reactivity during experimental schistosomal mansoni infection, characterized by an increase of lymphocytes, macrophages, plasmocytes, mast cells, neutrophils and expression of eosinophil metaplasia. Intraperitoneal injection of purified Schistosoma mansoni (Sm) eggs provoked a rise in the number and size of MS, which developed the sessile marginal and pedunculated types. The authors conclude that egg antigens are, at least partially, responsible for MS reactivity during Sm infection.

Standardization of the dot enzyme-lynked immunosorbent assay (dot-ELISA) for experimental plague

A dot enzyme linked immunosorbent assay (dot-ELISA) was previously developed to detect specific antibodies in rabbits sera immunized against FIA protein obtained from Yersina pestis. This antigen was covalently linked onto the surface of dacron (polyethyleneterephthalate). Here, standard conditions are described for the optimization of this procedure: an amount of 20 ng of FIA protein was fixed onto dacron; anti-rabbit IgG peroxidase conjugate diluted 1:8,000 and 30% non-fat instant milk as blocking substance were used throughout the method. This procedure was compared with that employing nitrocellulose as solid-phase which showed to be more sensitive. However, the method based on dacron did not show false positive reactions against non-immunized rabbits sera at low antigen amount and diluted anti-IgG peroxidase conjugate.

The use of polyvinyl alcohol glutaraldehyde as solid-phase in ELISA for plague

Discs of polyvinyl alcohol cross-linked with glutaraldehyde were synthesized under acid catalysis (H2SO4). Then, the antigen F1 purified from Yersinia pestis was covalently linked to this modified polymer. Afterwards, an enzyme-linked immunosorbent assay (ELISA) was established for the diagnosis of plague in rabbit and human. The best conditions for the method were achieved by using 1.3 ¼g of F1 prepared in 0.067 M phosphate buffer, pH 7.2, containing 1 M NaCl (PBS); anti-IgG peroxidase conjugate diluted 6,000 times and as a blocking agent 3% w/v skim milk in PBS. The titration of positive rabbit serum according to this procedure detected antibody concentrations up to 1:12,800 times. The present method, the conventional ELISA and passive haemagglutination assay are compared.

Morphometric Analysis of McCoy Cells Inoculated with Cerebrospinal Fluid from Patients with Rabies

To demonstrate the potential of McCoy cells for the isolation of rabies virus from the cerebrospinal (CSF) fluid of a patient with a diagnosis of rabies, McCoy cells were inoculated with CSF from a patient with a clinical diagnosis of rabies and investigated in terms of morphometric aspect using the JAVA analysis system for the quantification of the increased size of infected cells compared to noninfected cells. The cells were also examined in terms of specific staining for the diagnosis of rabies by the method of Sellers for the observation of intracytoplasmic inclusions and by specific immunofluorescence staining for rabies virus. Infected cells showed changes in cell permeability and morphologic modifications which differed significantly compared to normal cells (P<0.001) when analyzed by the Mann-Whitney and Kruskal-Wallis tests. Intense activity of the endoplasmic reticulum was also observed, as indicated by the presence of intracytoplasmic inclusions visualized by specific staining. The present study demonstrated the isolation of rabies virus from the CSF of a patient with rabies, showing that McCoy cells can be used for the laboratory diagnosis of patients suspected to have rabies.

Cellulose acetate as solid phase in ELISA for plague

Antigen from Yersinia pestis was adsorbed on cellulose acetate discs (0.5 cm of diameter) which were obtained from dialysis membrane by using a paper punch. ELISA for human plague diagnosis was carried out employing this matrix and was capable to detect amount of 1.3 µg of antigen, 3,200 times diluted positive serum using human anti-IgG conjugate diluted 1:4,000. No relevant antigen lixiviation from the cellulose acetate was observed even after washing the discs 15 times. The discs were impregnated by the coloured products from the ELISA development allowing its use in dot-ELISA. Furthermore, cellulose acetate showed a better performance than the conventional PVC plates.