Composite skin contact smears in multibacillary leprosy patients

"Composite skin contact smears" technique was used in 20 multibacillary leprosy patients to find out the presence of AFB on their intact skin. One hundred and twenty one AFB were found in 930 sq.cm. area of skin surface examined. Seventy-five AFB were solidly staining and 46 were granular. The bacilli were found in all the sites examined. Immunofluorescent staining showed M. leprae in one out of four patients. The number of bacilli discharged from the intact skin seems adequate for the transmission of leprosy considering the large surface area of the skin and evidence in mouse experiments that multiplication of M. leprae can be obtained after inoculation of very small number of bacilli.     

Inter- and intra-laboratory variation in the reporting of skin smears in leprosy

This paper defines the variations in the reporting of skin smears between a base and field laboratory in a leprosy control program. Ten percent of all slides read by the field laboratory in a control area were re-read by the base laboratory. There was almost no variation in the reporting of negative slides, but a variation of 1 + was present in approximately 92% of positive slides. Thus, there was agreement in approximately 8% of positive slides. This paper also defines the variations in the reporting of positive slides under "ideal" conditions by describing the results of a study on intra- and inter-observer variations among technicians at the base laboratory. There was between 45% and 55% agreement within observers and about 36% agreement between observers. The results of both studies are compared. Simple guidelines are derived to monitor the reporting of skin smears in leprosy control programs.     

"Nose-blow" smears in multibacillary leprosy patients

332 nose-blow specimens have been examined from 73 untreated multibacillary patients of leprosy before and periodically after they were put on a maximal, minimal or an intermediate multi-drug regimen. 80% of these specimens were found to be positive for acid fast bacilli initially. Bacillary positivity rate was more in samples containing pus or blood. Bacilli were seen in LL, LI as well as BL patients. Nearly half of the cases became negative for AFB in their nose-blow specimens within 3 months of initiation of treatment whereas none of these patients has become negative in skin smears. However, a few cases have continued to discharge bacilli in their nasal secretions even after 12 months of multi-drug regimen therapy.     

qPCR detection of Mycobacterium leprae in biopsies and slit skin smear of different leprosy clinical forms

Leprosy, whose etiological agent is Mycobacterium leprae, is a chronic infectious disease that mainly affects the skin and peripheral nervous system. The diagnosis of leprosy is based on clinical evaluation, whereas histopathological analysis and bacilloscopy are complementary diagnostic tools. Quantitative PCR (qPCR), a current useful tool for diagnosis of infectious diseases, has been used to detect several pathogens including Mycobacterium leprae. The validation of this technique in a robust set of samples comprising the different clinical forms of leprosy is still necessary. Thus, in this study samples from 126 skin biopsies (collected from patients on all clinical forms and reactional states of leprosy) and 25 slit skin smear of leprosy patients were comparatively analyzed by qPCR (performed with primers for the RLEP region of M. leprae DNA) and routine bacilloscopy performed in histological sections or in slit skin smear. Considering clinical diagnostic as the gold standard, 84.9% of the leprosy patients were qPCR positive in skin biopsies, resulting in 84.92% sensitivity, with 84.92 and 61.22% positive (PPV) and negative (NPV) predictive values, respectively. Concerning bacilloscopy of histological sections (BI/H), the sensitivity was 80.15% and the PPV and NPV were 80.15 and 44.44%, respectively. The concordance between qPCR and BI/H was 87.30%. Regarding the slit skin smear, 84% of the samples tested positive in the qPCR. Additionally, qPCR showed 100% specificity, since all samples from different mycobacteria, from healthy individuals, and from other granulomatous diseases presented negative results. In conclusion, the qPCR technique for detection of M. leprae using RLEP primers proved to be specific and sensitive, and qPCR can be used as a complementary test to diagnose leprosy irrespective of the clinical form of disease.     

Rosette-forming cells in patients with treated leprosy

White blood cell counts and the percentages and absolute numbers of lymphocytes in the peripheral blood of active lepromatous (BT, BB, BL) leprosy patients, patients with borderline leprosy, and normal controls were determined. Lepromatous patients showed decreased leukocyte counts and elevated percentages of lymphocytes, resulting in normal absolute lymphocyte counts. The proportion of peripheral blood mononuclear cells forming "active" rosettes, standard (4 degrees C overnight) rosettes, and "high affinity" (29 degrees C for 1 hr) rosettes with sheep erythrocytes, and rosettes with EAC were determined. Lepromatous patients, compared with normal controls, had decreased "active" rosettes, standard rosettes, and "high affinity" rosettes with sheep erythrocytes with an increase in the nonrosetting proportion. Both lepromatous and borderline leprosy patients showed increased percentages of EAC rosettes compared with normal controls.     

Immunological profile of treated lepromatous leprosy patients

The immune responses of 19 treated lepromatous patients who had remained smear negative for a long period were assessed for specific cell-mediated immunity (CMI), anti-Mycobacterium leprae antibodies and cytokine release in response to challenge with M. leprae soluble antigen (MLSA). All of these patients remained anergic to Mitsuda lepromin. Lymphoproliferation in response to M. leprae antigen was noted in only two patients. Significant reduction in the phenolic glycolipid I (PGL-I) antibody response in treated patients with no difference in the M. leprae 35-kDa antibody response was observed when these responses were compared with those of active lepromatous patients. More treated patients produced interleukin-2 (IL-2) and interferon gamma (IFN-gamma) than did active patients. On the other hand, fewer treated patients produced IL-10 than did active patients. These limited findings suggest that the host immune response makes an attempt toward upregulation of CMI in some treated LL/BL patients.     

Food intolerance and skin prick test in treated and untreated irritable bowel syndrome

AIM: To correlate the clinical features of treated and untreated patients with irritable bowel syndrome (IBS) to the results of skin prick test (SPT) for food and inhalant allergens. METHODS: We recruited 105 subjects to form three different target groups: treated group (n=44) undergoing treatment for IBS, untreated group (n=31) meeting the RomeⅡcriteria without treatment for IBS, control group (n=30) with no IBS symptoms. RESULTS: SPT results were different among the three groups in which SPT was positive in 17 (38.6%) treated patients, in 5 (16.1%) untreated patients and in 1 (3.3%) control (P<0.01). The number of positive SPTs was greater in the IBS group than in the control group (P< 0.001). The number of positive food SPTs was higher in the treated IBS group than in the untreated IBS group (P=0.03). CONCLUSION: Positive food SPT is higher in IBS patients than in controls.     

Unfavorable cardiovascular risk profiles in untreated and treated psoriasis patients

Psoriasis is a chronic inflammatory skin disease that is associated with an increased cardiovascular risk profile. The systemic inflammation present in psoriasis, various systemic treatments for psoriasis and an increased prevalence of unhealthy life style factors may all contribute to this unfavorable risk profile. The purpose of this article is to provide an overview of what is known about these risk factors in psoriasis, the way they influence the cardiovascular risk of psoriasis patients, and what can be done to reduce this risk. Genetic studies demonstrate that psoriasis and cardiovascular disease share common pathogenic features in which, for example inflammatory cytokines like TNF-alpha and IL-1 play an important role. The chronic inflammation in psoriasis has an unfavorable effect on the cardiovascular risk profile. Multiple cardiovascular risk factors seem to be influenced; the blood pressure, oxidative stress, dyslipidemia, endothelial cell dysfunction, homocysteine levels and blood platelet adhesion. Moreover, classic cardiovascular risk factors like smoking and obesity that have an increased prevalence among patients with psoriasis, indirectly also worsen the cardiovascular risk profile by stimulating the psoriasis activity. Systemic treatments in psoriasis reduce the cardiovascular risk by diminishing the inflammation, but it should be taken into account that most therapies also have adverse cardiovascular effects like dyslipidemia, hyperhomocysteinemia and hypertension. As a consequence preventive measures may be indicated at least during long-term treatments. Prospective research is warranted to accurately estimate the increased cardiovascular risk in psoriasis, to determine the underlying processes and to consider preventive measures according to the absolute risk of cardiovascular disease. The present overview provides data to advice health care providers to pay more attention to the cardiovascular risk profile in psoriasis patients.     

Antroduodenojejunal motor activity in untreated and treated celiac disease patients

Background and Aim:  Patients with celiac disease may present with abnormal upper gut motor activity. However, it is not known if these abnormalities persist after the introduction of a gluten-free diet. The present study aimed to compare antroduodenojejunal motor variables recorded in untreated celiac patients with those of celiac patients given a gluten-free diet and healthy volunteers.
Methods:  Eleven untreated celiac disease patients, 12 age- and sex-matched celiac patients on a gluten-free diet (at least 12 months), and 33 controls entered the study. Antroduodenojejunal motility was recorded for 6 h during fasting and for 3 h after a standard meal by means of a perfused, multiple lumen catheter.
Results:  More than 80% of untreated celiac patients had discrete motor abnormalities of the upper gut, in both fasting and fed recordings, compared to the other subjects. Patients on a gluten-free diet also showed motor abnormalities, albeit to a lesser extent. In these patients histological evaluation showed the persistence of mild mucosal abnormalities.
Conclusions:  Upper gut motor abnormalities are frequent in patients with celiac disease, even in those on a gluten-free diet. In the latter group, these abnormalities may suggest an incomplete adherence to the dietary regimen.     

Anaemia, iron-related measurements and erythropoietin levels in untreated patients with active leprosy.

The mechanisms responsible for anaemia in leprosy were studied prior to the institution of therapy in 56 patients with active disease. Haematological indices, iron-related measurements, inflammatory markers and erythropoietin levels were assessed, with bone-marrow studies being performed on anaemic patients. Anaemia was more common in the patients with lepromatous leprosy (85.7%) than it was in the rest of the group (19%). The lepromatous group exhibited the disordered iron transport of the anaemia of chronic disorders in that they had a significantly lower mean serum iron level (P less than 0.05), and a mildly raised serum ferritin concentration. Anaemic lepromatous patients also showed a blunted erythropoietin response compared with controls with non-inflammatory anaemia. A subgroup of five anaemic subjects displayed apparently adequate transport of iron to the erythroid marrow (normal percentage transferrin saturations and appropriate sideroblast counts) and the blunted erythropoietin response appeared to be the dominant factor in the pathogenesis of their anaemia. Analysis of inflammatory markers revealed that while the erythrocyte sedimentation rate was very high in the lepromatous subjects, there was no concomitant rise in C-reactive protein concentration. This suggests the presence of a disordered cytokine-mediated acute phase response in the condition.     

Chromosomal aberrations in cultures of skin fibroblasts of leprosy patients

A search for structural and numerical chromosomal aberrations was made on metaphases obtained by culturing in vitro skin fibroblasts of 16 leprosy patients (10 lepromatous, 2 borderline, and 4 tuberculoid cases) and 2 healthy individuals used as controls. The data were analyzed taking into account sex, age, race, form of leprosy, bacterial index, type of therapy, and age of the fibroblast cultures. Structural chromosomal aberrations are the only type of chromosomal abnormality that may be accepted as significantly increased in the skin fibroblasts of leprosy patients who are under treatment with dapsone alone or with combined therapy.     

A comparison of antineutrophil cytoplasmic antibody prevalence in patients treated and untreated for hyperthyroidism

We aimed to compare the prevalence of antineutrophil cytoplasmic antibody (ANCA) and its subgroups between on-treatment (with anti-thyroid drugs; propylthiouracil, methimazole) and untreated patients with hyperthyroidism in our unit. Overall 78 consecutive patients were enrolled in the study; 45 patients were on-treatment (female/male 31:14) and 33 were newly diagnosed (female/male 20:13). We have studied ANCA, perinuclear-ANCA (p-ANCA), cytoplasmic-ANCA (c-ANCA), myeloperoxidase-ANCA (mpo-ANCA), and proteinase 3-ANCA (pr3-ANCA) in sera of all the patients. The data about clinical status, laboratory tests, and physical examination and mean duration of treatment in treated group were recorded. There was no statistically significant difference between the two groups for ANCA, c-ANCA, and pr3-ANCA (P=0.13, P=0.07, and P=0.63 respectively). p-ANCA and mpo-ANCA prevalences were significantly higher in on-treatment group than in untreated group (P=0.04 and P=0.01, respectively). The mean duration of treatment was 17 months in on-treatment group. The use of antithyroid drugs (propylthiouracil, methimazole) seems to be correlated with increased prevalence of ANCA. These drugs may especially increase p-ANCA and mpo-ANCA positivity.     

Secondary hyperparathyroidism in patients with untreated and treated congestive heart failure

BACKGROUND: The congestive heart failure syndrome includes a systemic illness with wasting of soft tissues and bone. We hypothesized secondary hyperparathyroidism (HPT) would be found in hospitalized patients with decompensated congestive heart failure (CHF), where secondary aldosteronism is expected, and who were either untreated or treated medically. METHODS: In 9 consecutive patients (7 males, 2 females; 8 African-American, 1 Caucasian; 33-60 yrs) admitted to the Regional Medical Center during a 28-day period with chronic left ventricular systolic dysfunction (EF<35%) and decompensated CHF (5 untreated; 4 treated with an angiotensin converting enzyme inhibitor, furosemide, and small-dose spironolactone), we measured: plasma parathyroid hormone (PTH); serum calcium corrected for albumin, magnesium, and phosphorus; serum creatinine and calculated creatinine clearance. RESULTS: Plasma PTH was elevated above the normal range (6-65 pg/mL) in both untreated and treated patients with CHF (204+/-60 and 134+/-14 pg/mL, respectively). Serum corrected calcium was normal (8.4-10.2 mg/dL) in both untreated and treated CHF (9.7+/-0.l and 9.1+/-0.2 mg/dL, respectively) as were serum magnesium and phosphorus. Calculated creatinine clearance did not differ between untreated and treated patients (74+/-15 and 83+/-21 mL/min, respectively). CONCLUSIONS: Secondary HPT was found in 5 untreated and 4 treated patients consecutively hospitalized over a 28-day period with decompensated CHF. Corrected serum calcium was normal. Plasmaionized calcium, a determinant of PTH secretion, was not measured. Although vitamin D levels were not assessed, the presence of hypovitaminosis D in these housebound patients with symptomatic CHF cannot be discounted. HPT may contribute to the systemic illness that accompanies CHF, including bone wasting.     

Study of apparently uninvolved skin in leprosy as regards bacillary population at various sites.

Slit smears form 16 LL and 4 BL patients were taken from scalp, axilla, inguinal regions and apparently involved skin patch. The bacilli were found in 100% LL and 75% BL patients at all sites. Scalp showed AFB in all LL and 3 out of 4 BL cases. No lesions were seen on the scalp. Bacterial morphology showed no uniform pattern. Contrary to belief, no immune zones were found on the skin as judged by results of bacteriological examination. Our studies do not support the view that the leprosy bacillus has a predilection for sites with relatively low temperature as far as human leprosy is concerned.     

Vaccination and skin test studies on the children of leprosy patients

In an attempt to achieve maximal skin-test positivity to leprosin A in children of leprosy patients living in Baba Baghi Leprosy Sanatorium in Iranian Azerbaijan, two new vaccines have been employed. Children without scars of previous BCG and without response to leprosin A were given a vaccine containing 10(8) viable units of BCG Glaxo plus 10(7) killed Mycobacterium vaccae per dose (vaccine B). Children with BCG Pasteur (Teheran) scars but without response to leprosin A were given a vaccine containing 10(8) killed M. vaccae alone (vaccine D). Eight years later skin testing was repeated, and both new vaccines were found to have significantly increased the numbers of children responding to leprosin A above the level that would have been expected had they received BCG Pasteur alone. This increase was due in large part to increases in the proportions of individuals responding to group i (common mycobacterial) antigens, and known as category 1 responders. The use of suspensions of killed M. vaccae in conjunction with BCG may represent a considerable advance in inducing protection from multibacillary leprosy in close contacts of leprosy patients if leprosin A positivity is truly a correlate of protective immunity. A comparison, using the same criteria, with the other proposed vaccines for leprosy would be very interesting.