Protection of mice against gastric colonization of Helicobacter pylori by therapeutic immunization with systemic whole cell inactivated vaccines.

The protective effect of therapeutic immunization with heat inactivated Helicobacter pylori cells administered with aluminum phosphate as an adjuvant was evaluated with "Swiss albino mice" infected with H. pylori Sydney strain 1 (SS1). The presence of bacteria in histological sections of the stomach was evaluated to confirm the colonization of H. pylori. The infection dose was determined to be 1 × 108 cells which resulted to be the optimal concentration to sustain infection for required time. Systemic immunization of H. pylori 26695 and SS1 Whole cell heat inactivated vaccine were induced on mice. The IgG titer levels of high dose adjuvant vaccine of both strains were proportionate on the 7th and 14th day. Subsequently on the 21st and 28th day SS1 high dose adjuvant revealed a higher titer value. The Probability values were <0.0001 which is statistically significant. Moreover, therapeutic immunization with SS1 (WC) vaccine confers efficacious protection against H. pylori infection in mice. These results represent strong evidence for feasibility of therapeutic use of whole cell based vaccine formulations against H. pylori infection in animal model.

Detection and characterization of metallo-beta-lactamases in Pseudomonas aeruginosa by phenotypic and molecular methods from clinical samples in a tertiary care hospital.

AIMS: The aim of this study was to detect and characterize the presence of metallo-beta-lactamase (MBL) production in multidrug resistant (MDR) P. aeruginosa collected from clinical samples in a tertiary care hospital. METHODS AND MATERIALS: A total of 67 non-repetitive isolates of MDR P. aeruginosa recovered from various clinical specimens were screened for MBL production by IPM/MEM-EDTA combined disc test. Polymerase chain reaction was performed on all isolates using bla(IMP) and bla(VIM) consensus primers to characterize them genotypically. RESULTS: Among 67 P. aeruginosa isolates, 62.7% (42/67) and 70.1% (47/67) were resistant to imipenem and meropenem respectively and 47 (70.1%) were found to be MBL producers. Among this 47 MBL-producing isolates, 41 (61.1%) strains carried the bla(VIM) gene and 2 (3%) strains carried the bla(IMP) gene. Three strains were phenotypically negative but positive genotypically for bla(VIM) gene. One strain was resistant to both imipenem and meropenem but did not show phenotypic positivity. CONCLUSION: This study confirms the dissemination of bla(VIM) genes among MDR Pseudomonas aeruginosa and hence it is indispensible to identify and aptly control the threat of horizontal and vertical transfer.

Carriage of Candida species in oral cavities of HIV infected patients in South India.

Fifty-four patients with human immunodeficiency virus (HIV) infection were studied to assess the load of oral carriage of Candida spp. The mean oral Candida carriage density (30,305.93 +/- 56,643.93 CFU ml(-1)) in HIV patients was significantly higher than that seen in the control population (93.48 +/- 358.48 CFU ml(-1); P = 0.000). The mean Candida load in HIV patients with oral thrush (46,591.43 +/- 65,002.57 CFU ml(-1)) was significantly higher than in the HIV subjects without oral thrush (306.32 +/- 699.50 CFU ml(-1); P = 0.000). Non-C. albicans Candida species (56%) were more predominant than the C. albicans (44%) isolates. 25S rDNA PCR analysis of C. albicans revealed preponderance of genotype A strains. Interestingly, 42.6% of rinse specimens grew multiple Candida species, with the combination of C. albicans and C. krusei (39.1%) being the most frequent.

Comparison of cheiloscopy, odontometric, and facial index for sex determination in forensic dentistry.

BACKGROUND: The "gender determination" which is an important human identification procedure not only helps in establishing the biological profile from skeletal and dental remains but also in facial reconstruction of unidentified victims. AIM: The aim of this study is to analyze predominant types of lip prints (cheiloscopy), accuracy of mandibular canine index (MCI) (odontometric), and facial index in the study population and to identify whether any correlation among the above parameters could help forensic dentistry in solving crimes. MATERIALS AND METHODS: A pilot study was conducted in 100 individuals, 50 males and 50 females aged between 20 and 25 years. For each individual, the lip prints, MCI, and facial index measurements were recorded on the same day analyzed by two observers. All the analysis was done using SPSS version 14 assessed using t-test and Chi-square test. RESULTS: Type II pattern of lip prints is observed as common pattern among male and female. There is no significant difference in Odontometric analysis. The mean value of facial index analysis in both genders shows highly significant. CONCLUSION: A large-scale study is required in order to validate our results to arrive at definitive results and value.

Recent advances in immunodiagnosis of leprosy.

Although prevalence of leprosy is considerably reduced, the unabated emergence of about 300,000 cases worldwide indicates that the source of infection and transmission are not being addressed. Early diagnosis and treatment still remain the cornerstone of leprosy control. Many diagnostic issues hinder the correct and timely diagnosis and classification of leprosy. Delayed and missed diagnosis of infectious leprosy patients and the lack of tests to measure asymptomatic M. leprae infection in contacts also hamper the assessment of transmission of M. leprae infection. An important goal would be the development of improved diagnostic tools to diagnose difficult cases and to detect M. leprae infection before clinical manifestation. The search for an ideal immunodiagnostic tool for leprosy had gone through various phases and development over the years, with inherent limitations in the sensitivity and specificity of the immunodiagnostic tests for leprosy. With improvement in technology many modifications of previously used PGL-1 assay in the form of rapid and less expensive techniques, such as dipstick, ELISA, ML flow test, have been introduced. Many new skin test antigens with potential for improving their efficiency, such as MLSA LAM, MLCwA and their fractionates, have been studied. After the completion of genome sequencing of M. leprae in 2000, many genes that were studied in M. tuberculosis and found potential for the immunodiagnosis of tuberculosis, such as CFP-10 and ESAT-6 proteins, have been investigated in M. leprae also. Genes that are unique to M. leprae with no homologous in M. tuberculosis have been explored for novel M. leprae-specific antigens. In order to overcome the problem of cross-reactivity, a number of workers have synthesized overlapping short peptides of different M. leprae recombinant proteins and studied their sequence divergence and attempted to identify M. leprae-specific B- and T-cell epitopes. This review makes an effort to present an overview of all these developments in the field of immunodiagnosis of leprosy.

Protective role of vitamin E on the oxidative stress in Hansen's disease (Leprosy) patients.

BACKGROUND: A constellation of reactive oxygen species (ROS) capable of damaging cellular constituents generated in excess during the chronic, inflammatory, neurodegenerative disease process of leprosy. The consequences of this leads to enhanced oxidative stress and lower antioxidant status. Enzymatic antioxidants provide first line defense against ROS. We have measured the levels of oxidative stress indices like lipid peroxidation (LPO), protein carbonyls together with enzymatic antioxidants in the blood samples of control and leprosy patients. Nutritional rehabilitation by way of exogenous supplementation of functionally efficient antioxidants like vitamin E reactivates the enzymatic antioxidant system and guards against the insult caused by ROS during the pathogenesis of the disease and antileprosy chemotherapy. DESIGN: Untreated leprosy patients were selected on the basis of clinical examination and skin smear. All diagnosed untreated leprosy patients received multi drug therapy (MDT) consisting of rifampicin, dapsone and clofazimine as recommended by World Health Organization. A small number of untreated cases were selected for co-supplementation of vitamin E along with MDT. Oxidative stress indices, enzymatic and nonenzymatic antioxidant status were assayed in untreated, MDT treated and those supplemented vitamin E along with MDT. STATISTICAL METHODS: We have compared the significance in the mean+/-s.d. values of the oxidative stress indices and the levels of antioxidants using one way analysis of variance (ANOVA) between control, untreated, MDT treated and those supplemented vitamin E with MDT and the results were significant at P < 0.05. Statistical analysis of the results suggests that oral administration of vitamin E lowers oxidative stress and augments antioxidant status in affected individuals. RESULTS: Enhanced oxidative stress as evidenced by increased LPO and protein carbonyl in leprosy cases lowers the antioxidant status. Treatment with MDT has a limited impact on increased oxidative stress and decreased antioxidant status. Coadministration of vitamin E along with MDT decreases oxidative stress and activate the antioxidant status. DISCUSSION: The excess production of ROS as seen in leprosy cases could lead to degeneration of tissues and derangement of internal organs. The possible reason for the decreased antioxidant status in leprosy cases may be increased production of ROS, deranged liver function, and the free radical producing ability of drugs used in MDT of leprosy. Intervention with antioxidant supplementation like vitamin E prevents oxidative stress mediated through ROS and activates the net antioxidant status during the chronic course of the disease and antileprosy chemotherapy.

Expression of inducible nitric oxide synthase in carcinomas and sarcomas affecting the oral cavity.

CONTEXT: Inducible nitric oxide synthase (iNOS) is a cytoplasmic enzyme which plays a crucial role in the pathogenesis of oral carcinomas and sarcomas. AIMS: The objective of this study was to analyze the immunohistochemical expression of iNOS in carcinomas and sarcomas affecting the oral cavity in order to understand the possible role of iNOS in their biologic behavior and to correlate iNOS expression with lymph node metastasis in carcinomas and sarcomas. SETTINGS AND DESIGN: Patients, who attended the oral diagnosis department of Vinayaka Missions Sankarachariyar Dental College, were screened, for the purpose of the study. Besides these, paraffin-embedded tissue blocks were also retrieved from archives of the Oral and Maxillofacial Pathology Department. A total of 40 cases (20 carcinomas and 20 sarcomas) were selected for the study. SUBJECTS AND METHODS: A total of 40 cases (20 carcinomas and 20 sarcomas) were selected for the study. Five apparently normal tissues were obtained from the tumor adjacent normal tissue to be used as a control. These were subjected to immunohistochemical staining using antibody to iNOS and evaluated. STATISTICAL ANALYSIS USED: The results were analyzed using the Chi-square test. RESULTS: Among the 20 carcinomas 19 showed a positive immunoreactivity for iNOS and 1 case was negative. Among the 19 immunopositive iNOS cases of carcinomas, 15 cases showed positive lymph node metastasis. Among the sarcomas, positive immunoreactivity for iNOS was seen in 10 hard tissue sarcomas, while the remaining 10 soft tissue sarcomas were negative for iNOS expression. The results were analyzed using the Chi-square test. CONCLUSIONS: iNOS is a reliable marker for lymph node metastasis in carcinomas irrespective of the histologic grade. The high expression in carcinomas shows that the carcinomas elaborate more angiogenesis for growth compared with the sarcomas with the exception of hard tissue sarcomas.

Magnetic resonance imaging and the diagnosis of dementia.

We describe seven patients with clinical, laboratory, and CT evidence of primary degenerative dementia (Alzheimer's, Pick's). Magnetic resonance imaging demonstrated regions in the white matter consistent with cerebral infarction. MRI may be a sensitive way to differentiate multi-infarct dementia and primary degenerative dementia.

Aneurysmal compression of the anterior visual pathways.

Ten patients with aneurysmal compression of the anterior visual pathways had visual loss, unilateral in 4 and bilateral in 6. There was no typical clinical presentation. Visual loss was acute or gradual, acuity sometimes fluctuated, and visual field testing was highly variable. The aneurysms were supraclinoid (four patients), carotid-ophthalmic (two), anterior communicating-anterior cerebral (three), and intracavernous carotid (one). Nine patients had successful clipping of their aneurysm, and in one, ipsilateral common carotid ligation was performed. Postoperatively, visual acuity was improved in six cases, unchanged in three, and worse in one.

Pitfalls in computed tomographic evaluation of the cavernous sinus.

Computed tomography (CT) is the diagnostic procedure of choice in cavernous sinus evaluation. Two patients are reported with painful ophthalmoplegia due to metastatic carcinoma to the cavernous sinus in which initial CT scanning was considered to be normal. These cases illustrate the necessity of thin tomographic sections (5 mm or less), multiple projections (axial, coronal) and large doses of intravenous contrast medium (bolus and drip infusion). Careful technique is essential in order to thoroughly evaluate the parasellar region and avoid a false-negative CT examination.

Progressive multifocal leukoencephalopathy: three patients diagnosed by brain biopsy, with prolonged survival in two.

Since 1980, three immunocompromised patients have been proved to have progressive multifocal leukoencephalopathy (PML) by brain biopsy at the University of Alabama at Birmingham. Two patients presented with focal neurological findings, and the third presented with dementia. Computed tomography (CT) revealed white matter low density lesions in areas appropriate to the neurological abnormalities. Brain biopsy of areas that were abnormal on CT produced diagnostic tissue in all three patients. No patient suffered ill effects from the biopsy. Neuropathological findings on light microscopy were compatible with PML in each case, although there was diversity within the group. Involvement of gray and white matter was present in all biopsy specimens; oligodendrocytes, astrocytes, and neurons were affected. Electron microscopic demonstration of particles compatible with polyoma virus confirmed the diagnosis in each case. Immunosuppressive medication was discontinued in two of the patients; these two have survived more than 2 years after diagnosis. One of these two has gradually improved and is independent in simple activities of daily life. Brain stem and cerebellar involvement and seizure disorders have been present in all reported cases. PML can be accurately and rapidly diagnosed by brain biopsy, enabling therapeutic manipulations that may prolong survival.

Symptomatic spinal epidural lipomatosis as a complication of steroid immunosuppression in cardiac transplant patients. Report of two cases.

Patients with Cushing's syndrome may develop spinal epidural lipomatosis, an abnormal accumulation of fat in the spinal epidural space. This accumulation of fat may cause compression of the spinal cord or cauda equina with resulting neurological deficit. Two cases of symptomatic spinal lipomatosis are reported in cardiac transplant patients receiving chronic corticosteroid treatment. The literature is reviewed, and diagnostic and therapeutic considerations are discussed.

Brain tumors presenting as a seizure disorder in infants.

Seizures occur in 25% to 40% of children with supratentorial tumors and are the presenting complaint in 10% to 15%. However, when divided by age, only 2% of children with seizures as the presenting complaint of brain tumors were less than 1 year of age. Three children, ranging in age from 20 days to 7 months and seen within the past 2 years, form the basis of this report. The presenting complaint in all children was seizures. Computed tomographic (CT) scan was indicated in all children because of intractability to anticonvulsant drug therapy (one patient) and focal electroencephalographic (EEG) abnormality with clinical evidence of complex partial seizure activity (two patients). CT scan showed a contrast-enhancing mass in the medial temporal lobe in all patients. At surgery, a temporal lobe tumor was found and resected in all children. Histopathologic examination revealed a ganglioglioma, a fibrillary astrocytoma, and an anaplastic astrocytoma. All children did well postoperatively and are seizure free to date. Our experience suggests that supratentorial tumors should be considered as a cause of intractable and/or focal seizures in children under 1 year of age, and that such tumors should be attacked aggressively neurosurgically. Our experience is also in agreement with that of Tadmor et al, who have suggested that with the advent of CT scanning supratentorial tumors in this age group have been found to be more common than previously realized.

Evaluation of Mycobacterium leprae particle agglutination test, using eluates of filter paper blood spots.

A comparison of the ELISA test with the newly-developed MLPA test was carried out, using eluates of blood spots from filter paper for the detection of the anti-PGL-I antibody. A very good positive correlation was observed between these two tests. The concordance rate was found to be 92.6%, ranging from 71.4% to 100%. This nonconcordance was not found when freshly-collected samples were used. The MLPA test is simple and reliable. The use of eluates from blood spots collected on filter paper further simplifies the test in the collection and transportation of blood samples. This accurate and rapid method makes the MLPA test logistically feasible for large-scale screening. With our modification of MLPA with eluates more samples can be screened with the kit than with sera.

Indeterminate leprosy: a seroimmunological and histochemical evaluation.

An effort was made to differentiate indeterminate (IND) leprosy from other types of the paucibacillary (PB) group of leprosy and to identify among indeterminate leprosy cases those which may evolve to multibacillary (MB) leprosy, using serological, immunological and histochemical parameters. A total of 92 untreated, histologically classified (TT-19, BT-30, IND-32) patients, including 11 cases diagnosed as nonspecific dermatitis (NSD), which were clinically strongly suspected to be leprotic, were screened for antibodies against PGL-I, 35-kDa and LAM antigens. Lepromin tests and antigen demonstration in tissue by indirect immunoperoxidase staining were also carried out. Though a qualitative analysis did not differentiate, a quantitative analysis in terms of a cumulative index (CI) showed a higher antibody level amongst the indeterminate group of patients than the other groups included in PB leprosy. Also, the lepromin negative indeterminate group patients showed a higher CI than the lepromin positive cases, indicating that perhaps these may be the cases which may develop into MB leprosy. Thus, the semiquantification of antibody levels in the form of a CI may be a useful parameter to predict the possible evolution of a given case of indeterminate leprosy. Interestingly 64% of NSD cases had either antigen or antibody which indicated that they were probably cases of leprosy.

Ofloxacin-containing combined drug regimens in the treatment of lepromatous leprosy.

A total of 26 clinically diagnosed adult patients, with active untreated lepromatous leprosy, with a Bacteriological Index of 4+ or more, were admitted to the hospital of the Central Leprosy Teaching and Research institute, Chengalpattu, India, between 1989 and 1991. After prescribed investigations, the patients were randomly allocated in groups of 3 to 3 treatment regimens, namely: 1, clofazimine 50 mg daily and 300 mg once in 4 weeks + dapsone 100 mg daily (AA); 2, (AA)+ofloxacin 400 mg daily (BB); and 3, (AA)+ofloxacin 800 mg daily (CC). The drugs were administered for 56 days continuously under supervision. Sequential biopsy results on day 0, 7, 14, 28 and 56 in normal mouse footpad revealed no growth by day 28 and 56 in all patients treated with CC and BB regimens, respectively. Calculation of the proportion of viable Mycobacterium leprae through analysis of median infectious dose (ID50) showed significant differences on day 7 in the percentage of kill between the ofloxacin-containing regimens and the other. Moderate to marked clinical improvement has been observed in a significantly higher proportion of patients treated with ofloxacin-containing regimens. All the 3 regimens were well tolerated. No severe complications or side-effects to the drugs were noticed with any of the regimens that required any suspension of treatment or the administration of steroids. Addition of ofloxacin to the standard WHO recommended MDT regimen for multibacillary patients may reduce the present duration of therapy. Ofloxacin may also be considered as an alternative drug in rifampicin-resistant cases or where rifampicin is contraindicated.

Drug susceptibility of Mycobacterium leprae: a retrospective analysis of mouse footpad inoculation results from 1983 to 1997.

We analysed the results of mouse foot pad (MFP) tests performed between 1983 and 1997 in our laboratory for the cases referred with clinical suspicion of relapse/drug resistance. A total of 214 cases, with clinical suspicion of relapse/drug resistance were investigated for susceptibility to the drugs of MDT by MFP inoculation. Among 96 inoculations that showed conclusive results, 81 (84%) were fully sensitive to dapsone, suggesting that most of the clinically suspected relapse is due to drug susceptible Mycobacterium leprae. Of the remaining 15 strains (16%) found resistant to dapsone, 13 (87%) were of high grade resistance and one strain each of intermediate grade and low grade dapsone resistance, suggesting that most of the dapsone resistance is secondary in nature. No case of rifampicin resistance was found. Only one case of combined dapsone and unconfirmed clofazimine resistance was found. No other combined multidrug resistance was observed in our analysis.

Serological response of leprosy patients to Mycobacterium leprae specific and mycobacteria specific antigens: possibility of using these assays in combinations.

The serological response of 147 leprosy patients to 3 mycobacterial antigens, PGL-I, 35 kDa (Mycobacterium leprae-specific) and LAM (which is a common mycobacterial antigen) were analysed. A stronger serological response was seen amongst the MB patients than the PB patients in all the assays. The 3 antibody levels correlated positively with each other in both MB and PB cases. An overlap of seropositivity was seen between anti-PGL-I and anti-LAM (p > 0.05). A progressive increase in seropositivity and a significant difference of absorbance or titre in antibody levels in all 3 assays over increasing grades of BI were seen in the MB patients (p < 0.05). A significant difference in seropositivity between untreated and treated groups of patients was observed for anti PGL-I (p < 0.05) and antiLAM (p < 0.01) antibodies. The sensitivity, specificity and efficiency of antiPGL-I (50%; 99%; 70%), antiLAM (43%; 95%; 64%) and anti35 kDa (66%; 100%; 80%) assays taken individually were less than that of combinations of antiPGL-I/anti-35 kDa (74%; 99%; 84%) or antiPGL-I/anti-35 kDa/antiLAM (80%; 94%; 86%). The difference in the efficiency of both sets of combination of assays were not statistically significant (p > 0.05).

Non-organ specific & organ-specific auto-antibodies in hepatitis-B virus associated glomerulonephropathies.

Eighty eight patients of glomerulonephropathies (HBsAg positive 67; HBsAg negative 21) and 88 matched and healthy controls were screened for non-organ specific autoantibodies-ANA, AMA, ASMA and APCA by indirect immunofluorescent technique. The 2.3 per cent positivity in the test group and the 8 per cent positivity in the control group did not suggest the involvement of hepatitis-B virus (HBV), as an influencing or associated agent. When 48 patients with glomerulonephropathies and 23 controls were screened for liver cell membrane (LMA) and renal cell membrane antibodies (RMA) by indirect immunofluorescent technique using isolated rat hepatocytes and renal cells, 79.2 per cent LMA positivity was seen in the HBsAg positive group and 41.7 per cent in the negative group and RMA positivity was 58 per cent in the positive group and 25 per cent in the negative group. Simultaneous positivity for both LMA and RMA was recorded in 50 per cent of the HBsAg positive patients and 15.7 per cent of the negative ones. The results suggest the possibility of an organ specific autoimmune trigger more frequently in HBV associated glomerulonephropathy.

Cavernous hemangiomas of the internal auditory canal.

Cavernous hemangiomas are vascular malformations that are seldom found in the central nervous system. Four cavernous hemangiomas of the internal auditory canal and one of the cerebellopontine angle have been reported previously. We present a series of seven (six of which have never been reported) cavernous hemangiomas limited primarily to the internal auditory canal. Whereas all cases but one had marked unilateral sensorineural hearing loss, only two had preoperative facial nerve dysfunction. There are no specific symptoms or physical or audiometric findings to differentiate cavernous hemangiomas from acoustic neuromas. However, a hemangioma should be suspected when facial paralysis is present in conjunction with a small intracanalicular tumor as demonstrated by computed tomography or magnetic resonance imaging. The former reveals calcium stippling in the lesions, whereas magnetic resonance imaging reveals a high signal intensity on both T1- and T2-weighted images.