Enhancing identification and counseling skills of dental undergraduate students using a customized Tobacco Counseling Training Module (TCTM) - A piloting of the process using ADDIE framework.

BACKGROUND: Tobacco use has emerged as a major public health problem. But, most graduates in medical and dental schools receive limited systematic training. The objective of this education innovation project was to enhance dental undergraduate student's ability to identify tobacco users through oral manifestations and improve their counseling skills using a customized Tobacco Counseling Training Module (TCTM). METHODS: A TCTM for students of dentistry was developed using ADDIE framework as a guide. Content and construct validation of the module was done by six subject experts using Delphi technique for obtaining consensus. Pilot testing was done on 20 students of third year BDS. Pre- and post-intervention assessment of knowledge, attitude, self-confidence was done using learning outcomes questionnaire. Ability to correctly identify oral manifestations was assessed using extended item MCQs and tobacco counseling skills using a modified KEECC. The difference in mean scores were computed and subjected to further statistical analysis using SPSS version 22. RESULTS: There was a significant improvement in post intervention scores for mean knowledge (5.5 ± 1.4 to 13.2 ± 1.1), attitude (5.6 ± 0.9 and 8.5 ± 0.5), self-confidence (1.5 ± 0.5 and 3.1 ± 0.2), ability to correctly identify oral manifestations (5.2 ± 1.4 and 9.4 ± 0.8) and tobacco counseling skills. CONCLUSION: It is possible to introduce the module in the existing curriculum and its effectiveness evaluation shows benefit in terms of Kirkpatrick's Level 1, 2, 3 (improvement in knowledge, attitude, self-confidence, ability to identify oral manifestations, and tobacco counseling skills) of training effectiveness.

Molecular basis for the effects of zinc deficiency on spermatogenesis: An experimental study in the Sprague-dawley rat model.

INTRODUCTION: The objective of this study is to investigate the molecular mechanisms underlying the effects of zinc deficiency on spermatogenesis in the Sprague-Dawley (SD) rat. MATERIALS AND METHODS: Three groups of eight adult male SD rats were maintained for 4 weeks on a normal diet as control, zinc deficient diet and zinc deficient diet with zinc supplementation of 28 mg zinc/kg body weight respectively. Using standard techniques, the following parameters were compared between the three groups of experimental animals at the end of 4 weeks: (a) Serum zinc, magnesium (Mg), copper (Cu), selenium (Se) and cadmium (Cd), (b) serum sex hormones, malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase (GPX), (c) interleukin-4 (IL-4), tumor necrosis factor-alpha (TNF-α), Bcl-2, Bax and caspase-3 expression in the testes, (d) assessment of apoptosis of testicular cells using electron microscopy and (e) testicular volume and histology using the orchidometer and Johnsen score, respectively. RESULTS: The zinc deficient group showed a reduction of testicular volume, serum concentrations of Zn, Cu, Se, Mg, SOD, GPX, IL-4, Bcl-2 and testosterone (P < 0.05), as well as increased levels of serum Cd, MDA and tissue TNF-α, Bax, caspase-3 and apoptosis of the germ cells (P < 0.05) compared with control and zinc supplementation groups. CONCLUSION: Zinc deficiency is associated with impaired spermatogenesis because of reduced testosterone production, increased oxidative stress and apoptosis. These findings suggest that zinc has a role in male reproduction.

On-line capacity-building program on "analysis of data" for medical educators in the South Asia region: a qualitative exploration of our experience.

BACKGROUND AND OBJECTIVE: In medical education, using the World Wide Web is a new approach for building the capacity of faculty. However, there is little information available on medical education researchers' needs and their collective learning outcomes in such on-line environments. Hence, the present study attempted: 1)to identify needs for capacity-building of fellows in a faculty development program on the topic of data analysis; and 2) to describe, analyze and understand the collective learning outcomes of the fellows during this need-based on-line session. MATERIALS AND METHODS: The present research is based on quantitative (on-line survey for needs assessment) and qualitative (contents of e-mails exchanged in listserv discussion) data which were generated during the October 2009 Mentoring and Learning (M-L) Web discussion on the topic of data analysis. The data sources were shared e-mail responses during the process of planning and executing the M-L Web discussion. Content analysis was undertaken and the categories of discussion were presented as a simple non-hierarchical typology which represents the collective learning of the project fellows. RESULTS: We identified the types of learning needs on the topic 'Analysis of Data' to be addressed for faculty development in the field of education research. This need-based M-L Web discussion could then facilitate collective learning on such topics as 'basic concepts in statistics', tests of significance, Likert scale analysis, bivariate correlation, and simple regression analysis and content analysis of qualitative data. CONCLUSIONS: Steps like identifying the learning needs for an on-line M-L Web discussion, addressing the immediate needs of learners and creating a flexible reflective learning environment on the M-L Web facilitated the collective learning of the fellows on the topic of data analysis. Our outcomes can be useful in the design of on-line pedagogical strategies for supporting research in medical education.

Applications of fluorodeoxyglucose positron emission tomography in the diagnosis of infection.

The purpose of this study was to assess the accuracy of fluorodeoxyglucose positron emission tomography (FDG PET) in diagnosing infection in a large population of patients and in a variety of clinical circumstances where the performance of conventional imaging modalities has been questioned. We retrospectively analysed 167 FDG PET scans obtained to evaluate 175 anatomical sites for the presence of infection. The major indications for the scans were (1) complicated orthopaedic hardware (n=97), (2) chronic osteomyelitis (n=56), and (3) other (n=14: six fever of unknown origin, three vascular grafts, and five soft tissue). We assessed the overall diagnostic accuracy of FDG PET for each of these indications. In addition, we further analysed this modality's effectiveness by grouping the scans into specific clinical situations. A final diagnosis was made on the basis of surgical pathology and clinical follow-up for a minimum of 6 months. The overall accuracy of FDG PET in evaluating orthopaedic hardware was 96.2% for hip prosthesis, 81% for knee prosthesis, and 100% in 15 patients with other orthopaedic devices. Among the patients in our sample suspected of having chronic osteomyelitis, the accuracy was 91.2%. FDG PET was inaccurate in three cases of fever of unknown origin and accurate in all vascular graft and soft tissue infections. In 49 patients with a clinically apparent soft-tissue infection, FDG PET was able to detect or exclude underlying osteomyelitis with an accuracy of 92.3%. Among the 23 patients who had recent orthopaedic procedures, FDG PET imaging was accurate in 87% of cases. It is concluded that FDG PET is a highly effective imaging modality in the assessment of patients with suspected infection.

The importance of the location of fluorodeoxyglucose uptake in periprosthetic infection in painful hip prostheses.

Ten per cent of patients with hip replacement will eventually complain of significant pain after surgery, often requiring a revision arthroplasty. The majority of these patients experience aseptic loosening rather than infection. Despite significant advances made in diagnostic imaging, distinguishing infection from aseptic loosening remains a significant challenge. Imaging using fluorodeoxyglucose (FDG) positron emission tomography (PET) has been reported to have excellent sensitivity in detecting infections associated with hip prostheses. However, in some studies, a high rate of false positive results has been reported, especially when increased tracer uptake adjacent to the prosthesis (which is not surrounded by bone) is used as the sole criterion for diagnosing infection. The objective of this investigation was to determine the optimal criteria for diagnosing periprosthetic infection, thereby avoiding false positive results in this setting. A total of 41 total hip arthroplasties from 32 patients and for whom complete clinical follow-up was available were included in this analysis. The location and intensity of FDG uptake were determined for each scan. Final diagnosis was made by microbiology, histopathology, surgical findings and clinical follow-up. Patients who did not undergo surgery were followed up to at least 9 months. Twelve patients were proven eventually to have periprosthetic infection. Images from 11 of these patients displayed increased tracer uptake along the interface between bone and prosthesis. The intensity of the increased tracer uptake varied from mild to moderate, with standardized uptake values less than 2. In contrast, images from uninfected, loose hip prostheses revealed very intense uptake around the head or neck of the prosthesis with standardized uptake values as high as 7. It is concluded that the intensity of increased FDG uptake is less important than the location of increased FDG uptake when FDG PET is used to diagnose periprosthetic infection in patients with hip arthroplasty. Using increased uptake as the sole criterion for diagnosing infection will result in false positive results in this setting.

Expression and partial purification of a recombinant secretory form of human liver carboxylesterase.

Serine-dependent carboxylesterases (EC 3.1.1.1) are found in a variety of tissues with high activity detected in the liver. Carboxylesterases (CaE) hydrolyze aliphatic and aromatic esters, and aromatic amides, and play an important role in the detoxification of xenobiotic chemicals that contain organophosphate (OP) compounds. The detoxifying ability of CaE is limited by its low concentration in serum where it encounters OP compounds. Studies in our laboratory have shown that a pRC/CMV-hCaE plasmid construct, stably integrated into 293T cells, expresses a human liver CaE in culture. However, the enzyme remained inside the cell and reached a low steady-state level of expression. The goals of this study were to overexpress a functional human liver CaE from a recombinant cDNA in a human cell line and to isolate and purify the recombinant protein. To accomplish these goals, a single amino acid change was made in the C-terminal retrieval signal, HIEL (His-Ile-Glu-Leu), of human liver CaE. The mutation produced a unique Eco47III restriction site, which aided in clone selection. The recombinant plasmid, pRc/CMV-mhCaE, was isolated and stably integrated into human 293T cells. Expression of the altered cDNA resulted in secretion of an active CaE up to levels of 500 enzyme units per liter of growth medium. Secretory CaE displayed isoelectric focusing patterns similar to those of the native enzyme with no observable changes in activity. The secreted enzyme was partially purified by hydrophobic interaction chromatography and Cibacron blue affinity chromatography. Partial enzyme purification was achieved, and CaE retained a high level of enzymatic activity.

Prevalence and correlates of fissure foot in a rural area in Tamilnadu.

Fissuring of feet is a common but neglected problem in India. In this paper an attempt has been made to determine the prevalence of fissuring of feet in a rural village in Tamilnadu. In addition the relationship of fissuring to age, sex, occupation, non-use of footwear and weight is determined Over all prevalence is found to be 48% for age's 15 years and above. The prevalence is found to be higher in females (58.4%) than in males (33.3%) and it is seen more among the housewives (63.7%) and among the farmers (41.9%). Low weight and non-use of footwear are significantly associated with fissuring. 40% of the affected group felt that this is more acute during winter. We conclude that fissuring of feet is a significant problem.

Expression of myosin isoforms in smooth muscle cells in the corpus cavernosum penis.

Corpus cavernosum smooth muscle (CCSM) in the penis is unique in that it exhibits a high resting tone and, on stimulation, the muscle cells relax, allowing cavernous spaces to fill with blood, which results in an erection (tumescence). During detumescence, the muscle cells contract and return to the state of high resting tone. This study was undertaken to determine whether CCSM with these unique properties contains myosin isoforms typical of aorta or bladder smooth muscles, muscles that exhibit tonic and phasic characteristics, respectively. RT-PCR revealed that normal CCSM contains an SM2/SM1 mRNA ratio of 1.2:1 (similar to the rabbit aorta). Approximately 31% of the myosin heavy chain transcripts possess a 21-nt insert (predominant in bladder smooth muscle but not expressed in aorta) that encodes the seven-amino acid insert near the NH2-terminal ATP binding region in the head portion of the myosin molecule found in SMB, with the remaining mRNA being noninserted (SMA). Quantitative competitive RT-PCR revealed that the CCSM possesses approximately 4.5-fold less SMB than the bladder smooth muscle. Western blot analysis using an antibody specific for the seven-amino acid insert reveals that both SM1 and SM2 in the CCSM contain the seven-amino acid insert. Furthermore, SMB containing the seven-amino acid insert was localized in the CCSM by immunofluorescence microscopy using this highly specific antibody. The analysis of the expression of LC17 isoforms a and b in the CCSM revealed that it is similar to that of bladder smooth muscle. Thus the CCSM possesses an overall myosin isoform composition intermediate between aorta and bladder smooth muscles, which generally express tonic- and phasiclike characteristics, respectively. Two-dimensional gel electrophoresis showed a relatively low level (approximately 10%) of Ca2+-dependent light-chain (LC20) phosphorylation at the basal tone, which reaches approximately 23% in response to maximal stimulation. The presence of noninserted and inserted myosin isoforms with low and high levels of actin-activated ATPase activities, respectively, in the CCSM may contribute to the ability of the CCSM to remain in a state of high resting tone and to relax rapidly for normal penile function.

Differences in contractile protein content and isoforms in phasic and tonic smooth muscles.

The basis of tonic vs. phasic contractile phenotypes of visceral smooth muscles is poorly understood. We used gel electrophoresis and quantitative scanning densitometry to measure the content and isoform composition of contractile proteins in opossum lower esophageal sphincter (LES), to represent tonic muscle, and circular muscle of the esophageal body (EB), to represent phasic smooth muscle. The amount of protein in these two types of muscles is similar: approximately 27 mg/g of frozen tissue. There is no difference in the relative proportion of myosin, actin, calponin, and tropomyosin in the two muscle types. However, the EB contains approximately 2.4-times more caldesmon than the LES. The relative ratios of alpha- to gamma-contractile isoforms of actin are 0.9 in the LES and 0.3 in EB. The ratio between acidic (LC17a) and basic (LC17b) isoforms of the 17-kDa essential light chain of myosin is 0.7:1 in the LES, compared with 2.7:1 in the EB. There is no significant difference in the ratios of smooth muscle myosin SM1 and SM2 isoforms in the two muscle types. The level of the myosin heavy chain isoform, which contains the seven-amino acid insert in the myosin head, is about threefold higher in the EB compared with LES. In conclusion, the esophageal phasic muscle in contrast to the tonic LES contains proportionally more caldesmon, LC17a, and seven-amino acid-inserted myosin and proportionally less alpha-actin. These differences may provide a basis for functional differences between tonic and phasic smooth muscles.

Comparison of the Amplicor HIV-1 monitor test and the nucleic acid sequence-based amplification assay for quantitation of human immunodeficiency virus RNA in plasma, serum, and plasma subjected to freeze-thaw cycles.

The Amplicor HIV-1 Monitor test was compared to the nucleic acid sequence-based amplification (Nasba) assay system for the quantitation of human immunodeficiency virus (HIV) RNA in three different types of clinical samples: plasma, serum, and plasma subjected to freeze-and-thaw cycles. Each assay detected HIV RNA in the same 73 (90%) of 81 samples tested, and the quantitative results obtained with the two assays were significantly correlated. Both assays detected higher RNA levels in patients with CD4+ cell counts lower than 200 cells/mm3 than in patients with CD4+ cell counts higher than 200 cells/mm3. In addition, RNA levels in plasma higher than 5 logs predicted higher numbers of clinical events than did RNA levels in plasma lower than 5 logs. Quantitation of HIV RNA in paired plasma and serum samples showed lower HIV RNA content in serum than in the paired plasma sample, with mean differences between HIV RNA contents of plasma and serum of 0.54 and 0.28 log RNA copy/ml by the Nasba assay and the Amplicor HIV-1 Monitor assay, respectively. No significant loss of HIV RNA was detected with either assay in plasma samples subjected to multiple freeze-and-thaw cycles. These studies demonstrate that the Nasba and Amplicor assays perform similarly with plasma and serum samples. Further, the results indicate that freeze-and-thaw cycles do not result in significant loss of detectable HIV RNA.

Effect of stavudine on human immunodeficiency virus type 1 virus load as measured by quantitative mononuclear cell culture, plasma RNA, and immune complex-dissociated antigenemia.

The antiviral effect of stavudine (2', 3'-didehydro-3'-deoxythymidine) against human immunodeficiency virus (HIV) type 1 was measured in 15 HIV-infected patients at baseline and at weeks 4, 10, 22, 34, and 52 of therapy. Patients received 0.1, 0.5, 1.0, or 2.0 mg/kg/day of stavudine. At all time points examined during the 52 weeks of therapy, the median virus titers in peripheral blood mononuclear cells were decreased 1-2 logs, and median immune complex-dissociated antigen levels were reduced 37%-67% compared with baseline values. Plasma RNA content measured by polymerase chain reaction was reduced approximately 0.5 log from baseline median values at both time points examined (weeks 10 and 52). These data demonstrate that stavudine has a substantial and durable antiviral effect.

Stability of free and complexed human immunodeficiency virus type 1 antigen at 4 degrees C and at room temperature.

Free and immune-complex-dissociated (ICD) human immunodeficiency virus type 1 (HIV-1) antigenemias in serum specimens stored at room temperature (RT) and 4 degrees C for 1 to 35 days were evaluated. At all time points examined, there was no significant loss in detectable levels of ICD HIV-1 antigen at either RT or 4 degrees C. Free HIV-1 antigen was not stable in serum samples stored at RT for more than 2 days but was stable in samples stored at 4 degrees C for up to 4 days. Loss of free antigen occurred more rapidly in samples with high antigen content at baseline. Use of the ICD antigen assay allowed accurate quantitation of antigen in samples stored at RT or 4 degrees C for as long as 1 month.

Comparative performance of peptide-, recombinant protein-, and viral lysate-based enzyme immunosorbent assays for the detection of HIV-1 antibodies.

The performance of peptide-, recombinant protein-, and whole virus lysate-based enzyme immunosorbent assays (EIAs) for the detection of antibodies to the human immunodeficiency virus type 1 (HIV-1) was compared on a panel of 245 routine samples, a panel of low-positive samples, four seroconversion panels, and serial dilutions of five known positive samples. Of the 245 routine samples, 83 were confirmed to be HIV-1 antibody positive by Western blot and were reactive in the three EIAs used. Agreement between the three EIA tests was also 100% for all 162 negative samples. Although there was no significant difference in the performance of the three types of assays in seroconversion panels, the whole virus and recombinant protein-based EIAs detected 15 of 15 samples in the low-positive panel whereas only 10 of 15 samples were reactive when the peptide-based EIA was used. In addition, evaluation of diluted positive samples suggested that the virus lysate-based EIA was more sensitive than the peptide- and recombinant-based EIAs. These results show that although the three types of assays performed well on routine serum panels, differences in sensitivities were demonstrated when performance panels were evaluated. The data suggest that seroconversion and low-positive performance panels should be included in evaluations of new generations of EIAs for HIV-1 antibodies.

Comparison of enzyme-linked immunosorbent and indirect immunofluorescence assays for the detection of human T-cell lymphotropic virus type-I antibodies in sera from rural Haiti.

Serum samples were obtained from 340 healthy individuals without evidence of neurologic disease living in rural Haiti. Sera were screened for antibodies to human T-cell lymphotropic virus type I (HTLV-I) using two commercially available enzyme immunoassays (EIA) and by an indirect immunofluorescence assay (IFA) using a mixture of uninfected H9 cells and HTLV-I-infected MT-2 cells. Repeatedly positive samples were confirmed by Western blot (WB). Results with the two EIA systems were concordant and detected 13 positive samples, each of which was confirmed by WB. Only 9 (69%) of 13 WB-positive sera were detected by IFA, and four additional samples, positive by IFA, could not be confirmed by WB. The prevalence of HTLV-I seropositivity in this selected rural Haitian population was 3.8% (13 of 340).