Diagnosing melanoma: how do we assess how good we are?

BACKGROUND: Evaluating and improving diagnostic accuracy in identification of melanomas is important for both conservation of healthcare resources and reduction in patient morbidity. Useful indicators in assessing this accuracy include the number needed to treat (NNT) and the benign:malignant (B:M) ratio. Both of these methods lack sensitivity, as they do not account for the ability to detect early or in situ melanomas. AIM: To assess the NNT and B:M ratio for a busy hospital serving a population of 650,000 over a 5-year period, and to assess a new ratio of diagnostic accuracy by calculating the ratio of invasive (malignant) melanomas to melanoma in situ (MM:MMIS) as a marker of sensitivity. METHODS: This was a retrospective analysis of data on all melanocytic lesions excised during two separate years (2006 and 2011) with a 5-year interval between them. The lesions were divided into benign naevi (BN), dysplastic naevi (DN), MMIS and MM. RESULTS: In 2006, 650 melanocytic lesions were excised (462 BN/DN, 45 MMIS, 143 MM). The NNT was 3.46, the B:M ratio was 2.46 and the MM:MMIS ratio was 3.18. In 2011, 730 melanocytic lesions were excised (464 BN/DN, 99 MMIS, 167 MM). The NNT was 2.74, the B:M ratio was 1.74 and the MM:MMIS ratio was 1.69. CONCLUSIONS: The NNT and B:M ratios from our study compare favourably with those in the published literature. The fall in the MM:MMIS and B:M ratios over this 5-year study appears to be an indicator of the ability to detect early disease and is probably secondary to the changes to our skin cancer service. This study may encourage physicians to aim not only for low B:M ratios but also low MM:MMIS ratios.

Troponin I Elevation after Elective Percutaneous Coronary Interventions: Prevalence and Risk Factors.

Percutaneous coronary intervention (PCI) is one of the most important modalities of treatment for coronary artery disease (CAD). Minor extents of injury to the myocardium have been observed even after successful PCI. This peri-procedural injury might therefore reduce some of the beneficial effects of coronary revascularization. The objective of this hospital based comparative observational study was to determine the prevalence of post procedural Cardiac troponin I (cTnI) elevation after elective PCI and also to find out the relation with risk factors such as age, sex, body mass index (BMI), smoking, anemia, diabetes mellitus, hypertension, dyslipidemia, family history, left ventricular dysfunction, renal insufficiency, type of stent, number of stent and length of stent. This was a hospital based comparative observational study carried out in the Department of Cardiology, Chattogram Medical College Hospital (CMCH), Chattogram, Bangladesh from July 2018 to June 2019. A total of 50 patients who underwent elective PCI were included as sampled by purposive sampling method. Serum cTnI was measured by FIA8000 quantitative immunoassay analyzer with an analytical measurement before and at 24 hours of PCI. Value >1.0ng/ml was considered elevated. Univariate and multivariate analysis were applied to assess predictors for the occurrence of post-procedural elevation of cTnI. The mean±SD age of the study population was 54.96±9.1 years (range 35-74 years) and 34(68.0%) patients were male. Regarding cardiovascular risk factors, 17(34.0%) patients had diabetes mellitus, 27(54.0%) had dyslipidemia, 30(60.0%) had hypertension, 32(64.0%) were current or ex-smokers and 20(40.0%) had a family history of CAD. Eighteen patients (36.0%) had post-procedural cTnI elevation but only 8(16.0%) had significant (>1.0ng/ml) elevation. Change of cTnI before and at 24 hours of PCI was not significant (p=0.057). Cardiac Troponin I increase was related to age, pre-procedural serum creatinine and multi-vessel stenting. Minor elevation of cTnI was common following elective PCI and associated with few risk factors such as elderly patient (more than 50 years), raised serum creatinine and multi-vessel stenting. So, early detection of these risk factors, as well as effective intervention may help to prevent injury to cardiac tissue hence stop elevation of cardiac TnI following elective PCI.

Comparison of Risk Factors and Angiographic Profile between Younger and Older Patients with Acute Myocardial Infarction.

Acute myocardial infarction (AMI) in younger adults (≤40 years) is being increasingly encountered in recent years among the South Asian population. Data regarding the presentation, risk factors and angiographic findings on this important subset of patients is lacking in our country. The aim of this study was to compare the risk factors and pattern of Coronary artery involvement in younger patients presenting with AMI with that of the older age group. This was a cross-sectional observational study conducted during the period from October 2018 to June 2019. Seventy consecutive AMI patients age ≤40 years and another 70 consecutive AMI patients age >40 years undergoing Coronary Angiogram (CAG) were included in the study. After taking informed written consent; demographic, anthropometric, risk factors, CAG findings were recorded in a pre-designed case record form. The severity of Coronary Artery Disease (CAD) was calculated by using Gensini score. The mean age of the younger and older patient groups was 36.89±4.4 years and 57.00±8.4 years respectively. Among the risk factors, smoking (67.1% versus 45.7%, p=0.017), positive family history CAD (38.6% versus 22.9%, p=0.040) and obesity (34.3% versus 20.0%, p= 0.05) were more common in younger group. Whereas, Hypertension (41.4% versus 72.9%, p=0.010) and DM (28.6% versus 50.0%, p=0.024) were more common in older patients. Younger patients mainly presented with STEMI (60.0% versus 48.6%) and predominantly had single vessel disease (42.9%), whereas older patients readily presented with NSTEMI (51.4%) and had a higher incidence of double vessel disease (32.9%) and triple vessel disease (30.0%). The Median Gensini score was significantly higher among the older patients than in the younger age group. Patients in younger age group showed a different pattern of risk factors and coronary artery involvement in comparison to the older age group. Thus, offering younger individuals to make them aware of these risk factors and their early detection, as well as an effective intervention may help to prevent AMI in younger people.

Predictive Value of Waist Height Ratio, Waist Hip Ratio and Body Mass Index in Assessing Angiographic Severity of Coronary Artery Disease in Myocardial Infarction Patients.

Coronary artery disease (CAD) is the leading cause of mortality and morbidity both in developed and developing countries. The body mass index (BMI), waist hip ratio (WHR) and waist height ratio (WHtR) are some of the clinical tools enabling clinicians to assess obesity. Although for decades there have been controversy regarding the relationship between obesity and CAD; it has been assumed that high BMI is a risk factor for CAD. However, the findings of some recent studies were paradoxical. The aim of this study was to identify the best tool among BMI, WHtR and WHR to evaluate angiographically severe CAD in myocardial infarction patients. This was a cross-sectional analytical study carried out in the Department of Cardiology, Chattogram Medical College and Hospital (CMCH), Chattogram, Bangladesh from January 2017 to December 2017. Three hundred and thirty two consecutive MI patients undergoing CAG during the study period were included in the study as per inclusion and exclusion criteria. Severity of CAD was calculated by using Gensini score. Patients were categorized and compared according to anthropometric indices and CAD severity. The mean±SD of the age of study population was 53.62±10.36 years (range 25-92) and 276(83.1%) were male. Regarding cardiovascular risk factors, 113(34%) patients had diabetes mellitus, 108(32.5%) had dyslipidaemia, 137(41.3%) had hypertension, 205(61.7%) were current or ex-smokers and 59(17.8%) had a family history of CAD. The mean±SD of the patients' BMI was 24.05±3.24kg/m² (range 16.14-32.72), mean±SD of their WHR was 0.964±0.052 (range 0.823-1.125) and mean±SD of their WHtR was 0.546±0.059 (range 0.389-0.748). The mean±SD of the severity of CAD according to the Gensini score was 41.11±28.66 (ranged from 2 to 244). Study findings showed a positive correlation between the severity of CAD with WHtR and WHR but not with BMI, according to Gensini scores (p=0.004, p=0.023 and p=0.43 respectively). Receiver Operating Characteristics (ROC) curve analysis revealed that waist height ratio had the highest area under the curve (AUC) among the three anthropometric parameters for predicting presence of severe CAD. Study showed the superiority of WHtR over WHR and BMI for predicting angiographic severity of CAD in patients with MI. WHtR should therefore be considered as a screening tool.

8-Hydr-oxy-8-phenyl-2,3,7,8-tetra-hydro-6H-1,4-dioxino[2,3-f]isoindol-6-one.

In the title compound, C(16)H(13)NO(4), the indole system is essentially planar, whereas the dioxane ring adopts a twist conformation. The mol-ecules are linked into chains by -O- H⋯O=C- hydrogen bonds and these chains are linked into rods by means of N-H⋯O hydrogen bonds. Exept for weak C-H⋯O inter-actions between the rods, no other inter-molecular contacts of inter-est are present.

Mechanism of stimulation by human interferon of prostaglandin synthesis in human cell lines.

Human interferon beta (IFN-beta) stimulated the synthesis of prostaglandin E (PGE) and prostaglandin F2 alpha in IFN-sensitive RSa and GM258 cell lines, but not in IFN-resistant HEC-1 cells. IFN-beta at a concentration of 1000 units/ml elicited 2- to 3-fold increases in PGE production in these cell lines. In the presence of exogenous arachidonic acid (1 microgram/ml), IFN-pretreated cells produced 5-fold more PGE compared to the cell cultures in the absence of arachidonic acid. Prednisolone, an inhibitor of phospholipase A2, at a concentration of 2 micrograms/ml inhibited the enhanced synthesis of PGE by IFN-pretreated cells. Indomethacin (4 micrograms/ml), a potent fatty acid cyclooxygenase inhibitor, also inhibited the increased synthesis of PGE. IFN stimulated the release of [14C]arachidonic acid from phospholipids but did not stimulate the activity of fatty acid cyclooxygenase. These data suggest that IFN stimulates prostaglandin synthesis by promoting the release of arachidonic acid from phospholipids. Since cycloheximide and actinomycin D inhibited the stimulation of PGE synthesis, the stimulation of prostaglandin synthesis by IFN seemed to be due to de novo enzyme synthesis which catalyzes the release of fatty acid. Addition of exogenous PGE suppressed the growth of RSa and GM258 cells. Prednisolone and iodomethacin partially inhibited anti-cell growth activity of IFN, suggesting a possibility that IFN-inhibited cell growth was partly mediated by prostaglandin.

Induction of cyclooxygenase and suppression of 12-lipoxygenase in human erythroleukemia cells upon phorbol ester-induced differentiation.

When human erythroleukemia (HEL) cells were incubated with arachidonic acid, both fatty acid cyclooxygenase and arachidonate 12-lipoxygenase activities were exhibited. Subcellular localization of these enzymes were examined by differential centrifugation, and both the cyclooxygenase and the 12-lipoxygenase were present predominantly in the microsomes rather than the cytosol of the cells. The cyclooxygenase activity was stimulated by 12-O-tetradecanoylphorbol 13-acetate (TPA) in a dose-dependent and time-dependent manner. Actual increase in the cyclooxygenase protein by TPA was demonstrated by immunoprecipitation and Western blot analysis of the enzyme with the aid of an anti-cyclooxygenase antibody. Furthermore, the cyclooxygenase mRNA level increased as assessed by RNA blot analysis using a cDNA probe for the enzyme. In contrast, the TPA treatment reduced the activity of 12-lipoxygenase. By RNA blot analysis using a cDNA probe of HEL cell 12-lipoxygenase, the mRNA level of the enzyme was shown to decline by the TPA treatment. Taken together, the TPA treatment of HEL cells brought about the induction of cyclooxygenase and the suppression of 12-lipoxygenase.

Protein in the food prolongs DNA synthesis in the liver of rats after starvation.

After a period on protein-free diet and then starvation, high protein diet induces DNA synthesis in rat liver for a longer period than diet containing an equivalent amount of amino acids. The requirement for high protein diet is limited to an early part of the prereplicative period and administration of the high protein diet later did not prolong DNA synthesis.

Two-wavelength fundus autofluorescence and macular pigment optical density imaging in diabetic macular oedema.

PURPOSE: To evaluate the application of 488 and 514 nm fundus autofluorescence (FAF) and macular pigment optical density (MPOD) imaging in diabetic macular oedema (DMO) and to demonstrate the typical imaging features. PATIENTS AND METHODS: A hundred and twenty-five eyes of 71 consecutive patients with diabetic retinopathy who underwent examination at a specialist university clinic employing a modified Heidelberg Retina Angiograph, using two different light sources of 488 and 514 nm wavelength, were retrospectively reviewed. MPOD images were calculated using modified Heidelberg Eye Explorer software. All images were evaluated by two independent masked graders. Features from FAF and MPOD images were correlated with optical coherence tomography (OCT) imaging findings and inter-grader variability, sensitivity and specificity were calculated using OCT as reference. RESULTS: Sixty-seven eyes had DMO on OCT. The inter-grader variability was 0.84 for 488 nm FAF, 0.63 for 514 nm FAF and 0.79 for MPOD imaging. Sensitivity and specificity for detection of DMO were 80.6 and 89.7% for 488 nm FAF; 55.2 and 94.8% for 514 nm FAF; and 80.6 and 91.4% for MPOD imaging. In 488 nm FAF and MPOD imaging, DMO was better visualised in comparison with 514 nm FAF imaging, P<0.01. MPOD revealed displacement of macular pigment by intraretinal cysts. CONCLUSION: MPOD imaging, and particularly its combination with 488 nm and 514 nm FAF, provides a valuable addition to OCT in the evaluation of DMO and is clinically useful in rapid en-face assessment of the central macula.

Implementation of and experiences with new automation.

In an environment where cost, timeliness, and quality drives the business, it is essential to look for answers in technology where these challenges can be met. In the Novartis Pharmaceutical Quality Assurance Department, automation and robotics have become just the tools to meet these challenges. Although automation is a relatively new concept in our department, we have fully embraced it within just a few years. As our company went through a merger, there was a significant reduction in the workforce within the Quality Assurance Department through voluntary and involuntary separations. However the workload remained constant or in some cases actually increased. So even with reduction in laboratory personnel, we were challenged internally and from the headquarters in Basle to improve productivity while maintaining integrity in quality testing. Benchmark studies indicated the Suffern site to be the choice manufacturing site above other facilities. This is attributed to the Suffern facility employees' commitment to reduce cycle time, improve efficiency, and maintain high level of regulatory compliance. One of the stronger contributing factors was automation technology in the laboratoriess, and this technology will continue to help the site's status in the future. The Automation Group was originally formed about 2 years ago to meet the demands of high quality assurance testing throughput needs and to bring our testing group up to standard with the industry. Automation began with only two people in the group and now we have three people who are the next generation automation scientists. Even with such a small staff,we have made great strides in laboratory automation as we have worked extensively with each piece of equipment brought in. The implementation process of each project was often difficult because the second generation automation group came from the laboratory and without much automation experience. However, with the involvement from the users at 'get-go', we were able to successfully bring in many automation technologies. Our first experience with automation was SFA/SDAS, and then Zymark TPWII followed by Zymark Multi-dose. The future of product testing lies in automation, and we shall continue to explore the possibilities of improving the testing methodologies so that the chemists will be less burdened with repetitive and mundane daily tasks and be more focused on bringing quality into our products.

Eicosanoid effects on cell proliferation in vitro: relevance to atherosclerosis.

Several eicosanoids were tested for ability to inhibit proliferation of cells in culture. In rabbit aortic smooth muscle cells and mouse B16BL6 melanoma cells, order of potency was: 12-HETE greater than PGJ2 greater than PGA1 greater than or equal to PGE1 greater than PGE2 greater than or equal to PGD2 greater than or equal to PGA2. PGB1 was active in smooth muscle cells (greater than PGD2) but not in B16 cells. 5-HETE and Leukotriene B4 were weakly active in smooth muscle cells, and PGB2, PGF2 alpha and TXB2 were inactive in both cells types. In Swiss albino mouse 3T3 fibroblasts, PGJ2 and PGE1 showed much lower relative potency than in the other two cell lines, although the profile was otherwise similar. These findings may be relevant to the anti-atherosclerotic (and perhaps anti-tumor activity) of some eicosanoids.

Thromboxane synthesizing system in rat liver.

Thromboxane B2 (TXB2), along with other primary prostaglandins, was synthesized when rat liver microsomes were incubated with radioactive arachidonic acid. TXB2 was identified directly by chemical ionization mass spectrometry and indirectly by using specific inhibitors of TX synthetase, viz., imidazole and OKY-1555 ((E)-3(4-(3-pyridyl-methyl) phenyl)-2 methyl acrylic acid HCl). The supernatant fraction obtained after centrifugation at 105,000 X g for 60 min contained a possible regulatory component that suppressed thromboxane synthesis. The regulatory influence is lost after partial hepatectomy.

Purification and some properties of NADP+ -specific isocitrate dehydrogenase from an extreme thermophile, Thermus flavus AT-62.

Thermostable NADP+ -specific isocitrate dehydrogenase (EC 1.1.1.42) was purified from crude extract of an extremely thermophilic bacterium Thermus flavus AT-62 through DEAE-cellulose column, acetone fractionation, DEAE-Sephadex A-50 column and isoelectric focussing. The enzyme was purified about 500-folds in its specific activity and purity was found to be about 96%. The enzyme was not inactivated after 60 min at 70 degrees C, but 20 and 80% of the activity were lost after 60 min at 80 degrees and 90 degrees C, respectively. Oxalacetate plus glyoxylate (each 1 nM) demonstrated 75% inhibition of the activity in concerted manner. The degree of the inhibition and the affinity of the enzyme for isocitrate and NADP+ decreased with the rise of temperature, especially above 60 degrees C. The activation energy below and above 60 degrees C were 14,500 and 8,000 cal per mole respectively. In CD spectra negative bands at 210 and 220nm were observed and alpha-helix content was calculated to be about 26%. In the course of heating up to 60 degrees practically no change in CD bands are observed, but above 60 degrees the depth of CD bands decreased gradually and remarkably above 80 degrees C. The effect of temperature on kinetic parameters and secondary structures of the enzyme was discussed in relation to the temperature adaptation of the organism.

Prostaglandin D synthase in human megakaryoblastic cells.

The cytosol fraction of human platelets did not convert prostaglandin (PG) H2 to PGD2. However, a homogenate of human megakaryoblastic CMK cells (precursor cells of platelets) produced PGD2 from PGH2. The PGD synthase activity was localized in the cytosol of CMK cells, and absolutely required glutathione. The catalytic properties and Western and Northern blottings indicated that the enzyme was PGD synthase of the hematopoietic type rather than the lipocalin type. When CMK cells were differentiated to megakaryocytes with phorbol ester along with induction of cyclooxygenase-1, the PGD synthase activity increased about 2-fold for 2 days and then decreased. In another human megakaryoblastic cell line, Dami, the PGD synthase increased about 10-fold by the addition of phorbol ester. Thus, the PGD synthase, which was undetectable in platelets, appeared during differentiation of megakaryoblasts to megakaryocytes.