Enhancing learning objectives by use of simple virtual microscopic slides in cellular physiology and histology: impact and attitudes

The impact and perception of students on the use of a simple, low technology-driven version of a virtual microscope in teaching and assessments in cellular physiology and histology were studied. Its impact on the time and resources of the faculty were also assessed. Simple virtual slides and conventional microscopes were used to conduct the same examinations for the same students. Students performed significantly better in the examination with the virtual slide and also showed a significantly higher preference for virtual slides. The time and cost implications of conducting examinations using the simple virtual slides were reduced by >1,400%. The results reemphasize the need for the design and adoption of simple sustainable technological innovations in developing countries to bridge gaps in purposeful learning environments.     

Fatal hemolytic anemia associated with metformin: A case report

Introduction

Metformin is a widely prescribed biguanide antidiabetic drug that has been implicated as a cause of hemolytic anemia in three previous case reports. We report a case of rapidly fatal hemolysis that was temporally associated with the initiation of metformin treatment for diabetes. Clinicians need to be aware of this rare but potentially serious side effect of metformin.

Case presentation

A 56-year-old Caucasian man with type 2 diabetes mellitus was started on metformin to improve glycemic control. Shortly afterwards, he developed progressive fatigue, exertional dyspnea, cranberry-colored urine and jaundice. Laboratory studies showed severe hemolysis, with a drop in hemoglobin from 14.7 to 6.6 g/dl over 4 days, markedly elevated lactate dehydrogenase, bilirubin and reticulocyte counts, and a low haptoglobin level. A peripheral blood smear showed no schistocytes, and a direct Coombs test was positive for anti-IgG and negative for anti-C3. Despite corticosteroid treatment and transfusion of packed red blood cells, the patient developed increasing dyspnea, hypotension, further decline in hemoglobin to 3.3 g/dl, and fatal cardiorespiratory arrest 12 hours after admission.

Conclusion

The serologic findings in this case suggest an autoimmune hemolytic anemia, caused either by a drug-induced autoantibody or a warm autoantibody. Based on the temporal association with metformin and the lack of other clear precipitating causes, we propose that metformin-induced hemolysis with a drug-induced autoantibody is a strong possibility. This mechanism differs from a previously described case with a possible antibody to the erythrocyte-drug complex. It has been shown, however, that hemolysis may occur via multiple mechanisms from the same drug. Clinicians should consider the possibility of metformin-associated immune hemolytic anemia in patients with otherwise unexplained hemolysis.

     

Automated error-tolerant macromolecular structure determination from multidimensional nuclear Overhauser enhancement spectra and chemical shift assignments: improved robustness and performance of the PASD algorithm

We report substantial improvements to the previously introduced automated NOE assignment and structure determination protocol known as PASD (Kuszewski et al. (2004) J Am Chem Soc 26:6258-6273). The improved protocol includes extensive analysis of input spectral data to create a low-resolution contact map of residues expected to be close in space. This map is used to obtain reasonable initial guesses of NOE assignment likelihoods which are refined during subsequent structure calculations. Information in the contact map about which residues are predicted to not be close in space is applied via conservative repulsive distance restraints which are used in early phases of the structure calculations. In comparison with the previous protocol, the new protocol requires significantly less computation time. We show results of running the new PASD protocol on six proteins and demonstrate that useful assignment and structural information is extracted on proteins of more than 220 residues. We show that useful assignment information can be obtained even in the case in which a unique structure cannot be determined.     

Heavy Metals in Sediments of the Gold Mining Impacted Pra River Basin,Ghana, West Africa

Total concentrations of Hg, Al, Fe, As, Pb, Cu, Cr, Ni, Mn, Co, V, and Zn were determined in surface sediments collected from 21 locations within the gold mining impacted Pra River basin in southwestern Ghana. Samples were collected during both the rainy and dry seasons. We hypothesized that in the rural southwestern portion of Ghana, the lack of industrial activities makes artisanal gold mining (AGM) by Hg amalgamation the main source of water resource contamination with heavy metals. Therefore, metals showing concentration trends similar to that of Hg in the studied system are likely impacted by AGM. We found that total-Hg (THg) concentrations in riverine sediments are rather low as compared to other aquatic systems that are impacted by similar mining activities. Measured THg concentrations ranged from 0.018 to 2.917 mg/kg in samples collected in the rainy season and from about 0.01 to 0.043 mg/kg in those collected during the dry season. However, the determination of the enrichment factor (EF) calculated using shale data as reference background values showed signs of severe contamination in most of the sampled sites. In the dry season, THg concentrations correlated positively and significantly to the concentrations of As (r = 0.864, p < 0.01), Cu (r = 0.691, p < 0.05), and Ni (r = 0.579, p < 0.05). Based on our previously stated hypothesis, this could then be an indication of the impact of AGM on ambient levels of these 3 elements. However, the determined concentrations of Cu, and Ni co-varied significantly with Al, suggesting that natural sources do account for the observed levels. Accordingly, both AGM and metal inputs from weathered natural deposits are likely co-responsible for the observed levels of Cu and Ni. In contrast, the lack of correlation between As and Al tends to suggest an impact of AGM on As levels. Overall, our data suggest that besides Hg and to some extent As, the impact of AGM on ambient levels of investigated metals in the gold mining impacted Pra River remains negligible. Finally, the increase in metal concentrations from the dry to the rainy season underlines the impact of changes in hydrologic conditions on levels and fate of metals in this tropical aquatic system.     

Spectroscopic studies of perturbed T1 Cu sites in the multicopper oxidases Saccharomyces cerevisiae Fet3p and Rhus vernicifera laccase: allosteric coupling between the T1 and trinuclear Cu sites

The multicopper oxidases catalyze the 4e- reduction of O2 to H2O coupled to the 1e- oxidation of 4 equiv of substrate. This activity requires four Cu atoms, including T1, T2, and coupled binuclear T3 sites. The T2 and T3 sites form a trinuclear cluster (TNC) where O2 is reduced. The T1 is coupled to the TNC through a T1-Cys-His-T3 electron transfer (ET) pathway. In this study the two T3 Cu coordinating His residues which lie in this pathway in Fet3 have been mutated, H483Q, H483C, H485Q, and H485C, to study how perturbation at the TNC impacts the T1 Cu site. Spectroscopic methods, in particular resonance Raman (rR), show that the change from His to Gln to Cys increases the covalency of the T1 Cu-S Cys bond and decreases its redox potential. This study of T1-TNC interactions is then extended to Rhus vernicifera laccase where a number of well-defined species including the catalytically relevant native intermediate (NI) can be trapped for spectroscopic study. The T1 Cu-S covalency and potential do not change in these species relative to resting oxidized enzyme, but interestingly the differences in the structure of the TNC in these species do lead to changes in the T1 Cu rR spectrum. This helps to confirm that vibrations in the cysteine side chain of the T1 Cu site and the protein backbone couple to the Cu-S vibration. These changes in the side chain and backbone provide a possible mechanism for regulating intramolecular T1 to TNC ET in NI and partially reduced enzyme forms for efficient turnover.     

Synapsin Isoforms and Synaptic Vesicle Trafficking

Synapsins were the first presynaptic proteins identified and have served as the flagship of the presynaptic protein field. Here we review recent studies demonstrating that different members of the synapsin family play different roles at presynaptic terminals employing different types of synaptic vesicles. The structural underpinnings for these functions are just beginning to be understood and should provide a focus for future efforts.