Comparison of In Vivo and Ex Vivo MRI of the Human Hippocampal Formation in the Same Subjects.

Multiple techniques for quantification of hippocampal subfields from in vivo MRI have been proposed. Linking in vivo MRI to the underlying histology can help validate and improve these techniques. High-resolution ex vivo MRI can provide an intermediate modality to map information between these very different imaging modalities. This article evaluates the ability to match information between in vivo and ex vivo MRI in the same subjects. We perform rigid and deformable registration on 10 pairs of in vivo (3 T, 0.4 × 0.4 × 2.6 mm3) and ex vivo (9.4 T, 0.2 × 0.2 × 0.2 mm3) scans, and describe differences in MRI appearance between these modalities qualitatively and quantitatively. The feasibility of using this dataset to validate in vivo segmentation is evaluated by applying an automatic hippocampal subfield segmentation technique (ASHS) to in vivo scans and comparing SRLM (stratum/radiatum/lacunosum/moleculare) surface to manual tracing on corresponding ex vivo scans (and in 2 cases, histology). Regional increases in thickness are detected in ex vivo scans adjacent to the ventricles and were not related to scanner, resolution differences, or susceptibility artefacts. Satisfactory in vivo/ex vivo registration and subvoxel accuracy of ASHS segmentation of hippocampal SRLM demonstrate the feasibility of using this dataset for validation, and potentially, improvement of in vivo segmentation methods.

Haemagglutinin mutations and glycosylation changes shaped the 2012/13 influenza A(H3N2) epidemic, Houston, Texas.

While the early start and higher intensity of the 2012/13 influenza A virus (IAV) epidemic was not unprecedented, it was the first IAV epidemic season since the 2009 H1N1 influenza pandemic where the H3N2 subtype predominated. We directly sequenced the genomes of 154 H3N2 clinical specimens collected throughout the epidemic to better understand the evolution of H3N2 strains and to inform the H3N2 vaccine selection process. Phylogenetic analyses indicated that multiple co-circulating clades and continual antigenic drift in the haemagglutinin (HA) of clades 5, 3A, and 3C, with the evolution of a new 3C subgroup (3C-2012/13), were the driving causes of the epidemic. Drift variants contained HA substitutions and alterations in the potential N-linked glycosylation sites of HA. Antigenic analysis demonstrated that viruses in the emerging subclade 3C.3 and subgroup 3C-2012/13 were not well inhibited by antisera generated against the 3C.1 vaccine strains used for the 2012/13 (A/Victoria/361/2011) or 2013/14 (A/Texas/50/2012) seasons. Our data support updating the H3N2 vaccine strain to a clade 3C.2 or 3C.3-like strain or a subclade that has drifted further. They also underscore the challenges in vaccine strain selection, particularly regarding HA and neuraminidase substitutions derived during laboratory passage that may alter antigenic testing accuracy.

Frequency of Defecation and Form of Stool among General Bangladeshi Population.

Data on stool form and defecation frequency which are a prerequisite for defining normal bowel habit are lacking in Bangladesh. This observational cross sectional study was designed to find out defecation frequency and stool form among general population in Bangladesh. This study was performed in the Department of Gastroenterology, Shaheed Suhrawardy Medical College Hospital, Dhaka, Bangladesh from July 2017 to June 2018. Apparently healthy 1090 respondents were evaluated for predominant stool form (Bristol chart) and frequency. Data on demographic and life-style were collected. The study population consisted of 1090 respondents, among them, 65.13% male and 34.87% female and mean age of them was 40.20±12.39 years. Most of the people 874(80.2%) passed stool between 12-14 times per week followed by 111(10.2%) less than 3 stools per week, 95(8.7%) passed more than 14 stools per week and 10(0.9%) between 3-12 stools per week, p<0.001. Most people passed predominantly Bristol type IV stool- 610(56.0%); followed by type III- 274(25.1%). Other stool forms were: type I- 52(4.8%), type II- 59(5.4%), type V- 31(2.8%), type VI- 33(3.0%), type VII- 31(2.8%), p<0.001. In regard to the physical activity, most of the respondents (70.0%) are physically active whereas about 13.0% are sedentary and about 17.0% are physically intermediate between the two, p<0.001. In the case of dietary habit, most of the participants are non-vegetarian (82.5%) and the remaining are vegetarian (11.1%) and occasional non vegetarian (6.4%), p<0.001. Median stool frequency in the studied population was 14 per week and predominant form was Bristol type IV. Older age was associated with lesser stool frequency, particularly among female subjects.

Role of Nuclear Morphometry in the Cytologic Evaluation of Benign and Malignant Breast Lesions.

Breast cancer is the most common diagnosed cancer in female over the world. It is the most important cause of 'cancer death' among female. For diagnosis of breast masses the Fine needle aspiration cytology (FNAC) is applied as the primary tool. Though it is an easy, rapid and comparatively minimally invasive and inexpensive procedure for quick diagnosis, it is largely a subjective tool. Computer-assisted image morphometry provides a new influential method for high-precision measurement of nuclear features and can help to differentiate between benign and malignant breast aspirates. Therefore, the aim of study was to evaluate computer assisted semi-automatic nuclear morphometry on cytology of breast lesions to differentiate benign and malignant lesions of breast. Total 70 cases were included in this cross sectional study, performed in the department of Pathology, Mymensingh Medical College, Mymensingh, Bangladesh from March 2019 to February 2021. Morphometric analysis was done on images captured from FNAC slides of the selected cases. Image J Morphometric Software was used for image analysis. Statistical analysis was done by using SPSS 26.0 version. On analysis of morphometry, four nuclear size parameters, two nuclear shape parameters and one nuclear chromasia parameter were measured. The cut-off values with sensitivity and specificity between benign and malignant category for mean nuclear area were 61.54µm², 91.0%, 97.0%, for mean maximum feret diameter 10.89µm, 94.0%, 95.0%, for mean minimum feret diameter 7.71µm, 94.0%, 97.0% and for mean nuclear perimeter 33.32µm, 94.0%, 95.0% respectively. So, in this observation, morphometry is adjunctive tools which may overcome the limitations of inter observer agreement, improve diagnostic accuracy and avoid unnecessary repeat Fine Needle Aspiration (FNA) in breast aspirates.

Vascular Endothelial Growth Factor Expression in Colorectal Adenocarcinoma of Resected Samples and its Association with Histological Grade and Pathological Stage.

Colorectal carcinoma (CRC) is one of the major public health problems. Vascular endothelial growth factor (VEGF) is an effective angiogenic factor and plays a pivotal role in the development, progression and metastasis of CRC. It also could further help in selecting patients of high risk of disease progression for adjuvant therapy. Therefore, this study was undertaken to reveal the importance of angiogenic factor (VEGF) as an adjunctive tool with histologic parameters of CRC. Total 45 cases were included in this cross-sectional observational study, performed in the department of Pathology, Mymensingh Medical College, Mymensingh and Immunohistochemistry was done in the department of Pathology, Bangabandhu Sheikh Mujib Medical University, Dhaka, Bangladesh from March 2019 to February 2021. All the cases were evaluated for VEGF immunohistochemical expression. Majority of cases were low grade adenocarcinoma (86.9%) and most common stage was stage III (48.9%). Among 45 cases collectively 13 (4-negative + 9-weak positive) cases were low for VEGF expression and 32 (22-moderate positive + 10-strong positive) cases were high for VEGF expression. Statistically significant association was found with VEGF expression and increasing tumor stage as well as with lymph node metastasis (p<0.05). VEGF had positive significant correlation with stage (r=0.322; p=0.031) of tumor. However, no correlation with VEGF expression and grade (r=0.219; p=0.149) and other clinicopathological parameters of tumor was seen.

Pathological drivers of neurodegeneration in suspected non-Alzheimer's disease pathophysiology.

BACKGROUND: Little is known about the heterogeneous etiology of suspected non-Alzheimer's pathophysiology (SNAP), a group of subjects with neurodegeneration in the absence of ß-amyloid. Using antemortem MRI and pathological data, we investigated the etiology of SNAP and the association of neurodegenerative pathologies with structural medial temporal lobe (MTL) measures in ß-amyloid-negative subjects. METHODS: Subjects with antemortem MRI and autopsy data were selected from ADNI (n=63) and the University of Pennsylvania (n=156). Pathological diagnoses and semi-quantitative scores of MTL tau, neuritic plaques, α-synuclein, and TDP-43 pathology and MTL structural MRI measures from antemortem T1-weighted MRI scans were obtained. ß-amyloid status (A+/A-) was determined by CERAD score and neurodegeneration status (N+/N-) by hippocampal volume. RESULTS: SNAP reflects a heterogeneous group of pathological diagnoses. In ADNI, SNAP (A-N+) had significantly more neuropathological diagnoses than A+N+. In the A- group, tau pathology was associated with hippocampal, entorhinal cortex, and Brodmann area 35 volume/thickness and TDP-43 pathology with hippocampal volume. CONCLUSION: SNAP had a heterogeneous profile with more mixed pathologies than A+N+. Moreover, a role for TDP-43 and tau pathology in driving MTL neurodegeneration in the absence of ß-amyloid was supported.

Downstream effects of polypathology on neurodegeneration of medial temporal lobe subregions.

The medial temporal lobe (MTL) is a nidus for neurodegenerative pathologies and therefore an important region in which to study polypathology. We investigated associations between neurodegenerative pathologies and the thickness of different MTL subregions measured using high-resolution post-mortem MRI. Tau, TAR DNA-binding protein 43 (TDP-43), amyloid-ß and α-synuclein pathology were rated on a scale of 0 (absent)-3 (severe) in the hippocampus and entorhinal cortex (ERC) of 58 individuals with and without neurodegenerative diseases (median age 75.0 years, 60.3% male). Thickness measurements in ERC, Brodmann Area (BA) 35 and 36, parahippocampal cortex, subiculum, cornu ammonis (CA)1 and the stratum radiatum lacunosum moleculare (SRLM) were derived from 0.2 × 0.2 × 0.2 mm3 post-mortem MRI scans of excised MTL specimens from the contralateral hemisphere using a semi-automated approach. Spearman's rank correlations were performed between neurodegenerative pathologies and thickness, correcting for age, sex and hemisphere, including all four proteinopathies in the model. We found significant associations of (1) TDP-43 with thickness in all subregions (r = - 0.27 to r = - 0.46), and (2) tau with BA35 (r = - 0.31) and SRLM thickness (r = - 0.33). In amyloid-ß and TDP-43 negative cases, we found strong significant associations of tau with ERC (r = - 0.40), BA35 (r = - 0.55), subiculum (r = - 0.42) and CA1 thickness (r = - 0.47). This unique dataset shows widespread MTL atrophy in relation to TDP-43 pathology and atrophy in regions affected early in Braak stageing and tau pathology. Moreover, the strong association of tau with thickness in early Braak regions in the absence of amyloid-ß suggests a role of Primary Age-Related Tauopathy in neurodegeneration.

Work Ability among Nurses in a Public Medical College Hospital.

This cross sectional study was carried out 197 nurses of Shaheed Suhrawardy Medical College Hospital (ShSMCH), Dhaka, Bangladesh from 1st July 2015 to 30th June 2016. Purpose of the study was to assess work ability and its association with sociodemographic characteristics & work related variables which affect work ability among nurses in a Public Medical College Hospital. Sampling method was simple random sampling. By face to face interview data was collected with the help of semi-structured questionnaire and all data were analyzed with SPSS software version 21. For descriptive statistics means, SD and range were calculated as required. Data were presented in frequency table, pie diagram. Statistical test chi-square was used and p<0.05 was consider to be statistically significant. It was found that 136(69%) respondents were between the age of 21 to 39 years and mean age was 35.95 Years, 187(94.9%) were female, 156(79.2%) were Muslim, 166(84.3%) had Diploma, 180(91.4%) family income were equal to and more than 40,000 Tk. 189(95.5%) had done clinical work; 49(24.9%) worked at medicine. Majority 183(92.9%) respondents had good to excellent work ability and 14(7.1%) had less good work ability. In this study significant association (p<0.05) was found between work ability and gender, educational status and type of job. This research provides an initial step in understanding the work ability of nurses in a Public Medical College Hospital. The present study showed that nurses' work ability is at the good to excellent level. This study also reveals significant association between work ability and gender, educational status and type of job.

Defining SNAP by cross-sectional and longitudinal definitions of neurodegeneration.

Introduction: Suspected non-Alzheimer's pathophysiology (SNAP) is a biomarker driven designation that represents a heterogeneous group in terms of etiology and prognosis. SNAP has only been identified by cross-sectional neurodegeneration measures, whereas longitudinal measures might better reflect "active" neurodegeneration and might be more tightly linked to prognosis. We compare neurodegeneration defined by cross-sectional 'hippocampal volume' only (SNAP/L-) versus both cross-sectional and longitudinal 'hippocampal atrophy rate' (SNAP/L+) and investigate how these definitions impact prevalence and the clinical and biomarker profile of SNAP in Mild Cognitive Impairment (MCI). Methods: 276 MCI patients from ADNI-GO/2 were designated amyloid "positive" (A+) or "negative" (A-) based on their florbetapir scan and neurodegeneration 'positive' or 'negative' based on cross-sectional hippocampal volume and longitudinal hippocampal atrophy rate. Results: 74.1% of all SNAP participants defined by the cross-sectional definition of neurodegeneration also met the longitudinal definition of neurodegeneration, whereas 25.9% did not. SNAP/L+ displayed larger white matter hyperintensity volume, a higher conversion rate to dementia over 5 years and a steeper decline on cognitive tasks compared to SNAP/L- and the A- CN group. SNAP/L- had more abnormal values on neuroimaging markers and worse performance on cognitive tasks than the A- CN group, but did not show a difference in dementia conversion rate or longitudinal cognition. Discussion: Using a longitudinal definition of neurodegeneration in addition to a cross-sectional one identifies SNAP participants with significant cognitive decline and a worse clinical prognosis for which cerebrovascular disease may be an important driver.

Mortality prediction in adult cardiac surgery patients: comparison of two risk stratification models.

OBJECTIVE: To assess and compare the two commonly applied models--EuroSCORE and Parsonnet--in our local adult cardiac surgery patients, according to risk factor quantification related to mortality using a risk stratification protocol to assess the quality of cardiac surgical care. DESIGN: Prospective study. SETTING: Cardiac surgery centre in a regional hospital in Hong Kong. PATIENTS: All adult patients undergoing coronary artery bypass graft and heart valve surgery at the Grantham Hospital were evaluated prospectively from November 1999 to July 2005. MAIN OUTCOME MEASURES: In-hospital mortality was the defined end-point. Statistical analyses consisted of observed against expected mortality, Hosmer-Lemeshow goodness-of-fit test for calibration accuracy, and receiver operating characteristic curve for discrimination performance. RESULTS: During the study period, 1247 patients underwent coronary artery bypass graft surgery and 1406 underwent heart valve surgery. Observed mortality rates in these two patient groups were 2.9% and 4.8% respectively. The expected mortality rates as predicted by the EuroSCORE were (mean+/-standard deviation) 4.0+/-3.3% and 5.2+/-3.0% respectively, and by the Parsonnet model were 5.9+/-4.2% and 7.3+/-4.4% respectively. EuroSCORE performed better than the Parsonnet model at predicting in-hospital mortality assessed by the Hosmer-Lemeshow goodness-of-fit test. The areas under the receiver operating characteristic curves in coronary artery bypass graft surgery were 0.76 for EuroSCORE and 0.74 for Parsonnet. The receiver operating characteristic curve areas in valve surgery were 0.77 for EuroSCORE and 0.79 for Parsonnet. CONCLUSION: Despite significant geographic and demographic differences between European and Asian patients, in our local adult cardiac surgery patients, the EuroSCORE performed well with good calibration and discrimination in predicting mortality. There was a tendency for both models to over predict. However, the EuroSCORE can serve as a baseline for the development of a local risk model.

Mapping of pain circuitry in early post-natal development using manganese-enhanced MRI in rats.

Premature or ill full-term infants are subject to a number of noxious procedures as part of their necessary medical care. Although we know that human infants show neural changes in response to such procedures, we know little of the sensory or affective brain circuitry activated by pain. In rodent models, the focus has been on spinal cord and, more recently, midbrain and medulla. The present study assesses activation of brain circuits using manganese-enhanced magnetic resonance imaging (MEMRI). Uptake of manganese, a paramagnetic contrast agent that is transported across active synapses and along axons, was measured in response to a hindpaw injection of dilute formalin in 12-day-old rat pups, the age at which rats begin to show aversion learning and which is roughly the equivalent of full-term human infants. Formalin induced the oft-reported biphasic response at this age and induced a conditioned aversion to cues associated with its injection, thus demonstrating the aversiveness of the stimulation. Morphometric analyses, structural equation modeling and co-expression analysis showed that limbic and sensory paths were activated, the most prominent of which were the prefrontal and anterior cingulate cortices, nucleus accumbens, amygdala, hypothalamus, several brainstem structures, and the cerebellum. Therefore, both sensory and affective circuits, which are activated by pain in the adult, can also be activated by noxious stimulation in 12-day-old rat pups.

A protocol for manual segmentation of medial temporal lobe subregions in 7 Tesla MRI.

Recent advances in MRI and increasing knowledge on the characterization and anatomical variability of medial temporal lobe (MTL) anatomy have paved the way for more specific subdivisions of the MTL in humans. In addition, recent studies suggest that early changes in many neurodegenerative and neuropsychiatric diseases are better detected in smaller subregions of the MTL rather than with whole structure analyses. Here, we developed a new protocol using 7 Tesla (T) MRI incorporating novel anatomical findings for the manual segmentation of entorhinal cortex (ErC), perirhinal cortex (PrC; divided into area 35 and 36), parahippocampal cortex (PhC), and hippocampus; which includes the subfields subiculum (Sub), CA1, CA2, as well as CA3 and dentate gyrus (DG) which are separated by the endfolial pathway covering most of the long axis of the hippocampus. We provide detailed instructions alongside slice-by-slice segmentations to ease learning for the untrained but also more experienced raters. Twenty-two subjects were scanned (19-32 yrs, mean age = 26 years, 12 females) with a turbo spin echo (TSE) T2-weighted MRI sequence with high-resolution oblique coronal slices oriented orthogonal to the long axis of the hippocampus (in-plane resolution 0.44 × 0.44 mm2) and 1.0 mm slice thickness. The scans were manually delineated by two experienced raters, to assess intra- and inter-rater reliability. The Dice Similarity Index (DSI) was above 0.78 for all regions and the Intraclass Correlation Coefficients (ICC) were between 0.76 to 0.99 both for intra- and inter-rater reliability. In conclusion, this study presents a fine-grained and comprehensive segmentation protocol for MTL structures at 7 T MRI that closely follows recent knowledge from anatomical studies. More specific subdivisions (e.g. area 35 and 36 in PrC, and the separation of DG and CA3) may pave the way for more precise delineations thereby enabling the detection of early volumetric changes in dementia and neuropsychiatric diseases.

Automated Hippocampal Subfield Segmentation at 7T MRI.

BACKGROUND AND PURPOSE: High resolution 7T MRI is increasingly used to investigate hippocampal subfields in vivo, but most studies rely on manual segmentation which is labor intensive. We aimed to evaluate an automated technique to segment hippocampal subfields and the entorhinal cortex at 7T MRI. MATERIALS AND METHODS: The cornu ammonis (CA)1, CA2, CA3, dentate gyrus, subiculum, and entorhinal cortex were manually segmented, covering most of the long axis of the hippocampus on 0.70-mm(3) T2-weighted 7T images of 26 participants (59 ± 9 years, 46% men). The automated segmentation of hippocampal subfields approach was applied and evaluated by using leave-one-out cross-validation. RESULTS: Comparison of automated segmentations with corresponding manual segmentations yielded a Dice similarity coefficient of >0.75 for CA1, the dentate gyrus, subiculum, and entorhinal cortex and >0.54 for CA2 and CA3. Intraclass correlation coefficients were >0.74 for CA1, the dentate gyrus, and subiculum; and >0.43 for CA2, CA3, and the entorhinal cortex. Restricting the comparison of the entorhinal cortex segmentation to a smaller range along the anteroposterior axis improved both intraclass correlation coefficients (left: 0.71; right: 0.82) and Dice similarity coefficients (left: 0.78; right: 0.77). The accuracy of the automated segmentation versus a manual rater was lower, though only slightly for most subfields, than the intrarater reliability of an expert manual rater, but it was similar to or slightly higher than the accuracy of an expert-versus-manual rater with ∼170 hours of training for almost all subfields. CONCLUSIONS: This work demonstrates the feasibility of using a computational technique to automatically label hippocampal subfields and the entorhinal cortex at 7T MRI, with a high accuracy for most subfields that is competitive with the labor-intensive manual segmentation. The software and atlas are publicly available: http://www.nitrc.org/projects/ashs/.

Compact panicle architecture is detrimental for growth as well as sucrose synthase activity of developing rice kernels.

The increase of spikelet number in the panicles of modern super rice has made the architecture compact, as the extra spikelets are accommodated mostly on secondary branches than on primary branches. However, the grain yield did not improve because of poor grain filling, which was more visible in the basal spikelets than apical spikelets. The objective of this study was to examine the effect of the compactness and positional difference of spikelets in the panicle on grain filling by comparing the activity and genetic expression of starch synthesising enzymes in the developing kernels of lax-(Upahar and CR3856-45-11-2-7-2-5 (CR-45)) and compact-(Mahalaxmi and CR3856-29-14-2-1-1-1 (CR-29)) panicle cultivars. Upahar and Mahalaxmi are genetically related, whereas CR-45 and CR-29 are recombinant inbred lines. The grain carbohydrate concentration and activity of sucrose synthase (SUS) enzyme were estimated during the active period of grain filling. Further, expression of isoforms of SUS, ADP glucose pyrophosphorylase (APL and APS for large and small units respectively) and starch synthase (SS and GBSS for soluble and granule bound starch synthases respectively) were also assayed through PCR studies. The genotype approach used revealed grain SUS activity and starch concentration high and sugar concentration low in the lax- compared with compact-panicle cultivars and in the apical spikelets compared with basal ones. The margin of variation between apical and basal spikelets was higher in the compact- than the lax-panicle cultivars. Genetic expression of most of the isoforms of the enzymes was higher in the lax- than the compact-panicle cultivars as seen in RT-PCR studies. A quantitative appraisal of transcript levels of isoforms in the qRT-PCR identified greater expression of SUS3 in the basal spikelets of Upahar than that in Mahalaxmi and in CR-45 over CR-29, most prominently during the active period of grain filling. We conclude that proximal location as well as increased density of spikelets on panicles affected SUS3 expression in the basal spikelets. The metabolic dominance of a spikelet in rice panicle is dependent on the expression of the genes for different isoforms of starch synthesising enzymes, but the expression of SUS3 could be more specific than the others. SUS3 expression is most active during grain filling of the lax-panicle cultivars, but its dominance is reduced significantly in the kernels of the compact-panicle cultivars.

A large-scale radiometric micro-quantitative complement fixation test for serum antibody titration.

A micro-quantitative complement fixation (CF) procedure based on 51Cr release is described. The method employs 50% hemolysis as end point and the alternation equation to calculate the amount of complement involved in the hemolytic reaction. Compared to the conventional CF tests, the radiometric procedure described here is very precise and consistently reproducible. Also, since only 3 4-fold dilutions of sera are used for the titration of antibodies over a wide range of concentrations, the test is very concise and is economical to perform. Its format is amenable to automation and computerization. This radiometric CF procedure is thus most useful for large-scale immunological research and epidemiological surveillance studies.

Pathogenicity and antigenic components of excysted Giardia lamblia isolated from patients in Riyadh, Saudi Arabia.

Giardia lamblia cysts were isolated from patients in Riyadh, Saudi Arabia. Cysts and trophozoites (from axenically excysted cysts) were given orally by gavage to mice to establish the pathogenicity of the Riyadh isolate. There was no effect of varying the dose of administered parasite on parasite excretion or morbidity. A typical pathologic pattern of giardiasis was demonstrated by histologic methods and electron microscopy. Antigenic components of the Riyadh isolate were compared with the Portland strain and with Entamoeba histolytica by gel diffusion immunoprecipitation and immunoblotting. There were few antigens in common between Riyadh isolate and the Portland strain, and little cross-reactivity of the Riyadh isolate with Entamoeba histolytica was observed.

5'-Nor carbocyclic 5'-deoxy-5'-(isobutylthio)adenosine and a 2',3'-dideoxy-2',3'-didehydro derivative.

The inhibition of biochemical processes requiring S-adenosylmethionine as a co-factor have led to many nucleoside-based medicinal agents. Included in this group are 5'-deoxy-5'-(isobutylthio)adenosine (SIBA), a nucleoside with antiparasitic, antiviral and antiproliferative effects, and 5'-noraristeromycin, a carbocyclic-derived nucleoside with potent antiviral properties. This report brings together the structural components of these two compounds by describing both enantiomers of carbocyclic 5-nor SIBA (3 and 4). Owing to the recent interest in 2',3'-dideoxy-2',3'-didehydro nucleosides as antiviral agents, this derivative of 3 (5) is also described. All three compounds were screened against a variety of viruses and were found to be inactive at high concentrations or at limiting concentrations for the screening methods. The viruses subjected to 3-5 were herpes simplex virus types 1 and 2, human cytomegalovirus, vaccinia virus, vesicular stomatitis virus, respiratory syncytial virus, varicelIa zoster virus, coxsackie virus, parainfluenza-3 virus, sindbis virus, punta toro virus, reovirus-1, human immunodeficiency virus, influenza virus types A and B, adenovirus type 1 and measles virus. These results suggest that the C-5' methylene of the C-5' thio-based carbocyclic nucleosides is important for their antiviral properties.

Inhibition of measles virus replication by 5'-nor carbocyclic adenosine analogues.

Despite intense efforts to increase vaccine coverage, measles virus (MV) still causes significant morbidity and mortality in the world, sometimes as the result of severe, chronic, lethal disease. In an effort to develop therapies to supplement immunization strategies, a number of 5'-nor carbocyclic adenosine analogues were evaluated for anti-MV activity in CV-1 monkey kidney cells. Of those compounds tested, those either unsubstituted at C4 or possessing a hydroxyl, azido or amino substituent at that position were the most active, with particularly significant inhibition of MV, strain Chicago-1. The EC50 values against this strain ranged from <0.1 to 1 mg/ml, as determined by cytopathic effect reduction assay, and confirmed by neutral red uptake. By virus yield reduction assay (+)-(1S,25,3R,4S)-4-(6'-amino-9'H-purin-9'-yl)cyclopentane-1,2,3-triol (2) (-)-(1R,2S,3R)-1-(6'-amino-9'H-purin-9'-yl)-2,3-dihydoxycyclopent-4-ene (3) (-)-(1R,2S,3R)-1-(6'-amino-9'H-purin-9'-yl)cyclopentane-2,3-dihydoxycyclopentane (5) and (-)-(1R,2R,3R,4S)-4-amino-1-(6'-amino-9'H-purin-9'-yl)cyclopentane-2,3-diol (8) were the most potent compounds tested, all with EC90 values of < or =0.4 mg/ml. Compounds 3 and 5 were also tested against other MV strains, and similarly inhibited those strains except for four designated as Bil, Edmonston, SA and X-1108. Compound 8 did not potently inhibit these other MV strains. In addition, 3, 5 and 8 demonstrated synergistic (additive) inhibition of MV replication in combination with ribavirin at several concentrations. Compounds 3, 5 and 8 were also potent MV inhibitors even when added to infected cells 24 h after virus exposure. None of these three compounds was virucidal at concentrations that inhibited viral replication as determined by virus yield reduction assay. Most compounds tested were also not toxic at concentrations >100 mg/ml in actively growing and stationary-phase cells. Results suggest that these compounds may be clinically useful anti-MV virus agents.

Isoenzyme pattern of clones of Entamoeba histolytica.

The isoenzyme pattern of clones of E. histolytica isolated from symptomatic and asymptomatic patients of amoebiasis were determined by starch gel electrophoresis. Ten parent cultures (uncloned), 5 from symptomatic and 5 from asymptomatic patients were isolated and established in Robinsons medium as xenic cultures. A total of 100 clones were isolated from parent cultures but only 65 could be established. Starch gel electrophoresis was carried out on the uncloned and cloned cultures. The analysis showed that the 5 uncloned culture from symptomatic patients had 4 isolates with pathogenic zymodeme and 1 isolate with non-pathogenic zymodeme pattern. The presence of beta band and absence of alpha band for phosphoglucomutase (PGM) and a fast moving Hexokinase (HK) band were taken as markers of pathogenicity. The 5 cloned cultures from asymptomatic patients had three isolates with pathogenic and 2 with non-pathogenic pattern. The cloned isolates from symptomatic patients (35) had 20 pathogenic zymodemes and asymptomatic patients (30) had 15 pathogenic zymodemes. Non-pathogenic zymodeme obtained in the present study were I, IV, V, X and XIII and the pathogenic zymodemes obtained were II and XIV. The data of the study indicated that (i) asymptomatic patients also harbour pathogenic zymodemes; and (ii) zymodeme II also coexsist with zymodeme XIV in the Indian population.

Characterization of potentially pathogenic free-living amoebae in sewage samples of Calcutta, India.

1. It is widely accepted that foul or polluted environments are the principal sources of potentially pathogenic species of free-living amoebae. The present paper is the first report of occurrence of potentially pathogenic free-living amoebae in sewage samples of Calcutta, India. 2. We describe the occurrence, isolation, specific identification and comparative mouse pathogenicity test of two pathogenic amoebae, viz., Naegleria fowleri (N. aerobia) Carter, 1970, causing human meningoencephalitis and Acanthamoeba castellanii Douglas, 1930, causing granulomatous amoebic encephalitis, and one non-pathogenic amoeba, viz., A. astronyxis Ray and Hayes, 1959, in sewage samples of Calcutta, India. 3. The existence of both pathogenic and non-pathogenic amoebae living side by side is of considerable epidemiological relevance.