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Biomedical subjects

R Krishnan

Publications and source records attributed to R Krishnan.

At least 73 records · Page 4Linked to original sources

Accuracy and reproducibility of brain and tissue volumes using a magnetic resonance segmentation method.

Magnetic resonance (MR) imaging now allows the qualitative and quantitative assessment of the human brain in vivo. As MR imaging resolution has improved, precise measurement of small brain structures has become possible. Methods of measuring brain regions from MR images include both manual and semiautomated methods. Despite the development of numerous volumetric methods, there have been only limited attempts so far to evaluate the accuracy and reproducibility of these methods. In this study we used phantoms to assess the accuracy of the segmentation process. Our results with simple and complex phantoms indicate an error of 3-5% using either manual or semiautomated techniques. We subsequently used manual and semiautomated volumetric methodologies to study human brain structures in vivo in five normal subjects. Supervised segmentation is a semiautomated method that accomplishes the division of MR images into several tissue types based on differences in signal intensity. This technique requires the operator to manually identify points on the MR images that characterize each tissue type, a process known as seeding. However, the use of supervised segmentation to assess the volumes of gray and white matter is subject to pitfalls. Inhomogeneities of the radiofrequency or magnetic fields can result in misclassification of tissue points during the tissue seeding process, limiting the accuracy and reliability of the segmentation process. We used a structured seeding protocol that allowed for field inhomogeneity that produced reduced variation in measured tissue volumes. We used repeated segmentations to assess intra- and inter-rater reliability, and were able to measure small and large regions of interest with a small degree of variation. In addition, we demonstrated that measurements are reproducible with repeat MR acquisitions, with minimal interscan variability. Segmentation methods can accurately and reliably measure subtle morphometric changes, and will prove a boon to the study of neuropsychiatric disorders.

Adult↗

Tandem placement of a coronavirus promoter results in enhanced mRNA synthesis from the downstream-most initiation site.

Insertion of the 17-nucleotide promoter region for the bovine coronavirus N gene as part of a 27-nucleotide cassette into the open reading frame of a cloned synthetic defective-interfering (DI) RNA resulted in synthesis of subDI RNA transcripts from the replicating DI RNA genome. Duplicating and triplicating the promoter sequence in tandem caused a progressive increase in the efficiency of subgenomic mRNA synthesis despite a concurrent decrease in the rate of DI RNA accumulation that was not specific to the promoter sequences being added. Although initiation of transcription (leader fusion) occurred at each of the three promoter sites in the tandem construct, almost all of the transcripts were found as a product of the most downstream (3'-most on the genome) promoter. These results show that enhancement of subgenomic mRNA synthesis is a property that can reside within sequence situated near the promoter. A possible role for the plus strand in the downstream promoter choice is suggested.

Animals↗

Synthesis, structure, and structure-activity relationships of divalent thrombin inhibitors containing an alpha-keto-amide transition-state mimetic.

A new class of divalent thrombin inhibitors is described that contains an alpha-keto-amide transition-state mimetic linking an active site binding group and a group that binds to the fibrinogen-binding exosite. The X-ray crystallographic structure of the most potent member of this new class, CVS995, shows many features in common with other divalent thrombin inhibitors and clearly defines the transition-state-like binding of the alpha-keto-amide group. The structure of the active site part of the inhibitor shows a network of water molecules connecting both the side-chain and backbone atoms of thrombin and the inhibitor. Direct peptide analogues of the new transition-state-containing divalent thrombin inhibitors were compared using in vitro assays of thrombin inhibition. There was no direct correlation between the binding constants of the peptides and their alpha-keto-amide counterparts. The most potent alpha-keto-amide inhibitor, CVS995, with a Ki = 1 pM, did not correspond to the most potent divalent peptide and contained a single amino acid deletion in the exosite binding region with respect to the equivalent region of the natural thrombin inhibitor hirudin. The interaction energies of the active site, transition state, and exosite binding regions of these new divalent thrombin inhibitors are not additive.

Amides↗

Crystal structures of thrombin with thiazole-containing inhibitors: probes of the S1' binding site.

Structures of the blood clotting enzyme thrombin complexed with hirugen and two active site inhibitors, RWJ-50353 10080(N-methyl-D-phenylalanyl-N-[5-[(aminoiminomethyl)amino]-1- [[(2-benzothiazolyl)carbonyl]butyl]-L-prolinamide trifluoroacetate hydrate) and RWJ-50215 (N-[4-(aminoiminomethyl)amino-1-[2- (thiazol-2-ylcarbonylethyl)piperidin- 1-ylcarbonyl]butyl]-5-(dimethylamino)naphthalenesulfonamide trifluoroacetate hydrate), were determined by x-ray crystallography. The refinements converged at R values of 0.158 in the 7.0-2.3-A range for RWJ-50353 and 0.155 in the 7.0-1.8-A range for RWJ-50215. Interactions between the protein and the thiazole rings of the two inhibitors provide new valuable information about the S1' binding site of thrombin. The RWJ-50353 inhibitor consists of an S1'-binding benzothiazole group linked to the D-Phe-Pro-Arg chloromethyl ketone motif. Interactions with the S1-S3 sites are similar to the D-phenylalanyl-prolyl-arginyl chloromethylketone structure. In RWJ-50215, a S1'-binding 2-ketothiazole group was added to the thrombin inhibitor-like framework of dansylarginine N-(3-ethyl-1,5-pentanediyl)amide. The geometry at the S1-S3 sites here is also similar to that of the parent compound. The benzothiazole and 2-ketothiazole groups bind in a cavity surrounded by His57, Tyr60A, Trp60D, and Lys60F. This location of the S1' binding site is consistent with previous structures of thrombin complexes with hirulog-3, CVS-995, and hirutonin-2 and -6. The ring nitrogen of the RWJ-50353 benzothiazole forms a hydrogen bond with His57, and Lys60F reorients because of close contacts. The oxygen and nitrogen of the ketothiazole of RWJ-50215 hydrogen bond with the NZ atom of Lys60F.

Binding Sites↗

Human infection with Dirofilaria repens in Malaysia.

Human dirofilariasis is a rare infection in Malaysia. Thus far, only two human cases have been reported viz. Dirofilaria immitis and D. (Nochtiella) repens and in both instances, adult worms were recovered from infected patients. The two cases reported in the present study, one from Melaka and the other from Penang, were diagnosed histologically. Based on the diagnostic criteria for identifying Dirofilaria in tissue sections, the parasites were identified as D. (Nochtiella) repens.

Adult↗

The UCUAAAC promoter motif is not required for high-frequency leader recombination in bovine coronavirus defective interfering RNA.

The 65-nucleotide leader on the cloned bovine coronavirus defective interfering (DI) RNA, when marked by mutations, has been shown to rapidly convert to the wild-type leader of the helper virus following DI RNA transfection into helper virus-infected cells. A model of leader-primed transcription in which free leader supplied in trans by the helper virus interacts by way of its flanking 5'UCUAAAC3' sequence element with the 3'-proximal 3'AGAUUUG5' promoter on the DI RNA minus strand to prime RNA replication has been used to explain this phenomenon. To test this model, the UCUAAAC element which occurs only once in the BCV 5' untranslated region was either deleted or completely substituted in input DI RNA template, and evidence of leader conversion was sought. In both cases, leader conversion occurred rapidly, indicating that this element is not required on input RNA for the conversion event. Substitution mutations mapped the crossover region to a 24-nucleotide segment that begins within the UCUAAAC sequence and extends downstream. Although structure probing of the bovine coronavirus 5' untranslated region indicated that the UCUAAAC element is in the loop of a prominent stem and thus theoretically available for base pair-directed priming, no evidence of an unattached leader early in infection that might have served as a primer for transcription was found by RNase protection studies. These results together suggest that leader conversion on the DI RNA 5' terminus is not guided by the UCUAAAC element and might arise instead from a high-frequency, region-specific, homologous recombination event perhaps during minus-strand synthesis rather than by leader priming during plus-strand synthesis.

Animals↗

Pharmacokinetics of erythropoietin following single-dose subcutaneous administration in preterm infants.

The pharmacokinetics of recombinant human erythropoietin was studied in 12 very low birth weight preterm infants < 32 weeks of gestation after subcutaneous administration of 300 IU/kg at a postconceptional age of 34 (32-37) weeks and a weight of 1,505 (1330-1,740)g (median and range). The administration of recombinant human erythropoietin produced a rapid increase in serum erythropoietin levels with a peak level of 362.8 mIU/ml at 8.9 h. The area under the curve was 8,177.5 (4,597.1-15,453.0) mIU/ml/h, the absorption half-life was 5.5 (1.6-6.6) h, the elimination half-life was 7.9 (5.6-19.4) h, and the residence time was 19.6 (5.1-32.6) h (all values reflect median and range). There was no significant correlation between absorption and elimination half-life of erythropoietin and birth weight, gestational age, sex, and age and weight of the infants at the time of administration of erythropoietin. Based on absorption and elimination kinetics, the dosing interval for subcutaneous administration must not be < 48 h.

Absorption↗

A 3.5-kb DNA fragment contains the cis-regulatory elements for retina-specific expression and partial dosage compensation of the Arrestin B (ArrB) gene of Drosophila miranda.

A 3.5-kb genomic DNA fragment containing the X1R chromosome-linked retina-specific Arrestin B gene (also called Arrestin 2) of Drosophila miranda (ArrB-mr) was introduced into the Drosophila melanogaster genome via germ-line transformation. The results showed that the ArrB-mr transgene was expressed only in the retina of the transformed flies. The transgene also showed male-specific transcriptional hyperactivation or dosage compensation, but the level was found to be 71-79% of the expected value. The endogenous autosomal ArrB-ml gene of D. melanogaster, as expected, was not found to be dosage compensated in the transformants. Thus, while the cis-regulatory elements for retina-specific expression are fully present in the 3.5-kb DNA, part of the sequences needed for full dosage compensation are missing in this DNA. Nucleotide (nt) sequence comparison of the upstream DNA revealed that both ArrB-mr and ArrB-ml possess a putative TATA box at -32 nt. They also have an 11-bp motif (5'-AATCCAGTTAG) similar to the photoreceptor conserved element I (PCE I) believed to play an important role in retina-specific expression of the D. melanogaster opsin and mouse Arr genes. In addition, both genes contain a TGACCT motif which is known to bind a transcription factor activated by retinoic acid and vitamin D3. However, five tandem repeats of a heptanucleotide sequence (TGGGCNR) and a 29-bp sequence with the potential to form a stem-loop structure, are found only in the ArrB-ml gene. These sequences may play an important role in dosage compensation of the X1R-linked ArrB-mr gene.

Animals↗

Some epidemiological aspects and economic costs of injuries in Malaysia.

Injuries are one of the leading causes of death in the world. In Malaysia, injuries form one of the three main causes of mortality. They are also an important cause of permanent and temporary disability and work absenteeism in the productive age group. Increasing affluence and industrialization coupled with growing population and transportation needs in rapidly developing countries like Malaysia have resulted in a surge of road and occupational injuries. Three quarters of fatalities due to road, occupational, drowning and home injuries occur in those below 45 years of age. A majority of injuries in these categories are attributed to "human" factors and therefore can be prevented by public education and enforced training of workers. The total annual economic loss due to all types of injuries is estimated to be 2 billion Malaysian Ringgit (US$1 = MR2.76 approximately). The government is currently in the process of setting up full-time departments for road safety and occupational health and safety.

Accident Prevention↗

MR imaging of diseases of the limbic system.

The limbic system refers to the part of the brain that is involved in emotional expression and in cognitive and somatomotor control systems; it most likely also has a significant role in the pathogenesis of certain dementias, neuropsychiatric disturbances, and seizure disorders. This system, as originally defined, has no clear anatomic boundaries. Limbic regions of the telencephalon include a continuous medial zone of cortical regions, including the hippocampal formation; a perihippocampal zone, including the cingulate gyrus, prefrontal region, and perirhinal region; along with subcortical areas, including the septum pellucidum and the amygdala. A distinguishing characteristic of the limbic region is that it is highly interconnected and appears to form the only major route for information transfer between the neocortex and the hypothalamus. With the multiplanar capability and resolution of MR imaging, the individual parts of the limbic system and the diseases that affect them can be studied. In this review, we discuss MR imaging of the diseases that affect this system.

Brain Diseases↗

Does myocardial perfusion scintigraphy demonstrate clinical usefulness in patients with markedly positive exercise tests? An assessment of the method in a high-risk subset.

To evaluate the clinical utility of perfusion scintigraphy in patients with markedly positive exercise ECGs, we studied 94 consecutive patients with markedly positive exercise ECGs; 74 of them were also studied with scintigraphy. Patients undergoing scintigraphy had an intermediate pretest likelihood of coronary disease and were divided into two groups: those with reversible defects involving one complete area or aspects of multiple vascular areas (group 1, 38 patients), and those with normal scintigrams and reversible defects in a limited aspect of one vascular area, isolated fixed defects, or both (group 2, 36 patients). Among all demographic and exercise variables, only a hypotensive or blunted blood pressure response and scintigraphic lung uptake or cavitary dilation, although insensitive, were more frequent in group 1 (all p < 0.05). All 36 patients in group 1 and 14 of 18 in group 2 who underwent coronary angiography had significant coronary lesions; 31 in group 1 but only seven in group 2 had multivessel disease (p < 0.05). Subsequently 32 patients in group 1 had revascularization compared with only two patients in group 2. Only one cardiac event was noted among 34 patients in group 2 who were followed for a mean of 38 months while they were being treated medically. However, four of nine patients in group 1 initially treated medically required late revascularization because of clinical progression of disease, and one patient died (p < 0.05). Compared with patients having scintigraphy, patients not imaged had a higher pretest likelihood of coronary disease, a higher incidence of angina, unstable angina, and induced angina, with a lower exercise time and time to ST depression (p < 0.05). All underwent angiography, and 16 had multivessel disease. Not all patients with markedly positive exercise ECGs were at similarly high coronary risk. Some with high-risk coronary anatomy were identified without the use of scintigraphy. In others, where diagnosis and prognosis were less clear, scintigraphy aided in the diagnosis and accurately identified a low-risk subgroup as did no other parameter.

Aged↗

Myocardial perfusion scintigraphy in left bundle branch block: a perspective on the issue from image analysis in a clinical context.

Among selected study populations, myocardial perfusion scintigraphy (MPS) in patients with left bundle branch block (LBBB) has been reported to show a low specificity for the diagnosis of coronary disease. However, the stress electrocardiogram (ECG) is nondiagnostic in this setting. To place this method in its appropriate clinical context, we evaluated MPS in all 69 consecutive patients with LBBB studied with scintigraphy for clinical reasons during a 4-year period. Among 32 patients who underwent coronary angiography for clinical indications, per patient sensitivity, 96%; per vessel sensitivity, 84%, 50%, and 100% for left anterior descending (LAD), left circumflex (LCX), and right coronary artery (RCA) involvement, respectively; and per vessel specificity, 95% and 68% for LCX and RCA disease, respectively, were not significantly different from those previously published for the method in patients without LBBB. Although per patient specificity, 38%, and specificity, 39%, for LAD disease were low, the predictive value of a positive test remained relatively high (83%) owing to the small number of patients selected for angiography, in part based on scintigraphic findings, with normal coronary anatomy. In addition to a possible specific pathophysiologic cause related to LBBB, apparent perfusion abnormalities in the LAD distribution may relate to generic conditions that can make scintigraphic interpretation ambiguous, often in the anterior distribution, regardless of the clinical setting. Additionally, the apparent lack of scintigraphic specificity in the LAD distribution could relate in part to a selection bias toward catheterization of patients with induced scintigraphic abnormalities, especially in the LAD distribution.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗