Biomedical subjects

G Sundaresan

Publications and source records attributed to G Sundaresan.

7 recordsLinked to original sources

PubMed2005

Quantum dots for live cells, in vivo imaging, and diagnostics.

Research on fluorescent semiconductor nanocrystals (also known as quantum dots or qdots) has evolved over the past two decades from electronic materials science to biological applications. We review current approaches to the synthesis, solubilization, and functionalization of qdots and their applications to cell and animal biology. Recent examples of their experimental use include the observation of diffusion of individual glycine receptors in living neurons and the identification of lymph nodes in live animals by near-infrared emission during surgery. The new generations of qdots have far-reaching potential for the study of intracellular processes at the single-molecule level, high-resolution cellular imaging, long-term in vivo observation of cell trafficking, tumor targeting, and diagnostics.

Animals↗

PubMed2004

MicroPET imaging of Cre-loxP-mediated conditional activation of a herpes simplex virus type 1 thymidine kinase reporter gene.

Site-specific recombination tools such as the Cre-loxP system are used to create animal models where conditional gene deletion/activation studies are required. In the current proof of principle study, we have demonstrated that a PET reporter gene (PRG), the herpes simplex virus type 1 thymidine kinase (HSV1-tk), can be made to remain silent and can be activated by Cre-loxP-mediated recombination in cell culture and in living mice. An adenovirus carrying a silent HSV1-tk was tail-vein injected (1 x 10(9) PFU) in six transgenic mice that express Cre recombinase in their liver (Cre+) and in four control mice (Cre-). The liver-specific expression of the PRG in Cre+ mice was detected in the microPET following injection of the reporter probe, 9-[4-fluoro-3-(hydroxymethyl)butyl]guanine ([(18)F]-FHBG). The [(18)F]-FHBG accumulation in the liver in terms of percent-injected dose per gram of tissue was 7.72+/-1.13 for the Cre+ mice and 0.10+/-0.02 for the Cre- mice (P<0.05) 48 h after adenoviral injection. These results were further validated by quantitative RT-PCR, western blotting and by in vitro assays for herpes simplex virus type 1 thymidine kinase enzyme activity. Thus by using the Cre-loxP system it is possible to modulate a PRG and noninvasively monitor the extent of Cre-loxP-mediated gene activation by imaging in a microPET scanner.

Adenoviridae↗

PubMed2001

Noninvasive optical imaging of firefly luciferase reporter gene expression in skeletal muscles of living mice.

The ability to monitor reporter gene expression noninvasively offers significant advantages over current techniques such as postmortem tissue staining or enzyme activity assays. Here we demonstrate a novel method of repetitively tracking in vivo gene expression of firefly luciferase (FL) in skeletal muscles of mice using a cooled charged coupled device (CCD) camera. We first show that the cooled CCD camera provides consistent and reproducible results within +/-8% standard deviation from mean values, and a detection sensitivity (range tested: 1 x 10(4) - 1 x 10(9) plaque form-ing units (pfu)) of 1 x 10(6) pfu of E1-deleted adenovirus expressing FL driven by a cytomegalovirus promoter (Ad-CMV-FL). The duration and magnitude of adenoviral mediated (1 x 10(9) pfu) FL gene expression were then followed over time. FL gene expression in immunocompetent Swiss Webster mice peaks within the first 48 hours, falls by 98% after 20 days, and persists for >150 days. In contrast, FL activity in nude mice remains elevated for >110 days. Finally, transduced Swiss Webster and nude mice were sacrificed to show that the in vivo CCD signals correlate well with in vitro luciferase enzyme assays (r(2)=0.91 and 0.96, respectively). Our findings demonstrate the ability of the cooled CCD camera to sensitively and noninvasively track the location, magnitude, and persistence of FL gene expression. Monitoring of gene therapy studies in small animals may be aided considerably with further extensions of this technique.

Adenoviridae↗

PubMed2001

Superior cervical ganglionectomy leads to dampening of amplitude of temperature rhythm in rats.

The circadian rhythm of body temperature (CRT) is a robust marker of the endogenous pacemaker function and is one of the most frequently studied rhythms. Melatonin, the main secretion of the pineal gland seems to have more of a thermomodulatory role in controlling the body temperature than having a direct role in thermoregulation. The sympathetic innervation to the pineal via the superior cervical ganglion determines the melatonin secretion, and superior cervical ganglionectomy (SCGx) decreases the secretion of melatonin. The present study was conducted on the Wistar rats (n = 12) to determine the role of melatonin in modulation of CRT. Adult male rats were either ganglionectomised (n = 6) or sham ganglionectomised (n = 6). Rectal temperature was recorded for CRT analysis. Cosinar analysis of the temperature record was done to get the acrophase, amplitude and mesor. Our results show that SCGx decreases the amplitude to the rhythm but has no effect on the mesor of the rhythm. Our study confirms that melatonin has little role to play in the thermoregulation and its role is mainly in thermomodulation.

Animals↗

PubMed2001

Short-term REM sleep deprivation effect on temperature rhythm of rats.

Circadian rhythm of body temperature (CRT) is altered in endogenous depression and many psychiatric disorders. Even the sleep pattern is disrupted. Sleep deprivation alleviates symptoms in depression. The present study was conducted to find the role of noradrenergic innervation to the pineal gland in bringing about the effect of REM sleep deprivation (REMSD) on the CRT. Adult male Wistar rats (n = 12) divided into 2 groups were used for the study. The group I rats (n = 6) underwent superior cervical ganglionectomy and the group II rats (n = 6) were sham ganglionectomised. After recovery rats were given REMSD for 48 hours. The CRT was measured in three periods of the study i.e. basal, post operative and post REMSD. The results indicated REMSD increased the Amplitude and Mesor of the CRT in both the groups which was shortlasting and reversible thus suggesting non sympathetic mediation of the pineal in bringing about the circadian rhythm alteration due to REMSD.

Animals↗

PubMed1990

Certain immunological parameters in subacute cold stress.

The effect of subacute cold swimming stress on the immune system of albino rats was investigated. Subacute cold stressed animals showed an increase in total WBC count, eosinophils and basophils. Phagocytic index and avidity index were also increased indicating hyperactive phagocytic process. On the other hand NBT reduction and soluble immune complex levels decreased significantly in stressed animals. There were no significant changes in the weight of the lymphoid organs.

Animals↗

PubMedDate not supplied

Genetic analyses of cis-acting sequences controlling expression of human immunodeficiency virus type 1 coreceptor-CCR5 gene in rabbits and CXCR4 gene in monkeys.

INTRODUCTION: Human immunodeficiency virus type 1 (HIV-1), HIV-2, and simian immunodeficiency virus all use chemokine receptors (CCR5, CXCR4, and minor receptors) to gain entry into a susceptible cell and establish infection successfully by way of membrane fusion. Many such chemokine receptors that can act as entry cofactors under in vitro conditions have been identified, but the roles of CCR5 and CXCR4 chemokine receptors in infection, tropism, and pathogenesis have been studied in greater detail. The promoter region of CCR5 gene is quite polymorphic in humans, and mutations that affect the progression of HIV-1 have been identified. STUDY DESIGN/METHODS: We studied the nature of mutations in the CCR5 promoter region in rabbits. Large number of mutations, deletions, substitutions, and point mutations were observed all along the 400 base pair region of the promoter. RESULTS: We show that rabbit CCR5 promoter possesses features common to both humans and monkeys and lacks the second highly polymorphic region B in the CCR5 promoter that was previously identified in monkeys. Besides providing important evolutionary information, our findings can directly make an impact on the known expression levels of CCR5 protein that can modulate the progression of HIV-1 in rabbits. The CXCR4 promoter of monkeys showed polymorphisms that were largely caused by single nucleotide changes when compared with humans. CONCLUSIONS: This distinctly different evolutionary pattern suggests a more important role for chemokine receptor-CCR5 in the host defense.

Animals↗