Biomedical subjects

R P Misra

Publications and source records attributed to R P Misra.

At least 19 recordsLinked to original sources

PubMed1996

Expression of the serum response factor gene is regulated by serum response factor binding sites.

The serum response factor (SRF) is a ubiquitous transcription factor that plays a central role in the transcriptional response of mammalian cells to a variety of extracellular signals. Notably, SRF has been found to be a key regulator of members of a class of cellular response genes termed immediate-early genes (IEGs), many of which are believed to be involved in regulating cell growth and differentiation. The mechanism by which SRF activates transcription of IEGs in response to mitogenic agents has been extensively studied. Significantly less is known about how expression of the SRF gene itself is mediated. We and others have previously shown that the SRF gene is itself transiently induced by a variety of mitogenic agents and belongs to a class of "delayed" early response genes. We have cloned the SRF promoter and in the present study have analyzed the upstream regulatory sequences involved in mediating serum responsiveness of the SRF gene. Our analysis indicates that inducible SRF expression requires both SRF binding sites located within the first 63 nucleotides upstream from the start site of transcriptional initiation and an Sp1 site located 83 nucleotides upstream from the start site. Maximal transcriptional activity of the promoter also requires two CCAATT box sites located 90 and 123 nucleotides upstream of the start site.

3T3 Cells↗

PubMed1996

Observer variation in AgNOR counts in neoplastic breast lesions.

OBJECTIVE: To determine interobserver and intraobserver variability of AgNOR quantitation in neoplastic lesions of the breast. STUDY DESIGN: Forty-five cases, 20 benign and 25 malignant lesions, were included in the study. Counts were performed on one slide from each case within a pre-marked area of about 1 cm2 in a standardized manner by two observers blind to the histopathologic diagnosis and independent of each other and repeated after two weeks. Interobserver and intraobserver agreement was assessed using the Bland-Altman method. RESULTS: Our results showed small mean interobserver and intraobserver differences but wide limits of agreement. CONCLUSION: Observer variation in AgNOR counts is too high for the method to be of any diagnostic or prognostic relevance.

Breast Neoplasms↗

PubMed1994

L-type voltage-sensitive calcium channel activation stimulates gene expression by a serum response factor-dependent pathway.

A mechanism by which calcium-induced signals are transduced to the nucleus to activate transcription of the c-fos proto-oncogene has been characterized. The serum response element (SRE), a region of the c-fos gene which controls growth factor-induced transcription, is now shown to mediate c-fos transcription in response to activation of L-type voltage-sensitive calcium channels. Calcium-dependent transcriptional activation through the SRE is mediated by the serum response factor (SRF). Membrane depolarization induces phosphorylation of SRF at Ser-103, an event shown to enhance the ability of SRF to bind the SRE. Ca(2+)-induced SRF phosphorylation occurs via a pathway that may involve Ca2+/calmodulin-dependent kinases.

Animals↗

PubMed1994

Capnocytophaga keratitis. A clinicopathologic study of three patients, including electron microscopic observations.

BACKGROUND: Histopathologic studies of this unusual keratitis caused by Capnocytophaga species have not been reported previously. METHODS: The authors report the light microscopic and ultrastructural findings of three patients with a distinctive necrotizing keratitis caused by an anaerobic gram-negative bacillus. In three patients, ages 19, 81, and 91 years, a necrotizing stromal keratitis developed; two of these patients had a previous penetrating keratoplasty for pseudophakic bullous keratopathy. The first patient did not have ocular surgery previously and was treated initially for presumed Acanthamoeba keratitis. RESULTS: By light microscopy, all three keratectomy specimens were strikingly similar and showed a necrotizing and/or suppurative stromal keratitis displaying myriad slender, fusiform, gram-negative bacilli located anterior to Descemet's membrane and extending into the deep corneal stroma, assuming a "picket fence" appearance. Cultures of the cornea in case 1 grew Capnocytophaga ochracea. For the remaining two patients, a diagnosis presumptively was made based on characteristic histopathologic features. Results of electron microscopic examination showed numerous bacilli that were mostly extracellular; occasional organisms were phagocytosed by macrophages. CONCLUSION: The histopathologic features of Capnocytophaga keratitis are unique; therefore, a presumptive diagnosis can be made based on the morphology and location of the bacilli in the keratectomy specimens. To the authors' knowledge, this is the first study describing the typical histopathologic and electron microscopic findings of Capnocytophaga keratitis.

Adult↗

PubMed1993

A growth factor-induced kinase phosphorylates the serum response factor at a site that regulates its DNA-binding activity.

A signaling pathway by which growth factors may induce transcription of the c-fos proto-oncogene has been characterized. Growth factor stimulation of quiescent fibroblasts activates a protein kinase cascade that leads to the rapid and transient phosphorylation of the serum response factor (SRF), a regulator of c-fos transcription. The in vivo kinetics of SRF phosphorylation and dephosphorylation parallel the activation and subsequent repression of c-fos transcription, suggesting that this phosphorylation event plays a critical role in the control of c-fos expression. The ribosomal S6 kinase pp90rsk, a growth factor-inducible kinase, phosphorylates SRF in vitro at serine 103, the site that becomes newly phosphorylated upon growth factor stimulation in vivo. Phosphorylation of serine 103 significantly enhances the affinity and rate with which SRF associates with its binding site, the serum response element, within the c-fos promoter. These results suggest a model in which the growth factor-induced phosphorylation of SRF at serine 103 contributes to the activation of c-fos transcription by facilitating the formation of an active transcription complex at the serum response element.

3T3 Cells↗

PubMed1992

Bipolar cautery and internal thermal sclerostomy in a rabbit model.

Internal thermal sclerostomy (ITS) was performed unilaterally in 35 adult New Zealand rabbits using a pinpoint bipolar cautery probe and radio-frequency power supply, with the nonoperated eyes serving as controls. Standard trabeculectomy using a limbal-based flap was also performed on 10 additional rabbits, and served as a second bench mark for comparison with the ITS technique. Intraocular pressure (IOP) was measured in all eyes preoperatively and on postoperative days 2, 4, 6, and 8. A significant (P = .005) difference between the reduction in IOP in the ITS eyes and in the control eyes was found up to postoperative day 8; on that day the reduction in IOP was 5.2 mm Hg. IOP in the eyes undergoing standard trabeculectomy was significantly (P = .05) reduced up to postoperative day 2, and gradually decreased, to 2.5 mm Hg, on postoperative day 8. The greatest reduction in IOP (2.9 mm Hg) for these eyes also occurred on postoperative day 2. Complications of ITS included iris burn (23%), peripheral corneal edema (17%), and iritis (9%). No ruptured blebs, flat anterior chambers, hyphemas, or lens damage occurred. The potential advantages of the ITS procedure using the bipolar cautery probe include a decreased risk of cataract formation because of the curved probe design. The procedure is also technically simple to perform and requires only inexpensive and readily-available equipment.

Animals↗

PubMed1991

The serum response factor is extensively modified by phosphorylation following its synthesis in serum-stimulated fibroblasts.

Growth factor regulation of c-fos proto-oncogene transcription is mediated by a 20-bp region of dyad symmetry, termed the serum response element. The inner core of this element binds a 67-kDa phosphoprotein, the serum response factor (SRF), that is thought to play a pivotal role in the c-fos transcriptional response. To investigate the mechanism by which SRF regulates c-fos expression, we generated polyclonal anti-SRF antibodies and used these antibodies to analyze the biochemical properties of SRF. These studies indicate that the synthesis of SRF is transient, occurring within 30 min to 4 h after serum stimulation of quiescent fibroblasts. Newly synthesized SRF is transported to the nucleus, where it is increasingly modified by phosphorylation during progression through the cell cycle. Within 2 h of serum stimulation, differentially modified forms of SRF can be distinguished on the basis of the ability to bind a synthetic serum response element. SRF protein exhibits a half-life of greater than 12 h and is predominantly nuclear, with no change occurring in its localization upon serum stimulation. We find that the induction of SRF synthesis is regulated at the transcriptional level and that cytoplasmic SRF mRNA is transiently expressed with somewhat delayed kinetics compared with c-fos mRNA expression. These features of SRF expression suggest a model whereby newly synthesized SRF functions in the shutoff of c-fos transcription.

Animals↗

PubMed1990

Carcinoid tumorlet of the bronchus in a child.

We have reported a case of bronchial carcinoid tumorlet in a 10-year-old boy. Because this type of tumor rarely occurs in children under the age of 16 years, is relatively benign in its biologic behavior, and may be misdiagnosed as cancer, occurrences of such a tumor warrant close consultation with the pathologist and radiologist as well as a conservative approach in surgical management.

Bronchial Neoplasms↗

PubMed1990

Contact lens wear enhances adherence of Pseudomonas aeruginosa and binding of lectins to the cornea.

Extended wear soft contact lenses are associated with an increased incidence of Pseudomonas aeruginosa keratitis. Because the first step in the pathogenesis of this disease is adherence of the microorganism to the corneal surface, we studied the effect of soft contact lens wear on the adherence of P. aeruginosa to the cornea. Rabbits were fitted for extended wear soft contact lenses in the left eye, and the right eye served as a control. Both eyes were then closed with a partial tarsorrhaphy. After 1-5 days of wear, the lenses were removed and the corneas of the left and right eye were removed. Differences in the number of adherent Pseudomonas and in lectin binding to lens-wearing corneas and non-lens-wearing corneas were determined. After 1, 3, and 5 days of soft contact lens wear, there was a significant increase in the number of P. aeruginosa adherent to the lens-wearing cornea. Three to eight times as many bacteria adhered to the lens-wearing eye as compared with the control eye (p less than 0.05). In addition, a soft contact lens placed in the eye followed by the immediate application of P. aeruginosa resulted in an eightfold increase in adherence of bacteria to the lens-wearing cornea (p less than 0.05). Lens wear also led to an increase in binding of concanavalin A (Con A), wheat germ agglutinin (WGA), and Maclura pomifera agglutinin (MPA) to surface epithelium covered by the lens. These corneal epithelial changes induced by extended wear soft contact lenses may provide some insight as to why soft contact lens wearers are predisposed to Pseudomonas keratitis.

Animals↗

PubMed1990

Use of carbon fibers for repair of abdominal-wall defects in rats.

Carbon in the form of 8-micron fibers induces growth of connective tissue. The purpose of this study was to measure and histologically characterize tissue ingrowth occurring in carbon fibers implanted for up to 12 months in abdominal-wall defects in rats, compared with polypropylene mesh. Carbon fibers induced significantly more tissue ingrowth than polypropylene mesh at 6 to 12 months postoperatively. The predominant tissues associated with carbon fibers and polypropylene mesh were dense connective tissue and fat, respectively. Fragmentation of the implants did not occur, and implant debris was not found in the regional lymph nodes. Carbon fibers are potentially useful for reinforcing abdominal-wall defects.

Abdominal Muscles↗

PubMed1989

Aggregated platelets enhance adherence of Candida yeasts to endothelium.

The adherence of Candida albicans yeasts to human umbilical vein endothelium to subendothelial extracellular matrix (ECM) was investigated. Yeasts added to confluent endothelium in citrated platelet-poor plasma adhered on the average of 1 colony forming unit (cfu) per culture well. When platelets were added as platelet-rich plasma, a significant increase of yeast adherence was not seen. However, when endothelium was contracted by treatment with 2 mM EDTA, resulting in exposure of ECM, yeast adherence was increased to 10 cfu/well. When platelets were added with these yeasts, the number of adhering yeasts was further increased to 23 cfu/well (P less than .01). This represented an increase in adherence of yeasts of 230%. When the endothelial cells were completely removed and ECM exposed, platelets were found to likewise augment yeast adherence. Platelets, when added to the ECM, formed aggregates to which the yeasts firmly adhered. Likewise, when platelets were aggregated by adenosine diphosphate and mixed with yeasts, yeasts were shown to bind avidly to aggregated platelets, whereas yeasts did not adhere to unactivated, discoid platelets. Thus, exposed subendothelial ECM induces the aggregation of platelets and yeasts bind avidly to these platelet aggregates.

Blood Platelets↗

PubMed1989

The contribution of bacterial surface hydrophobicity to the process of adherence of Pseudomonas aeruginosa to hydrophilic contact lenses.

Ten isolates of Pseudomonas aeruginosa obtained from the corneas of patients with Pseudomonas keratitis adhered to soft contact lenses in significantly greater numbers than did six isolates from other body sites (P less than .05). However, there was no predominant serotype among the 10 corneal isolates tested. Isolates grown statically in broth at 37 degrees C formed a pellicle and adhered two times as much to contact lenses as did isolates grown in broth while shaking which did not form a pellicle (P less than .01). The more adherent isolates (grown at 37 degrees C) were shown to be more hydrophobic than the less adherent bacteria (grown at 26 degrees C) by their propensity to accumulate at the interface between hexadecane and saline and their movement into polyethylene glycol from dextran. These corneal isolates agglutinated erythrocytes, a process that was inhibited by dilute solutions (as low as 0.01%) of three commonly used surfactants. These same surfactants inhibited the adherence of Pseudomonas aeruginosa to soft contact lens surfaces by as much as 52%. It is concluded that hydrophobic interactions may significantly contribute to the ability of Pseudomonas aeruginosa to adhere to contact lenses.

Bacterial Adhesion↗

PubMed1989

Isolation of a herpes simplex virus type 2 that is retinovirulent in mice.

The virulence of a herpes simplex virus type 2 (HSV-2) isolated from the urine of a patient (SL) with acquired immunodeficiency syndrome (AIDS) and bilateral acute retinal necrosis (ARN), was investigated in mice. The ratio of plaque forming units (PFU) in fibroblasts to the 50% lethal dose (LD50) of HSV-2(SL) in mice was 10 fold more than the PFU to LD50 ratio of a neurovirulent HSV-2, strain 186. Further, HSV-2(SL) caused retinitis with and without lethal encephalitis in mice inoculated intracranially (i.c.). In contrast, mice inoculated with HSV-2(186) died of encephalitis without ocular disease. HSV-2(SL) was isolated from eye and/or brain tissue 1 to 15 days post i.c. inoculation. Ocular disease progressed from an initial mild chorioretinitis on day 8 to total retinal necrosis with panuveitis by day 11 in mice given 10 PFU of HSV-2(SL) i.c. HSV antigen was detected initially in the cells of the optic nerve and spread into the ganglial cells of the nerve fiber layer, the neurosensory cells of the inner nuclear layer, and the cells of the retinal pigment epithelium (RPE) between days 8 and 10. Thus, this study supports the concept that HSV neurovirulence varies between strains and presents a HSV-2 neurotransmission animal model of ARN.

Acquired Immunodeficiency Syndrome↗

PubMed1989

A partial-thickness epithelial defect increases the adherence of Pseudomonas aeruginosa to the cornea.

Some patients with infectious keratitis have no clinically demonstrable corneal abrasion predisposing them to infection. Subtle, undetectable corneal injuries may facilitate bacterial adherence to the cornea, eventually leading to keratitis. To study this concept, we have developed a rabbit model in which a partial-thickness corneal epithelial defect was induced by filter paper impression on the cornea that removed one to two layers of corneal epithelium. Following this injury, the corneas were incubated with Pseudomonas aeruginosa, washed, and the number of bacteria adhering to the injured corneas as well as to control corneas was quantitated. Corneas treated with filter paper, either ex vivo or in vivo, allowed 20 times more bacteria to adhere than did the untreated control corneas (P less than 0.01). This superficial epithelial defect increased Pseudomonas adherence to the cornea for up to 72 hr after injury. When corneal injury was extended to the stroma, the adherence of Pseudomonas was further augmented as compared to adherence to the superficially injured cornea. Thus, we conclude that a clinically subtle, partial-thickness corneal epithelial injury can markedly facilitate the adherence of Pseudomonas aeruginosa, which may be an important predisposing factor for infectious keratitis.

Animals↗

PubMed1987

Carbohydrate deposits on the surfaces of worn extended-wear soft contact lenses.

Three different commercial extended-wear soft contact lenses worn continuously by patients for at least 28 days were stained with fluorescein isothiocyanate-labeled lectins. These lectins detected the presence of alpha-linked or beta-linked D-mannose, D-glucose, D-galactose, L-fucose, N-acetyl-D-glucosamine, N-acetyl-D-galactosamine, and N-acetyl neuraminic acid (sialic acid) on the surfaces of the contact lenses. These saccharides are bound to other sugars that likely account for an integral part of glycoprotein and/or glycolipid deposits on lens surfaces. These tear deposits may contribute to the chemical spoilage of the lens and, furthermore, may serve as specific receptors for pathogenic microorganisms commonly implicated in extended-wear soft contact lens-associated infectious keratitis.

Contact Lenses, Extended-Wear↗

PubMed1987

Factors influencing survival of patients with adenoid cystic carcinoma of the salivary glands.

The uncertain long-term cure of adenoid cystic carcinoma of the salivary glands was determined in 30 treated patients seen between 1952 to 1984. Ten of 25 patients received elective postoperative irradiation (SEPI) after surgical extirpation of the primary/recurrent lesion; five patients had definitive radiotherapy alone. Overall, survival rates were 60%, 30%, and 7% at five, 10, and 20 years, respectively. The incidence of local recurrence was 37%, regional recurrence 3%, and distant metastasis 37%. Fewer recurrences and distant metastases were in the SEPI group than the surgery alone group. The majority of failures occurred within 10 years after initial diagnosis of primary malignancy; most of the patients died within five years after failure(s) detection. Correlations of prognosis to site of origin, and local control to total radiotherapy dose were observed. It is concluded that cure of disease can be achieved with the use of appropriate therapy.

Adult↗

PubMed1987

The adherence of Pseudomonas aeruginosa to soft contact lenses.

The adherence of Pseudomonas aeruginosa to extended-wear soft contact lenses (EWSCLs) may be an important initial step in the pathogenesis of EWSCL-associated infectious keratitis. P. aeruginosa tend to adhere more to worn EWSCLs than unworn EWSCLs (P less than 0.05). Normal tear components such as aqueous solutions of albumin, lysozyme, and lactoferrin all significantly enhance adherence of P. aeruginosa to unworn EWSCLs often by as much as 300%. The presence of a 1% solution of sialic acid in the bathing medium significantly reduces the adherence of P. aeruginosa to both unworn and worn lenses. Inhibition of bacterial adherence could also be achieved with the addition of mucin (which contains terminal sialic acid residues in its major sugar chains). Therefore, selective adherence by P. aeruginosa to a specific sugar (sialic acid) may be important in the initial attachment of the bacterium to soft contact lenses.

Bacterial Adhesion↗

PubMed1986

Blastomycotic meningitis.

A difficult and tragic case of central nervous system blastomycosis is presented as the basis for a review of the diagnostic criteria required to establish this diagnosis and to present a scheme for the diagnosis and therapy of chronic meningitis. The diagnosis in our case was complicated by preexisting inadequately treated tuberculosis; a prepontine mass; and a cervical intradural, extramedullary, circumferential mass. This exceptional case of chronic basilar meningitis with cervical myelopathy was caused by Blastomyces dermatitidis.

Adult↗