Prevention of selenite-induced cataractogenesis by Origanum vulgare extract.

The present study sought to assess antioxidant effect of Origanum vulgare extract in preventing selenite-induced cataractogenesis. This study was performed on Young white rats received sodium selenite (30 nmol g(-1) birth weight) subcutaneously on day 13 post partum during two months in 2009. Cataract formation and intensity was detected and measured by slit-lamp. Origanum vulgare (Ov) extract (2 g kg(-1)) was given (1-2 times) intraperitoneal at different times with respect to the selenite administration lens opacification was analyzed in selenite, selenite-Ov, Ov and control groups on day 7 after selenite administration. Ov extract have revealed a significant protective effect against selenite induced cataract when injected 1 and 2 day (2 times) before selenite injection. There is a protective effect of Ov against selenite induced cataract formation. It is supposed that the anticataract effect of Ov extract could be based on direct or indirect antioxidant mechanisms.

A comparison of cold knife, CO2 laser, and electrosurgical loop conization in the treatment of cervical intraepithelial neoplasia.

Thirty patients with histologically confirmed high-grade squamous intraepithelial lesions (SIL) were treated by either cold knife, laser, or electrosurgical loop conization, all of which were performed under general anesthesia. The three methods were compared with respect to the immediate surgical complications, ease of performance, delayed complications, and quality of histologic specimens. The electrosurgical loop conization had decreased blood loss and reduced operative time and proved to be tissue sparing. There were no significant differences in the three groups in the persistence rates of cervical intraepithelial neoplasia (CN) after treatment. Histologic analysis revealed comparable coagulation artifact in the laser and electrosurgical loop groups that the cold knife group did not have. The endocervical component of the electrocautery showed extensive denudation and coagulation artifact that made recognition of CIN extremely difficult. We conclude that the electrocautery should be used only as an excisional method of the transformation zone for lesions well defined on the ectocervix, since it is unreliable if the lesion extends into the endocervix.

Leiomyomatosis peritonealis disseminata. A report of two cases.

Leiomyomatosis peritonealis disseminata (LPD), also known as diffuse peritoneal leiomyomatosis, is characterized by the presence of multiple, small nodules scattered over the abdominopelvic viscera and peritoneum. These nodules are composed of benign smooth muscle cells. As shown in this report of two cases, the disorder occurs mostly in women of reproductive age and rarely in postmenopausal ones. LPD is a benign condition for which conservative management is indicated.

Structural features of Plasmodium cytochrome b that may underlie susceptibility to 8-aminoquinolines and hydroxynaphthoquinones.

Appropriate functioning of mitochondria is critical for survival and growth of erythrocytic stages of malarial parasites, making it an attractive target for antimalarial drugs which may take advantage of unique features of parasite mitochondrial metabolism. We have sequenced the presumptive mitochondrial DNA, the 6-kb element, of Plasmodium falciparum, permitting an analysis of the predicted structure of parasite electron transport proteins. Although the overall structures of the 3 polypeptides, cytochrome c oxidase subunit 1, cytochrome c oxidase subunit 3, and cytochrome b (cyt b), were similar to those from other species, some striking differences were observed, especially for the cyt b. Analysis of the cyt b structure showed that the critical quinone binding sites of the protein are quite divergent from those of other species. Comparative analysis suggests that these changes are the likely cause for the resistance of parasite cytochrome bc1 complex to antimycin and related inhibitors. We suggest that the same features are responsible for increased affinity of the parasite cyt b for antimalarial compounds of class 8-aminoquinolines and hydroxynaphthoquinones, explaining the therapeutic value of these drugs.

Primary biliary carcinoma with metastasis to the ovary.

The ovary is a relatively frequent site of metastasis from malignant neoplasia arising elsewhere in the body, the majority of these originating from the GI tract. The best known tumor of this type is signet-ring cell adenocarcinoma (Krukenberg tumor) of gastric origin. The gall bladder and bile ducts are rare sources of these metastases. Asymptomatic carcinoma of the cystic duct metastasizing to the ovary is extremely rare. We are reporting such a case in which the patient presented with no GI or hepatic symptoms. The cystic duct carcinoma was an incidental finding from routine and careful examination of the abdominal viscera.

Regulation of JCVL promoter function: evidence that a pentanucleotide "silencer" repeat sequence AGGGAAGGGA down-regulates transcription of the JC virus late promoter.

The human neurotropic papovavirus JCV contains sequences within the two 98-bp tandem repeats which play a key role in glial-specific transcription of the viral early and late promoters. Previous analysis of the 98-bp sequence has delineated several protein-binding domains that are recognized by nuclear factors present in human brain cells. In the present study, by deletion mutation analysis, we have identified a region within each 98-bp repeat that reduces transcriptional activity of the JCV late promoter (JCVL). Using synthetic oligonucleotides spanning this region, designated "OP," we demonstrate that down-regulation of the JCVL promoter is associated with a pentanucleotide repeat sequence (AGGGAAGGGA) juxtaposed to the poly(dA) tract within the 98-bp tandem repeats. The OP sequence interacts specifically with a protein derived from glial nuclear extract and forms a major 56- to 60-kDa complex. Methylation interference experiment indicates that the three G residues proximal to the poly(dA) tract make major groove contacts with the protein. Single-base-pair substitution of these residues suggests that the complex can form in the presence of two of the three guanosyl residues. The possible role of this protein in regulating the JCV lytic cycle in concert with nearby regulatory elements within JCV promoter region is discussed.

Nuclear proteins in mouse brain cells bind specifically to the myelin basic protein regulatory region.

Expression of myelin basic protein (MBP) in mice is regulated in a cell- and stage-specific manner during brain development. The MBP control region contains multiple cis-acting elements, shown by in vivo and in vitro assays, which are responsible for its unique pattern of transcription. Using synthetic DNA fragments spanning the MBP control region, we have analyzed nuclear proteins obtained from newborn (2-3 d), young adult (18-30 d), and adult (60 d) animals; these nuclear proteins form DNA-protein complexes with the MBP regulatory region. Brain extracts from young adult and adult mice showed enhanced binding activities with the sequences supporting transcriptional activation in glial cells. Deletion analysis of the proximal activating sequence located at position -14 to -50 with respect to the RNA initiation site resulted in identification of a small region, located between nucleotides -14 to -37, which is required for formation of the complexes. Southwestern assay revealed a major 39-kD protein from young adult brain extract that recognizes the sequences between nucleotides -14 to -37. An additional minor 37-kD protein, derived from young adult brain extract, was also found to be associated with this proximal activating region. Of particular interest is the observation that the minor 37-kD protein became more abundant in the extract derived from adult brain, whereas the major 39-kD protein became less abundant. The possible role of these proteins in cell/stage-specific transcription of MBP is discussed.

Myelin basic protein gene transcription. Identification of proximal and distal cis-acting regulatory elements.

Myelin basic proteins (MBPs) represent a major component of the myelin membrane which are exclusively expressed by glial cells in the nervous system. The cell type-specific expression of MBP is controlled preferentially at the level of RNA synthesis. To investigate the mechanisms by which the MBP gene is regulated, we analyzed transcriptional regulation of this gene in glial and non-glial cells. We have demonstrated that the 320 base pairs upstream of the MBP transcriptional start site contain regulatory elements that preferentially stimulate transcription of MBPs in glial cells. Using a test vector containing the simian virus 40 (SV40) early promoter placed upstream of the bacterial chloramphenicol acetyltransferase gene, we localized three major promoter elements within the 5'-upstream sequence. These elements, designated MB1, MB4, and MB7, spanning proximal (-14 to -50) and distal (-130 to -169 and -249 to -288) positions with respect to the RNA initiation site, activated SV40 promoter transcription more than 40-fold in glial cells. The promoter distal elements, MB4 and MB7, enhanced SV40 promoter activity 2- and 8-fold, respectively, in L cells. Using the gel mobility shift assay, we have demonstrated that the MBP activators (MB1, MB4, and MB7) interact with multiple proteins derived from glial and L cell extract and result in the formation of several complexes. Comparison of band intensity of these complexes implies that these cells contain both unique and ubiquitous DNA binding proteins that recognize the DNA sequences within these activators. These studies suggest that the MBP promoter consists of several regulatory sequences in which the proximal element, MB1, and one of the distal elements, MB4, are selectively more active in glial cells than in L cells. Thus, these novel regulatory elements, in concert with other sequences, appear to stimulate MBP promoter transcription in glial cells.

Trans-activation of the JC virus late promoter by the tat protein of type 1 human immunodeficiency virus in glial cells.

Progressive multifocal leukoencephalopathy (PML) is a demyelinating disease of the central nervous system caused by the JC virus (JCV), a human papovavirus. PML is a relatively rare disease seen predominantly in immunocompromised individuals and is a frequent complication observed in AIDS patients. The significantly higher incidence of PML in AIDS patients than in other immunosuppressive disorders has suggested that the presence of human immunodeficiency virus type 1 (HIV-1) in the brain may directly or indirectly contribute to the pathogenesis of this disease. In the present study we have examined the expression of the JCV genome in both glial and non-glial cells in the presence of HIV-1 regulatory proteins. We find that the HIV-1-encoded trans-regulatory protein tat increases the basal activity of the JCV late promoter, JCVL, in glial cells. In a reciprocal experiment, the JCV early protein, the large tumor antigen, stimulates expression from JCVL and HIV-1 long terminal repeat promoter in both glial and non-glial cells. This trans-activation occurs at the level of RNA synthesis, as measured by the rate of transcription, stability of the message, and translation. We conclude that the presence of the HIV-1-encoded tat protein may positively affect the JCV lytic cycle in glial cells by stimulating JCV gene expression. Our results suggest a mechanism for the relatively high incidence of PML in AIDS patients than in other immunosuppressive disorders. Furthermore, our findings indicate that the HIV-1 regulatory protein tat may stimulate other viral and perhaps cellular promoters, in addition to its own.

Regulation of JCVL promoter function: transactivation of JCVL promoter by JCV and SV40 early proteins.

To better understand the basis of cell type specificity of JCV replication, we have analyzed the expression of the viral late promoter in glial cells. Using transient transfection procedures, we show that the late gene expression, like that of the early gene, is restricted to glial cells. However, cotransfection with a plasmid producing the JCV early protein, T-antigen, stimulates expression from the JCV late promoter in both glial and non-glial cells. The SV40-encoded T-antigen acts similarly on transcription of JCV late promoter in both cell types. This transacting effect occurs at the level of RNA synthesis, as measured by the rate of transcription, stability of the message, and translation. These results indicate that basal JCV late promoter activity is restricted to glial cells, whereas in the presence of viral early protein this promoter functions in both glial and non-glial cells.

Cell type-specific expression of JC virus early promoter is determined by positive and negative regulation.

We analyzed control sequences of the human papovavirus JC virus (JCV) to define the cis-acting elements that regulate specific expression of the viral early region genes in glial cells. Nuclear run-on transcription, S1 analysis, and chloramphenicol acetyltransferase enzyme activity in a transient transfection assay established that the cell type-specific expression of JCV early genes is determined at the transcriptional level. Using DNase footprinting analysis of nuclear proteins prepared from glial and nonglial cells, we located four regions within the JCV control sequences that specifically interacted with the proteins. In glial cells, all four domains contributed to the specific expression of a heterologous promoter, whereas in nonglial cells, two protein-binding regions showed no effect on basal transcriptional activity and the other two domains significantly downregulated transcription of the promoter. We conclude that cell type-specific transcription of the JCV early promoter is under both positive and negative regulation in eucaryotic cells.

Suppression of matrix protein synthesis by herpes simplex virus in bovine smooth muscle cells.

We investigated the effect of herpes simplex virus type 1 infection on the synthesis of extracellular matrix proteins by bovine smooth muscle cells. The cells supported viral replication, which reached maximum at 24 hrs post-infection. Uninfected and infected (multiplicity of infection of 5 or 20) cultures of smooth muscle cells were labeled with [14C]proline at five hrs post-infection. Incorporation of radioactivity into non-dializable protein was determined following electrophoresis of the cell-matrix and medium fractions. The synthesis of fibronectin and collagen types I and III was almost completely suppressed. The data suggest that HSV-1 infection of smooth muscle cells in vitro alters extracellular matrix synthesis.

Excision of Bartholin duct cysts using the CO2 laser.

Many methods have been used to treat cystic enlargement of the duct of the Bartholins gland. Widely used methods have the disadvantage of recurrence, hemorrhage, persistent drainage, or considerable scarring. The carbon dioxide laser offers a simple and effective means of treating cystic Bartholin ducts. Ten patients were treated with the CO2 laser. The cyst was incised with the laser and the cyst wall was vaporized from the inside. In a follow-up period of one to four years, there were only two recurrences, and these responded to a second treatment. Surgery was rapid and uncomplicated in all cases, and healing occurred without scar formation. The CO2 laser offers several advantages over conventional methods for treating cysts of the Bartholin duct.

Serological comparison and antigenic relationships of seven serotypes of infectious bronchitis virus using the hemagglutination-inhibition test.

The antigenic relationships, antigenic spectrum, and immunogenicity of seven isolates of infectious bronchitis virus (IBV) were examined using the hemagglutination-inhibition (HI) test. Because there was a discontinuity of antigenic relationships and a high degree of cross-reactivity among serotypes of IBV in cross-hemagglutination-inhibition tests, the range of antigenic spectrum used to group the serotypes with the HI test should be wider than the limits suggested by the plaque-reduction test. The HI test may provide valuable information in monitoring the immune status of a flock following vaccination when the area has a history of infectious bronchitis infection. It may also be used as a rapid diagnostic test if a flock is experiencing an outbreak of a disease caused by emergence of a new type of IBV. Interpretation of HI titers in evaluating immune status of chickens following infection with IBV depends on further cross-challenge and cross-protection studies of various types of IBV.

Determination of the antigenic relationships within the Massachusetts (M41) type of infectious bronchitis virus using the hemagglutination-inhibition test.

The antigenic relationships, antigenic spectrum, and immunogenicity of seven IBV-Massachusetts-41 isolates were investigated using the hemagglutination-inhibition (HI) test. HI titers equal to 32 are considered suspicious, titers lower than 32 are considered negative, and titers higher than 32 are considered positive immune responses to infectious bronchitis virus (IBV). Some isolates of Massachusetts-41 ( M41 ) were capable of inducing large quantities of antibodies in chickens following inoculation and demonstrated a wider antigenic spectrum than others. Variations in antigenic spectrum observed within M41 isolates in this study are in agreement with previous reports. This variation is of importance in selecting a proper vaccine strain for a successful immunization program for IBV.

Preparing hemagglutinating antigen from isolates of infectious bronchitis virus.

The hemagglutinating (HA) activity of 14 strains of infectious bronchitis virus (IBV) was investigated. The optimal conditions for IBV antigen preparation include inoculation of 10- or 11-day-old specific pathogen-free embryonated eggs and incubation for 30 hours at 37 C. Embryos were inoculated via the allantoic cavity with 0.1 ml of a low embryonic passage of the virus (10(7) to 10(8) EID50/ml). Allantoic fluid was harvested and pooled, and a 100-fold concentration of virus particles was achieved by centrifugation for 3 hours at 30,000 x g. Virus pellets were resuspended in Tris-hydrochloride buffer containing 3 units of phospholipase-C (type-1) enzyme/ml and incubated for 2 hours at 37 C. All IBV strains tested demonstrated positive HA activity with chicken red blood cells. The antigen was stored in liquid state or lyophilized at 4 C.

Acute inversion of the uterus.

Acute inversion of the uterus is a rare emergency that occurs during the third stage of labor. Three cases of this complication are reported. General anesthesia, fundal pressure on a soft uterus, fundal implantation of the placenta, and adherent placenta were identified as the possible causes. Prompt recognition and replacement are essential in the management to preempt morbidities and mortality. The prevention, etiology, and management of this serious complication are discussed.