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Biomedical subjects

J W Ross

Publications and source records attributed to J W Ross.

At least 19 recordsLinked to original sources

Porcine conceptus Oct-4 mRNA expression during peri-implantation development.

OCT-4 is a transcriptional regulator of pluripotent cells throughout embryonic and germ cell lineage development prior to cellular differentiation during murine, bovine and porcine peri-implantation development. In contrast to murine OCT-4 expression, bovine and porcine expression is detected in both the inner cell mass and trophoblast. Delayed down regulation of OCT-4 gene expression in farm species may be a consequence of the lengthened period of peri-implantation. Expression of OCT-4 mRNA has not been characterized during conceptus attachment to the uterine surface in the pig. The objective of the present study was to determine conceptus OCT-4 mRNA expression during porcine peri-implantation development from days 10-17 of gestation. Total RNA was extracted from multiple pools of conceptuses collected on days 10, 12, 13, 15 and 17 of pregnancy. Quantitative RT-PCR was utilized to assay conceptus OCT-4 mRNA synthesis. Day of conceptus development significantly affected (p < 0.001) OCT-4 mRNA expression. Conceptus expression of OCT-4 was greatest on days 10 and 12 of pregnancy being approximately 7.7- and 11.6-fold greater compared to expression on days 15 and 17, respectively. Results from the present study suggest that down regulation of OCT-4 may be critical in trophoblastic expansion and uterine epithelial attachment during establishment of pregnancy in the pig.

Animals↗

Porcine endometrial and conceptus tissue kallikrein 1, 4, 11, and 14 gene expression.

Previous studies have suggested that the porcine endometrium may express several tissue kallikreins during the estrous cycle and early pregnancy. The present study investigated porcine endometrial and conceptus tissue kallikrein 1, 4, 11, and 14 mRNA expression during the estrous cycle and early pregnancy. Tissue kallikrein (KLK) gene expression was evaluated using quantitative RT-PCR and in situ hybridization. KLK1 expression was similar across the estrous cycle and early pregnancy, and localized to the endometrial luminal (L) and glandular (G) epithelium. KLK4 endometrial mRNA expression was greatest on days 0, 5, and 10 when compared with days 12, 15, and 17 of the estrous cycle and greater in cyclic compared with pregnant gilts. Expression of KLK4 was more intense in the stroma and uterine epithelium from days 0 to 10 of the estrous cycle. Endometrial KLK11 mRNA was not different between cyclic and pregnant gilts but the expression was greatest on days 10 and 12 compared with all other days evaluated. There was an increased intensity of KLK11 gene expression in the stratum compactum on day 10 of the estrous cycle and early pregnancy. Endometrial KLK14 mRNA expression was not detectable on days 5 and 10 but was expressed on days 0, 12, 15, and 17 of the estrous cycle and pregnancy. KLK14 expression was localized in the uterine L and G epithelium, and stroma throughout the endometrium after day 10. Conceptus KLK1 mRNA did not change from days 10 to 17 of gestation. However, conceptus KLK4, and 14 mRNA expression was greatest on day 10 with expression declining after day 14 of gestation. Expression of the various tissue kallikreins in the endometrium and conceptus during the estrous cycle and early pregnancy in the pig can serve in the activation of growth factors and tissue remodeling during the establishment of pregnancy.

Animals↗

Maternal recognition of pregnancy signal or endocrine disruptor: the two faces of oestrogen during establishment of pregnancy in the pig.

Timing of conceptus growth and attachment to the uterine luminal epithelium is regulated by progesterone secretion from the corpus luteum and by expression of progesterone receptor in the uterine epithelia and stroma. Conceptus growth and uterine attachment are temporally associated with the disappearance of progesterone receptors from uterine epithelia. While the loss of progesterone receptor from the endometrial epithelia on day 10 of the oestrous cycle and pregnancy has been well documented, the factors involved with cell specific down-regulation of progesterone receptor are yet to be established. We propose that several progesterone stimulated factors activate nuclear factor kappa B (NF-kB) within the uterine epithelia, which leads to inhibition of progesterone receptor and concomitant stimulation of endometrial genes expressed during early conceptus development. Although oestrogens secreted by pig conceptuses function to establish pregnancy, timing of endometrial exposure to oestrogen is critical. Early oestrogen administration alters the pattern of gene expression through the NF-kB system desynchronising the uterine environment for conceptus implantation resulting in later embryonic loss.

Animals↗

Endocrine disruption of uterine insulin-like growth factor expression in the pregnant gilt.

Early exposure of pregnant gilts to oestrogen, prior to the normal period of porcine conceptus oestrogen secretion, disrupts the uterine environment resulting in complete embryonic mortality during the period of placental attachment to the uterine surface. The current study evaluates the uterine insulin-like growth factor (IGF) system following endocrine disruption of early pregnancy in gilts through exposure to exogenous oestrogen on Days 9 and 10 of gestation. Endometrial IGF gene and protein expression, IGF-I receptor (IGF-IR) gene expression, and uterine lumenal content of IGF binding proteins (IGFBPs) were evaluated in control and oestrogen-treated gilts on Days 10, 12, 13, 15 and 17 of gestation. Oestrogen treatment altered endometrial IGF-I and IGF-IR gene expression on Days 12 and 13 of gestation. Uterine content of IGF-I and IGF-II in control gilts was greatest on Days 10, 12, and 13 followed by a four- to sixfold decrease on Day 15 of gestation. Oestrogen treatment caused a premature proteolysis of IGFBPs within the pregnant pig uterus on Day 10 of gestation, and an earlier decline in uterine lumenal IGF-I content. Results demonstrate that early exposure of pregnant gilts to oestrogen causes premature loss of uterine IGFs during the period of conceptus elongation. Timing for the release of uterine IGFs during early porcine conceptus development may play an important function in the ability of the conceptus to attach and survive during the establishment of pregnancy.

Animals↗

Effects of grazing program and subsequent finishing on gene expression in different adipose tissue depots in beef steers.

This experiment was conducted to examine the effects of grazing program and subsequent finishing on gene expression in adipose tissue from steers. Twenty Angus x Angus-Hereford steer calves (initial BW = 231 +/- 25 kg) were allotted randomly to one of two winter grazing treatments: 1) grazing winter wheat pasture to achieve a high rate of BW gain (HGW); or 2) grazing dormant tallgrass native range (NR). Steers in the NR treatment were provided 0.91 kg.steer(-1).d(-1) of a 41% CP (as-fed basis) cottonseed meal supplement. Following the grazing period, steers were assigned randomly to feedlot pens. Steers were fed to a common endpoint of 1.27 cm of backfat between the 12th and 13th rib. Four steers from each treatment were slaughtered at the end of the grazing period, and the remaining steers from each treatment (n = 6) were slaughtered at the predetermined compositional endpoint. Intramuscular and s.c. fat samples were collected from LM sections of each steer at the 12th-/13th-rib interface on the left side. Pools of RNA were prepared for HGW and NR s.c. adipose tissue from steers slaughtered immediately after grazing. Suppression subtractive hybridization was performed followed by dot-blot hybridization screening to confirm differential expression of subtracted transcripts. Transcripts confirmed to be differentially expressed were subjected to dideoxy chain-termination sequencing. Quantitative reverse transcription PCR was performed on three differentially expressed clones: osteonectin, ferritin heavy chain, and decorin. Osteonectin, ferritin heavy chain, and decorin gene expression was greater (P < 0.05) in s.c. than in i.m. adipose tissue of finished steers. A depot x background interaction for osteonectin (P < 0.01) and ferritin heavy chain (P = 0.03) gene expression was observed for steers slaughtered after grazing, indicating that nutritional management can affect gene expression in adipose tissue depots differently. No differences resulting from prefinishing nutritional background (HGW or NR) were noted in osteonectin, ferritin heavy chain, or decorin gene expression in i.m. adipose tissue collected from finished steers, which might have resulted from feeding steers to the same compositional endpoint. Our data suggest that nutritional background alters gene expression in adipose depots, and that depots are influenced differently.

Adipose Tissue↗

Porcine endometrial expression of kininogen, factor XII, and plasma kallikrein in cyclic and pregnant gilts.

Establishment of pregnancy in the pig is accompanied by a localized uterine acute inflammatory response and increase in uterine blood flow. Following rapid trophoblast elongation on Day 12 of pregnancy there is an increase in tissue kallikrein activity and release of bradykinin into the uterine lumen, suggesting the kallikrein-kininogen-kinin system is active in the porcine uterus. The present study investigated endometrial expression and presence of the various factors of the kallikrein-kininogen-kinin system. Endometrial L- and H-kininogen gene expression as well as presence of kininogens in the uterine flushings was evaluated throughout the estrous cycle and early pregnancy in the pig. The possible involvement of plasma kallikrein and Factor XII, activators of the kallikrein-kininogen-kinin system, were evaluated through analysis of gene expression in endometrial and conceptus tissues. Gene expression for plasma kallikrein, Factor XII, and H-kininogen were detected in endometrium but not early conceptus tissues. Factor XII and H-kininogen gene expression were similar across the days of the estrous cycle and early pregnancy. Endometrial plasma kallikrein gene expression was low but increased on Day 15 of the estrous cycle, whereas expression was similar across the days of early pregnancy. In comparison to cyclic gilts, endometrial L-kininogen gene expression increased fourfold on Days 15 and 18 of pregnancy. Both L- and H-kininogen were detected in the uterine flushings of cyclic and pregnant gilts. Presence of L- and H-kininogen in the porcine uterus and endometrial gene expression of plasma kallikrein and Factor XII provide evidence that the kallikrein-kininogen-kinin system is biologically active during establishment of pregnancy in the pig.

Animals↗

Adherence to highly active antiretroviral therapy predicts virologic outcome at an inner-city human immunodeficiency virus clinic.

This study's hypothesis is that human immunodeficiency virus-infected patients in the inner city (predominantly injection drug users and ethnic minorities) do not take highly active antiretroviral therapy (HAART) as prescribed and that nonadherence leads to virologic failure. A prospective, observational, 3-month study of adherence to HAART was undertaken at an inner-city clinic. There were 40 subjects [110 subject-months]; 30 were male, 10 were female, 75% were Hispanic, 23% were African American, 68% were injection drug users, and 68% were receiving triple therapy. At 3 months, adherence, which was determined by use of the Medication Event Monitoring System (Aprex) was significantly associated with virologic success: lower virus loads were associated with a rate of adherence of >80% (P<.05). Although nonadherence predicted virologic failure, virologic success was not always predicted by adherence: 11 (27.5%) of 40 subjects with suboptimal adherence rates (<90%) had complete virologic suppression.

Adult↗

Numerous indications for office flexible minihysteroscopy.

STUDY OBJECTIVE: To evaluate the feasibility of diagnostic office flexible minihysteroscopy for general gynecology, infertility, oncology, and urogynecology examinations. DESIGN: Prospective cohort analysis (Canadian Task Force classification II-2). SETTING: Private clinic. PATIENTS: Five hundred thirty-eight women. INTERVENTION: Diagnostic hysteroscopy or cystourethroscopy. MEASUREMENTS AND MAIN RESULTS: Office flexible minihysteroscopes 2.5 and 3.5 mm were successfully used in 384 patients to evaluate abnormal and postmenopausal bleeding, intrauterine device location, secondary amenorrhea, severe dysmenorrhea, infertility, tubal patency, and selective tubal insemination. Of these, 382 women (99.5%) were examined without local anesthesia or cervical dilatation. Biopsies with 3F instruments and tubal cannulations were performed without difficulty where indicated. In 225 patients, flexible minihysteroscopes were used to evaluate the genitourinary tract for symptoms of urethritis, interstitial cystitis, and urinary incontinence. Dynamic cystourethroscopy was done to examine the integrity of the bladder neck with hold command, cough, and maximum Valsalva maneuver. There were no unsuccessful hysteroscopic or cystoscopic examinations. CONCLUSION: Flexible minihysteroscopes can be successfully used in an office setting for both gynecologyic and urogynecologyic indications with high patient acceptance. The procedure is rapid and saves physician time.

Adult↗

Treatment decisions for newborns at the threshold of viability: an ethical dilemma.

The major ethical issues involved in decision-making in the care of extremely low birth weight newborns are analyzed here. We propose a schema for assessment and management of these infants that is consistent with ethical principles broadly accepted by the pediatric community, and which takes into account mortality risk at any given institution rather than arbitrary weight limits, with a major decision-making role for the infant's parents. When possible, the decision of whether or not to resuscitate should be made before delivery; when not possible, delivery room resuscitation is recommended, and the decision to continue or withdraw care should be made subsequently based on likelihood of survival and the wishes of the parents.

Critical Illness↗

Unusual presentation of "extracavitary" primary effusion lymphoma in previously unknown HIV disease.

Human herpes virus, type 8, also called Kaposi's sarcoma-associated virus, is associated with primary effusion lymphoma, an uncommon and unusual subset of acquired immunodeficiency syndrome-related lymphomas mostly confined to body cavities, which primarily affects human immunodeficiency virus-positive men. We report the case of a 40-year-old male with primary effusion lymphoma that presented initially with generalized lymphadenopathy and hepatosplenomegaly, followed by pericardial effusion and cardiac tamponade, in a previously undiagnosed human immunodeficiency virus patient. Cytomorphological studies disclosed a large-cell lymphoma with a population of cells demonstrating intermediate CD45 expression and partial coexpression of CD20 and CD23 markers, as well as universal expression of HLA-DR, CD71, CD38, and CD-30. Molecular studies showed clonal B-cell gene rearrangements and molecular evidence of human herpes virus, type 8. This case stresses the necessity, even in the absence of the 'classical clinical features,' of molecular testing for human herpes virus, type 8 in a subset of patients with high risk for human herpes virus, type 8-associated lymphomas.

Adult↗

Development of flexible culdoscopy.

STUDY OBJECTIVE: To assess correlation between findings at standard laparoscopy and office flexible culdoscopy. DESIGN: Observational study (Canadian Task Force classification II-1). SETTING: University-affiliated hospital. PATIENTS: Eleven women undergoing diagnostic laparoscopy to evaluate infertility. INTERVENTIONS: Microlaparoscopy and flexible culdoscopy. In the first six cases laparoscopy was performed first followed by culdoscopy; in the last five cases the order was reversed. MEASUREMENTS AND MAIN RESULTS: The number of endometriotic implants or adhesions was counted for both operative techniques. The flexible 2.2-mm culdoscope provided a view of the pelvis similar to the laparoscope. Correlation between the procedures was 100%. The flexible scope achieved better visualization than the standard rigid posterior cul-de-sac culdoscope. CONCLUSION: Flexible culdoscopy is a excellent procedure for diagnostic endoscopy. It correlates well with office laparoscopy, requires less anesthesia, and is better tolerated by patients. Several operative procedures are currently being tested through this mode of access.

Culdoscopy↗

The use of monoclonal antibody R92F6 and polymerase chain reaction to confirm the presence of parvovirus B19 in bone marrow specimens of patients with acquired immunodeficiency syndrome.

BACKGROUND: Parvovirus B19 infection is a cause of chronic anemia and red cell aplasia in patients with acquired immunodeficiency syndrome (AIDS) and in other immunocompromised hosts. Anemia in AIDS patients has a multifactorial etiology, with parvovirus B19 infection being an infrequent but nevertheless treatable cause. Therapy with intravenous immune globulin can result in rapid improvement of parvovirus-induced anemia. This treatment is expensive, therefore accurate and rapid confirmation of parvovirus infection is important in providing appropriate and cost-effective therapy. METHODS: Bone marrow samples from 2 AIDS patients with severe anemia and reticulocytopenia were studied. Bone marrow morphology and serologic studies were evaluated for parvovirus B19 infection. An immunohistochemical method using a monoclonal antibody, R92F6, to B19 capsid proteins was utilized on decalcified, B5-fixed, paraffin-embedded bone marrow biopsies. Bone marrow aspirate cells were examined by electron microscopy for evidence of viral particles. In addition, polymerase chain reaction (PCR) studies using a nested PCR assay to the parvovirus B19 viral genome were performed in a case for which fresh cells were available. RESULTS: Bone marrow findings included marked erythroid hypoplasia with characteristic giant pronormoblasts and intranuclear inclusions. Serologic studies were negative in one case, while the second case showed positive parvovirus B19 immunoglobulin M antibody. Immunohistochemical studies for parvovirus B19 were positive in both cases. The presence of intranuclear virions was demonstrated by electron microscopy and was confirmed by PCR analysis. Both patients were treated with intravenous immune globulin, and subsequent improvement was noted. CONCLUSIONS: Both immunohistochemistry and PCR studies on bone marrow specimens from AIDS patients with anemia are rapid and sensitive methods for the confirmation of parvovirus B19 infection. They are valuable tools, particularly when serologic studies are negative. When PCR is not available, immunohistochemical methods can be useful. The rapid confirmation of parvovirus B19 infection will allow for early and cost-effective therapy.

Acquired Immunodeficiency Syndrome↗

Pushing proficiency testing results to their limits.

PT is a valuable component of laboratory quality control and management practice. PT results are useful to determine over time that a method is performing in a laboratory according to the manufacturer's design parameters, as exhibited by the performance characteristic of a large group of peer laboratories. PT peer group results give a good measure of the interlaboratory imprecision of a method, and summary reports document the state-of-the-art in user preference. PT results typically cannot be used to validate the accuracy of a method or its clinical utility or to compare the accuracy of various methods. These limitations are caused by the common and unpredictable matrix interferences with PT materials and most routine laboratory methods. In situations where a laboratory adjusts the calibration of one method to make patient results agree with those of another method, PT specimens results must be reported without calibration adjustments to allow those results to be compared to an appropriate peer group mean target value.

Humans↗

Multichannel urodynamic evaluation of laparoscopic Burch colposuspension for genuine stress incontinence.

OBJECTIVE: To evaluate the 2-year outcome of laparoscopic Burch colposuspension in treating genuine stress incontinence, and to evaluate the prevention of subsequent vaginal vault prolapse by prophylactic apical vault repair. METHODS: This is a prospective study of 48 consecutive cases diagnosed with genuine stress incontinence and followed for 2 years or longer (range 30 to 41 months) after correction by laparoscopic Burch colposuspension. All patients had a routine urogynecology evaluation, including preoperative and postoperative multichannel urodynamics. Follow-up testing was done at 6 weeks and 1 and 2 years. RESULTS: At 6 weeks, 1 year, and 2 years, the cure rate for genuine stress incontinence was 98, 93, and 89%, respectively. There was a significant decrease in urethral hypermobility and an increase in urethral pressure transmission ratios. Thirty-nine (81%) patients voided spontaneously in less than 24 hours. There was no clinically significant pelvic organ prolapse at 2 years. CONCLUSION: The 2-year cure rate and multichannel urodynamics findings for laparoscopic Burch repair are similar to those reported for laparotomy. Early results of prophylactic apical vault suspension suggest that the incidence of prolapse observed after routine retropubic urethropexies may be reduced.

Adult↗