Hematopoietic stem and progenitor cell populations in MoMuLV-ts-1 induced lymphoma in a murine model.

During self-renewal of the hematopoietic stem cells there is a potential for these new cells to develop into abnormal malignant cells due to environmental, chemical or microbial challenges. This may lead to blood malignancies, including leukemia and lymphoma. Our laboratory was the first to develop a mouse model to study the development of lymphoma among pups of retrovirus infected mothers. We have established that if the newborn pups suckle breast milk from the mothers infected with a temperature sensitive Moloney Murine Leukemia Virus (MoMuLV-ts1), they develop lymphoma. Stem and progenitor cell populations were determined from blood, spleen, and thymus samples in infected mice with and without lymphoma using flow cytometry and specific protein markers, including CD117+/CD34+; CD117+/Sca-1+; CD135+/Sca-1+; Ter119+/CD71+ and CD117+/CD45+. Some stem cell populations were increased in infected mice with lymphoma. These stem cells appear to be involved in the development of lymphoma in our mouse model.

Retrovirus-induced lymphomagenesis: a correlation between disease pathogenesis and flow cytometric analysis.

Perinatal infection with a temperature-sensitive mutant (ts-1) of Moloney murine leukemia virus (MoMuLV) results in massive splenomegaly and thymomegaly in mice and development of lymphoma in >55 % of infected pups. Previous flow cytometry studies showed a decrease in CD4(+) cells in perinatally infected pups, but cell population changes in infected animals with lymphoma compared with infected animals without lymphoma has not yet been reported. In the current study, BALB/c mice were infected with ts-1 through breast milk transmission and observed until development of clinical signs and symptoms of lymphoma and/or symptomatic ts-1 infection. Flow cytometry studies were performed on blood, spleen and thymus samples and correlated with gross morphology and histological changes, resulting from the development of lymphoma. Infected animals with lymphoma had significant decreases in CD4(+) and CD8(+) cell counts in blood and spleen compared with controls. The spleens of infected animals without lymphoma showed a decrease in CD4(+) and CD8(+) cell counts, but this was not significant compared with controls. In the thymus, CD4(+) and CD8(+) cell counts also decreased, but this was not significant in infected animals with and without lymphoma compared with controls. Markers of myeloid cell dysfunction increased in the thymus of animals with infection with and without lymphoma compared with controls. Thus, immunosuppression and CD4(+)/CD8(+) cell decreases in the spleen and thymus are associated with malignant transformation and development of lymphoma in this animal model.

Adherence to antiretroviral therapy: a survey of factors associated with medication usage.

The use of highly active antiretroviral therapy (HAART) has decreased morbidity and mortality for people living with HIV/AIDS, but adherence to HAART is a critical factor in successful treatment. Adherence to medication is a complex and poorly understood behavior. A survey was undertaken to evaluate subjective correlates of adherence and non-adherence based on previously distilled themes from a qualitative study of adherence. A 60-question survey was completed by patients in the outpatient clinic setting regarding demographics, CD4 cell count, viral load, adherence, and screening questions about medication usage and attitudes toward HIV. Ninety-eight adherent and 34 non-adherent patients completed the survey. After logistic regression analysis, several questions appeared to be the main predictors of non-adherence: Have you ever thought of having HIV as a "punishment?" Do you feel that your medicines are hard to take? Do you believe the medicines for HIV that you take are working for you? These questions may be helpful in the development of a clinically useful screening tool to assess patients at risk for non-adherence.

Retroviral gene insertion in breast milk mediated lymphomagenesis.

We have demonstrated breast milk transmitted MoMuLV-ts1 retrovirus infection and subsequent lymphoma development in offspring of uninfected mothers suckled by infected surrogate mothers. Additionally, we have shown that the lymphoma development occurs as a result of viral gene integration into host genome. A total of 146 pups from Balb/C mice were divided into 5 groups; one control and 4 experimental. All offspring suckled from surrogate infected or control mothers, except one group of infected pups left with their biological mothers. Thirteen of 91 infected pups developed lymphoma. Inverse-PCR, DNA cloning, and quantitative real-time PCR (qRT-PCR) were used to study the virus integration sites (VIS) and alterations in gene expression. VIS were randomly distributed throughout the genome. The majority of insertion sites were found in chromosomes 10, 12 and 13. A total of 209 proviral genomic insertion sites were located with 52 intragenic and 157 intergenic sites. We have identified 29 target genes. Four genes including Tacc3, Aurka, Gfi1 and Ahi1 showed the maximum upregulation of mRNA expression. These four genes can be considered as candidate genes based on their association with cancer. Upregulation of these genes may be involved in this type of lymphoma development. This model provides an important opportunity to gain insight into the relationship of viral gene insertion into host genome and development of lymphoma via natural transmission route such as breast milk.

Right varicocele and hypoxia, crucial factors in male infertility: fluid mechanics analysis of the impaired testicular drainage system.

Varicocele is considered a predominantly unilateral left-sided disease. However, since male fertility is preserved with only one healthy testis, infertility perforce represents bilateral testicular dysfunction. It was hypothesized that: (i) right varicocele cannot be diagnosed by palpation and therefore has not been treated in the past by the traditional treatment, and (ii) right varicocele causes impaired oxygen supply in the right testicular microcirculation, leading to germ cell degeneration. This study performed venographies of both right and left internal spermatic veins during the treatment of 840 infertile men with varicocele and analysed the results using tools of fluid mechanics. Histopathology of the right testis revealed stagnation of blood flow and degenerative changes attributed to lack of adequate oxygenation in all testicular cell types. Right varicocele was found in the vast majority of the patients. We found that due to the destruction of one-way valves, pathologic hydrostatic pressure is produced in the testicular venous microcirculatory system about five times higher than normal, exceeding arteriolar pressure. The pressure gradient between the arterioles and venules in the testicular tissue is therefore reversed, leading to persistent hypoxia. Right varicocele, although undetected, is prevalent in infertile men with varicocele, hence only bilateral occlusion of the internal spermatic veins, including the associated bypasses, eliminating the pathologic hydrostatic pressure will lead to resumption of arterial blood flow in the testicular microcirculation.

Transmission of Moloney murine leukemia virus (ts-1) by breast milk.

A murine model has been developed to study maternal transmission of the temperature-sensitive Moloney murine leukemia virus (ts-1). The goal of this study was to confirm early and late mother-to-offspring transmission of the virus and demonstrate transmission via breast milk. A series of six experiments was performed using six groups of BALB/c mice. Group 1 consisted of pups born to ts-1-infected mothers removed at birth to suckle from surrogate uninfected mothers. Groups 2 and 5 consisted of pups born to ts-1-infected mothers that suckled from ts-1-infected mothers (surrogate and biological). Group 3 consisted of non-infected pups removed at birth to suckle from ts-1-infected mothers. Groups 4 and 6 consisted of non-infected pups suckled from non-infected mothers. The combined in utero, intrapartum and breast-milk infection rate was 100 % to the offspring (groups 2 and 5). The in utero to early post-partum group (group 1) had an infection rate of 78 %. Breast milk alone (group 3) resulted in a 97 % infection rate. Control groups (groups 4 and 6) had a 0 % infection rate. The relative frequency of maternal CD4(+) cells in peripheral blood mononuclear cells was consistently lower in infected mothers, whilst offspring did not show a significant decrease in CD4(+) frequency. Pups infected via breast milk had a lower CD4(+) frequency (group 3) than those infected by the uterine and/or intrapartum route (group 1). Breast milk from ts-1-infected mothers appears to be highly infectious for neonatal BALB/c mice.

Varicocele, hypoxia and male infertility. Fluid Mechanics analysis of the impaired testicular venous drainage system.

BACKGROUND: Varicocele is a bilateral vascular disease, involving a network of collaterals and small, retroperitoneal bypasses. The right and the left testicular venous drainage systems are complex and not identical to each other. It was considered a predominantly unilateral (left-sided) disease. Its pathophysiology has not been clearly delineated and the treatments offered do not seem to be effective. The medical literature is replete with articles demonstrating inconsistent and even contradictory results which have led clinicians to dissociate varicocele from male infertility. Since male fertility is preserved with only one healthy testis, male infertility perforce represents bilateral testicular dysfunction. This poses an enigma to clinicians: How can left-sided varicocele causes bilateral testicular dysfunction? METHODS: We investigated the internal spermatic veins by venography to understand testicular damage due to varicocele. A total of 740 venographies of the internal spermatic veins (ISVs) were performed, with sclerotherapy of the ISV as treatment for varicocele. Epon-embedded testicular tissue sections were used to identify blood stagnation in the testis. RESULTS: Varicocele is predominantly a bilateral disease in 84% of cases, associated with collaterals and retroperitoneal venous bypasses in 70% in the left side and 75% in the right side. Histopathology demonstrate stagnation in the testicular microcirculation and hypoxic-ischaemic degenerative changes in all cells' types in the sperms' production site. CONCLUSION: Based on our findings (i) varicocele is a bilateral disease; (ii) the disease is expressed earlier in the left side and is more intense because the blood column is longer in the left side than the right; (iii) partial treatment to the left side only and ignoring bypasses is not adequate to correct the problem; (iv) hypoxia leading to ischaemic damage to both testes is the effect of varicocele due to hydrostatic pressures in the impaired venous drainage system, which exceeds the pressures in the testicular arterial microcirculation due to blood columns produced in the disease; (v) hydrostatic pressure does not depend on vein diameter but on blood column height, only; and (vi) thermography alone or combined with ultrasonography with special attention to the bilaterality of the disease are the best non-invasive tools for its detection.

Vertical transmission of a murine retrovirus, ts1.

Mechanism of maternal retroviral transmission remains an unsolved problem. The current investigation is a part of our ongoing research on vertical transmission of MoMuLV-TB ts1 in BALB/c mice. A total of 270 adult mice and 165 fetuses were used. Forty-four experimental mice were injected with 0.1 mL of 4.0 x 10(6) ffu/mL of ts1 virus at 72 h after birth; 24 controls were injected with DMEM. Almost half of the females went through two rounds of pregnancies. In the first round, 135 experimental and 57 control pups were produced. Forty-three experimental and 20 control pups were followed until they developed clinical symptoms. The second round of pregnancy produced a total of 46 mid-gestational and 119 full-term fetuses. PCR, and light and electron microscopy were performed to evaluate viral transmission. Overall, 99% vertical transmission occurred in pups of infected mothers. Twelve percent of mid-gestational and 39% full-term fetuses were PCR positive. We have established that, if mothers are infected with ts1 virus at 72 h after birth, then nearly 100% vertical transmission occurs, via in utero, intrapartum, or breast milk. Thirty-nine percent transmission occurred in utero alone. This is an excellent model to study the transplacental and post-gestational transmission of retroviruses, such as ts1.

Tamm-Horsfall protein in patients with kidney damage and diabetes.

Tamm-Horsfall protein (THP) is a glycoprotein present abundantly in human urine. It is localized in the thick ascending limb of the loop of Henle (TAL) and the early distal convoluted tubule (DCT). The rate of urinary excretion of THP has been studied in various diabetic groups. It has been postulated that urinary THP may be a useful marker for renal damage. The aim of this study was to compare directly the immunogold localization of THP in diabetic and control kidney tissue specimens with or without kidney damage. Immunogold labeling was performed on archival tissue samples of 34 diabetic and 18 control human kidneys at the light microscope level. Slides were ranked as having a high, moderate or low degree of reaction. The majority of diabetic samples had a slightly lower degree of THP, while patients with known renal dysfunction had lowest THP. Previous studies have found a decreased excretion of urinary THP in diabetics. Our results show that decreased gold labeling is associated with known renal damage and may indicate damage to the thick ascending limb of the loop of Henle and the early distal convoluted tubule, irrespective of presence or absence of diabetes.

HIV/AIDS knowledge among female migrant farm workers in the midwest.

The rate of HIV infection in the migrant farm worker community is 10 times the national average. A survey was conducted of 106 female migrant farm workers in rural Northwest Ohio to assess HIV knowledge. The average participant's age was 28.7 years, 78 spoke Spanish, and 47 had an < or =8th- grade education. Fifty-six women received their information on HIV/AIDS from television. Eighty-seven women identified sexual contact as the major source of HIV transmission and 54 women identified the combination of sex, use of needles, and blood contact as the important routes. Sixty-nine women identified both homosexual and heterosexual intercourse as risk factors. Only 58 women identified perinatal infection as a route of HIV transmission and 59 women knew that treatment was available to prevent perinatal transmission. Although the majority of women had a good general knowledge of HIV transmission, further prevention education on perinatal transmission is needed among this population.

A small animal model for mother-to-fetus transmission of ts1, a murine retrovirus.

Infection with a murine retrovirus, MoMuLV-TB, ts1 in BALB/c mice has been established as a small animal model for retroviral neurodegenerative disease as shown with infections such as HIV. However, mother-to-pup transmission has never been demonstrated in this model. The current investigation examines vertical transmission of ts1 in this mouse model. A total of 15 females were used to produce 59 pups (16 were used for control, and 43 were used as experimental animals). For experiment 1, 24 5-day-old mice were injected with [0.2 mL of 2.0 x 10(6) ffu/mL ts1] virus. For experiment 2, 19 48-h-old mice were injected with [0.1 mL of 4 x 10(6) ffu/mL ts1] virus. Control groups were injected with DMEM only. PCR and electron microscopy were performed to determine the presence of virus. All mice from experiment 1 injected with ts1 showed viral infection, and retained 100% reproductive capacity. Three out of 102 pups produced by these infected females were infected with ts1. Nine percent of the pups from experiment 2 injected with ts1 retained normal reproductive capacity, and two out of eight (25%) pups had viral infection. Vertical transmission of this unique retrovirus occurs and is dependent, in part, on the timing of maternal infection.

Survey of physician attitudes toward HIV testing in pregnant women in Ohio.

HIV infection among women of childbearing age is still increasing in the United States. In most states, HIV testing of women or neonates during pregnancy is not mandatory. The current study assessed HIV prenatal testing practices among obstetrician-gynecologists and primary care physicians listed in a regional physician referral data base in a predominantly rural region. Between December 2000 and March 2001 a 20-question survey was sent by mail to regional physicians in obstetrics/gynecology and primary care regarding physician practice demographics and prenatal HIV testing practices. Of 1116 surveys sent, 431 were returned (38.6% response). Only 42% of physicians offered universal HIV prenatal testing. Factors associated with universal testing (p < 0.5) included obstetrics/gynecology as the practice specialty (90%) physicians' age younger than 50 years, and a practice with predominantly Medicaid or African American patients. Further educational and public health initiatives may be needed to increase nonselective, universal HIV testing in pregnant women.