Current trends in Rwanda's HIV/AIDS epidemic.

OBJECTIVE: To review the trajectory of Rwanda's HIV epidemic, including long term trends and more recent trends in HIV prevalence, markers of HIV incidence, and behavioural indicators. METHODS: This paper reviews the history of HIV serological and behavioural surveillance efforts in Rwanda, dating back to the early 1980s, synthesising findings from surveillance, research, and other relevant HIV programmatic data. The documentation reviewed includes published findings, conference abstracts, and unpublished analyses. Special emphasis is given to more recent sentinel surveillance results and data collected using known, documented methods. Recent trends in HIV prevalence were assessed among sites participating in the three most recent consecutive rounds of antenatal clinic sentinel surveillance. RESULTS: Early HIV surveillance in Rwanda documented high HIV prevalence in urban areas with HIV widely disseminated into rural areas by 1986. Between 1988 and 1996, HIV prevalence among pregnant women ranged from 21% to 33% in Kigali, from 8% to 22% in other urban settings, and from 2% to 12% in rural settings. More recent surveillance among pregnant women has demonstrated more moderate prevalence, with urban/rural differences narrowing slightly. Between 1998 and 2003, HIV prevalence may have declined in urban areas, whereas rural areas appear to have remained stable. Age at first sexual intercourse is relatively late in Rwanda (20 years for both males and females) and has remained stable since at least 1992. CONCLUSIONS: The present analysis suggests that Rwanda may have experienced declines over the long term in HIV prevalence in urban areas, especially in Kigali, and may have stable or slightly rising HIV prevalence in rural areas. The limited behavioural data available suggest that, on the national level, Rwanda may benefit from a unique combination of low numbers of partners and late sexual debut, which may have had a mitigating effect on HIV prevalence.

The receptor activator of nuclear factor-kappaB ligand inhibitor osteoprotegerin is a bone-protective agent in a rat model of chronic renal insufficiency and hyperparathyroidism.

Osteoprotegerin (OPG) acts by neutralizing the receptor activator of nuclear factor-kappaB ligand (RANKL), the primary mediator of osteoclast differentiation, function, and survival. We examined whether OPG could affect the bone loss associated with chronic kidney disease (CKD) in a rodent model of CKD and secondary hyperparathyroidism (SHPT). SHPT was induced in rats by 5/6 nephrectomy (5/6 Nx) and a 1.2% P/0.6% Ca(2+) diet. Starting 1 week after 5/6 Nx, rats were treated with vehicle (veh) or OPG-Fc (3 mg/kg, intravenously) every 2 weeks for 9 weeks. At baseline, 3, 6, and 9 weeks, blood was taken and bone mineral density (BMD) and bone mineral content (BMC) were assessed by dual-energy X-ray absorptiometry. Serum parathyroid hormone (sPTH) levels reached 912 pg/ml in 5/6 Nx rats vs. 97 pg/ml in shams at 9 weeks. OPG-Fc had no effect on sPTH or Ca(2+) levels throughout the 9-week study, indicating that SHPT was a renal effect independent of bone changes. At 3 weeks, 5/6 Nx-veh rats had osteopenia compared with sham-veh rats and 5/6 Nx-OPG-Fc rats had significantly higher percent changes in whole-body BMC, leg BMD, and lumbar BMD versus 5/6 Nx-veh rats. By 6-9 weeks, elevated sPTH was associated with reversal of bone loss and osteitis fibrosa in the proximal tibial metaphysis. OPG-Fc decreased this sPTH-driven high bone turnover, resulting in augmented thickness of proximal tibial trabeculae in 5/6 Nx rats. Thus, RANKL inhibition with OPG-Fc can block the deleterious effects of continuously elevated sPTH on bone, suggesting that RANKL may be an important therapeutic target for protecting bone in patients with CKD and SHPT.

Statistical tools for setting in-process acceptance criteria.

Establishing meaningful and reasonable acceptance criteria for process validation or continual monitoring is crucial to approval and successful manufacturing. The limits should be based on statistical analysis of historical data when possible. The control limits of "mean +/- 3 standard deviations" is one industry standard. However, the limits may be artificially constraining if the standard deviation does not reflect the true variance of the process. Under-estimation of process variance is common with small data sets. This paper presents three methods for correcting underestimated variance, allowing the setting of acceptance criteria that are slightly larger than +/- 3 standard deviations. These limits are more meaningful in that they account for true process variability and will signal process deviations due only to a specific cause.

Anti-interleukin-1 and anti-tumor necrosis factor-alpha synergistically inhibit adjuvant arthritis in Lewis rats.

Interleukin-1 (IL-1) and tumor necrosis factor-alpha (TNF-alpha) play dominant roles in mediating the progression of many inflammatory joint diseases, including rheumatoid arthritis in humans, collagen-induced arthritis in mice and rats, and adjuvant arthritis in rats. Blockade of either cytokine partially controls these diseases. The present study investigated the value of combination anti-cytokine therapy in arthritis: the efficacy of IL-1 receptor antagonist (IL-1ra) and 30 kDa polyethylene glycol (PEG)-conjugated soluble TNF receptor type I (PEG sTNF-RI) given together was assessed in Lewis rats with adjuvant arthritis. Administration of either IL-1ra or PEG sTNF-RI partially alleviated joint inflammation, loss of bone mineral density, and loss of body weight. In contrast, combination of these anti-cytokine treatments exhibited a synergistic capacity to inhibit these changes, even when combining doses of IL-1ra and PEG sTNF-RI that did not affect lesion severity when used alone. Statistical analysis of these adjuvant arthritis data using the isobologram method proved that IL-1ra and PEG sTNF-RI were clearly synergistic in inhibiting inflammation, loss of bone mineral density, loss of body weight, and histopathologic parameters of inflammation and joint destruction. These results suggest that treating autoimmune arthritic diseases with combinations of anti-IL-1 and anti-TNF molecules will achieve superior efficacy compared to the use of a single class of anti-cytokine agent and may allow for dose reductions that could prove useful in minimizing potential side effects.

The prolonged hematologic effects of a single injection of PEG-rHuMGDF in normal and thrombocytopenic mice.

A single injection of > or =10 microg/kg PEG-rHuMGDF in mice causes a dose-dependent increase in circulating platelets beginning on day 3 and peaking on days 5-6. The mean platelet volume and platelet distribution width at doses > or =100 microg/kg initially increase in a dose-dependent fashion and later decrease. However, the mean platelet volume does not change when platelets are incubated with PEG-rHuMGDF in vitro. The number of marrow megakaryocytes increases in a dose-dependent fashion as early as day 1 and peaks on day 3. Marrow megakaryocyte colony-forming units (CFU-Meg) do not increase on days 1-3 at a dose of 100 microg/kg (a dose that increases platelet numbers two- to threefold and may be clinically relevant), but the relative frequency of high ploidy megakaryocytes and the proportion of large marrow megakaryocytes (29-50 microm in diameter) increases. After a dose of 1,000 microg/kg the percentage of megakaryocytes in mitosis peaks at 24-48 hours and the percentage of megakaryocytes incorporating BrdU is maximal at 48 hours, the relatively delayed peak of BrdU incorporation most likely representing endomitosis. The relative frequency of type II and III megakaryocytes peaks on days 3 and 4, respectively. Pharmacokinetic analysis of PEG-rHuMGDF shows peak serum concentrations at 2-4 hours and a terminal half-life of 11.4+/-2.5 hours. A single injection of PEG-rHuMGDF ameliorates carboplatin-induced megakaryocytopenia and thrombocytopenia in a dose-response dependent fashion. In conclusion, a single injection of PEG-rHuMGDF increases megakaryocyte and platelet production in normal and myelo-suppressed mice.

Systemic effects of pegylated recombinant human megakaryocyte growth and development factor in combination with recombinant murine granulocyte colony-stimulating factor in a murine model of myelosuppression.

Megakaryocyte growth and development factor (MGDF) stimulates megakaryopoiesis and thrombopoiesis in vivo. Previous studies indicate that administration of pegylated recombinant human (PEG-rHu) MGDF in combination with recombinant murine granulocyte colony-stimulating factor (rMuG-CSF) prevented lethality and reduced hematotoxicity in carboplatin-treated/irradiated mice, a disease-state animal model of radio-chemotherapy. In the current study we have further characterized the effects of PEG-rHuMGDF in combination with rMuG-CSF with respect to clinical chemistry, hematology variables, and histologic evaluations to determine whether any potential toxicological interaction exists both in normal and myelosuppressed mice. Myelosuppression and subsequent thrombocytopenia in mice was induced with a combination of a single intraperitoneal injection of 1.25 mg carboplatin followed 4 h later with sublethal gamma irradiation exposure of 500 rad. Both normal and carboplatin-treated/irradiated mice were administered daily subcutaneous injections of 50 micrograms/kg PEG-rHuMGDF alone and in combination with 10 micrograms/kg rMuG-CSF for 21 consecutive days. Administration of PEG-rHuMGDF alone or in combination with rMuG-CSF to carboplatin-treated/irradiated mice increased survival 70 and 100%, respectively, and accelerated platelet recovery. Microscopic examination of nonhematopoietic organs showed no evidence of any morphological changes in normal and carboplatin-treated/irradiated animals. In hematopoietic organs clinically significantly increased granulopoiesis and megakaryopoiesis, as well as extramedullary granulopoiesis within the mandibular and mesenteric lymph nodes, were present. The erythroid line was unaffected by cytokine treatment. In normal, non-carboplatin-treated/irradiated mice, platelet counts increased 6 and 12-fold above baseline in the groups administered PEG-rHuMGDF alone or in combination with rMuG-CSF, respectively. The results of this study provide a basis for coadministration of PEG-rHuMGDF with Filgrastim (rHuG-CSF) in the clinical treatment of myelosuppression induced by radiation and chemotherapy.

Combined treatment of acute EAE in Lewis rats with TNF-binding protein and interleukin-1 receptor antagonist.

Experimental autoimmune encephalomyelitis (EAE) is a term given to describe a collection of animal models representing the human disease multiple sclerosis (MS). Although not fully understood, the involvement of cytokines and the immune system in either EAE or human MS is well established. Past efforts have shown that inhibition of proinflammatory cytokines tumor necrosis factor (TNF-alpha) or interleukin-1 (IL-1) result in amelioration of acute EAE in Lewis rats. The present study examined this model for the effect of concomitant inhibition of both TNF-alpha and IL-1, which resulted in a modest but significant therapeutic effect that was superior to inhibition of either single agent alone with respect to four of the five variables used to follow the progression of disease in this model, i.e., clinical severity, frequency of disease, loss of body weight, and day of onset. These results are in accordance with the idea that combination treatments are likely to prove superior to single agent therapy in the treatment of autoimmune inflammatory disease.

Effect of bovine somatotropin administered to periparturient dairy cows on the incidence of metabolic disease.

Thirty-eight dry, pregnant Jersey cows were assigned to diet and bST treatment in a 2 x 2 factorial design. During the dry period, half of the cows were fed a normal TMR (0.4% Ca; 0.3 to 0.4% P), and half of the cows were fed a high Ca TMR (1.5 to 1.6% Ca; 0.4 to 0.7% P). The high Ca diets were designed to induce milk fever and were relatively cationic (194 to 293 meq/kg) compared with the normal diets (-131 to 30 meq/kg). A standard dairy diet was fed to all cows postcalving. Cows received subcutaneous injections of either an oil-based excipient or 500 mg of bST in an oil-based excipient every 14 d from 28 d before expected calving until approximately 14 d postcalving. Peripartal bST treatment decreased the incidence of clinical mastitis, did not affect incidence of milk fever, and increased the duration, but not the incidence, of ketosis in mature Jersey cows. Blood data confirmed the clinical responses and indicated that treated cows mobilized more bone Ca than did controls, as was evidenced by increased hydroxyproline concentrations. Treatment with bST did not affect blood concentrations of 1,25-dihydroxyvitamin D, Ca, or Mg. High Ca diets increased the incidence of milk fever and downer cow syndrome compared with normal diets. The effect of bST on mastitis and milk production must be considered as preliminary given the small size of the study. Although bST treatment increased Ca mobilization, the effect was insufficient to prevent milk fever in this model.

Effect of supplemental dietary fat or protein on the short-term milk production response to bovine somatotropin.

Effects of supplemental energy or protein on the milk production response to bST administration were examined in two separate trials. In trial 1, 40 cows were used in a 2 x 2 factorial, completely randomized design to determine the effects of bST and fat supplementation. The study consisted of a 7-d pretreatment period and a 42-d treatment period. Fat was top-dressed at 3.0 Mcal/d of NEL, and bST was administered. Supplemental fat had no effect on milk production, and NEL intakes were unaffected. Administration of bST increased milk production by 7.1 kg/d, and the milk production response was unaffected by supplemental fat. In trial 2, 4 cows were used in four periods with a 2 x 2 factorial arrangement in which water or casein was infused into the abomasum of cows fed for ad libitum intake or at 80% of their requirements. Diets and infusions were initiated simultaneously and continued for 11 d. All cows were given bST during the last 5 d. Infusion of water or casein did not alter the milk production response to bST, but restricted feeding reduced the bST response (3.2 vs. 7.2 kg/d). Concentrations of IGF-I in plasma were increased by bST administration, and the increase was greatest for cows fed for ad libitum intake. The milk production response to bST was not increased by additional energy or protein offered to cows fed well-balanced diets.

Performance, clinical chemistry, and carcass responses of finishing lambs to formulated sometribove (methionyl bovine somatotropin).

Formulated zinc methionyl bST (sometribove, 50, 100, or 150 mg) was administered as a single treatment once every 2 wk or as two equal treatments once/week to evaluate the efficacy of prolonged release delivery of bovine somatotropin (bST) in finishing lambs. Feed conversion during the 6-wk treatment period was improved 9 and 19% in lambs that received treatments once and twice/2 wk, respectively (P < .05), and the responses to differing doses were similar within a dosing frequency (P > .05). Carcass muscle:fat ratio indicators generally were affected in a dose-related manner and were independent of frequency of administration. For example, fat thickness was 17, 30, and 42% lower than control in lambs that received 50, 100, and 150 mg of formulated sometribove/2 wk, respectively (P < .05). Percentages of muscle were higher and of fat were lower with increasing dose of formulated sometribove, but weight of only fat was significantly affected (P < .05). Clinical chemistry indices of metabolic effects of bST (e.g., circulating bST, IGF-I, insulin, glucose, and urinary nitrogen concentrations) were affected in directions similar to those observed with bST administered by daily injection. The results of this study demonstrate the growth performance and carcass composition advantages of a formulation designed to deliver bST over a 2-wk period.

Effect of heat-stress on bovine embryo development in vitro.

Chronic elevation of uterine temperature has long been known to increase embryo mortality in dairy cattle. Short-term elevation in temperature of mouse embryos to 43 degrees C (acute) has been shown to induce intracellular production of heat-shock proteins. In this study, in vitro development of bovine embryos was assessed during short-term (60 h) coculture with oviduct epithelial cells at 38.6 degrees C (T1), 40 degrees C (T2), 38.6 degrees C after a prior pulse treatment (20 min) at 43 degrees C with 5% CO2 (T3), or 38.6 degrees C after a prior pulse treatment (20 min) at 43 degrees C with 100% CO2 (T4). During incubation, embryos cocultured at 40 degrees C had a greater (P < .05) mean embryo development score at 36 h than embryos cocultured at 38.6 degrees C. At 60 h of incubation, embryo development scores were greater (P < .05) for embryos cultured at 38.6 degrees C than for those cocultured at 40 degrees C. The number of embryos hatched at 60 h was similar after coculture at 38.6 degrees C (T1) or a prior pulse treatment with 5% CO2 and 43 degrees C (T3), but the embryo development score at 60 h was greater (P < .05) for the pulse-treated embryos. Embryos in T4 had greater (P < .05) embryo development scores than did T1 embryos from 36 through 60 h. Pulse treatment (T4) resulted in a greater (P < .05) number of hatched embryos at 60 h than T1, T2, and T3. These results indicate a detrimental effect of a chronic elevation in temperature that was evident shortly after embryo hatching.(ABSTRACT TRUNCATED AT 250 WORDS)

Stimulation of milk yield and feed intake by bovine placental lactogen in the dairy cow.

A 6 x 6 Latin square design was used to test the effects of recombinant bovine placental lactogen on milk yield, milk composition, feed intake, and blood hormone and metabolite levels in nonpregnant lactating cows. The six treatments (5, 10, 20, and 40 mg/d of placental lactogen, water as negative control, and 20 mg/d of bST as positive control) were administered by subcutaneous injection twice daily for 9 d. Blood samples were taken during the last 5 d of the treatment period. The three highest doses of placental lactogen increased milk yield, and there was a linear dose effect, although placental lactogen was less potent than bST. Milk concentrations of lactose, protein, and fat were not altered by any of the treatments. Dry matter intake was increased by two of the doses of placental lactogen, but not by bST. Blood urea N concentration was decreased in a dose-dependent manner by placental lactogen and was also decreased by bST. Similarly, serum insulin-like growth factor-I was increased in a dose-dependent manner by placental lactogen and was also increased by bST. Plasma concentrations of NEFA and glucose were increased by bST, but placental lactogen had little or no effect on either of these parameters. Thus, placental lactogen appears to act, in part, as a weak somatotropin agonist; however, it also appears to have specific activities, e.g., stimulating feed intake.

Evaluating two different evaporative cooling management systems for dairy cows in a hot, dry climate.

Milk production, rectal temperature, live weight gain, reproductive performance, and weather data were obtained on 150 Holstein cows managed under two cooling systems on a large dairy farm in Saudi Arabia during the summer months. Cows were paired at the onset of the trial according to days postpartum, lactation number, and current milk production. Females were then allocated either to a system that forced air, precooled by evaporative cooling, over the cows or to a system that alternately showered a fine mist onto the surface of the cows and then forced air at ambient temperature over them. The cows receiving evaporative cooling and those with spray and fan cooling were on sand and on slatted concrete floor, respectively, during the periods of cooling. The onset of estrus was observed during the night when the cows preferred the unshaded corral. For the 120-d trial period, 84% (62 of 75) of the cows receiving evaporative cooling and 60% (44 of 75) of the cows receiving spray and fan cooling became pregnant. In the evaporative cooling system, the pregnancy rate per insemination was 35.2% (179 inseminations) versus 23.2% (194 inseminations) for spray and fan cooling. The mean postpartum interval to pregnancy was 117.6 d for the evaporative cooling cows and 146.7 d for spray and fan cooling cows. The evaporative cooling system, with its open shades and sand bedding, enhanced reproductive performance and milk production compared with that of cows cooled with a spray and fan system with slatted flooring in this hot climate.

In-vitro development and implantation of human embryos following culture on fetal bovine uterine fibroblast cells.

Human zygotes resulting from IVF were placed in two different culture systems to evaluate in-vitro development and to establish pregnancies in patients following embryo replacement. Treatment A (control) consisted of culturing zygotes in a modified Earle's Balanced Salt solution while treatment B consisted of culturing zygotes on a monolayer of fetal bovine uterine fibroblasts in this same culture medium. At the time of embryo replacement, embryos within treatments A and B had 3.7 and 4.3 blastomeres present, respectively. After 24 h in culture, the cellular fragmentation rate for treatment A embryos was 0.85 which was greater (P less than 0.05) than the fragmentation rate of 0.40 for embryos within treatment B. The incidence of implantation for patients whose embryos were given treatment A was 17.0% which was lower (P less than 0.05) than 35% for those given treatment B. Implantation rates increased with time in culture (43%) for treatment B embryos. Culture by treatment B of three-pronucleate zygotes resulted in 7/9 and 4/9 reaching the blastocyst and expanded blastocyst stages, respectively, whereas only 1/26 three-pronucleate zygotes cultured using treatment A reached either of these stages.