Canine Visceral Leishmaniasis (CVL) shares many aspects with the human disease and dogs are considered the main urban reservoir of L. infantum in zoonotic VL. Infected dogs develop progressive disease with a large clinical spectrum. A complex balance between the parasite and the genetic/immunological background of the host are decisive for infection evolution and clinical outcome. This study comprised 92 Leishmania infected mongrel dogs of various ages from Mato Grosso, Brazil. Spleen samples were collected for determining parasite load, humoral response, cytokine mRNA expression and histopathology alterations. By real-time PCR for the ssrRNA Leishmania gene, two groups were defined; a low (lowP, n = 46) and a high parasite load groups (highP, n = 42). When comparing these groups, results show variable individual humoral immune response with higher specific IgG production in infected animals but with a notable difference in CVL rapid test optical densities (DPP) between highP and lowP groups. Splenic architecture disruption was characterized by disorganization of white pulp, more evident in animals with high parasitism. All cytokine transcripts in spleen were less expressed in highP than lowP groups with a large heterogeneous variation in response. Individual correlation analysis between cytokine expression and parasite load revealed a negative correlation for both pro-inflammatory cytokines: IFNγ, IL-12, IL-6; and anti-inflammatory cytokines: IL-10 and TGFβ. TNF showed the best negative correlation (r2 = 0.231; p<0.001). Herein we describe impairment on mRNA cytokine expression in leishmania infected dogs with high parasite load associated with a structural modification in the splenic lymphoid micro-architecture. We also discuss the possible mechanism responsible for the uncontrolled parasite growth and clinical outcome.

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BackgroundThe repair of large bone defects is a major orthopedic challenge because autologous bone grafts are not available in large amounts and because harvesting is often associated with donor-site morbidity. Considering that bone marrow stromal cells (BMSC) are responsible for the maintenance of bone turnover throughout life, we investigated bone repair at a site of a critically sized segmental defect in sheep tibia treated with BMSCs loaded onto allografts. The defect was created in the mid-portion of the tibial diaphysis of eight adult sheep, and the sheep were treated with ex-vivo expanded autologous BMSCs isolated from marrow aspirates and loaded onto cortical allografts (n = 4). The treated sheep were compared with control sheep that had been treated with cell-free allografts (n = 4) obtained from donors of the same breed as the receptor sheep.ResultsThe healing response was monitored by radiographs monthly and by computed tomography and histology at six, ten, fourteen, and eighteen weeks after surgery. For the cell-loaded allografts, union was established more rapidly at the interface between the host bone and the allograft, and the healing process was more conspicuous. Remodeling of the allograft was complete at 18 weeks in the cell-treated animals. Histologically, the marrow cavity was reestablished, with intertrabecular spaces being filled with adipose marrow and with evidence of focal hematopoiesis.ConclusionsAllografts cellularized with AOCs (allografts of osteoprogenitor cells) can generate great clinical outcomes to noncellularized allografts to consolidate, reshape, structurally and morphologically reconstruct bone and bone marrow in a relatively short period of time. These features make this strategy very attractive for clinical use in orthopedic bioengineering.

-The objective of this study was to verify the effects of GnRH on ovulation and pregnancy of ewes subjected to a short-term synchronization of estrus. Santa Inês and crossbred Santa Inês/Dorper ewes received 60 mg MAP sponges during 6 days plus 300 IU eCG and 30 μg d-cloprostenol 24 h prior to sponge withdrawal (SW). Ewes were assigned to receive 0.9% NaCl solution (T control ; n = 32) or 25 μg GnRH (licerelin, T GnRH ; n = 34) 24 hours after SW. Each group was assigned to intrauterine insemination by laparoscopy (n = 25) or to natural mating (n = 41). Artificial insemination was performed with a single dose of fresh semen. For controlled mating, females were exposed to males 12, 24, 36 and 48 hours after SW. Ten females per treatment were subjected to transrectal ultrasound examination at 12-hour intervals (SW to 60 hours after). Estrous response (100.0% vs 95.2%), interval from SW to estrus (32.9±7.4 vs 29.8±6.9 hours), estrous length (37.4±9.0 vs 31.5±10.4 hours), pregnancy rates (57.0% vs 41.0%), ovulation rate (100.0% vs 90.0%), number of ovulations/ewe (1.1±0.3 vs 1.2±0.4), maximum follicular diameter (6.4±0.7 vs 6.1±0.6 mm), interval from SW to ovulation (59.1±3.5 vs 58.4±3.5 hours) did not differ between T control and T GnRH , respectively. Administration of GnRH 24 hours after SW does not improve ovulation or pregnancy rate in estrous synchronization in ewes.

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Introduction: Nonunion is a challenging condition in orthopaedics as its etiology is not fully understood. Clinical interventions currently aim to stimulate both the biological and mechanical aspects of the bone healing process by using bone autografts and surgical fixation. However, recent observations showed that atrophic nonunion tissues contain putative osteoprogenitors, raising the hypothesis that its reactivation could be explored to achieve bone repair. Methods: Here we characterized atrophic nonunion stromal cells (NUSC) in vitro, using bone marrow stromal cells (BMSC) and osteoblasts as controls cells of the osteoblastic lineage, and evaluated its ability to form bone in vivo. Results: NUSC had proliferative and senescence rates comparable to BMSC and osteoblasts, and homogeneously expressed the osteolineage markers CD90 and CD73. Regarding CD105 and CD146 expression, NUSC were closely related to osteoblasts, both with an inferior percentage of CD105 + /CD146 + cells as compared to BMSC. Despite this, NUSC differentiated along the osteogenic and adipogenic lineages in vitro; and when transplanted subcutaneously into immunocompromised mice, new bone formation and hematopoietic marrow were established. Conclusions: This study demonstrates that NUSC are osteogenically competent, supporting the hypothesis that their endogenous reactivation could be a strategy to stimulate the bone formation while reducing the amount of bone autograft requirements.

The aim of this study was to verify the efficacy of reusing intravaginal progesterone (P4) devices on the reproductive parameters in Santa Inês ewes. Females received intravaginal P4 devices for their first, second or third use for five days plus 300 IU eCG IM and 5mg dinoprost laterovulvar 24h before device removal. Blood was collected at different moments. Transrectal ultrasonography was performed from device removal to ovulation. Part of the ewes were submitted to artificial insemination by laparoscopy (IAL -n=55) with fresh semen, whereas the rest were bred by fertile rams (n=41). On the initial 18 h, ewes that received devices for the first time showed higher P4 concentrations (5.1±1.8 vs 3.5±1.4 vs 2.4±1.1 -P<0.05). However, after the first 48h no difference was observed among all treatments and P4 supraluteal concentrations were detected in all ewes upon device removal. Estrous response, interval from device removal to estrus, rate of ovulating animals, number of ovulations, time from device removal to ovulation and average conception rates after IAL or natural mating were similar among all 3 groups. Intravaginal progesterone devices can be used up to three times without altering reproductive parameters in Santa Inês ewes.Keywords: sheep, artificial insemination, CIDR, intravaginal progesterone insert, ovulation Palavras-chave: ovino, inseminação artificial, CIDR, dispositivo intravaginal de progesterona, ovulação RESUMO Avaliou-se a eficácia da reutilização de dispositivos intravaginais de progesterona (P4) sobre características reprodutivas em ovelhas Santa Inês. As fêmeas receberam dispositivos intravaginais contendo P4 para o seu primeiro, segundo ou terceiro uso por cinco dias, associado a 300 UI eCG IM e 5mg dinoprost laterovulvar 24h antes da remoção dos dispositivos. Ultrassonografia transretal foi realizada da remoção dos dispositivos até a ovulação. Parte das ovelhas foi submetida à inseminação artificial laparoscópica (IAL-n=55) com sêmen a fresco, enquanto outra parte foi acasalada por machos férteis (n=41). Nas 18 h iniciais, as ovelhas que receberam dispositivos pela primeira vez