Microsurgery and external fixation in orthoplastic reconstruction of tibial injuries

Background Orthoplastics is a relatively new approach to lower limb reconstruction, where an integration of both plastic and orthopedic expertise is required, together with the availability of well-equipped facilities. Acute shortening and long-term frames for lengthening are generally considered alternatives to length preservation and soft tissue microsurgical reconstruction, but an integration of external fixation and reconstructive microsurgery is gaining an increasing role with refinements of joint approaches. Material and methods Data on sixteen patients who underwent microsurgical lower limb reconstruction and external fixation with an orthoplastic approach, following acute or chronic tibial injury, were retrospectively reviewed. All patients presented a post traumatic soft tissue defect associated with a Gustilo III tibial fracture or a tibial septic pseudarthrosis. Data on type and timing of bone and soft tissue reconstruction, outcomes, complications and need for re-operation were extrapolated and compared to an historic group of patients treated with an orthopedic-based approach. Results In the orthoplastic group, soft tissues were reconstructed with an ALT flap in most cases; a muscle-sparing VL or ALT-VL chimeric flap was necessary in cases with a very extensive defect. In the orthopedic group, soft tissues were left to heal by second intention or patients were lately referred to plastic surgeons. Statistical comparison between the two groups has showed significant differences on the following data: Time for soft tissue healing, time to bone union, number of reinterventions, post-operative deep infection rate, time to return to work. Conclusion The orthoplastic approach to complex leg defects yields shorter treatment time and better functional results compared to the orthopedic-based approach. External fixation and microsurgical reconstruction are not necessarily alternative procedures but can integrate in an orthoplastic path to address at best both soft tissue and bone reconstruction

Fertility ontrol by GnRH analogues in dogs.

GnRH plays a pivotal role in reproduction by stimulating the release of gonadotrophins. Chemical substitutions in the GnRH molecule lead to analogues possessing antagonist or agonist activity (Paramo RM et al. 1993 J. Reprod. Fertil. Suppl. 47, 387–397). The highly potent agonist analogue, Buserelin, with up to 20 times of potency, by increasing binding affinity, desensitizing competitive receptors, and resisting metabolic degradation, shuts down rather than stimulates reproductive function (Bertschinger HJ et al. 2001 J. Reprod. Fertil. Suppl. 57, 275–283). In man, Buserelin is employed in several gonadal hormone-dependent diseases and for prostatic cancers. We suppress gonadal function in male dogs using Buserelin. Eight intact male German sheep dogs 20 months old were divided into two groups; A, 4 subjects treated for pharmacological castration (Buserelin acetate, 0.3 mg/each, s.c., every 8 h for 30 days) (Suprefact-Aventis Pharma, Italy); B, 4 subjects treated with placebo (NaCl, 0.9%, s.c., every 8 h for 30 days). Plasma testosterone concentrations were measured twice a week by RIA using commercial kits (Coat-A-Count, Los Angeles, USA). Clinical examination of the male genital tract was conducted by ultrasound monitoring. Before and after the pharmacological treatment, semen was collected and evaluated for macroscopic and microscopic parameters. After treatment, testicular specimens were collected by orchiectomy, fixed in Bouin's solution, and embedded in paraffin wax. Thin sections were cut and stained with hematoxylin/eosin. The presence of germ cells (spermatogonia to spermatozoa, Sertoli and Leydig cells number) were analyzed. Randomly selected fields of transverse and longitudinal sections of seminiferous tubules were observed and analyzed using a computer assisted image analyzer (MONO system, Italy). The images acquired were segmented and binarized in order to obtain the masks of the tubular profiles; the mean values of the area, major and minor axes, mean diameter, and perimeter occupied by the testicular tubules were calculated automatically. Data were analyzed by ANOVA test. After Buserelin, all dogs (group A) showed a reduction in testicular and prostatic diameters compared to group B. Azoospermia was observed in group A. Histological examination revealed a statistically significant cell reduction of the germinal line (spermatogonia and spermatocytes, P < 0.05; spermatids and spermatozoa, P < 0.001). GnRH pharmacological treatment induced a cessation of normal spermatogenesis at the spermatocyte level while no statistical difference in morphometric parameters of seminiferous tubules were observed. The basic testosterone level (3.2 ± 1.3 ng/mL) rose to 12 ± 3.7 ng/mL (21° day) and than shut down to 0.5 ± 0.3 ng/mL (30° day), giving a long-term suppression. The present study demonstrates that Buserelin is an anti-fertility agent that gives suppression of reproductive function in male dogs. The method may have a clinical application. The utilization of a Buserelin depot will be a successive step

Notch signaling regulates tubular morphogenesis during repair from biliary damage in mice.

BACKGROUND & AIMS: Repair from biliary damages requires the biliary specification of hepatic progenitor cells and the remodeling of ductular reactive structures into branching biliary tubules. We hypothesized that the morphogenetic role of Notch signaling is maintained during the repair process and have addressed this hypothesis using pharmacologic and genetic models of defective Notch signaling. METHODS: Treatment with DDC (3,5-diethoxycarbonyl-1,4-dihydrocollidine) or ANIT (alpha-naphthyl-isothiocyanate) was used to induce biliary damage in wild type mice and in mice with a liver specific defect in the Notch-2 receptor (Notch-2-cKO) or in RPB-Jk. Hepatic progenitor cells, ductular reaction, and mature ductules were quantified using K19 and SOX-9. RESULTS: In DDC treated wild type mice, pharmacologic Notch inhibition with dibenzazepine decreased the number of both ductular reaction and hepatic progenitor cells. Notch-2-cKO mice treated with DDC or ANIT accumulated hepatic progenitor cells that failed to progress into mature ducts. In RBP-Jk-cKO mice, mature ducts and hepatic progenitor cells were both significantly reduced with respect to similarly treated wild type mice. The mouse progenitor cell line BMOL cultured on matrigel, formed a tubular network allowing the study of tubule formation in vitro; \u3b3-secretase inhibitor treatment and siRNAs silencing of Notch-1, Notch-2 or Jagged-1 significantly reduced both the length and number of tubular branches. CONCLUSIONS: These data demonstrate that Notch signaling plays an essential role in biliary repair. Lack of Notch-2 prevents biliary tubule formation, both in vivo and in vitro. Lack of RBP-Jk inhibits the generation of biliary-committed precursors and tubule formation

The cystic fibrosis transmembrane conductance regulator controls biliary epithelial inflammation and permeability by regulating Src tyrosine kinase activity

In the liver, the cystic fibrosis (CF) transmembrane conductance regulator (CFTR) regulates bile secretion and other functions at the apical membrane of biliary epithelial cells (i.e., cholangiocytes). CF-related liver disease is a major cause of death in patients with CF. CFTR dysfunction affects innate immune pathways, generating a para-inflammatory status in the liver and other epithelia. This study investigates the mechanisms linking CFTR to toll-like receptor 4 activity. We found that CFTR is associated with a multiprotein complex at the apical membrane of normal mouse cholangiocytes, with proteins that negatively control Rous sarcoma oncogene cellular homolog (Src) activity. In CFTR-defective cholangiocytes, Src tyrosine kinase self-activates and phosphorylates toll-like receptor 4, resulting in activation of nuclear factor kappa-light-chain-enhancer of activated B cells and increased proinflammatory cytokine production in response to endotoxins. This Src/nuclear factor kappa-light-chain-enhancer of activated B cells-dependent inflammatory process attracts inflammatory cells but also generates changes in the apical junctional complex and loss of epithelial barrier function. Inhibition of Src decreased the inflammatory response of CF cholangiocytes to lipopolysaccharide, rescued the junctional defect in vitro, and significantly attenuated endotoxin-induced biliary damage and inflammation in vivo (Cftr knockout mice). Conclusion: These findings reveal a novel function of CFTR as a regulator of toll-like receptor 4 responses and cell polarity in biliary epithelial cells; this mechanism is pathogenetic, as shown by the protective effects of Src inhibition in vivo, and may be a novel therapeutic target in CF-related liver disease and other inflammatory cholangiopathies. (Hepatology 2016;64:2118-2134)