Spontaneous differentiation of adult rat marrow stromal cells in a long-term culture

It is well recognized that bone marrow stromal cells (MSCs) can differentiate into neuron-like cells when supplemented with growth factors and/or chemical treatments. We demonstrated that primary MSCs obtained from adult rats could spontaneously differentiate into neural precursor cells after long-term culture. During the outset of in vitro culture, less than 0.1% of adult rat primary MSCs expressed nestin, the common protein of neural precursors. These MSCs didn't show neuronal morphology nor express neuronal antigens. In contrast, after continuous maintenance for 6 weeks, a significant subpopulation of MSCs formed cellular clumps and expressed nestin (32.3 +/- 6.3%). Less than 0.1% of cells expressing immature neuron marker betaIII-tubulin could be detected in these prolonged cultured MSCs. After serum deprivation and growth factor supplement, these nestin-positive cells could express neuron-like morphology and neuron-specific markers NF-H, betaIII-tubulin, tau, and neurotransmitter GABA. In contrast, the MSCs without prolonged culture didn't show neuronal morphology nor neuronal markers even after serum withdrawal and growth factors stimulation. These results demonstrated that neural precursors could be obtained from long-term cultured MSCs, and suggested that MSCs should be useful as a potential source for treatment of neurological disease.     

Genetic variation of viral protein 1 genes of field strains of waterfowl parvoviruses and their attenuated derivatives

To understand the genetic variations between the field strains of waterfowl parvoviruses and their attenuated derivatives, we analyzed the complete nucleotide sequences of the viral protein 1 (VP1) genes of nine field strains and two vaccine strains of waterfowl parvoviruses. Sequence comparison of the VP1 proteins showed that these viruses could be divided into goose parvovirus (GPV) related and Muscovy duck parvovirus (MDPV) related groups. The amino acid difference between GPV- and MDPV-related groups ranged from 13.1% to 15.8%, and the most variable region resided in the N terminus of VP2. The vaccine strains of GPV and MDPV exhibited only 1.2% and 0.3% difference in amino acid when compared with their parental field strains, and most of these differences resided in residues 497-575 of VP1, suggesting that these residues might be important for the attenuation of GPV and MDPV. When the GPV strains isolated in 1982 (the strain 82-0308) and in 2001 (the strain 01-1001) were compared, only 0.3% difference in amino acid was found, while MDPV strains isolated in 1990 (the strain 90-0219) and 1997 (the strain 97-0104) showed only 0.4% difference in amino acid. The result indicates that the genome of waterfowl parvovirus had remained highly stable in the field.     

A comparative study of the interaction of Bartonella henselae strains with human endothelial cells

Bartonella henselae can cause a wide range of clinical outcomes and may lead to severe disease, especially in patients with acquired immunodeficiency syndrome. It is well-known that B. henselae-induced cell proliferation is mediated by anti-apoptotic activity; however, the detailed mechanism is still unclear. In this study, the cellular responses of endothelial cells after infection with four B. henselae strains were compared and protein candidates that may be involved in the interaction between cells and bacteria were determined. The Houston-1 strain elicited the fastest response in terms of stimulating endothelial cell proliferation, and the JK-40 strain had the strongest ability to induce cell proliferation. By Western blot analysis, it was demonstrated that B. henselae-induced cell proliferation involved the mitochondria intrinsic apoptotic pathway. In addition, the adhesion abilities of the U-4 and JK-40 strains were much greater than those of the Houston-1 and JK-47 strains; however, the ability of Houston-1 to invade host cells was high. By two-dimensional gel electrophoresis analysis, it was found that succinyl-CoA synthetase subunit beta, phage-related protein, and ATP synthase subunit alpha might be involved in the invasion process. The expression of superoxide dismutase [Cu-Zn] precursor increased with infection time for all four strains but was significantly higher in the Houston-1 strain, which may increase the competitive advantage of Houston-1 in terms of survival in host cells and render it successful in invading host cells and stimulating cell proliferation. Our data suggest that the interaction of B. henselae and endothelial cells differed between strains, and the results indicated possible candidate proteins that may play a role in the pathogenesis of B. henselae infection.     

Cytotoxic sesterterpenoids from a sponge Hippospongia sp

One new pentacyclic sesterterpene, hippospongide A (1), and one new scalarane sesterterpenoid, hippospongide B (2), along with six previously reported known scalarane-type sesterterpenes (3-8), were isolated from a sponge Hippospongia sp. The structures of these compounds were elucidated on the basis of their spectroscopic data and comparison of the NMR data with those of known analogues. These metabolites are the first pentacyclic sesterterpene and scalarane-type sesterterpenes to be reported from this genus. Compounds 3-5 exhibited significant cytotoxicity against DLD-1, HCT-116, T-47D and K562 cancer cell lines.     

Use of vacuum-assisted closure for management of a large skin wound in a cat

CASE DESCRIPTION: A 9-month-old domestic shorthair cat was evaluated after being struck by a car. CLINICAL FINDINGS: The cat had a fractured tibia and avulsion of the tail base. Motor and deep pain sensation were absent from the tail. The fractured tibia was repaired 2 days after the trauma. On the third day, the cat developed tachypnea, dyspnea, high serum urea nitrogen and total bilirubin concentrations, epistaxis, persistent hypotension, and oliguria. The cat recovered with supportive care but developed extensive necrosis of the skin on the dorsum by 9 days after the initial trauma. TREATMENT AND OUTCOME: The skin was debrided from the caudal portion of the scapula to the anus and down each pelvic limb to the level of the distal portion of the femur. The tail was amputated. Wet-to-dry bandages were applied to the wound for 3 days. Approximately 50% of the wound underwent delayed primary closure, and the remainder was managed with vacuum-assisted closure. A healthy granulation bed was quickly established. Vacuum-assisted closure was also applied after graft application. Graft acceptance was 100%, and use of the vacuum-assisted closure bandage was not associated with the complications associated with the traditional bandage. CLINICAL RELEVANCE: Vacuum-assisted closure is a useful, easily applicable technique for open and grafted wounds, even when wounds are in challenging anatomic locations.