Nondébridement of laser char after two carbon dioxide laser passes results in faster reepithelialization

Skin repair following laser injury can be accelerated by using techniques that promote rapid reepithelialization. In this article, the benefit of intraoperative nondébridement of laser debris after two laser passes is discussed. After carbon dioxide laser resurfacing of the face, skin specimens were examined using indirect immunofluorescence with antibodies to specific epidermal and basement membrane proteins. Biopsy specimens obtained immediately after resurfacing showed a greater injury to epidermal and basement membrane proteins when skin was wiped with saline-soaked gauze after laser passes than when there was no débridement after two passes. Later examination of skin specimens obtained from nine patients 2 days after carbon dioxide resurfacing showed that nondébrided, occluded skin had faster reepithelialization than the other treatments. Nondébridement of the skin at the time of resurfacing along with the use of postoperative occlusive dressings led to the rapid reestablishment of a multilayered epidermis only 2 days after resurfacing. Nondébridement along with occlusive dressings results in rapid reepithelialization of the skin after two carbon dioxide laser passes for skin rejuvenation.     

Enzymatic liberation of lycotetraose from the Solanum glycoalkaloid alpha-tomatine

The branched tetrasaccharide, O-β-d-glucopyranosyl-(1 → 2)-O-[β-d-xylopyranosyl-(1 → 3)]-O-β-d-glucopyranosyl-(1 → 4)-d-galactose (lycotetraose) is a key constituent of many biologically interesting natural products. Described herein is a convenient enzymatic preparation of lycotetraose from the readily available Solanum glycoalkaloid α-tomatine. The preparation makes use of the recombinant endo-glycosidase, tomatinase, from the plant pathogen Fusarium oxysporum f. sp. lycopersici.     

Activation of sperm motility in striped bass via a cAMP-independent pathway

The objective of the present study was to identify the effect of osmolality, ions (K+, H+, Ca2+, Mg2+) and cAMP on the initiation of sperm motility in striped bass (Morone saxatilis). Striped bass spermatozoa remained motile in solutions isotonic to seminal plasma (350 mOsm/kg) until osmolality reached 600 mOsm/kg. K+ (0-100 mM) had no effect ( p>0.05 ) on sperm motility, and sperm displayed a high percentage of motility over a wide range of pH (6.0-8.5). Sperm motility could be initiated in Ca2+-free solutions. In contrast, sperm motility was inhibited (P<0.01) by solutions containing > or =10 mM Ca2+, and sperm could not be reactivated by a Ca2+-free solution. This Ca2+ inhibition was not affected by verapamil, a Ca2+ channel blocker. However, if sperm motility was first initiated in a Ca2+-free solution, the addition of Ca2+ solutions, up to 80 mM, failed to inhibit sperm motility, suggesting that Ca2+ inhibited the initiation of motility, but had no control of motile spermatozoa. Mg2+ solutions had similar inhibitory effects on sperm motility as Ca2+ solutions. Therefore, initiation of motility in striped bass sperm may be related to voltage-gated channels across the cell's plasma membrane. Membrane permeable cAMP did not initiate motility of quiescent, intact striped bass spermatozoa, and motility of demembranated sperm could be activated in the absence of cAMP.     

Adenosine A2A-dopamine D2 receptor-receptor heteromerization: qualitative and quantitative assessment by fluorescence and bioluminescence energy transfer

There is evidence for strong functional antagonistic interactions between adenosine A2A receptors (A2ARs) and dopamine D2 receptors (D2Rs). Although a close physical interaction between both receptors has recently been shown using co-immunoprecipitation and co-localization assays, the existence of a A2AR-D2R protein-protein interaction still had to be demonstrated in intact living cells. In the present work, fluorescence resonance energy transfer (FRET) and bioluminescence resonance energy transfer (BRET) techniques were used to confirm the occurrence of A2AR-D2R interactions in co-transfected cells. The degree of A2AR-D2R heteromerization, measured by BRET, did not vary after receptor activation with selective agonists, alone or in combination. BRET competition experiments were performed using a chimeric D2R-D1R in which helices 5 and 6, the third intracellular loop (I3), and the third extracellular loop (E3) of the D2R were replaced by those of the dopamine D1 receptor (D1R). Although the wild type D2R was able to decrease the BRET signal, the chimera failed to achieve any effect. This suggests that the helix 5-I3-helix 6-E3 portion of D2R holds the site(s) for interaction with A2AR. Modeling of A2AR and D2R using a modified rhodopsin template followed by molecular dynamics and docking simulations gave essentially two different possible modes of interaction between D2R and A2AR. In the most probable one, helix 5 and/or helix 6 and the N-terminal portion of I3 from D2R approached helix 4 and the C-terminal portion of the C-tail from the A2AR, respectively.     

Differential expression of Smad7 transcripts identifies the CD4+CD45RChigh regulatory T cells that mediate type V collagen-induced tolerance to lung allografts

Regulatory T cells (Tregs) induced by oral tolerance may suppress immunity by production of TGF-beta that could also enhance Treg activity. However, all cells that are phenotypically Tregs in rats (CD4(+)CD45RC(high)-RC(high)) may not have regulatory function. Because Smad7 expression in T cells is associated with inflammation and autoimmunity, then lack of Smad7 may identify those cells that function as Tregs. We reported that feeding type V collagen (col(V)) to WKY rats (RT1(l)) induces oral tolerance to lung allografts (F344-RT1(lvl)) by T cells that produce TGF-beta. The purpose of the current study was to identify the Tregs that mediate col(V)-induced tolerance, and determine Smad7 expression in these cells. RC(high) cells from tolerant rats were unresponsive to allogeneic stimulation and abrogated rejection after adoptive transfer. In contrast, CD4(+)CD45RC(low) (RC(low)) cells from tolerant rats and RC(high) or RC(low) cells from normal rats or untreated allograft recipients proliferated vigorously in response to donor Ags, and did not suppress rejection after adoptive transfer. TGF-beta enhanced proliferation in response to col(V) presented to tolerant RC(high), but not other cells. In contrast to other cells, only RC(high) cells from tolerant rats did not express Smad7. Collectively, these data show that the Tregs that mediate col(V)-induced tolerance to lung allografts do not express SMAD7 and, therefore, are permissive to TGF-beta-mediated signaling.     

Seasonal changes in the physiology of male Virginia opossums (Didelphis virginiana): signs of the Dasyurid semelparity syndrome?

Semelparity, which is multiplying once in a lifetime, is a rare reproductive strategy among mammals. Several species of the marsupial family Dasyuridae experience 100% male mortality following an intense mating period. We investigated seasonal physiological changes that may be associated with early mortality in the male Virginia opossum (Didelphis virginiana; Didelphidae) and compared these changes with those of semelparous, male dasyurids. Free-ranging male Virginia opossums (n=36) were collected during 2001 at the Oklahoma State University Cross Timbers Experimental Range. Seasonal data were collected on hematological, morphological, and helminth parameters of these individuals. We used one-way ANOVA to determine whether there were seasonal differences among means for each parameter. It appeared that male Virginia opossums experienced some physiological changes similar to those of male dasyurids exhibiting semelparity. All males collected in summer (August) were juveniles of the year. Lack of adult males in August suggests high mortality of this cohort during the breeding season. Opossum characteristics exhibiting the dasyurid semelparity syndrome included packed cell volume, adrenal mass, and helminth numbers. Minor lymphocytopenia, neutrophilia, and testosterone concentrations also were similar to semelparous dasyurids. However, a lack of change in serum cortisol concentration and body mass and dynamics in immunoglobulin protein, serum protein, and testes mass were not consistent with previous reports of semelparous dasyurid physiology. Evolutionary divergence and differences in breeding behavior between dasyurids and didelphids may be responsible for the lack of consistency between the taxa.     

Antibody response to sheep red blood cells in platypus and echidna

There is limited information regarding the kinetics of antibody responses exhibited by the platypus and the echidna in response to a T cell dependent antigen. In this preliminary study a platypus, an echidna and a rabbit were inoculated with sheep red blood cells to compare their antibody responses and kinetics. The antibody titres, produced by the platypus and echidna, were less than those elicited in the rabbit. Furthermore, the echidna and platypus exhibited a weak secondary response. This was most likely due to a failure of the platypus and echidna to undergo the characteristic IgM to IgG isotype switch following second antigen exposure. The conformational structure of these antibodies may differ from eutherian antibodies. This was further supported by a heat sensitivity experiment that indicated that these antibodies are more labile than rabbit immunoglobulins and therefore structurally less stable.