Induction of matrix metalloproteinases and collagenolysis in chick embryonic membranes before hatching

In NIH 3T3 fibroblasts and several other cellular systems, ethanol (50-80 mM) was previously shown to greatly enhance the mitogenic effects of insulin particularly in the presence of zinc. Here we report that in NIH 3T3 fibroblasts the combined stimulatory effects of ethanol and insulin on DNA synthesis can be further increased by bombesin both in the absence and presence of zinc. Bombesin also enhanced insulin-plus-ethanol-induced DNA synthesis in mouse Swiss 3T3 and Balb/c 3T3 fibroblasts, but in these cells bombesin was effective only in the presence of zinc. In NIH 3T3 fibroblasts, the potentiating effects of ethanol on insulin-induced DNA synthesis by the zinc-dependent and bombesin-dependent mechanisms were additive. Wortmannin, an inhibitor of phosphatidylinositol 3'-kinase (PI3K), prevented the comitogenic effect of ethanol in the presence of bombesin but not in the presence of zinc. Furthermore, bombesin, but not ethanol, was found to enhance the stimulatory effect of insulin on PI3K activity. Rapamycin, an indirect inhibitor of p70 S6 kinase actions, inhibited the comitogenic effects of ethanol in the presence of both zinc and bombesin. However, only ethanol, but not bombesin, enhanced the stimulatory effect of insulin on p70 S6 kinase activity; this effect of ethanol was zinc-dependent. Neither ethanol nor bombesin enhanced the stimulatory effects of insulin on the phosphorylation (activation) of p38/p42/p44 mitogen-activated protein kinases. The results suggest that in mouse fibroblasts maximal stimulation of DNA synthesis by physiologically relevant concentrations of ethanol occurs if both PI3K and p70 S6 kinase are activated. These data suggest a mechanism by which ethanol may affect growth in affected human tissues during its tumor promoting actions.     

Absorption, excretion, and tissue deposition of titanium in sheep

Titanium metabolism was measured in three 18 kg lambs each fed 450 g chopped hay daily. Two of the lambs were dosed orally and one intravenously with 3 muCi titanium-44 each. Clearance of the intravenous dose was extremely slow; after oral administration, however, no titanium-44 was detected in blood plasma for 48 h. Over 96% of the oral dose was recovered in feces and digestive tract contents. Titanium-44 absorption, estimated from total carcass recovery and by comparison of concentrations in internal organs of orally and intravenously dosed lambs, was less than .5%. Fecal titanium could be a satisfactory index of soil ingestion by grazing ruminants.     

Distribution, tolerability and tissue compatibility of intrathecal tizanidine in the sheep

Allelic variants for the HIV-1 co-receptors chemokine receptor 5 (CCR5) and CCR2, as well as the ligand for the co-receptor CXCR4, stromal-derived factor (SDF-1), have been associated with a delay in disease progression. We began this study to test whether polymorphisms in the CCR5 regulatory regions influence the course of HIV-1 disease, as well as to examine the role of the previously identified allelic variants in 1,090 HIV-1 infected individuals. Here we describe the evolutionary relationships between the phenotypically important CCR5 alleles, define precisely the CCR5 regulatory sequences that are linked to the CCR5-delta32 and CCR2-641 polymorphisms, and identify genotypes associated with altered rates of HIV-1 disease progression. The disease-retarding effects of the CCR2-641 allele were found in African Americans but not in Caucasians, and the SDF1-3'A/3'A genotype was associated with an accelerated progression to death. In contrast, the CCR5-delta32 allele and a CCR5 promoter mutation with which it is tightly linked were associated with limited disease-retarding effects. Collectively, these findings draw attention to a complex array of genetic determinants in the HIV-host interplay.     

Current practices in antepartum and intrapartum fetal monitoring

Survey forms were sent to 360 Ob./Gyn, residency programs in the United States and Canada with responses from 279 concerning usage of various types of fetal monitoring. The results are presented and discussed.     

Perforin-secreting killer cell infiltration in the aortic tissue of patients with atherosclerotic aortic aneurysm

Cell-mediated immunity has been implicated in the pathogenesis of vascular cell injury in patients with atherosclerotic aortic aneurysms. To clarify the immunologic mechanisms involved, we examined the expression of a cytolytic factor, perforin, in infiltrating cells from aortic tissue samples taken from 6 patients with atherosclerotic aortic aneurysms. Immunohistochemical studies showed that the infiltrating cells consisted mainly of macrophages, natural killer (NK) cells, cytotoxic T lymphocytes (CTLs), and T helper cells, and that perforin was expressed in NK cells and CTLs. Immunoelectron microscopic studies demonstrated that the infiltrating cells released massive amounts of perforin directly on to the surface of arterial vascular cells. These findings provide the first direct evidence that some of the infiltrating cells in the aortic tissue consist of killer cells, and strongly suggest that these killer cells, especially NK cells and CTLs, may play a critical role in the vascular cell injury caused by atherosclerotic aortic aneurysm by releasing perforin.     

An in vivo method for studying afferent fibre activity from cervical paravertebral tissue during vertebral motion in anaesthetised cats

PURPOSE: The effect of ethanol and formate radicals on the major proteins of human erythrocyte membranes has been investigated. MATERIALS AND METHODS: Human erythrocyte ghosts and of erythrocyte ghosts stripped of peripheric proteins were irradiated in phosphate buffer with 100 mmol dm(-3) ethanol or 100 mmol dm(-3) formate under N2 or N2O. The alterations of the proteins were investigated by SDS-polyacrylamide gel electrophoresis and high-performance gel permeation chromatography. RESULTS: In contrast to previous results on ribonuclease and on serum albumin the ethanol radicals were found to have a higher efficiency to damage erythrocyte membrane proteins than the formate radicals. Spectrin (Bands 1 and 2) and capnophorin (Band 3) showed the highest radiation-induced loss of all membrane proteins. When cysteamine or dithiothreitol were added to the erythrocyte ghosts with a similar OH-scavenging capacity as ethanol or formate, no degradation or aggregation of the membrane proteins could be observed even after a dose as high as 1800 Gy. CONCLUSIONS: The results of this study confirm the high radiosensitivity of spectrin and capnophorin to primary radicals. Similarly to soluble proteins, membrane-associated proteins are more significantly damaged by ethanol radicals than by formate radicals.     

Leukocyte polarization in cell migration and immune interactions

Cell migration plays a key role in a wide variety of biological phenomena. This process is particularly important for leukocyte function and the inflammatory response. Prior to migration leukocytes undergo polarization, with the formation of a lamellipodium at the leading edge and a uropod at the trailing edge. This cell shape allows them to convert cytoskeletal forces into net cell-body displacement. Leukocyte chemoattractants, including chemokines, provide directional cues for leukocyte motility, and concomitantly induce polarization. Chemoattractant receptors, integrins and other adhesion molecules, cytoskeletal proteins and intracellular regulatory molecules change their cellular localization during cell polarization. A complex system of signal transduction molecules, including tyrosine kinases, lipid kinases, second messengers and members of the Rho family of small GTPases is thought to regulate the cytoskeletal rearrangements underlying leukocyte polarization and migration. The elucidation of the mechanisms and signals that control this complex reorganization will lead to a better understanding of critical questions in cell biology of leukocyte migration and polarity.     

Phenotype, and migration properties of three major subsets of tissue homing T cells in sheep

T cells show a bias in their migration pathways: some T cells preferentially migrate to peripheral lymph nodes (LN), some to mucosal tissues, and some to peripheral tissues such as skin. These recirculation pathways were examined in sheep by collecting lymph draining into and out of peripheral and intestinal LN, and using fluorescent dyes to trace the recirculation of the lymph cells. Monoclonal antibodies (mAb) to alpha 4, beta 1, and beta 7 integrins, and L-selectin, were used to define three major populations of recirculating T cells. Naive-type T cells (L-selectin+, alpha 4 beta 1lo beta 7lo) migrated preferentially through peripheral LN. Two memory populations could be defined: alpha 4 beta 1hi beta 7- and alpha 4 beta 7hi beta 1lo. alpha 4 beta 1hi beta 7- T cells were present in lymph draining from the skin. T cells migrating preferentially through intestinal LN were alpha 4 beta 7hi beta 1lo. Consistent with this migration pattern, the endothelial receptor for alpha 4 beta 7, mucosal addressin cell adhesion molecule-1 (MAdCAM-1) was detected on high endothelial venules within intestinal LN and Peyer's patches, but only weakly on high endothelial venules within peripheral LN. Thus, there are at least three easily definable subsets of T cells, based on integrin expression, which show distinct migration preferences.     

Gingival recessions and noncarious cervical lesions: a soft and hard tissue challenge

Gingival recessions and noncarious cervical lesions are frequent and closely related defects which can be successfully treated by mucogingival surgery and restorative therapy techniques either used alone or in combination depending on the defect characteristics. Proper treatment planning is guided by careful clinical diagnosis. A selection of treatment modalities are presented and useful hints proposed: the long term success is always strictly bound to the identification and elimination of the etiologic factors, to a correct treatment and a meticulous maintenance program.     

Leukocyte recruitment and turnover in an experimental inflammatory exudate

The turn-over of leukocytes at sites of inflammation in vivo is to a large extent uninvestigated, mainly due to the technical difficulties associated with sampling and analysis of the inflammatory exudate. This paper investigates the immigration of fluorescently labeled granulocytes into exudate chambers at 8 h and at 1, 3, and 6 days after implantation into abdominal muscle of rat. In each experiment, the circulating granulocytes were labeled by intravenous administration of the DNA-labeling fluorochrome Hoechst 33342 and allowed to migrate into the chamber during 6 h before harvesting the chamber exudate. The rate of granulocyte immigration into the chamber varied considerably over time, showing a minimum at 3 days after implantation. The resulting kinetic pattern of granulocyte numbers in the exudate showed a two-step appearance, different from that of earlier determinations in soft tissue. A comparison between the calculated rates of granulocyte immigration and the total number of granulocytes present in the exudate at different times indicated that all immigrated cells survived in the chamber for the entire observation period of 6 days.     

Significant injury of the neck including the cervical spine and soft tissue in suicidal strangulation

A 46-year-old man committed suicide by jumping from a height of app. 4.5 m with a running noose round his neck. The neck was nearly cut through by the tightened ligature, so that there was only a small "bridge" of soft tissue between head and trunk.     

Antigen retrieval on epoxy sections based on tissue infiltration with a moderately increased amount of accelerator to detect immune complex deposits in glomerular tissue

OBJECTIVE: The purpose of this article is to describe MR findings in surgically proven solitary intraductal papillomas of the breast, including findings on MR galactography. CONCLUSION: Women with spontaneous nipple discharge and solitary intraductal papilloma show, on MR imaging, dilated ducts with an associated enhancing, well-circumscribed mass.     

Hypomethylation in cervical tissue: is there a correlation with folate status?

5-Aminolevulinic acid (ALA) is the first intermediate substrate in the heme synthetic pathway and is the substrate of aminolevulinic acid dehydratase (ALAD, porphobilinogen synthase). Because lead effectively inhibits ALAD activity, resulting in accumulation of ALA in urine and blood, urinary ALA (ALAU) has been used as a biomarker for lead exposure or early biologic effect of lead. Intraindividual variation in urinary excretion of ALA requires the use of 24-hour urine samples or adjustment of single urine samples by other normalizing variables, such as urinary creatinine concentration. Previous studies of ALAU concentration have used various adjustment methods; however, few have compared creatinine-adjusted ALAU concentration with ALA concentration in plasma (ALAP) from subjects with low (< 30 micrograms/dL) to moderate (< 60 micrograms/dL) levels of blood lead. To determine if creatinine-adjusted ALAU is associated with ALAP, we measured ALAU, ALAP, and urinary creatinine in 65 Korean lead workers with blood lead concentrations in the range of 14-60 micrograms/dL. ALAU, ALAU/creatinine, or ALAU/log creatinine all correlated with ALAP. However, ALAU/creatinine correlated more closely with ALAP based on Spearman's r (rs = 0.40, P, = 0.0009), supporting the use of ALA/creatinine in single urine samples as a surrogate for ALAP.     

Absorption of L-carnosine, L-methionine, and L-methionylglycine by isolated sheep ruminal and omasal epithelial tissue

The absorption of carnosine, methionine, and methionylglycine (using 35S-methionine and 35S-methionylglycine as representative markers) across ruminal and omasal epithelia collected from four (carnosine) and seven sheep (methionine and methionylglycine) were studied using parabiotic chambers that were repeatedly sampled over a 240-min incubation. The quantity of all substrates transferred was linearly (P < .01) dependent on initial substrate concentration and time. More (P < .01) carnosine, methionine, and methionylglycine was transferred across omasal than across ruminal epithelia. Carnosine was absorbed across both tissues without hydrolysis. Methionylglycine was transferred intact across both tissues. A greater (P < .01) quantity was hydrolyzed by omasal than by ruminal epithelia, after 240 min of incubation. Greater (P < .06) quantities of methionine and methionylglycine accumulated in ruminal tissue after 240 min. Total absorption of methionine and methionylglycine did not differ within tissues, but total absorption of both substrates was greater (P < .01) by omasal tissues. There was little evidence for saturable absorption. These results indicate that omasal epithelial tissue possesses a greater ability to absorb both free and peptide-bound amino acids than does ruminal epithelial tissues.     

Differentiation of staphylococci from sheep and goat milk samples

A total of 447 micrococcaceae strains isolated from 88 ewe and 359 goat milk samples from cases of chronic mastitis were differentiated by means of the ATB 32 STAPH-test. Of these strains 389 (= 87%) could be identified. Fourteen strains were sensitive in the bacitracin-resistance-test and therefore classified as Micrococcus spp. In ewe milk following Staphylococcus spp. were found: S. epidermidis, S. aureus, S. lentus, S. xylosus, S. warneri, S. equorum, S. haemolyticus, S. simulans, S. hominis and S. saprophyticus. Staphylococcus spp. identified in goat milk samples were: S. epidermidis, S. aureus, S. caprae, S. lentus, S. simulans, S. capitis, S. lugdunensis, S. xylosus, S. chromogenes, S. hominis, S. arlettae, S. warneri, S. sciuri, and S. saprophyticus. Highest cell counts in the milk of both animals species, and the highest incidence of clinical udder alterations were caused by S. aureus. Increases in milk cell counts as well as pathological udder findings were observed in coagulase-negative staphylococcal infections for novobiocin-sensitive Staphylococcus spp. (S. epidermidis, S. warneri, S. simulans, S. lugdunensis, and S. chromogenes) and several S. lentus strains.     

Kinematics of cervical spine discectomy with and without bone grafting: quantitative evaluation of late fusion in a sheep model

OBJECTIVE: This study was conducted to evaluate the kinematic response of late fusion results for cervical spine discectomies with and without bone grafting. MATERIALS AND METHODS: Fifteen Barbados Black Belly sheep underwent sham operations (Group A, n = 5), C2-C3 discectomies only (Group B, n = 5), and C2-C3 discectomies with autologous iliac bone grafting (Group C, n = 5). Ten months after surgery, the animals were killed. Fresh ligamentous spines (C1-C5) were subjected to the relevantly applied loads through a loading frame attached to the C1. Each vertebra (from C2 to C4) was attached with a set of three infrared light-emitting diodes to record the spatial location relating to each load application using a Selspot II system (Selcom Selective Electronics, Inc., Valdese, NC). The load-deformation data of the C2-C3 and C3-C4 motion segments were recorded and analyzed for the three groups. RESULTS: At the C2-C3 motion segment, the results indicated that Group B displayed larger motion ranges of rotation and lateral bending loads than did the other two groups. Significantly larger motion ranges of rotation loads were found in Group B than in Group C (P<0.05, for both comparisons). In contrast, Group C had the smallest motion ranges of flexion, lateral bending, and rotation loads. At the C3-C4 motion segment, both groups that had undergone discectomies had a significantly larger motion range of flexion load compared with Group A (P<0.05, for both comparisons). A significant increase in the motion range of right axial rotation was found in Group B (P<0.05), but not in Group C, compared with Group A. Group B exhibited larger motion ranges responding to all six tested loads than did Group C. CONCLUSION: The results indicate that anterior fusion after C2-C3 cervical discectomies, regardless of the presence or absence of bone grafting, decreases the motion range of flexion load at the C2-C3 motion segment, and contrary data were seen at the C3-C4 motion segment. For axial rotation loads, discectomies without bone grafting resulted in increased motion ranges of both C2-C3 and C3-C4 motion segments whereas discectomies with bone grafting did not. The data may have clinical relevance regarding the role of bone grafting in cases of cervical spine disease.