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Infections by respiratory syncytial virus (RSV) are more severe in patients with cystic fibrosis (CF), and many CF units use palivizumab as prophylaxis; however, information about palivizumab efficacy in CF patients is almost lacking.

Chronic wounds are of high socio-economic importance. Accurate and efficient treatment includes the use of outcomes measures in clinical care and in research. To date, consented standards for documentation of chronic wounds, agreements on interpretation of wound outcomes and harmonisation about standardised goal setting in wound treatment are lacking.

To examine the association between HIV RNA levels, patterns of antiretroviral resistance, and neurologic status.

Actinoplanic acids A and B are macrocyclic polycarboxylic acids that are potent reversible inhibitors of farnesyl-protein transferase. Actinoplanic acids A and B were isolated from Actinoplanes sp. MA 7066 while actinoplanic acid B was isolated from both MA 7066 and Streptomycessp. MA 7099. Actinoplanic acids A and B are competitive with respect to farnesyl diphosphate and are selective inhibitors of farnesyl-protein transferase because they do not inhibit geranylgeranyl-protein transferase type 1 or squalene synthase. MA 7066 is believed to be a novel species of actinomycetes while MA 7099 is believed to be a novel strain of Streptomyces violaceusnigeron the basis of morphological, biochemical and chemotaxonomic characteristics as well as its production of actinoplanic acids.

A method has been described for the long-term culture of human bone marrow cells in liquid medium. Hematopoiesis, as measured by the production of granulocytic-macrophage progenitor cells (CFUc), continued for at least 20 weeks and was dependent upon the presence of a marrow-derived adherent layer of cells. As in the case of murine marrow liquid cultures, the adherent layer consisted of mononuclear phagocytic cells, endothelial cells, and lipid-laden adipocytes, the latter being essential for long-term hematopoiesis. Optimal growth conditions included McCoy's medium supplemented with fetal bovine serum, horse serum, and hydrocortisone and incubation at 33 degrees C. Horse serum in conjunction with hydrocortisone appeared essential for the growth of adipocytes.

Several gentle hills close to the crest of the East Pacific Rise are covered with extensive Fe-Mn-rich deposists. The setting of the sediments precludes the possibility that turbidite flows, slumping or downhill winnowing of material can have been responsible for the accumulation of these sediments, as is also indicated by the lack of reworked faunas or crushed shells.Palaeontological dating and the magnetic anomalies of the crustal substratum show that the sediments must be very young, that is, the sedimentation rates are very high; thus, 8-10-m-thick sediments were formed in 600,000 yeas. The accumulation rates on these hills for several elements, such as Fe, Mn, Cu and Ba, are much higher than in ordinary pelagic regions. The element deposition is best described as an exhalative-sedimentary process. The areal extent of these sediments is considerably larger than that of any other currently forming analogous deposits. The results have far-reaching implications for the discussion of how sedimentary-exhalative iron-manganese ores originate.

Abstract: The resistance structure of a Turkish population of the clonal, apomictic composite Chondrilla juncea and the pathotypic structure of a co-occurring population of its obligate rust pathogen, Puccinia chondrillina, was determined by sequential inoculation of 19 host lines with 15 pathogen isolates each derived from single pustules collected from separate plants among the host population. The resultant matrix of resistant and susceptible reactions provides strong circumstantial evidence for a gene-for-gene interaction. Seven distinct pathotypes were detected in the pathogen population. One of these comprised 53% of the population, a second comprised 13%, while the remaining five pathotypes were each detected only once. The host population was similarly diverse, being composed of eight resistance phenotypes, only two of which were represented by more than one host line. Although C. juncea is apomictic, there was only 58% congruence between host resistance and multi-locus isozyme phenotype categories within this population. Pathotypic phenotypes of 13 other isolates of P. chondrillina collected from ten other Turkish and three more distant populations of C. juncea were markedly different from those found in the population studied in detail. There was no obvious relationship between the degree of geographic separation of pathotypes and their ability to attack particular C. juncea lines in this or three other populations represented by single host lines.

BALB/c mice thymectomized at 3 to 5 days of age were studied to determine if this procedure would result in enhanced susceptibility to infection with Mycobacterium leprae and, if so, whether or not administration of antilymphocyte serum would further increase this susceptibility. The plateau for growth in the footpads of intact mice occurred 4 months after inoculation, whereas in the thymectomized and thymetocomized plus antilymphocyte serum-treated groups the plateau occurred between months 11 and 12 after inoculation. Thymectomy resulted in at least a 10-fold increase in the number of M. leprae found in the footpads. Antilymphocyte serum did not appear to further enhance the M. leprae infection in the thymectomized mice. Although growth of M. leprae in the testes of both intact and thymectomized mice was erratic, the number of organisms reached a higher ceiling in the thymectomized groups. M. leprae harvested from all groups was passaged into intact mice at various intervals after inoculation to test for viability. Viable M. leprae were found at all intervals tested including 22 months after infection in the intact mice, suggesting that a chronic infection occurred that probably lasted during the entire life of the animals.

The susceptibility of highly purified human CD34+ cells to monocytotropic (Ba-L) and lymphotropic (A018-post) strains of human immunodeficiency virus-1 (HIV-1) was examined. Liquid cultures initiated with fresh immunomagnetically purified CD34+ cells using the K6.1 CD34 monoclonal antibody (MoAb) (K6.1/CD34+) were positive for HIV expression 2 weeks after exposure to HIV-1 Ba-L. These cells were initially greater than 90% CD34+ and had undetectable monocyte contamination by flow-cytometric staining and side-scatter analyses, respectively, and undetectable T-cell contamination by CD3 polymerase chain reaction (PCR) analysis. However, secondary CD34+ liquid cultures reselected from the primary liquid cultures 24 hours after HIV exposure by panning with the ICH3 CD34 MoAb (ICH3/CD34+) and maintained for an additional 14 days were negative for HIV expression. The ICH3-unbound cells were positive for both spliced and unspliced HIV RNA when exposed to HIV-1 Ba-L, and were DNA PCR positive when exposed to either monocytotropic or lymphotropic HIV-1. To further test that CD34+ cells were not infectible by HIV-1, we exposed K6.1/CD34+ cells continuously to HIV-1 in a culture system capable of maintaining and expanding primitive CD34+ cells. HIV-exposed K6.1/CD34+ cells proliferated and expanded as efficiently as uninfected cultures. However, when reselected magnetically using the K6.1 CD34 MoAb after expansion for 7 days, bound K6.1/CD34+ cells were again negative for HIV-1 expression, whereas unbound cells were positive for HIV-1 expression. These findings suggest that a sequential CD34+ cell-selection process, in which the two selections are separated by a brief culture period, can yield a population of CD34+ cells that are not infected with HIV-1. This process may be useful in the design of stem or progenitor cell- based transplantation therapies for HIV infection.

Nine first-appearance datums (FADs), twenty-three last-appearance datums (LADs), and three other micropaleontological datums are related to the magnetic-reversal, oxygen-isotope, and calcite-dissolution/coarse-fraction time scales to provide a preliminary basis for subdivision of the Quaternary in deep-sea sediments. The magnetic-reversal, oxygen-isotope, and calcite-dissolution/coarse-fraction scales have been correlated by determination on the same core materials, and absolute dates applied by 40K40Aror 14C dating of materials in known positions on one or another of these scales. FADS and LADs have been determined in cores for which either a magnetic-reversal, oxygen-isotope, or calcite-dissolution/coarse-fraction scale has also been available. Altogether 3 FADs and 5 LADs based on diatoms, 4 FADs and 5 LADs based on calcareous nannoplankton, 1 FAD and 8 LADs based on radiolarians, 1 FAD and 5 LADs based on planktonic foraminifers, 2 acme datums, and 1 ratio reversal datum have been determined, and absolute dates inferred by interpolation from known dates on the reference time scales. Some of the FADs and LADs apply or are synchronous only over limited areas of the oceans; others appear to be synchronous throughout the oceans. The base of the Quaternary is set at the top of the Olduvai event at 1.7 my. Four FADs, twelve LADs, two acme datums, and one ratio reversal datum occur above the base of the Quaternary at an average rate of about 1 per 100,000 yr. Five FADs and twelve LADs are recognized in the 0.8-my interval between the top of the Olduvai event and the Gauss/Matuyama Boundary at 2.5 my at an average incidence of about 1 per 50,000 yr.

Decline in blood CD4+ lymphocytes during primary symptomatic infections with HIV is usually attributed to viral killing, and has not been considered in terms of altered lymphocyte migration and sequestration. We therefore sought to examine whether CD4+ cell loss from blood of macaques undergoing an acute primary SIV infection might be due to increased synthesis of cytokines, known to profoundly affect lymphocyte trafficking, rather than to direct lymphocyte destruction by virus. The findings indicate that rapid lymphocyte depletion following acute infection is not selective for CD4+ cells, correlates precisely with increased plasma IFN-gamma and tumor necrosis factor-alpha levels, and is reversible. CD4/CD8 ratios in lymph nodes with high viral burdens remain relatively unchanged despite lymphocyte loss from blood. Levels of cytokine mRNA measured in lymphoid organs reflect neither cytokine plasma levels nor their potential to induce sequestration. These results support a model of cytokine-induced lymphocyte extravasation to account for the acute HIV/SIV-induced CD4+ cell lymphopenia and raise questions regarding the extent to which altered lymphocyte migration plays a role in the gradual CD4+ cell depletion throughout infection.

Dipyridamole (DPM) is commonly used as a coronary vasodilator and inhibitor of platelet aggregation in the treatment of cardiovascular diseases. We report here that DPM potentiates the inhibitory effects of 3'-azido-3'-deoxythymidine (AZT) and 2',3'-dideoxycytidine against human immunodeficiency virus type 1 (HIV-1) in human monocyte-macrophages. At the same concentrations, DPM does not potentiate the toxic effects of AZT on these cells or on human bone marrow (granulocyte-monocyte) progenitor cells. Since monocyte-macrophage lineage cells appear to be the major reservoir for HIV-1 in vivo, these findings suggest the possibility of using DPM or its analogues in combination chemotherapy of HIV infections.

Primary RNA transcripts from the human immunodeficiency virus type 1 (HIV-1) are processed into mature mRNA by a complex series of splicing events. Viral structural proteins and reverse transcriptase are translated from unspliced or singly spliced transcripts. Proteins which control virus replication, including tat, rev, and nef, are translated from transcripts which are the product of multiple splicing. We have analyzed the composition and relative abundance of the latter transcripts in long-term infected cell lines and in acutely infected peripheral blood cells by amplification with the polymerase chain reaction (PCR) followed by Southern blot, molecular cloning, and DNA sequence analyses. In H9 cells chronically infected with the HIV-1 strain HTLV-IIIB, the predominant of the three kinds of transcripts is those coding for nef. Transcripts with coding potential for rev constituted an intermediate fraction of those analyzed, while those for tat accounted for only a small minority. A similar pattern was observed with Southern blots of PCR-amplified transcripts from peripheral blood lymphocytes acutely infected with HTLV-IIIB. The same general pattern was also observed with PCR-amplified transcripts from peripheral blood monocyte-macrophages infected with an HIV-1 strain (BA-L) able to grow to high titers in macrophages. In these cells, however, the apparent major form of nef transcript contained only the first and third exons of the multiply spliced transcripts and appeared to be generated by either a single or a triple splicing mechanism. As with lymphocytes, tat-specific mRNAs were by far the least abundant. It thus appears that different cell types infected with different strains of HIV-1 maintain a similar balance of expression in which transcripts for nef vastly predominate over those for tat and that those for rev are intermediate in abundance.

Because of the probable role of HIV-infected monocyte/macrophages in the pathogenesis and progression of AIDS, it is essential that antiretroviral therapy address viral replication in cells of this lineage. Several dideoxynucleosides have been shown to have potent in vitro and, in the case of 3'-azido-2',3'-dideoxythymidine (AZT) and 2',3'-dideoxycytidine (ddC), in vivo activity against HIV. However, because these compounds must be phosphorylated (activated) in target cells, and because monocyte/macrophages may have levels of kinases that differ from those in lymphocytes, we investigated the capacity of these drugs to suppress HIV replication in monocyte/macrophages using HIV-1/HTLV-IIIBa-L (a monocytotropic isolate). In the present study, we observed that HTLV-IIIBa-L replication in fresh human peripheral blood monocyte/macrophages was suppressed by each of three dideoxynucleosides: 3'-azido-2',3'-dideoxythymidine (AZT), 2',3'-dideoxycytidine (ddC), and 2',3'-dideoxyadenosine (ddA). Similar results were observed in 5-d-cultured monocyte/macrophages, although higher concentrations of the drugs were required. We then studied the metabolism of AZT and ddC in such cells. The phosphorylation of ddC to a triphosphate moiety was somewhat decreased in monocyte/macrophages as compared with H9 T cells. On the other hand, the phosphorylation of AZT in monocyte/macrophages was markedly decreased to 25% or less of the level in T cells. However, when we examined the level of the normal endogenous 2'-deoxynucleoside triphosphate pools, which compete with 2',3'-dideoxynucleoside triphosphate for viral reverse transcriptase, we found that the level of 2'-deoxycytidine-triphosphate (dCTP) was six- to eightfold reduced, and that of 2'-deoxythymidine-triphosphate (dTTP) was only a small fraction of that found in T cell lines. These results suggest that the ratio of dideoxynucleoside triphosphate to normal deoxynucleoside triphosphate is a crucial factor in determining the antiviral activity of dideoxynucleosides in HIV target cells, and that the lower levels of dTTP may account for the antiretroviral activity of AZT in the face of inefficient phosphorylation of this compound.

A type C RNA virus has been detected in the culture fluids of the SU-DHL-1 human histiocytic lymphoma cell line previously established in this laboratory. In electron micrographs, the virus closely resembled other typical mammalian type C RNA tumor viruses in size and morphology. Viral RNA-dependent DNA polymerase activity has been demonstrated in particles (densities of 1.15 and 1.22 g/ml) in the microsomal cytoplasmic fraction and in pellets of culture fluids. The enzyme is partially inhibited by antibodies to the RNA-dependent DNA polymerases of simian sarcoma virus and RD-114 virus but not by antibody to the polymerase of murine leukemia virus, suggesting some degree of relatedness to type C viruses of subhuman primate origin. Typical syncytial microplaques were induced when SU-DHL-1 cells were cocultivated with rat XC cells. Although no focus formation was noted in similarly cocultivated mouse UC1-B cell cultures, the numbers of foci induced in rat embryo fibroblasts by murine sarcoma virus were significantly increased by coinfection with the virus from SU-DHL-1 cell culture fluids. No other evidence of infectivity, inducibility, or capacity for helper rescue of defective murine sarcoma virus genomes has been detected to date in cocultivation studies with a spectrum of fibroblastic and other nonlymphoid indicator cell lines of human and other species of origin.

Several aspects of the myocardial metabolism of cholesterol and its esters have been investigated using isolated perfused rat heart and heart homogenates. Between 35-50% of tracer amounts of circulating cholesterol-4-14C and cholesterol palmitate-1-14C were extracted by the heart during 45-min perfusion, but there was no indication of cholesterol ester synthesis or hydrolysis either in the medium or in the heart. Similar studies using heart homogenates revealed the lack of detectable enzymatic hydrolysis of cholesterol oleate and absence of both low- and high-energy-requiring ester synthesizing systems. Further evidence for the inability of heart tissue to split sterol esters was based on the finding that 14CO2was not produced from the palmitate-1-14C moiety of the ester. When diluted rat lymph containing 87% cholesterol-4-14C ester was perfused, about 5% of the circulating label was recovered in the myocardium, and of this, 80% was still esterified sterol.

Mobilization of endogenous lipid and glycogen by the isolated rat heart in vitrohas been studied under a variety of conditions. Recirculating perfusion of hearts with Krebs bicarbonate buffer (45 min) led to a marked depletion (66%) of cardiac glycogen, while cardiac triglyceride was decreased by 24%. There were no changes in the levels of phospholipid, diglyceride, cholesterol ester, or free fatty acid. After perfusion for 90 min, in the absence of added substrates, endogenous glycogen and triglyceride were depleted to 16% and 38%, respectively, of the levels in the unperfused controls. During this second 45-min period of perfusion, a small but significant depletion of phospholipid also occurred, indicating that this lipid class is only used for energy after marked depletion of the other endogenous energy sources.Perfusions with epinephrine (10–5M) or dibutyryl adenosine 3′,5′-monophosphate (dibutyryl-cAMP) for 45 min produced marked decreases in the levels of cardiac glycogen and triglyceride, but under these conditions, there was no apparent utilization of phospholipid fatty acids.The depletion of cardiac glycogen during perfusion with buffer alone (45 min) was partially prevented, and depletion of cardiac triglyceride was completely abolished when glucose (9.2 mM) or albumin-bound palmitate (0.5 mM) were added to the perfusate. Perfusion with heparin resulted in a decrease in cardiac glycogen but did not affect triglycerides. The results suggest that cardiac triglycerides provide a secondary endogenous source of energy to the heart in vitro, only after marked depletion of glycogen; phospholipid fatty acids may be utilized following exhaustion of these other endogenous energy sources.

Although lead and SQ20881are potent in vitro inhibitors of kininase II activity, SQ20881does not alter the sensitivity of rats to endotoxin. These results indicate that marked changes in plasma kininase activity do not contribute to endotoxin morbility and that kininase inhibition is not the mechanism whereby lead ions sensitize rats to endotoxin.

The induction of interferon (IFN) by human immunodeficiency virus type 1 (HIV-1) in primary, nonstimulated monocyte-macrophage cultures was studied. HIV-1 infection, as confirmed by p24 antigen levels in the cell supernatant, led to the production of alpha interferon (IFN-alpha) over 7 to 21 days following infection. In two of seven experiments, the IFN detected was acid labile. Coupled reverse transcription-polymerase chain reaction analysis confirmed the induction of IFN-alpha mRNA in cells of HIV-1-infected cultures.