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Biomedical subjects

J Bruszewski

Publications and source records attributed to J Bruszewski.

18 recordsLinked to original sources

[Evaluation of pulmonary function tests in a population of young men from different regions of Poland].

The aim of study was the evaluation results of pulmonary function test (PFT) relationships between air pollution particular ozone concentration and smoking habit. PFT 417 volunteers (men aged 18-23) living in two regions of Poland were studied. These men were divided into 2 groups according to their living place: Warsaw and Gizycko (north-agricultural region of Poland). Inside each group smokers and non-smokers were separated. PNEUMOBIL-mobile PFT laboratory Compact Lab-Jaeger was used. Mean values of PFT between these two groups were not statistically significant. However the percent of population with low values of FEF50 (< 70%) living in Warsaw was five times higher then in population living in Gizycko (10.1% vs 1.7%). It possible that these results were associated with high concentration of ozone in atmosphere in Warsaw.

Adult↗

[Magnetic resonance imaging and arthroscopy in meniscal and ligamentous injuries of the knee].

A series of 37 patients after knee injury underwent magnetic resonance imaging, in 21 cases subsequent arthroscopy has been performed. All patients had previous clinical diagnosis of meniscal and/or anterior cruciate injury. The changes within the menisci and anterior cruciate ligament have been confirmed by MRI in 76 per cent and 81 per cent respectively. In conclusion magnetic resonance imaging and arthroscopy are complementary methods in the knee joint diagnostics.

Anterior Cruciate Ligament Injuries↗

Germline transmission of exogenous genes in the chicken.

Difficulties associated with in vitro manipulation and culture of the early chicken embryo have restricted generation of transgenic chickens to approaches that use replication-competent retroviruses. The need to produce transgenic chickens in the absence of replicating virus prompted development of a new method of gene transfer into the chicken. Microinjection of the replication-defective reticuloendotheliosis virus (REV) vector ME111 beneath unincubated chicken embryo blastoderms results in infection of germline stem cells. This vector contains genetic information exogenous to the chicken genome, including both the herpes simplex virus type 1 thymidine kinase gene and the Tn5 neomycin phosphotransferase gene. About 8 percent of male birds hatched from injected embryos contained vector DNA in their semen. All four positive males tested passed vector sequences onto their progeny. Analysis of G1 offspring showed that gonads of G0 male birds were mosaic with respect to insertion of vector provirus. Thus, primordial germ cells present in the unincubated chicken embryo blastoderm are susceptible to infection by defective REV vectors.

Animals↗

Replication-defective vectors of reticuloendotheliosis virus transduce exogenous genes into somatic stem cells of the unincubated chicken embryo.

Replication-defective vectors derived from reticuloendotheliosis virus were used to transduce exogenous genes into early somatic stem cells of the chicken embryo. One of these vectors transduced and expressed the chicken growth hormone coding sequence. The helper cell line, C3, was used to generate stocks of vector containing about 10(4) transducing units per ml. Injection of 5- to 20-microliters volumes of vector directly beneath the blastoderm of unincubated chicken embryos led to infection of somatic stem cells. Infected embryos and adults contained unrearranged integrated proviral DNAs. Embryos expressed the transduced chicken growth hormone gene and contained high levels of serum growth hormone. Blood, brain, muscle, testis, and semen contained from individuals injected as embryos contained vector DNA. Replication-defective vectors of the reticuloendotheliosis virus transduced exogenous genes into chicken embryonic stem cells in vivo.

Animals↗

Generation of competent virus in the REV helper cell line C3.

We have studied the generation of replication-competent virus in cultures of the helper cell line C3, which harbors a packaging-defective provirus derived from reticuloendotheliosis virus. We transfected the C3 line with a defective provirus encoding the chicken growth hormone gene and carrying the HSV-1 tk gene. The appearance of competent virus was assayed by infection of Spafas C/E CEF, monitoring reverse transcriptase activity, and rescue of the TKTU assayed on BRLtk- cells. Our results indicate that cultures of C3 producing TKTU can release viruses which have variable growth characteristics and which can remain latent in culture for extended periods of time.

Animals↗

Replication-defective chimeric helper proviruses and factors affecting generation of competent virus: expression of Moloney murine leukemia virus structural genes via the metallothionein promoter.

Two chimeric helper proviruses were derived from the provirus of the ecotropic Moloney murine leukemia virus by replacing the 5'long terminal repeat and adjacent proviral sequences with the mouse metallothionein I promoter. One of these chimeric proviruses was designed to express the gag-pol genes of the virus, whereas the other was designed to express only the env gene. When transfected into NIH 3T3 cells, these helper proviruses failed to generate competent virus but did express Zn2+-inducible trans-acting viral functions needed to assemble infectious vectors. One helper cell line (clone 32) supported vector assembly at levels comparable to those supported by the Psi-2 and PA317 cell lines transfected with the same vector. Defective proviruses which carry the neomycin phosphotransferase gene and which lack overlapping sequence homology with the 5' end of the chimeric helper proviruses could be transfected into the helper cell line without generation of replication-competent virus. Mass cultures of transfected helper cells produced titers of about 10(4) G418r CFU/ml, whereas individual clones produced titers between 0 and 2.6 X 10(4) CFU/ml. In contrast, defective proviruses which share homologous overlapping viral sequences with the 5' end of the chimeric helper proviruses readily generated infectious virus when transfected into the helper cell line. The deletion of multiple cis-acting functions from the helper provirus and elimination of sequence homology overlapping at the 5' ends of helper and vector proviruses both contribute to the increased genetic stability of this system.

Animals↗

Recombinant human granulocyte colony-stimulating factor: effects on normal and leukemic myeloid cells.

Experiments were conducted to isolate and characterize the gene and gene product of a human hematopoietic colony-stimulating factor with pluripotent biological activities. This factor has the ability to induce differentiation of a murine myelomonocytic leukemia cell line WEHI-3B(D+) and cells from patients with newly diagnosed acute nonlymphocytic leukemia (ANLL). A complementary DNA copy of the gene encoding a pluripotent human granulocyte colony-stimulating factor (hG-CSF) was cloned and expressed in Escherichia coli. The recombinant form of hG-CSF is capable of supporting neutrophil proliferation in a CFU-GM assay. In addition, recombinant hG-CSF can support early erythroid colonies and mixed colony formation. Competitive binding studies done with 125I-labeled hG-CSF and cell samples from two patients with newly diagnosed human leukemias as well as WEHI-3B(D+) cells showed that one of the human leukemias (ANLL, classified as M4) and the WEHI-3B(D+) cells have receptors for hG-CSF. Furthermore, the murine WEHI-3B(D+) cells and human leukemic cells classified as M2, M3, and M4 were induced by recombinant hG-CSF to undergo terminal differentiation to macrophages and granulocytes. The secreted form of the protein produced by the bladder carcinoma cell line 5637 was found to be O-glycosylated and to have a molecular weight of 19,600.

Animals↗

Studies of TGEV spike protein gp195 expressed in E. coli and by a TGE-vaccinia virus recombinant.

The gene coding for the surface spike protein gp195 of TGEV has been cloned and expressed in E. coli in the form of fusion proteins. These proteins were isolated and used to immunize laboratory animals. All animals developed antibodies cross-reacting with the TGEV virion but failed to neutralize the virus. The entire gp195 gene was also inserted into vaccinia to generate a TGEV-vaccinia recombinant virus (vTGE) that expressed TGEV gp195. Animals vaccinated with vTGE produced neutralizing antibodies against both TGEV and vaccinia. These results suggest the potential use of the recombinant vTGE as a vaccine against TGEV infection.

Animals↗

Spontaneous release of endogenous ecotropic type C virus from rat embryo cultures.

Type C viruses were isolated from embryo cultures of two different rat strains, Sprague-Dawley and Fischer. Both viruses (termed rat leukemia virus, RaLV) were released spontaneously from rat embryo cells, have a density of 1.14 to 1.15 g/cm(3) based on equilibrium sedimentation in sucrose gradients, contain 60-70S RNA, RNA-directed DNA polymerase, and rat type C virus-specific 30,000 molecular-weight-protein determinants. Molecular hybridization studies using the Sprague-Dawley RaLV 60-70S RNA show that the virus-specific nucleotide sequences are present in the DNA of rat embryos. Both Sprague-Dawley and Fischer RaLV can rescue the murine sarcoma virus genome from Kirsten murine sarcoma virus-transformed nonproducer cells and are neutralized by antisera to the RPL strain of RaLV. In contrast to previous RaLV's, these viruses propagate in their own cells of origin as well as in cells of heterologous rat strains.

Animals↗