Enhancement of seminiferous tubular growth and spermatogenesis in testes of rats recovering from early hypothyroidism: a quantitative study

Testicular weight and DNA content were markedly reduced (63 and 69%) in weanling Long-Evans rat pups rendered hypothyroid from birth by administration of propylthiouracil (PTU), a reversible goitrogen. These growth deficits worsened to >80% by continuing hypothyroidism beyond weaning, to days 50 and 90. Recovery of thyroid function, brought about by discontinuing PTU at weaning, resulted in a paradoxical stimulation of testis growth, amounting to increased weight (40%), DNA content (60%) and size by 90 days, compared to age-matched controls. In the 25-day or older hypothyroid rats, testicular structure was immature and spermatogenesis markedly delayed, as evident by closed lumen and significantly reduced diameter of seminiferous tubules (38%), thickness of germinal layer (70%), and number of primary spermatocytes (86%), compared to control. Hypothyroidism did not alter the number of tubules per testis cross section. In the 90-day recovery rats, numbers of seminiferous tubules were unchanged but tubular diameter was significantly (20%) larger than in controls and spermatogenesis appeared very active as indicated by significantly increased germinal layer thickness (22%) and total number and density of primary spermatocytes (55% and 40%). The results show that although postnatal hypothyroidism is deleterious for testicular growth and spermatogenesis, recovery from this condition leads to enhanced seminiferous tubular growth and spermatogenesis.     

Immortalization of antigen specific, helper T cell lines by transformation with the radiation leukemia virus (RadLV)

Antigen-specific immune T lymphocytes of male C57BL/6 mice were enriched in vitro on monolayers of antigen-pulsed syngeneic macrophages. The cells were treated in vitro with RadLV and inoculated intrathymically into irradiated female C56BL/6 animals. Thymomas arising in the inoculated recipients were characterized as donor- (male) type according to their karyotype. In vivo and in vitro cell lines were established from the primary lymphomas, two of which (designated ROT/6.1 and ROT/6.2) were capable of providing antigen- (carrier) specific help in normal or preimmunized mice. None of the lymphomas could induce antigen-specific DTH reaction. Five months after their establishment, ROT/6.2 alone retained its carrier specificity. ROT/6.2 consisted mainly of Lyt-1+ cells, whereas the ROT/6.1 population was more heterogeneous and contained Lyt-1+, Lyt-2+, and Lyt-3+ cells. The carrier specificity of the latter may have been lost due to selection against the specific helper cells during prolonged passages.     

Differential effects of drugs upon hematopoiesis can be assessed in long-term bone marrow cultures established on nylon screens.

Hematotoxicity is associated with exposure to chemotherapeutic drugs and numerous other agents. Most measurements of the hematopoietic effects of prospective therapeutic drugs and environmental agents have been made in animal models. We tested the influence of various drugs on hematopoiesis in long-term cultures of Long-Evans rat bone marrow cells. These cultures were established on nylon screen-bone marrow stromal cell templates that were suspended in liquid medium. Previous phenotypic analyses of adherent zone cells of suspended nylon screen bone marrow cultures (NSBMC) using monoclonal antibodies and flow cytometry indicated that they maintain a multilineage character for extended periods in culture and display continuous proliferation of hematopoietic progenitors (colony-forming unit culture [CFU-C]). NSBMC of various ages were incubated for 21 hr with several concentrations of beta-D-cytosine arabinofuranoside, 5-fluorouracil, cyclophosphamide, or methotrexate. Adherent zone cells were dissociated enzymatically, phenotyped by flow cytometry, and assayed for colony-forming unit culture content. beta-D-cytosine arabinofuranoside, 5-fluorouracil, and methotrexate treatment of bone marrow cultures resulted in a dose-related diminution in colony-forming unit culture numbers in the adherent zones of NSBMC. Phenotypic analyses revealed similar trends but certain of these drugs manifested lineage specificities. Toxicity was also related to cyclophosphamide dose, but the presence of bone marrow stroma was necessary to demonstrate this effect in vitro. A subpopulation of these cells was found to metabolize ethoxyfluorescein ethyl ester to fluorescein after induction with 2,3,7,8-tetrachlorodibenzo-p-dioxin, an effect which was quantified by flow cytometry. NSBMC may be used to ascertain lineage-specific toxicities and evaluate the effects of drugs on the proliferation of hematopoietic progenitor cells.     

Route of antigen administration for tolerance production

Tolerance production is readily induced by inoculation of heterologous serum proteins in mice. The intravenous and intraperitoneal route are generally preferred because they allow rapid antigen equilibrium in the intra and extravascular spaces of the host. This report shows that the administration of particle-free antigen in the foot pad produces a greater number of tolerant animals at all concentrations used. The active participation of regional lymph nodes in tolerance production is postulated.     

Transient pain developers show increased abdominal muscle activity during prolonged sitting

BackgroundSitting is a commonly adopted posture during work and prolonged exposures may have detrimental effects. Little attention has been paid to the thoracic spine and/or multiple axes of motion during prolonged sitting. Accordingly, this study examined three-dimensional motion and muscle activity of the trunk during two hours of uninterrupted sitting.MethodsTen asymptomatic males sat during a simulated office task. Kinematics were analyzed from six segments (Neck, Upper-, Mid-, and Lower-thoracic, Lumbar, and Pelvis) and electromyography was recorded from eight muscles bilaterally.ResultsFour participants developed transient pain. These participants showed higher average muscle activations in the abdominal muscles. Additionally, the non-pain group showed less lateral bend positional change in the mid-thoracic region compared to the upper- and lower-thoracic regions. Weak-to-moderate positive correlations were also found between rated pain and low back muscle activation.DiscussionThe results provided further evidence of reduced movement in non-pain developers and altered muscle activation patterns in pain developers. Low-level, prolonged static contractions could lead to an increased risk of injury; and though the increased abdominal activity in the pain developers was not directly associated with increased rated pain scores, this could indicate a pre-disposition to, or enhancer of, transient pain development.     

Four new Psocoptera from Lebanese amber (Insecta: Psocomorpha: Trogiomorpha)

Four new trogiomorphan Psocoptera are described from the Lower Cretaceous Lebanese amber, viz. Bcharreglaris amunobi n. gen., n. sp., Setoglaris reemae n.gen., n. sp., Libanoglaris chehabi n.sp., and Libanoglaris randatae n. sp. These discoveries show that the Lower Cretaceous biodiversity of the Trogiomorpha was very high.     

New biting midges from the Cretaceous amber of Lebanon (Diptera: Ceratopogonidae)

New Ceratopogonidae from the Early Cretaceous amber of Hammana (Central Lebanon) are studied. Five new species (Lebanoculicoides daheri sp. nov., Protoculicoides krzeminskii sp. nov., Archiaustroconops annae sp. nov., Archiaustroconops hammanensis sp. nov., and Archiaustroconops dominiakae sp. nov.) are characterized, described, illustrated, and compared with other Cretaceous taxa. New keys for species of the three genera Lebanoculioides, Protoculicoides, and Archiaustroconops, but also for all genera of Lebanese fossil biting midges, are proposed.     

Metabolic engineering of <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis> for linalool production

Objectives

To engineer the yeast Saccharomyces cerevisiae for the heterologous production of linalool.

Results

Expression of linalool synthase gene from Lavandula angustifolia enabled heterologous production of linalool in S. cerevisiae. Downregulation of ERG9 gene, that encodes squalene synthase, by replacing its native promoter with the repressible MET3 promoter in the presence of methionine resulted in accumulation of 78 µg linalool l^?1 in the culture medium. This was more than twice that produced by the control strain. The highest linalool titer was obtained by combined repression of ERG9 and overexpression of tHMG1. The yeast strain harboring both modifications produced 95 μg linalool l^?1.

Conclusions

Although overexpression of tHMG1 and downregulation of ERG9 enhanced linalool titers threefold in the engineered yeast strain, alleviating linalool toxicity is necessary for further improvement of linalool biosynthesis in yeast.