"Cord formation" in smear specimen prepared from sputum for a more rapid method of presumptive identification of Mycobacterium tuberculosis

Cord formation of Mycobacterium tuberculosis complex is very uncommon in smear specimen prepared directly from sputum, although such a finding is well known in solid or liquid media and has recently been evaluated as a rapid method for presumptive identification in special liquid media (BACTEC or MGIT). We examined 308 (Mycobacterium tuberculosis 271 and Nontuberculous mycobacteria 37) positive smear specimens prepared directly from sputum in our hospital. These specimens all showed a "modified Gaffky scale" as +2 or more and this cord formation was found in four cases (five specimens). Each of these specimens was from a patient with severe lung tuberculosis showing cavity formation and each patient was complicated severe diabetes mellitus. The morphology of cord formation on smear specimens prepared directly from sputum was similar to that in liquid or solid media, and consequently the relevant bacilli were identified as Mycobacterium tuberculosis complex by PCR examination. In this study, we assessed "cord formation" in smear specimen prepared directly from sputum as a more rapid presumptive identification of Mycobacterium tuberculosis complex based on microscopic morphology, as well as cord formation in liquid or solid media.     

Novel radiator for carbon dioxide absorbents in low-flow anesthesia

During long-term low-flow sevoflurane anesthesia, dew formation and the generation of compound A are increased in the anesthesia circuit because of elevated soda lime temperature. The object of this study was to develop a novel radiator for carbon dioxide absorbents used for long durations of low-flow sevoflurane anesthesia. Eleven female swine were divided into two groups comprising a "radiator" group (n = 5) that used a novel radiator for carbon dioxide absorbents and a "control" group (n = 6) that used a conventional canister. Anesthesia was maintained with N2O, O2, and sevoflurane, and low-flow anesthesia was performed with fresh gas flow at 0.6 L/min for 12 hr. In the "control" group, the soda lime temperature reached more than 40 degrees C and soda lime dried up with severe dew formation in the inspiratory valve. In the "radiator" group, the temperature of soda lime stayed at 30 degrees C, and the water content of soda lime was retained with no dew formation in the inspiratory valve. In addition, compound A concentration was reduced. In conclusion, radiation of soda lime reduced the amounts of condensation formed and the concentration of compound A in the anesthetic circuit, and allowed long term low-flow anesthesia without equipment malfunction.     

Calcification preceding new bone formation induced by demineralized bone matrix gelatin.

Demineralized bone matrix gelatin (BMG) was implanted into the skeletal muscle of Sprague-Dawley (S.D.) rats, and histological changes were examined 3, 5, 7, 10 and 15 days later. Before bone formation, a specific calcification process was found in most of the BMG from day 5 and 7 after implantation. The heterotopic calcified sites were not always consistent with the sites of the alkaline phosphatase activity. It was considered that this calcification progresses without any cellular components, and we distinguished this type of calcification as "acellular mineral deposition" from the calcification which occurs in new bone formation. This "acellular mineral deposition" was first observed as small spherical calcified deposits in the BMG on day 7 after implantation; these deposits then gradually grew and fused with each other. Some multinucleated cells appeared near the site of calcification on day 7 after implantation, but osteoblasts or osteoblast-like cells were scarcely observed around the calcified deposits in BMG until day 7. Vascularization was often observed near the "acellular mineral deposition" and the new bone formation. Fourier transform infrared spectroscopy showed that the calcified deposits in BMG were composed of hydroxyapatite, carbonateapatite and other calcium phosphate components, and that the first two components became prominent with time. It is believed that the "acellular mineral deposition" is due to the deposition of calcium and phosphate into the BMG by a process of heterogenic nucleation that does not involve osteoblasts or matrix vesicles. Bone formation induced by the BMG occurred after the "acellular mineral deposition." The experimental calcification shown in this paper seems a useful model for the study of biocalcification.     

The role of hyaluronan-binding protein in assembly of pericellular matrices.

Hyaluronan-dependent pericellular matrices or "coats" are expressed by a variety of cell types in culture and modulation of their expression may be important in regulation of cell interactions in vivo during development. Monoclonal antibody IVd4, which recognizes hyaluronan-binding protein with the properties of a hyaluronan receptor, was shown to block formation of these coats by a variety of cells. Using rat fibrosarcoma cells, it was found that the antibody not only blocked initial formation of the coats but also caused their loss when added subsequent to formation. The loss of preformed coats in the presence of antibody occurred at 4 degrees and 37 degrees, implying that the function of hyaluronan-binding protein in coat formation is not in mediating metabolic processes. The antibody also had no significant effect on hyaluronan production by the fibrosarcoma cells. In addition, hyaluronan hexasaccharide, a competitive inhibitor of the interaction between polymeric hyaluronan and its cell surface receptor, was found to inhibit coat formation. Thus it is concluded that a hyaluronan-binding protein with the properties of a hyaluronan receptor is required for pericellular matrix formation.     

Hard tissue formation of STRO-1-selected rat dental pulp stem cells in vivo

The objective of this study was to examine hard tissue formation of STRO-1-selected rat dental pulp-derived stem cells, seeded into a calcium phosphate ceramic scaffold, and implanted subcutaneously in mice. Previously, STRO-1 selection was used to obtain a mesenchymal stem cell progenitor subpopulation from primary dental pulp-derived stem cells. In the current study, these cells were cultured with three different media: "BMP-plus" medium containing dexamethasone and 100 ng/mL of rhBMP-2, "odontogenic" medium containing dexamethasone, and "control" medium without supplements. The cell-scaffold complexes were cultured in these media for 1, 4, or 8 days before implantation. Histological analysis demonstrated that the cultures with BMP-plus and 4 days of culture gave the highest percentage of hard tissue formation per implant (36 +/- 9% of pore area). Real-time PCR confirmed these results. In conclusion, STRO-1-selected dental pulp stem cells show effective hard tissue formation in vivo, and a short in vitro culture period and addition of BMP-2 can enhance this effect.     

Die klinische Pathologie in der Hansestadt Lübeck

The formation of an university institute of pathology at the Hanseatic city of Lübeck preceded a long, changeful planning, accompanied by a chain of historical events. In the 19th century, Lübeck owned seven different places for clinical or forensic autopsies. Thus, in the first years of the 20th century it was generally called for the formation of a "Central Institute of Pathology" that was completed after all in the year 1927. Two years later, Eberhard von Praun became head of the institute and was immediately confronted with the "Lübeck-disaster". He was followed 1935 by Ernst Jeckeln, who identified the "Enteritis necroticans" which appeared in North Germany in the first years after the "Second World War". He called the disease "Darmbrand", a term that is since then internationally used. Jeckeln became the first "Ordinarius for Pathology" at the "Medical Academy Lübeck", which was associated to the "University of Kiel". His long-time colleague was Alfred Wegener, who identified the "Wegener's granulomatosis". In the year 1972 Alfred Gropp became head of the institute, an all-round scientist and pathologist, e.g. well-known in the field of developmental pathology. During his term, the "Medical Academy" got the university status and was named "Medizinische Hochschule Lübeck".     

The postnatal development of rat dentate gyrus and the effect of early thyroid hormone treatment

Summary The postnatal development of the dentate gyrus and the effect of 5 g/day triiodothyronine treatment was examined by means of tritiated thymidine autoradiography.The polymorph layer of the dentate gyrus is a secondary germinal layer and forms cells for the granular layer. The cell formation in the dentate gyrus was found to be nearly completed on the 21st day, but development ended only in adulthood.Thyroid-hormone treatment did not affect significantly the postnatal cell formation of the dentate gyrus, but resulted in the appearance of a great number of pycnotic cells in the granular layer.     

C5a promotes migration, proliferation, and vessel formation in endothelial cells

Objectives

The goal of this paper is to investigate the effects of activated complement C5a on vascular endothelium during vessel formation.

Methods

A human microvascular endothelial cell line (HMEC-1) derived from post-capillary venules in skin was used to measure DNA synthesis, proliferation and cell-cycle progression. In vitro ring-shaped formation by the cells was assessed by using type I collagen gel matrix and a cell-migration assay using the Chemotaxicell chamber. A Matrigel plug assay was performed to confirm the effect of C5a in vivo.

Results

C5a progressed the cell cycle of HMEC-1 into G2/M phases, and induced DNA synthesis and proliferation in a dose-dependent manner. C5a efficiently induced migration and ring-shaped structure formation both in vitro and in vivo. Furthermore, a C5a receptor antagonist (W-54011) suppressed all HMEC-1 activities including proliferation and migration.

Conclusions

Proliferation, migration, and ring-shaped formation by HMEC-1 cells was induced by C5a. The actions were efficiently inhibited by a specific antagonist against C5a. Our results implicated C5a in vessel formation and as a potent target for management of inflammatory diseases.     

Die thermische polymerisation von methylmethacrylat, 2. Bildung des ungesättigten dimeren

The spontaneous thermal polymerization of methyl methacrylate (MMA) is accompanied by the production of serveral oligomers among which a linear unsaturated dimer H-1 (dimethyl 1-hexene-2,5-dicarboxylate) is predominant. The reaction kinetics of this dimer formation was investigated in bulk and in solution. The temperature dependence of the second order dimerization constant was determined as Reaction mechanisms for the thermal dimer formation of MMA are discussed.     

Reverberation of Excitation in Living “Hippocampal Formation–Entorhinal Cortex” Slices from Rats. Optical Recording

A vital potential-dependent dye was used to conduct optical recording of the electrical activity of the hippocampal formation in living slices of the rat brain including the hippocampal formation and the entorhinal cortex. These studies showed that single electrical stimuli applied to the entorhinal cortex, subiculum, and dentate gyrus produced responses in which waves of excitation passed across the hippocampal formation sequentially from the dentate gyrus, through CA3, to the CA1 field of the hippocampus. When GABAergic inhibition was partially blocked with picrotoxin, the first wave of excitation was immediately followed by several further waves in all zones of the hippocampal formation, with a constant shift in latency, which increased from the dentate gyrus to CA3 and CA1. Reverberation of excitation in the hippocampal formation-entorhinal cortex structure is regarded as the most probable cause for the appearance of these sequences of waves.     

A study by the agar diffusion technique of precipitating antibody directed against blue tongue virus and its relation to homotypic neutralizing antibody

Summary The technique of agar diffusion was applied to a study of blue tongue virus antigens prepared from infected mouse brains, chicken embryos, and cell culture fluid. Antigens from these different sources, contributing to precipitate formation, appeared to be virus-specific, noninfectious and serologically indistinguishable one from another in the systems tested.The onset and production of circulating ovine precipitating antibody was correlated to corresponding data for homotypic virus infectivity neutralizing antibody. The onset of circulating precipitating antibody formation was detected about the same time as for neutralizing antibody but the precipitating antibody persisted longer. An anamnestic response was observed for neutralizing antibody but not for the precipitating antibody.An antigen-antibody system containing one reagent in low concentration precipitated if adjacent to a precipitating control system but not if placed by itself or an angle >90° to the control system. This phenomenon, termed the recruiting effect, was found to influence quantitation of precipitating antibody.     

Binding of Matrix Metalloproteinase 9 to Fibrin is Mediated by Amorphous Calcium-Phosphate

In our previous study we demonstrated selective, dose-dependent binding of matrix metalloproteinase-9 (MMP-9), a neutrophil collagenase, to fibrin. Here we investigated the mechanism of this interaction. We found that MMP-9 to fibrin was dependent on formation of a calcium-phosphate intermediate. The intermediate was precipitable by centrifugation and contained a Ca/P ratio of 1.52–1.54, consistent with amorphous calcium-phosphate (ACP). ACP formation exhibited a temperature optimum at 37°C. Gelatin zymography revealed that interaction of ACP with MMP-9 resulted in formation of a high molecular weight ACP : MMP-9 complex which was required for MMP-9 binding to fibrin. Complex formation was dependent on the generation of viable ACP that required both calcium (7.5–10 mM) and phosphate (225–250 M) (Ca × P product range, 1.7–2.5 mM²). Carbonate (CO₃) and sulfate (SO₄) were ineffective as calcium counteranions. Preformed ACP rapidly complexed MMP-9. Thus ACP formation was rate-limiting for MMP-9 fibrin binding activity. No MMP-9 fibrin binding activity was noted at 25°C, an observation consistent with lack of ACP production. The significance of these findings is discussed with respect to normal and pathologic wound healing.     

Monosynaptic innervation of facial motoneurones by neurones of the parvicellular reticular formation

In order to determine whether neurones in the parvicellular reticular formation are in direct synaptic contact with motoneurones innervating facial muscles, a combined retrograde and anterograde transport study was carried out in the rat. Animals received injections of the retrograde tracer cholera toxin B conjugated to horseradish peroxidase into facial muscles and of the anterograde tracer biocytin into the parvicellular reticular formation. The facial motor nucleus was then examined for anterograde and retrograde labelling in the light and electron microscopes. Retrogradely labelled neurones were found in the facial motor nucleus with a distribution that was dependent on the muscles injected. Terminals anterogradely labelled with biocytin from the parvicellular reticular formation were observed in the motor nucleus amongst the retrogradely labelled neurones. At the electron microscope, the retrogradely labelled cells were found to receive input from unlabelled terminals and from terminals that were anterogradely labelled from the injections of biocytin in the parvicellular reticular formation. The labelled terminals were 1–2 m in diameter at the active zone and packed with spherical vesicles. They formed both symmetrical and asymmetrical synapses with their labelled or unlabelled targets. It is concluded that neurones in the parvicellular reticular formation form direct synaptic contact with motoneurones of facial muslces. This may represent a pathway by which the basal ganglia can directly influence orofacial movement, as the substantia nigra is known to project to that part of the reticular formation.     

Tracheal Wall Thickening Is Associated with the Granulation Tissue Formation Around Silicone Stents in Patients with Post-Tuberculosis Tracheal Stenosis

Purpose

Tracheal restenosis due to excessive granulation tissue around a silicone stent requires repeated bronchoscopic interventions in patients with post-tuberculosis tracheal stenosis (PTTS). The current study was conducted to identify the risk factors for granulation tissue formation after silicone stenting in PTTS patients.

Materials and Methods

A retrospective study was conducted between January 1998 and December 2010. Forty-two PTTS patients with silicone stenting were selected. Clinical and radiological variables were retrospectively collected and analyzed.

Results

Tracheal restenosis due to granulation tissue formation were found in 20 patients (47.6%), and repeated bronchoscopic interventions were conducted. In multivariate analysis, tracheal wall thickness, measured on axial computed tomography scan, was independently associated with granulation tissue formation after silicone stenting. Furthermore, the degree of tracheal wall thickness was well correlated with the degree of granulation tissue formation.

Conclusion

Tracheal wall thickening was associated with granulation tissue formation around silicone stents in patients with post-tuberculosis tracheal stenosis.     

Secretory IgA and mucin-mediated biofilm formation by environmental strains of Escherichia coli: role of type 1 pili

Recent studies suggest the importance of secretory IgA (SIgA) and mucin in the mediation of biofilm formation by commensal bacteria within the mammalian gut. Studies using a variety of strains of Escherichia coli have indicated that the interaction between E. coli and SIgA is dependent on the type 1 pilus. In this study, the importance of the pilus in SIgA-mediated biofilm formation by a laboratory strain (MG1655) and environmental (fecal) strains of E. coli was evaluated. Transient expression of the type 1 pilus by the laboratory strain of E. coli failed to facilitate SIgA-mediated biofilm formation, whereas constitutive expression of the type 1 pilus by the laboratory strain was sufficient. In contrast, transient expression of the type 1 pilus was sufficient to facilitate SIgA-mediated biofilm formation by environmental isolates. The "threshold" for mucin-mediated biofilm formation appeared to be lower than that for SIgA-mediated biofilm formation, perhaps reflecting disparate roles of mucin and SIgA in mediating biofilm formation in the gut. These studies also provide the first procedures for the growth of bacterial biofilms on live epithelial cells in vitro, an important development that may facilitate future studies on the effects of bacterial biofilms on epithelial cells.     

Food hypermotivation syndromes in rabbits with hyperactivity of lateral hypothalamic structures

A focus of hyperactivity in the initiating centers for food motivation was created in experiments on unrestrained rabbits by blocking inhibitory mechanisms in the lateral hypothalamus (LH) by injection of tetanus toxin (group 1) or by chronic electrical stimulation of the same region of the brain (group 2). The syndromes thus evoked differed in the intensity of food hypermotivation. The syndrome evoked by injection of tetanus toxin was characterized by very rapid development, clear symptoms of hyperphagia, and a severe course. Marked epileptiform activity was recorded in LH, evidence of the formation of a generator of pathologically enhanced excitation in the pacemaker of food motivation. Mechanisms of adequate correction of behavior depending on the result obtained were completely disturbed in the animals. The syndrome evoked by chronic electrical stimulation of LH was characterized by slower formation and a more favorable course. Relatively weak epileptiform activity was recorded in LH. Enhanced food-getting activity of the rabbits also was observed in the late periods after the beginning of electrical stimulation, but the animals' behavior did not lose its adaptive character in this case. The formation of the two syndromes of food hypermotivation described above was connected with the formation of two excitation generators, differing in the intensity of excitation produced by them.Laboratory of General Pathology of the Nervous System, Institute of General Pathology and Pathological Physiology, Academy of Medical Sciences of the USSR, Moscow. Department of Pathological Physiology, Leningrad San.-Gig. Medical Institute. Group for Neurophysiology of Motivations, Institute of Normal Physiology, Academy of Medical Sciences of the USSR, Moscow. Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 89, No. 3, pp. 281–284, March, 1980.     

Interaction of simian virus 40 large T-antigen with cellular DNA polymerase α: studies with various T-antigen mutants of SV40

Summary Simian virus 40 (SV40) large T antigen binds to host cell DNA polymerase and the T-antigen-DNA polymerase complex is implicated in the initiation of viral DNA replication. We have examined various SV40 T-antigen mutants to test the correspondence between viral DNA replication and T-antigen-DNA polymerase complex formation. The various SV40 T-antigen mutants were used to either infect or transfect African green monkey kidney cell line CV-1, and at different time intervals we measured the production of T-antigen and host cell DNA polymerase by radioimmunoassay, complex formation by a sandwich radioimmunoassay and the amount of viral DNA synthesis by dot-blot hybridization analysis. There was a good correlation between complex formation and viral DNA synthesis in lytic mutants of SV40. Poor complex formation and correspondingly lower DNA synthesis were observed in the non-viable mutants of SV40, even though significant amounts of T-antigen and DNA polymerase were present. Our results substantiate the earlier findings of T-antigen-DNA polymerase complex formation and establish the need for formation of this complex in promoting viral DNA synthesis.     

A systemic review of randomized controlled studies about prevention with pharmacologic agents of adhesion formation in the rat uterine horn model

Introduction

Evaluation of treatment attempts in postoperative adhesion formation is pivotal for the prevention of several morbidities including infertility, pelvic pain, bowel obstruction, and subsequent intraoperative complications. The purpose of this systemic review was to assess the literature on the rat uterine horn model for adhesion formation and treatment modalities to prevent adhesion in the most frequently used experimental animal model.

Material and methods

We performed a systemic review of publications from January 1^st 2000 to December 31^st 2013 via a PubMed search. A high number of agents were evaluated for the prevention of postoperative adhesion formation in the rat uterine horn model.

Results

According to most of the studies, adjuvants such as antiinflamatuars, antiestrogens, antioxidants were effective to prevent adhesion formation.

Conclusions

Prevention of adhesion formation is pivotal and numerous types of agents were described in the literature were summarized in this review.