Immunohistochemical detection of Pneumocystis carinii in transbronchial lung biopsy specimens: antigen difference between human and rat Pneumocystis carinii.

Transbronchial lung biopsy specimens, from three patients with non-AIDS-related Pneumocystis carinii pneumonia (PCP) and rat lung tissue in which PCP was induced by the administration of dexamethasone, were studied to determine the diagnostic usefulness of an immunohistochemical method using commercially available anti-Pneumocystis monoclonal antibody, 3F6, on formalin-fixed, paraffin-embedded tissue. PC was consistently stained a bright red color and unambiguously identified in all three human lung specimens, but not stained in lung tissues at autopsy from patients with various fungal pneumonias. In contrast, PC was weakly stained in PCP-induced rat lungs. The present study indicates human PC and rat PC to be antigenically different in terms of the existence of the 82 kilo-dalton (kD) antigen against which 3F6 is directed.     

A case of dural arteriovenous malformation with bilateral thalamic infarction

A 59-year-old man was admitted because of frequent vomiting and obtundation in February 1982. Neurological examination on admission revealed only slight impairment of consciousness. Papilledema, meningeal irritation sign and paralysis were not elicited. The plain CT scan was normal, but the CT scan with contrast material showed patchy enhancement in the left temporal lobe and around the third ventricle. Cerebral angiography showed a dural arteriovenous malformation (dural AVM) in the left transverse sinus fed by the left occipital artery, and the retrograde flow into the straight sinus. By the third day following admission, the level of consciousness became alert. The patient did not complain of headache, bruit and visual disturbance. He showed mild disorientation and memory disturbance. But his ordinary daily-living was independent. In August 1982, the patient gradually became inactive and apathetic. At times he lay in bed with moving his eyes, swallowing foods. At other times, he lay in bed with closing his eyes, immobile, and unresponsive except to strong painful stimuli. The patient was incontinent and required nursing care. During three month periods, the patient progressively became somnolent, speechless and immobile. Eventually, he was in a state of akinetic mutism. The patient became unresponsive. The state of consciousness fluctuated within a narrow range. The pupils were isocoric and did not react to light. He sometimes moved his eyes horizontally, but the vertical eye movement was limited. Deep tendon reflexes were hyperactive with Babinski reflex bilaterally. Passive mobilization of extremities revealed hypertonic. The CT scan disclosed the bilateral symmetrical infarction of the thalamus.(ABSTRACT TRUNCATED AT 250 WORDS)     

Theoretical basis for herbal medicines, Tokishakuyaku-San and Sairei-To, in the treatment of recurrent abortion: enhancing the production of granulocyte-macrophage colony-stimulating factor in decidual stromal cells

PROBLEM: To get insight into the basis for the empirical usage of herbal medicines, such as Tokishakuyaku-san (Toki) and Sairei-to (Sai) in the treatment of recurrent abortion and intrauterine growth restriction, we examined whether these medicines modulate the production of granulocyte-macrophage colony-stimulating factor (GM-CSF), a cytokine working as an important mediator for intercellular communication in the embryonic development, in decidual stromal cells (DSCs). METHOD OF STUDY: Human DSCs were cultured with either Toki or Sai at several different concentrations. The effect on cell proliferation was assessed by WST-8 assay. GM-CSF released into culture medium was analyzed using enzyme-linked immunosorbent assay, and semi-quantitative polymerase chain reaction was carried out to see GM-CSF mRNA expression in DSCs. RESULTS: Sai inhibited the proliferation of cultured DSCs, while no interference was observed in the presence of Toki. Both Toki and Sai enhanced the release of GM-CSF into culture medium. The amount of GM-CSF mRNA in cultured DSCs was as well increased by either Toki or Sai. CONCLUSION: Considering the significance of GM-CSF in embryonic development, clinical benefit of these herbal medicines in the treatment of recurrent abortion might be based on the shown pharmacological reaction related to GM-CSF.     

Synovial stem cells are regionally specified according to local microenvironments after implantation for cartilage regeneration

We previously demonstrated that synovium-derived MSCs had greater in vitro chondrogenic ability than other mesenchymal tissues, suggesting a superior cell source for cartilage regeneration. Here, we transplanted undifferentiated synovium-derived MSCs into a full-thickness articular cartilage defect of adult rabbits and defined the cellular events to elucidate the mechanisms that govern multilineage differentiation of MSCs. Full-thickness osteochondral defects were created in the knee; the defects were filled with 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate-labeled MSCs and covered with periosteum. After 4 weeks, although the cell density decreased, transplanted MSCs produced a great amount of cartilage matrix extensively. The periosteum became thinner, and chondroprogenitors in the periosteum produced a small amount of cartilage matrix. In the deeper zone, transplanted MSCs progressed to the hypertrophic chondrocyte-like cells. In the deep zone, some transplanted cells differentiated into bone cells and were replaced with host cells thereafter. In the next phase, the border between bone and cartilage moved upwards. In addition, integrations between native cartilage and regenerated tissue were improved. Chondrocyte-like cells derived from the transplanted MSCs still remained at least after 24 weeks. Histological scores of the MSC group improved continuously and were always better than those of two other control groups. Immunohistological analyses and transmission electron microscopy confirmed that the MSCs produced abundant cartilage matrix. We demonstrated that transplanted synovium-derived MSCs were altered over a time course according to the microenvironments. Our results will advance MSC-based therapeutic strategies for cartilage injury and provide the clues for the mechanisms that govern multilineage differentiation of MSCs.     

Passive mechanical properties of large intestine under in vivo and in vitro compression

This paper presents experimental data obtained from both in vivo and in vitro compression of the large intestine of goat. In vivo experimental data were obtained from compression tests on the large intestine of an anesthetized goat using force-displacement acquisition equipment. In vitro experimental data were also obtained from tissue excised after the in vivo experiments, and two types of data were then compared. The results demonstrated that the stress values had a strong dependence on the compressive rate in the in vivo experiments, although such effect was not distinct in the in vitro experiments. Additionally, at a lower compression rate, the intestinal tissues were found to be stiffer in the in vitro experiments than in the in vivo ones. This paper is a preliminary report on the mechanical properties of the large intestine based on in vivo and in vitro experimental data.     

Crystal structure of human DAAM1 formin homology 2 domain

Reorganization of the actin filament is an essential process for cell motility, cell-cell attachment and intracellular transport. Formin proteins promote nucleation and elongation of the actin filament, and thus are key regulators for this process. The formin homology 2 (FH2) domain forms a head-to-tail ring-shaped dimer, and processively moves towards the barbed end. Dishevelled-associated activator of morphogenesis (DAAM) is a Rho-regulated formin implicated in neuronal development. Here, we present the crystal structure of human DAAM1 FH2 dimer at 2.8 A resolution. This is the first dimeric structure of the mammalian formin. The core structure of human DAAM1 is similar to those of mouse mDia1 and yeast Bni1p, whereas the orientations of the FH2 dimeric rings are different between human DAAM1 and yeast Bni1p, despite their similar dimer interactions. This difference supports the previous prediction that the dimer architecture of the formin is highly flexible in the actin-free state. The results of the actin assembly assays using the DAAM1 mutants demonstrated that the length of the linker connecting the N-terminal domain and the core region is crucial for the activity.     

Molecular cloning and functional characterization of porcine MyD88 essential for TLR signaling

We isolated cDNA encoding porcine MyD88 （poMyD88） from Peyer＇s patches （Pps） of GALT. The complete open reading frame （ORF） of poMyD88 contains 879 bp encoding a deduced 293 aa residues. The amino acid sequence of poMyD88 was characterized by N-terminal death, intermediate and C-terminal Toll/IL-1 receptor （TIR） domains. The putative poMyD88 protein shares a higher level of homology with its human （87.2% amino acid identity） than with its mouse （77.4% amino acid identity） counterpart. Overexpression of poMyD88 participated in the further enhanced activation of NF-w.B in human embryonic kidney （HEK） 293 cells expressing porcine TLR2 and porcine TLR4/MD-2, but not porcine RP105/MD-1 after stimulation with the corresponding ligands. The expression levels of MyD88 were highest in the spleen and mesenteric lymph nodes （MLNs）, and lower in digestive tissues of newborn swine. In adult swine, the expression levels in the digestive tissues were lower than those in MLNs and the spleen. These results suggest that an MyD88-dependent signaling pathway is present in newborn as well as in adult swine and that it is involved in the innate immune system of these animals.     

A prospective study of 3-day versus 2-week immobilization period after anterior cruciate ligament reconstruction

BACKGROUND: Aggressive early rehabilitation is recommended after reconstruction of the anterior cruciate ligament (ACL) using a patellar tendon-bone graft. However, rehabilitation after reconstruction of ACL using the multistranded hamstring is controversial. In this study, we compared the clinical results of 3-day immobilization after reconstruction of ACL using the multistranded hamstring, with those of 2-week immobilization prospectively. METHODS: A total of 30 consecutive patients with ruptured ACL underwent single-socket ACL reconstruction. 10 male and 5 female patients (mean age of 29.2+/-10.0 years) were prospectively randomized for a 3-day immobilization period, and 6 male and 9 female patients (mean age of 27.3+/-10.9 years) were randomized for a 2-week immobilization period. We measured the anterior laxity with a KT-2000 arthrometer, joint position sense, and thigh muscle strength at 3, 6 and 12 months after surgery. RESULTS AND CONCLUSIONS: There were no statistically significant differences between the two groups. We concluded that the post-operative 3-day immobilization period is preferable to the longer 2-week period in our rehabilitation program from the viewpoint of psychological load to the patients and the duration of hospitalization after ACL reconstruction using multistranded hamstring tendons.     

Epidermal growth factor receptor as a molecular determinant of glioblastoma response to dopamine receptor D2 inhibitors

BackgroundThere are ongoing clinical trials exploring the efficacy of dopamine receptor D2 (DRD2) inhibition against glioblastomas, the most common primary brain tumor. Here we examine potential molecular determinants of this efficacy.MethodsThe Cancer Genome Atlas glioblastoma database and other published mRNA profiles were used to analyze the DRD2 and epidermal growth factor receptor (EGFR) expression pattern. In vitro and in vivo responses to DRD2 inhibitors were determined using patient-derived xenograft (PDX) glioblastoma models. Immunohistochemical studies were performed on clinically annotated glioblastoma samples derived from patients treated with ONC201.ResultsAnalysis of clinical glioblastoma specimens derived from independent patient cohorts revealed an inverse correlation between EGFR and DRD2 mRNA expression, with implication that signaling mediated by these proteins shares overlapping functions. In independent panels of PDX glioblastoma lines, high EGFR expression was associated with poor in vitro and in vivo response to DRD2 inhibitors, including haloperidol and ONC201. Moreover, ectopic expression of a constitutively active EGFR, variant (v)III, suppressed glioblastoma sensitivity to ONC201. DRD2 expression positively correlated with expression of rate-limiting enzymes for dopamine synthesis as well as dopamine secretion, suggesting contribution of autocrine DRD2 signaling. Analysis of specimens from patients treated with ONC201 (n = 15) showed an inverse correlation between the intensity of EGFR staining and clinical response. The median overall survival for patients with high and low EGFR staining was 162 and 373 days, respectively (0.037).ConclusionsHigh EGFR expression is a determinant of poor glioblastoma response to DRD2. This finding should inform future clinical trial designs.