Evaluation of therapy with methanol extraction residue of BCG (MER)

Summary The current status of therapy with the methanol extraction residue of BCG (MER) is reviewed. We have identified 41 evaluable clinical trials of MER therapy, involving approximately 3,000 patients with malignant disease. The diagnoses have included lung, colon, and breast cancer, malignant melanoma, acute leukemia, and a small number of other malignancies. MER has been used as an adjunct to therapy for advanced disease and as prophylaxis against recurrence after surgery. Most studies have used the intradermal route but subcutaneous, intralesional, and intravenous routes have also been explored. The major local toxicity is pain and sterile abscess formation. The major systemic toxicity with administration by the intravenous route includes fever, malaise, and the development of pulmonary infiltrates. With the intradermal route little activity has been observed and there is no confirmed example of an increased remission rate, remission duration, or survival induced by MER therapy. When given by the intralesional route MER can cause regression of metastatic malignant melanoma nodules, and when given by the intravenous route MER is a potent immunoadjuvant. Antibody-dependent cellular cytotoxicity and natural killer cell activity were both boosted after one dose of intravenous MER. In rare patients receiving either intradermal or intravenous MER alone, without other therapy, tumor regression has been noted. These cases have included gastrointestinal cancer, lymphoma, and leukemia. Overall, the data indicate that the future of therapy with mycobacterial fractions awaits the development of more potent, less toxic fractions that can be administered systemically.     

Effect of treatment with the MER tubercle bacillus fraction on syngeneic plasma cell tumors of BALB/c mice

Summary MER administered prophylactically prolonged the survival of BALB/c mice challenged with transplants of syngeneic plasmacytomas to a moderate but significant extent. In contrast, MER exerted little therapeutic action when given alone at or after tumor implantation.Combined treatment, with MER introduced prophylactically and cyclophopshamide (CY) after tumor implantation, decreased the incidence of recurrence of one of the tumors tested (MPC-11 NP, a non-myeloma protein producer) and prolonged host survival significantly as compared with animals subjected to CY therapy alone. When, instead, MER was introduced at the time of tumor challenge or thereafter to animals also treated with CY, the therapeutic response was not appreciably different from that of mice under therapy with CY only. With regard to the second plasmacytoma (MPC-11 P, a myeloma protein producer), mice treated with CY and given MER prior to or after challenge showed similar responses to animals given chemotherapy only; when MER was injected at the time of challenge and CY thereafter, the chemoimmunotherapy was somewhat inferior to chemotherapy alone.This work was supported by US Public Health Service Contract NO1-CM-12127, and by grants from Mrs. J. H. Hazen, Ruth Estrin Goldberg Memorial for Cancer Research, Leukemia Research Foundation, Inc., Concern Foundation, Inc., and Mr. and Mrs. M. Gordon     

Effects of feed intake level on efficiency of microbial protein synthesis and nitrogen balance in Boran steers consuming tropical poor-quality forage

This study aimed at evaluating the effects of feed intake level on the efficiency of rumen microbial protein synthesis (EMPS), nitrogen (N) excretion, and N balance in twelve 18-months old Boran (Bos indicus) steers with initial average liveweight of 183 kg (standard deviation (SD) 15.2). The experiment followed a 4 × 4 complete Latin Square design with four dietary treatments tested in four periods. Each period ran for 5 weeks with 3 weeks of adaptation and 2 weeks of sample collection; separated by 2 weeks of re-feeding. Steers were fed at 100%, 80%, 60%, and 40% of their metabolisable energy requirement for maintenance (MER, referred to as MER100, MER80, MER60, and MER40, respectively). Steers receiving MER80, MER60, and MER40 were only fed Rhodes grass hay. MER100 steers were offered Rhodes grass hay at 80% of their MER and cottonseed meal and sugarcane molasses at each 10% of MER. Mean daily dry matter intake differed between treatments (p < 0.001) and ranged between 2.1 kg/animal (SD 0.13) in MER40 and 4.5 kg/animal (SD 0.31) in MER100. Urinary N excretion and N balance did not differ between MER80, MER60, and MER40. According to contrast test, declining feed intake level from MER80 to MER40 reduced duodenal microbial crude protein flow (p < 0.001), but did not alter the EMPS (g microbial N/kg digestible organic matter intake). Yet, if scaled to N intake, EMPS increased (p < 0.049), whereas total N and faecal N excretions decreased linearly with declining intake level (p < 0.001 for both variables). At similar grass hay intake, duodenal microbial crude protein flow was 41% higher in MER100 than in MER80 steers (p < 0.001). In cattle offered poor-quality tropical forage below their MER, the very low EMPS and thus microbial protein supply aggravate the negative effects of low dietary nutrient and energy intakes in periods of feed shortage.     

MER1, a yeast gene required for chromosome pairing and genetic recombination, is induced in meiosis.

The yeast MER1 gene is required for the production of viable meiotic products and for meiotic recombination. Cytological analysis of chromosome spreads from a mer1 mutant indicates that the MER1 gene product is also required for normal chromosome pairing. mer1 strains make axial elements, precursors to the synaptonemal complex; however, the chromosomes in most nuclei do not become fully synapsed. The DNA sequence of the MER1 coding region was determined; the MER1 open reading frame encodes a 270-amino-acid protein with a molecular mass of 31.1 kilodaltons. The MER1 protein shows limited sequence similarity to calmodulin. Expression of the MER1 gene was examined by RNA blot hybridization analysis and through the construction and analysis of mer1::lacZ fusion genes. Expression of the MER1 gene is meiotically induced and required the IME1 gene product. Thus, expression of the MER1 gene early in meiosis is required for proper chromosome pairing and meiotic recombination.     

The MER3 helicase involved in meiotic crossing over is stimulated by single-stranded DNA-binding proteins and unwinds DNA in the 3' to 5' direction

The meiosis-specific MER3 protein of Saccharomyces cerevisiae is required for crossing over, which ensures faithful segregation of homologous chromosomes at the first meiotic division. The predicted sequence of the MER3 protein contains the seven motifs characteristic of the DExH-box type of DNA/RNA helicases. The purified MER3 protein is a DNA helicase, which can displace a 50-nucleotide fragment annealed to a single-stranded circular DNA. MER3 was found to have ATPase activity, which was stimulated either by single- or double-stranded DNA. The turnover rate, k(cat), of ATP hydrolysis was approximately 500/min in the presence of either DNA. MER3 was able to efficiently displace relatively long 631-nucleotide fragments from single-stranded circular DNA only in the presence of the S. cerevisiae single-stranded DNA-binding protein, RPA (replication protein A). It appears that RPA inhibits re-annealing of the single-stranded products of the MER3 helicase. The MER3 helicase was found to unwind DNA in the 3' to 5' direction relative to single-stranded regions in the DNA substrates. Possible roles for the MER3 helicase in meiotic crossing over are discussed.     

Identification of differentially expressed genomic repeats in primary hepatocellular carcinoma and their potential links to biological processes and survival

Hepatocellular carcinoma (HCC) is one of the deadliest cancers. Research on HCC so far primarily focused on genes and provided limited information on genomic repeats, which constitute more than half of the human genome and contribute to genomic stability. In line with this, repeat dysregulation was significantly shown to be pathological in various cancers and other diseases. In this study, we aimed to determine the full repeat expression profile of HCC for the first time. We utilised two independent RNA-seq datasets obtained from primary HCC tumours with matched normal tissues of 20 and 17 HCC patients, respectively. We quantified repeat expressions and analysed their differential expression. We also identified repeats that are cooperatively expressed with genes by constructing a gene coexpression network. Our results indicated that HCC tumours in both datasets harbour 24 differentially expressed repeats and even more elements were coexpressed with genes involved in various metabolic pathways. We discovered that two L1 elements (L1M3b, L1M3de) were downregulated and a handful of HERV subfamily repeats (HERV-Fc1-int, HERV3-int, HERVE_a-int, HERVK11D-int, HERVK14C-int, HERVL18-int) were upregulated with the exception of HERV1_LTRc, which was downregulated. Various LTR elements (LTR32, LTR9, LTR4, LTR52-int, LTR70) and MER elements (MER11C, MER11D, MER57C1, MER9a1, MER74C) were implicated along with few other subtypes including Charlie12, MLT2A2, Tigger15a, Tigger 17b. The only satellite repeat differentially expressed in both datasets was GSATII, whose expression was upregulated in 33 (>90%) out of 37 patients. Notably, GSATII expression correlated with HCC survival genes. Elements discovered here promise future studies to be considered for biomarker and HCC therapy research. The coexpression pattern of the GSATII satellite with HCC survival genes and the fact that it has been upregulated in the vast majority of patients make this repeat particularly stand out for HCC.     

Yeast Mer1 Mutants Display Reduced Levels of Meiotic Recombination

Mutations at the MER1 locus were identified in a search for meiotic mutants defective in chromosome segregation. mer1 strains show decreased levels of inter- and intrachromosomal meiotic recombination and produce inviable spores. The MER1 gene was cloned by complementation of the spore inviability phenotype. Strains carrying disruptions of the MER1 gene are mitotically viable. The epistatic relationships between MER1 and previously characterized meiotic genes are described.     

肺淋巴瘤的CT表现

目的：肺部淋巴瘤可为原发,亦可为其他部位淋巴瘤的肺部浸润,临床上很少见。准确的影像学诊断对于采取及时的治疗措施具有重要意义,然而该病影像学表现复杂多变,容易与多种其他肺部病变混淆,以往文献对于肺部淋巴瘤的CT影像特征性表现报道不多,本文旨在探讨肺部淋巴瘤的特征性CT表现,以提高该痛影像诊断的准确率。方法：回顾性分析17例病理证实的肺淋巴瘤患者CT资料,分析特征性影像表现。结果：17例中多发者13例,单发者4例。叶段性或斑片状实变影13例,多发结节影13例,肿块状实变影8例,具有两种以上病灶的12例。特征性的征象有：病灶密度均匀（88．2％,15／17）、病灶边缘模糊（82．3％,14／17）、支气管气相（64．7％,11／17）、支气管血管柬增厚（58．8％,10／17）和CT血管造影征（41．2％,7／17）,对诊断有较大意义。结论：肺部淋巴瘤CT表现以多发、多形态病变居多,具有一定特征性影像表现,密度均匀、边缘模糊的实变、团块或结节影,具有支气管气相、支气管血管束增厚、增强扫描可见CT血管造影征等表现可高度提示本病的可能,结合病史、临床表现以及实验室检查,可提高本病的诊断准确率,对于及时开展有效的治疗措施、改善患者预后具有重要的临床意义。     

Antisense RNA of the latent period gene (MER5) inhibits the differentiation of murine erythroleukemia cells

The MER5 cDNA was cloned from RNA preferentially synthesized in murine erythroleukemia (MEL) cells during the early period of MEL cell differentiation. To understand the role of the MER5 gene in the differentiation, we have transferred the MER5 cDNA into MEL cells in both sense and antisense orientations under control of the promoter of the human metallothionein gene. Only in the transformants with the antisense MER5 cDNA, did their elevated expression inhibit differentiation. The result suggests that the MER5 gene product may promote early events in the differentiation of MEL cells.     

DNA content as a prognostic marker of oral lichen planus with a risk of cancer development

OBJECTIVE: To assess the presence of aneuploidy in oral lichen planus (OLP) and its usefulness as a prognostic marker. STUDY DESIGN: Eighty-one formalin-fixed, paraffin-embedded biopsy samples taken from atrophic-erosive OLP from 70 patients were studied. Approximately 150 random nuclei in basal and/or parabasal epithelia were analyzed with static cytometry. RESULTS: Aneuploidy was detected in 41% of samples. OLPs with ulcerations or location in the tongue had significantly higher values, respectively, for the 2.5c exceeding rate (ER) (p<0.001 and 0.001) and proliferation index (PI) (p = 0.012 and 0.013) than did lesions without ulcerations or at other locations. 2.5c ER was significantly higher in dysplastic OLP lesions (p < 0.001), and the significant value (p = 0.001)for 2.5c ER discriminating DNA aneuploidy was 15.3%. In multivariate analysis only the G2/M ER (G2/MER) was a significant independent predictor of developing cancer in OLP (OR 2.349, 95% CI 1.39-3.97, p = 0.001). CONCLUSION: Ulcerated atrophic-erosive OLPs of the tongue and with dysplasia are at increased risk of cancer development. 2.5c ER, PI and G2/MER might be useful in prognosticating the increased risk of malignancy in OLP.     

Characterization of the primate-specific repetitive DNA element MER1.

Computer analyses of the 3'-flanking DNA sequence of the human elastase I gene revealed a significant degree of similarity with seven human gene sequences in the GenBank and EMBL databases. Genomic Southern analysis indicates that the shared nucleotide sequences are a primate-specific family of short interspersed elements. These elements are members of MER1 sequences (medium reiteration frequency sequences). The consensus sequence of MER1 repeats spans 543 nucleotides and contains several inverted repeats. Since the copy number of MER1 elements seems to be much smaller than that of Alu and L1 repeats, MER1 elements may provide useful landmarks marks for human genome mapping.     

Mycosis fungoides. The importance of pulmonary cytology in the diagnosis of a case with systemic involvement

A patient with cutaneous mycosis fungoides developed pulmonary lesions while under radiation therapy. Bronchial cytologic specimens demonstrated malignant lymphocytes, which open lung biopsy confirmed to be mycosis cells. We believe this is the first report to document pulmonary involvement of mycosis fungoides by the use of bronchial cytology.     

Increased susceptibility of MER5 (peroxiredoxin III) knockout mice to LPS-induced oxidative stress

MER5 (also called peroxiredoxin III, PrxIII) is a member of peroxiredoxin family that has antioxidant activity. The present study was performed to investigate its in vivo function using MER5 knockout mice. MER5 knockout mice were born in normal frequency and could grow to maturity, but we found that intracellular ROS levels are significantly higher in the macrophages of the knockout mice. We examined roles of MER5 function for the oxidative stress responses by intratracheal inoculation of lipopolysaccharide (LPS) to the mice. Lung inflammation such as inflammatory cell infiltration and airway wall thickening was more severely detected in the knockout mice. At the same time, oxidative damage on DNA and proteins was more strongly detected in lung tissues of the knockout mice, including 8-hydroxy-2'-deoxyguanosine (8-OHdG) formation and protein carbonylation. The degrees of lung inflammation and oxidative damage were positively related with LPS doses. Our results indicate that MER5 knockout mice accumulated higher intracellular ROS levels, which cause LPS-induced lung injury more severely, and thus, suggested that MER5 acts as an important scavenger of reactive oxygen species (ROS) under oxidative stress.     

Unique histological changes in lung metastases of osteosarcoma patients following therapy with liposomal muramyl tripeptide (CGP 19835A lipid)

Summary We have recently begun a phase II trial in patients with osteosarcoma who developed pulmonary metastases during adjuvant chemotherapy or who presented with pulmonary metastases that persisted despite chemotherapy. Eligible patients were rendered free of visible disease by surgery. Liposome-encapsulated muramyl tripeptide phosphatidylethanolamine (MTP-PE, CGP 19835A lipid) (2 mg/m²) was infused twice weekly for 3 months. In five patients, a single tumor nodule recurred within 6 weeks after completion of therapy. These lesions were resected and submitted for pathological examination. Tissue specimens obtained after therapy were compared to those obtained before therapy. All the patients showed a histological change in the characteristics of the pulmonary tumors. In three patients, peripheral fibrosis surrounded the tumor and inflammatory cell infiltration and neovascularization were present. This is in contrast to central necrosis, with viable peripheral tumor cells and no inflammatory response observed in lesions resected following chemotherapy. In a fourth case, evidence of early fibrotic changes was found. This and the fifth case showed a change in malignant characteristics, from high grade before liposomal therapy to low grade after therapy. The present study provides evidence for a biological effect of liposomal MTP-PE.     

Potentiation and modulation of the immune response of guinea pigs to poorly immunogenic protein-hapten conjugates by pretreatment with the MER fraction of attenuated tubercle bacilli

The MER fraction of attenuated tubercle bacilli of the BCG strain was shown capable of stimulating and modulating the immunological responsiveness of guinea pigs to immunization with DNP conjugates of allogeneic globulin (DNP-GPG) and xenogeneic albumin (DNP-HSA). These antigens are very poorly immunogenic and fail to evoke detectable immune responses following single administration alone.When incorporated in incomplete Freund's adjuvant (IFA) together with the conjugates, MER could substitute for whole tubercle bacilli in the adjuvant mixture, and cause the conjugates to evoke both cellular and humoral reactivity, the former indicated by the development of skin reactions of delayed type (DH) to test injections of the antigens, the latter by the formation of humoral antibodies detected by an indirect hemagglutination (HA) test. When administered in saline together with antigen, MER was ineffective.Pretreatment with MER by any of several different routes 7 or 14 days prior to sensitization enabled a large number of the animals to respond with either DH, circulating antibody formation, or both. Under similar circumstances, pretreatment with Freund's complete adjuvant (FCA) elicited no such preparatory effect. In order to be efficacious in pretreatment, MER had to be given in a saline suspension; activity was lost when it was applied in IFA.MER pretreatment modulated the immune response to subsequent sensitization with the conjugates preferentially towards DH or antibody production, depending on the parameters of treatment and specific immunization. It appeared that when the specific immunogenic stimulus was weak, pretreatment with MER strongly favored DH.     

Clinical and radiologic manifestations of pleural and pulmonary complications in sepsis]

Proceeding from the investigation and therapy of 106 patients with pleuropulmonary complications of sepsis the authors described clinical and x-ray features of different types of their course, complications and outcomes. In 58% of cases pulmonary lesions were detected at the stage of pyodestruction and pleural empyema; interstitial-focal and infiltrative pneumonias of considerable spreading with the development of a respiratory type of septic shock were observed less frequently. Pulmonary lesions in 24% were the only sign of septicopyemia, in 58% they prevailed in the clinical picture of polyorganic lesions; pleural complications developed in 32% of the patients. Convalescence was observed in 88 (83%) patients; they had residual bullous-sclerotic pulmonary changes. The lethality rate was 17%.     

IgG and IgM responses in high and low antibody-selected lines of chickens

Kinetics of IgG and IgM as measured by 2-mercaptoethanol-resistant (MER) and susceptible (MES) antibodies to sheep erythrocytes, respectively, were determined as correlated responses in lines of chickens selected for high (HA) and low (LA) antibody response to sheep erythrocytes. Primary response patterns for total, MER, and MES antibody differed according to the genetic line. Total antibody increased rapidly, peaked, and persisted at moderate levels in line HA, whereas both peak and persistency were lower in line LA. Levels of MES peaked and then declined in line HA chickens but persisted at low levels throughout in line LA. Titers of MER antibody were considerably greater in line HA than in line LA both on an absolute basis and as a proportion of total antibody titer. Secondary total titers were greater at five days after injection than at three days and greater for line HA than for line LA chicks. The pattern observed for MER and MES in line LA was similar to that for total antibody, as was MER in line HA. For MES the pattern was reversed in line HA.