Thermodynamic stoichiometries of participation of water, cations and anions in specific and non-specific binding of lac repressor to DNA. Possible thermodynamic origins of the "glutamate effect" on protein-DNA interactions. The objective of this study is to quantify the contributions of cations, anions and water to stability and specificity of the interaction of lac repressor (lac R) protein with the strong-binding symmetric lac operator (Osym) DNA site. To this end, binding constants Kobs and their power dependences on univalent salt (MX) concentration (SKobs = d log Kobs/d log[MX]) have been determined for the interactions of lac R with Osym operator and with non-operator DNA using filter binding and DNA cellulose chromatography, respectively. For both specific and non-specific binding of lac R, Kobs at fixed salt concentration [KX] increases when chloride (Cl-) is replaced by the physiological anion glutamate (Glu-). At 0.25 M-KX, the increase in Kobs for Osym is observed to be approximately 40-fold, whereas for non-operator DNA the increase in Kobs is estimated by extrapolation to be approximately 300-fold. For non-operator DNA, SKobsRD is independent of salt concentration within experimental uncertainty, and is similar in KCl (SKobs,RDKCl = -9.8(+/- 1.0) between 0.13 M and 0.18 M-KCl) and KGlu (SKobs,RDKGlu = -9.3(+/- 0.7) between 0.23 M and 0.36 M-KGlu). For Osym DNA, SKobsRO varies significantly with the nature of the anion, and, at least in KGlu appears to decrease in magnitude with increasing [KGlu]. Average magnitudes of SKobsRO are less than SKobsRD, and, for specific binding decrease in the order [SKobsRO,KCl[>[SKobsRO,KAc[>[SKobsRO,KGlu[ . Neither KobsRO nor SKobsRO is affected by the choice of univalent cation M+ (Na+, K+, NH4+, or mixtures thereof, all as the chloride salt), and SKobsRO is independent of [MCl] in the range examined (0.125 to 0.3 M). This behavior of SKobsRO is consistent with that expected for a binding process with a large contribution from the polyelectrolyte effect. However, the lack of an effect of the nature of the cation on the magnitude of KobsRO at a fixed [MX] is somewhat unexpected, in view of the order of preference of cations for the immediate vicinity of DNA (NH4+ > K+ > Na+) observed by 23Na nuclear magnetic resonance. For both specific and non-specific binding, the large stoichiometry of cation release from the DNA polyelectrolyte is the dominant contribution to SKobs. To interpret these data, we propose that Glu- is an inert anion, whereas Ac- and Cl- compete with DNA phosphate groups in binding to lac repressor. A thermodynamic estimate of the minimum stoichiometry of water release from lac repressor and Osym operator (210(+/- 30) H2O) is determined from analysis of the apparently significant reduction in [SKobsRO,KGlu[ with increasing [KGlu] in the range 0.25 to 0.9 M. According to this analysis, SKobs values of specific and non-specific binding in KGlu differ primarily because of the release of water in specific binding. In KAc and KCl, we deduce that anion competition affects Kobs and SKobs to an extent which differs for different anions and for the different binding modes.


Key residues in the allosteric transition of Bacillus stearothermophilus pyruvate kinase identified by site-directed mutagenesis. The structural gene for pyruvate kinase from Bacillus stearothermophilus has been cloned in Escherichia coli and sequenced. The open reading frame from the ATG start codon to the TAG stop codon is 1482 base-pairs and encodes a peptide of relative molecular mass 52,967. In the expression vector pKK223-3, containing the synthetic tac promoter, the gene is overexpressed in E. coli cells to an estimated level of 30% total soluble cell protein. A purification procedure for the overexpressed protein has been established. The construction and characterization of a pair of mutant proteins has given insight into the structural basis of allosteric regulation in the tetrameric enzyme. Substituting tryptophan for tyrosine at position 466 (mutant Trp466-->Tyr) resulted in an activated form of the enzyme, having a reduced K1/2 for the substrate phosphoenolpyruvate. We propose that the characteristics of this mutant might be the result of bulk removal releasing steric inhibition to the formation of an interdomain salt bridge between Asp356 and Arg444. The regulatory behaviour of the double mutant produced by making the additional substitution aspartate for glutamate at position 356 (Trp466-->Tyr/Asp356-->Glu) corroborates this. The position of the salt bridge is such that it might be pivotal to the conformation of a pocket that is proposed to open up when the active R-conformation is adopted. We suggest that the mechanism of activation of B. stearothermophilus pyruvate kinase by ribose-5-phosphate might hinge on an interaction with, or indirectly through, residue Trp466, removing it from the vicinity of the potential salt bridge between Asp356 and Arg444 and thus effecting a closing together of the protein structure concomitant with an opening up of the pocket region.


Crystallization and preliminary X-ray investigation of phosphoribosylaminoimidazolesuccinocarboxamide synthase from the yeast Saccharomyces cerevisiae. Crystals of phosphoribosylaminoimidazolesuccinocarboxamide synthase (EC 6.3.2.6) from the yeast Saccharomyces cerevisiae were grown by the vapor diffusion hanging-drop technique, using ammonium sulfate as the precipitant. The crystals had dimensions up to 1.2 mm. X-ray diffraction experiments indicated a space group of P2(1)2(1)2(1) and unit cell parameters of a = 62.3 A, b = 63.5 A and c = 80.9 A, with one molecule in the asymmetric unit. Native data have been collected to 2.5 A resolution.


Cloning and analysis of the entire Escherichia coli ams gene. ams is identical to hmp1 and encodes a 114 kDa protein that migrates as a 180 kDa protein. We have used an antibody to a previously identified 180 kDa (Hmp1) protein in Escherichia coli to clone the corresponding gene, which encodes a polypeptide of 114 kDa that has a mobility equivalent to 180 kDa in SDS/PAGE. We have demonstrated that the 180 kDa polypeptide is the primary gene product and not due to aggregation with other molecules. Moreover, our data indicate that the highly charged C-terminal region of the protein is responsible for its anomalous behaviour when analysed by SDS/PAGE. The hmp1 gene is in fact identical to ams (abnormal mRNA stability), also designated rne (RnaseE), and reported to have an ORF of 91 kDa. This discrepancy with the data in this paper can be ascribed to the omission of two bases in the previously reported sequence, generating an apparent stop codon. We previously demonstrated that the 180 kDa Hmp1/Ams protein cross reacted with both a polyclonal antibody and a monoclonal antibody raised against a yeast heavy chain myosin. However, we could detect no homology with myosin genes in the ams/hmp1 sequence. From the DNA sequence data, we identified a putative nucleotide binding site and a transmembrane domain in the N-terminal half of the molecule. In the C-terminal half, which appears to constitute a separate domain dominated by proline and charged amino acids, we also identified a region homologous to the highly conserved 70 kDa snRNP protein, involved in RNA splicing in eukaryotes. This feature would be consistent with reports that ams encodes RNaseE, an enzyme required for the processing of several stable RNAs in E. coli.


Crystallization and preliminary crystallographic investigation of methanol dehydrogenase from Methylobacterium extorquens AM1. Single crystals of methanol dehydrogenase (MDH) from Methylobacterium extorquens AM1 have been grown by the vapour diffusion method. These crystals diffract to beyond 2 A resolution and are suitable for X-ray crystallography. They belong to the orthorhombic space group P2(1)2(1)2(1) and have the following unit cell parameters: a = 66.79 A, b = 108.9 A, c = 188.9 A. One asymmetric unit contains an alpha 2 beta 2 tetramer of MDH and the location of the non-crystallographic 2-fold symmetry axis of this tetramer is defined by the paired positions of the binding sites of heavy atoms in four MDH-derivatives.


The expression, purification and crystallization of the epsilon subunit of the F1 portion of the ATPase of Escherichia coli. The epsilon subunit of the F0F1-ATPase from Escherichia coli has been expressed in E. coli as a fusion protein with glutathione S-transferase from the parasitic helminth Schistosoma japonicum. The epsilon subunit released by thrombin treatment of the purified fusion protein carried two amino acid changes, A1G and M2S, and was obtained in a yield of about five milligrams per litre of cultured cells. The two amino acid changes were shown not to affect function. The protein has been crystallized in a form suitable for X-ray diffraction structure analysis. The crystals are hexagonal, space group P6(1)22 (or P6(5)22), with a = b = 94.9 A, c = 57.1 A and gamma = 120 degrees. The diffraction from small crystals extends to at least 2.9 A resolution.


Crystallization of the A-domain of the mannitol transport protein enzyme IImtl. The A-domain of the mannitol transport protein enzyme IImtl from Escherichia coli (relative molecular mass 16,300) was crystallized, both at room temperature and 4 degrees C, from 40% polyethylene glycol 6000 (pH 8.5 to 9.0) using the hanging-drop method of vapour diffusion. The crystals have the monoclinic space group P2(1), with unit cell dimensions a = 54.0 A, b = 67.0 A, c = 80.9 A and beta = 100.8 degrees. They diffract to 2.6 A resolution. A self-rotation function and self-Patterson suggest that there are four molecules in the asymmetric unit showing mmm symmetry.


Characterization of the nucleic acid-binding activities of the isolated amino-terminal head domain of the intermediate filament protein vimentin reveals its close relationship to the DNA-binding regions of some prokaryotic single-stranded DNA-binding proteins. In order to demonstrate that the nucleic acid-binding activities of vimentin are dictated by its Arg-rich N-terminal head domain, this was cut off at position Lys96 with lysine-specific endoproteinase and analysed for its capacity to associate with a variety of synthetic and naturally occurring nucleic acids. The isolated polypeptide (vim NT) showed a preference for single-stranded (ss) polynucleotides, particularly for ssDNAs of high G-content. A comparison of the sequence and predicted secondary structure of vim NT with that of two prokaryotic ssDNA-binding proteins, G5P and G32P of bacteriophages fd and T4, respectively, revealed that the nucleic acid-binding region of all three polypeptides is almost entirely in the beta-conformation and characterized by a very similar distribution of aromatic amino acid residues. A partial sequence of vim NT can be folded into the same beta-loop structure as the DNA-binding wing of G5P of bacteriophage fd and related viruses. As in the case of G5P, nitration of the Tyr residues with tetranitromethane was blocked by single-stranded nucleic acids. This and spectroscopic data indicate intercalation of the Tyr aromatic ring systems between the bases of the nucleic acids and thus the contribution of a stacking component to the binding reaction. The binding was accompanied by significant changes in the ultraviolet absorption spectra of both vim NT and single-stranded nucleic acids. Upon mixing of vim NT with nucleic acids, massive precipitation of the reactants occurred, followed by the quick rearrangement of the aggregates with the formation of specific and soluble association products. Even at very high ionic strengths, at which no electrostatic reaction should be expected, a distinct fraction of vim NT incorporated naturally occurring ssRNAs and ssDNAs into fast sedimenting complexes, suggesting co-operative interaction of the polypeptide with the nucleic acids. In electron microscopy, the complexes obtained from 28 S rRNA appeared as networks of extended nucleic acid strands densely covered with vim NT, in contrast to the compact random coils of uncomplexed RNA. The networks produced from fd DNA were heterogeneous in appearance and their nucleoprotein strands in rare cases were very similar to the rod-like structures of G5P-fd DNA complexes.


Genetic analysis of cosB, the binding site for terminase, the DNA packaging enzyme of bacteriophage lambda. cosB, the binding site for terminase, the DNA packaging enzyme of bacteriophage lambda, consists of three binding sites (called R3, R2 and R1) for gpNu1, the small subunit of terminase; and I1, a binding site for integration host factor (IHF), the DNA bending protein of Escherichia coli. cosB is located between cosN, the site where terminase introduces staggered nicks to generate cohesive ends, and the Nu1 gene; the order of sites is: cosN-R3-I1-R2-R1-Nu1. A series of lambda mutants have been constructed that have single base-pair C-to-T transition mutations in R3, R2 and R1. A single base-pair transition mutation within any one of the gpNul binding sites renders lambda dependent upon IHF for plaque formation. lambda phage with mutations in both R2 and R3 are incapable of plaque formation even in the presence of IHF. Phages that carry DNA insertions between R1 and R2, from 7 to 20 base-pairs long, are also IHF-dependent, demonstrating the requirement for a precise spacing of gpNu1 binding sites within cosB. The IHF-dependent phenotype of a lambda mutant carrying a deletion of the R1 sequence indicates that IHF obviates the need for terminase binding to the R1 site. In contrast, a lambda mutant deleted for R2 and R1 fails to form plaques on either IHF+ or IHF- cells, indicating terminase binding of R2 is involved in suppression of R mutants by IHF. A fourth R sequence, R4, is situated on the left side of cosN; a phage with a mutant R4 sequence shows a reduced burst size on both an IHF+ and an IHF- host. The inability of the R4- mutant to be suppressed by IHF, plus the fact that R4 does not bind gpNu1, suggests R4 is not part of cosB and may play a role in DNA packaging that is distinct from that of cosB.


Protein folding within the cell is influenced by controlled rates of polypeptide elongation. Previous studies have proposed that specific translational pauses have evolved to promote protein folding inside the cell by temporally separating the folding of specific regions of some polypeptide chains during their synthesis. Here we show that this is the case for a bifunctional protein in Saccharomyces cerevisiae. The yeast TRP3 gene contains a translational pause comprising ten contiguous non-preferred codons within its second functional domain (indoleglycerol phosphate synthase). Site-directed mutagenesis was used to remove this translational pause by increasing the codon bias of the region without changing the amino acid sequence of the protein (to create the gene TRP3pr: pause replaced). The TRP3pr gene was able to complement a trp3:: URA3 null mutation in yeast. No significant differences in the doubling times of TRP3 or TRP3pr yeast transformants were observed during growth at 25 degrees C, 30 degrees C or 37 degrees C, or in the presence of sublethal concentrations of the analogue, 5-methyltryptophan. However, further analysis of TRP3 and TRP3pr transformants revealed that the removal of the translational pause causes a 1.5-fold decrease in indoleglycerol phosphate synthase activity per TRP3 mRNA. This observation which is statistically significant (P < 0.05) and reproducible, suggests that translational pausing promotes the correct intracellular folding of the TRP3 protein.


Genetic analysis of mutations affecting terminase, the bacteriophage lambda DNA packaging enzyme, that suppress mutations in cosB, the terminase binding site. Terminase, the DNA packaging enzyme of phage lambda, binds to lambda DNA at a site called cosB, and introduces staggered nicks at an adjacent site, cosN, to generate the cohesive ends of virion lambda DNA molecules. Terminase also is involved in separation of the cohesive ends and in binding the prohead, the empty protein shell into which lambda DNA is packaged. Terminase is a DNA-dependent ATPase, and both subunits, gpNu1 and gpA, have ATPase activity. cosB contains a series of gpNu1 binding sites, R3, R2 and R1; between R3 and R2 is a binding site, I1, for integration host factor (IHF), the Escherichia coli DNA bending protein. In this work, a series of mutations in Nu1 have been isolated as suppressors of cosB mutations. One of the Nu1 mutations is identical to the previously described Nu1ms1/ohm1 mutation predicted to cause the change L40F in the 181 amino acid-long gpNu1. Three other Nu1 missense mutations, the Nu1ms2 (L40I), ms3 (Q97K) and ms4 (A92G) mutations, have been isolated; the relative strengths of suppression of cosB mutations by the Nu1ms mutations are: ms1 > ms2 > ms3 > ms4. The Nu1 missense mutations all affect amino acid residues that lie outside of the putative helix-turn-helix DNA binding motif of gpNu1. The Nu1ms1 and Nu1ms2 mutations alter an amino acid residue (L40) that lies directly between two segments of gpNu1 proposed to be involved in ATP binding and hydrolysis; thus these mutations are likely to alter the gpNu1 ATP-binding site. The Nu1ms3 and Nu1ms4 mutations both affect amino acid residues in the central region of gpNu1 that is predicted to form a hydrophilic alpha-helix. To explain how the Nu1ms mutations suppress cosB defects, models involving alterations of the DNA binding and/or catalytic properties of terminase are considered. The results also indicate that terminase occupancy of a single gpNu1 binding site (R3) is necessary and sufficient for the efficient initiation of DNA packaging; the Nu1ms1, ms2 and ms3 mutations permit IHF-independent plaque formation by a phage lacking R2 and R1.


Role of MotA transcription factor in bacteriophage T4 DNA replication. At least two bacteriophage T4 replication origins, ori(uvsY) and ori(34), contain a T4 middle-mode promoter that is necessary for origin function. We wanted to analyze the requirement of these two replication origins for the MotA protein, which is the phage-encoded activator of middle-mode promoters. To ensure the complete absence of MotA protein, we deleted the motA gene from the T4 genome. Unexpectedly, the deletion mutant was not viable unless the MotA protein was provided from a recombinant plasmid. Therefore, MotA is an essential protein for T4 growth. The motA delta mutation reduced the synthesis of several proteins that are encoded by genes with middle-mode promoters, delayed and reduced the synthesis of late proteins, and substantially reduced phage genomic replication. The motA delta mutation also reduced the replication of an ori(uvsY)-containing plasmid and virtually abolished replication of an ori(34)-containing plasmid. The replication defects of the two origins correlated with transcriptional defects: the motA delta mutation modestly reduced transcription from the plasmid-borne ori(uvsY) promoter and strongly reduced transcription from the ori(34) promoter. These results provide strong evidence that MotA protein is normally involved in origin-dependent replication. However, MotA is not required for origin-directed replication as long as transcription can occur from the origin promoter.


[A historical review of flow cytometers and characteristics of commercial instruments]. Instruments utilized in flow cytometry are called flow cytometers and they can be classified into two kinds, namely, cell analyzers and cell sorters. Important technological developments related to the flow cytometer and various kinds of laboratory instruments are reviewed historically. Commercial cell sorters and cell analyzers produced by several companies are compared and the characteristics of each are listed.


[Progress and application of multiple-parameter flow cytometric analysis]. Multiple-parameter flow cytometric analysis for the expression of cell surface markers has been generally used in various clinical and fundamental fields. This was made possible by the development of many monoclonal antibodies and new fluorescence dyes coordinately with the progress in hardwares. Especially, the development of new fluorescence dyes enabled us simultaneously analysis of more than two markers, not only surface markers but also intracellular events, such as Ca2+ concentration and cell cycle. As a result, fine characterization of cells of interest is now possible even with a limited number of cells within a shorter time. The high resolution and sensitivity of current multiple-parameter flow cytometer make possible the isolation of diverse and unique cell populations that can be obtained by no other way.


[Applications of flow cytometry to molecular biology and molecular genetics]. Flow cytometry has been utilized in the fields of immunology, hematology, cancer biology and somatic cell genetics. It is also being used for cell cycle and chromosome ploidy analysis. Furthermore, flow cytometry is powerful for karyotype analysis and chromosome sorting. Here, I will review recent advances of flow cytometry in molecular biology and molecular genetics, and discuss some of our recent works.


[The theory of flow cytometric analysis of the cell cycle and their applications in biology and oncology]. The clockwise concept of cell cycle kinetics has changed since molecular biology made rapid advances in this area and added somewhat to cell cycle analysis by DNA histogram. During the last decade, it has been made possible to measure many cell-cycle-regulating gene products flow cytometrically. Application of bromodeoxyuridine (BrdU) to cell cycle analysis contributes to scientific progression in many fields of biology and oncology. In this article, we attempt to summarize the outline of new concepts regarding the cell cycle by means of flow cytometric techniques and to describe their application in biology and oncology. For this purpose, probes, biological differences amongst the dyes used, DNA analysis using DNA histograms and their limitations are explained. Next, new methods to analyse cell cycle kinetics by means of BrdU, Ki-67 antigen, cyclin, etc. discussed. As far as application of flow cytometric cell cycle analysis are concerned, influences of nutrition, growth factors and hormones, relationships between DNA index or cell cycle and the prognosis of patients with cancer, and the effect of anticancer drugs on the cell cycle are discussed. Finally, we emphasize the importance of multiparameter analysis of cell cycle kinetics using flow cytometry.


[Detection of proliferative cells by DNA polymerase a as a proliferation associated marker]. Proliferative cell fractions were measured by flow cytometry in 20 patients with acute leukemia, 4 with chronic myelocytic leukemia in blastic crisis and 7 with malignant lymphoma. The cells were fixed with 2% paraformaldehyde followed by staining with fluorescein isothiocyanate conjugated monoclonal antibody against DNA polymerase a. The DNA polymerase a-positive population was widely distributed in leukemia, from 20.4% to 84.7% in peripheral blood and from 6.5% to 92.5% in the bone marrow. A positive correlation was found between the values in peripheral blood and bone marrow. The values ranged from 66.4% to 88.1% in cells from cases of malignant lymphoma. Cryopreserved cells may be available for measurement of DNA polymerase a because the result obtained in both frozen and fresh cells were essentially the same.


[AgNORs (Ag nucleolar organizer regions)]. A quick and quantitative evaluation technique, "AgNORs-FCM", was developed to analyze Nucleolar Organizer Regions (NORs) stained with silver impregnation followed by cytofluorometric analysis. Cell suspension is stained with AgNORs similarly as described elsewhere for tissue sections, but with a modification of using a much lower concentration of silver nitrate. Cytological general rules in AgNORs-FCM are: 1) Specific exaggeration of side scatters (90 LSs) can be detected according to the intensity of AgNORs staining. 2) Significantly brighter 90 LSs were observed when cultured leukemia cell-line was applied to AgNORs-FCM. 3) AgNORs-FCM provides more specifically brighter 90 LS for cells in G2M and S phase, when gated along the P.I.-fluorescence. AgNORs-FCM can be applied to analyze clinico-pathological samples including neoplastic tissues, more quickly and quantitatively, than regular tissue staining with AgNORs.


[Flow cytometric analysis, mechanism of action and evaluation of viability by antineoplastic agents]. Since the cell cycle of tumor cells is evaluable with the aid of flow cytometry, the mechanism of action of antineoplastic agents has readily been analyzed based on the changes of the cell cycle by antineoplastic agents. Propidium iodide had been a conventional staining method for such analysis, but after the introduction of DNA/BrdU double staining method using BrdU, it has become much easier. For evaluating antineoplastic agents, simultaneous measurement by FDA of the viability of the tumor cells is important. In vitro studies of the action mechanism and the antineoplastic effects of 5-FU, ACNU, IFN, PG, CDDP, PEP, and HCFU have been reported previously elsewhere. Similar action mechanism has been confirmed in clinical studies, in which optimal agent was selected by the sensitivity test using flow cytometry, and thus selected agent was actually given to tumor patients. In analyzing antineoplastic agents in vitro, concomitant and total evaluation of DNA-histogram and viability by flow cytometry is needed in the future.


[Flow cytometric analysis of proliferative characteristics of human leukemic cells in bone marrows in vivo using bromodeoxyuridine (BrdU) and its monoclonal antibody]. We developed a new method for analyzing the proliferative characteristics of human acute leukemic cells in bone marrows in vivo using BrdU and its monoclonal antibody. The thirty minutes after a bolus BrdU infusion, a 1st bone marrow aspiration (BMP) was performed to calculate BrdU-LI. Then, a 150-minute continuous infusion of BrdU was started. After the infusion, a 2nd BMP was done to calculate the increasing rate of the BrdU-LI for 3 hours. Each of the bone marrow cells was analyzed by FCM. A mean value of 10.4% for BrdU-LI in 22 previously untreated patients was obtained. In 3 of the patients, Ts and Tpot ranged from 13.5 to 33.9 hours and 6.8 to 10.7 days respectively.


[Chemo-sensitivity test for malignant brain tumors by DNA histogram, in vitro]. Application of flow cytometric DNA analysis was tried to determine the sensitivity of ACNU, one of nitrosourea derivatives which had been used very commonly for malignant brain tumors, to tumor cells. To evaluate the degree of chemo-sensitivity of ACNU, factor B was introduced, indicating the accumulated cells in SG2M phases after ACNU treatment as the percentage of cells that was previously in the SG2M phases and it revealed that ACNU sensitive cell clones gave much larger values of Factor B than ACNU resistant ones at the concentration of 10 micrograms/ml ACNU treatments. Twenty clinical materials obtained by operation were examined by measuring the degree of values of this Factor B, and the findings indicated that this method would be applicable as a clinical test for chemo-sensitivity of malignant brain tumors, after some improvement of the method.


[Flow cytometric bromodeoxyuridine (BrdU)/DNA analysis using fresh solid tumors and its clinical significance]. It is very important to determine the biological characteristics of individual cancers, not only in order to evaluate malignant potentials but also to decide on the most effective anticancer therapy. A method which combines BrdU labeling and flow cytometric bromodeoxyuridine (BrdU)/DNA analysis makes it possible to plot the DNA synthetic rate of cells against their DNA content. Hence, this method yields more useful information than the previous method of flow cytometric DNA analysis. When this method is performed correctly, it can be also applied to fresh solid tumors. In this paper, we will describe the results of our preliminary investigation and discuss its clinical significance.


[Flow cytometric analysis of the DNA ploidy pattern of primary lung cancer--its prognostic and biological implications]. In order to clarify the significance of the nuclear DNA ploidy pattern in primary lung cancer, recent reports of flow cytometric analysis of nuclear DNA are discussed. It has been reported in many institutes, that nuclear DNA ploidy pattern was a significant and an independent postsurgical prognostic factor in lung cancer, statistically. Diploidy tumors showed a better post-surgical outcome than aneuploid tumors. This fact is more evident in stage I tumor than in advanced cases and was more in squamous cell carcinomas than in adenocarcinomas. Some papers reported that tumors with a high DNA index showed more invasive character than that with a low DNA index with flow cytometric and histopathological examination. As the percentage of aneuploid tumor was much higher in primary lung cancer than in other cancers, more detailed characterization is believed to be needed in aneuploid tumors by means of other oncological analysis, e.g. growth factors or expression of oncogene products.


[DNA ploidy pattern as an indicator of degree of malignancy and prognosis in gastric cancer]. We used fresh materials and paraffin blocks of advanced gastric cancer to examine the relationship of DNA ploidy pattern from flow cytometry (FCM) to degree of malignancy and prognosis. No significant relationship was found between the frequency of DNA aneuploid (DA) and the clinicopathological factors (depth of invasion, degree of progression, histological type, lymph node metastasis, ly, v etc.). FCM examination using fresh materials revealed significant difference in prognosis between the non-DA and DA groups for non-curative stage III or IV cases (p < 0.05). On the other hand, in terms of the relationship between DNA ploidy patterns from paraffin blocks and prognosis for curative resection cases of ss-gastric cancer, prognosis was significantly better in the non-DA group than in the DA (p < 0.01). These results indicate that the clinicopathological prognostic factors for gastric cancer are connected in a complex way and that DNA ploidy patterns alone are not superior to the prognosis determining factors used in the past.


[Study of nuclear DNA content with flow cytometer in pancreatic diseases]. Benign and malignant diseases of the pancreas were compared with nuclear DNA content by flow cytometry, and the relationship between nuclear DNA content and histopathological findings was studied in ductal carcinoma of the pancreas. The nuclear DNA content in the malignant disease was higher than that in the benign disease. No significant correlation between nuclear DNA content and histopathological findings was observed in ductal carcinoma of the pancreas. A tendency for the nuclear DNA content to increase in proportion to the tumor size and the stage was, however, observed. Study of the nuclear DNA content is believed to be an effective adjuvant modality in differential diagnosis between benign and malignant disease of the pancreas.


[DNA ploidy pattern of flow cytometry as indicator of degrees of malignancy and prognosis in colorectal cancers]. Cellular DNA content of primary colorectal cancers was measured by flow cytometry and investigated on clinico-pathological features to elucidate the relationship between DNA ploidy patterns and outcomes. IN 144 colorectal carcinomas, DNA diploid carcinomas accounted for 23% and DNA aneuploid carcinomas for 77%. NO significant difference was observed between DNA ploidy pattern and tumor differentiation, lymph node metastasis or lympangial invasion. DNA aneuploid tumor had a tendency to invade vein and to lead to hematogenic metastasis. Patients with DNA aneuploid tumor showed a significantly poor disease-free and overall survival rate. Significantly increased incidence of hematogenic recurrence was demonstrated in the DNA aneuploid cases. These results suggest that the DNA ploidy pattern of colorectal cancers may prove to be of prognostic value.


[DNA ploidy pattern of flow cytometry as indicator of malignancy and prognosis in bladder cancer]. Flow cytometry (FCM) of bladder cancer has unique characteristics. First, bladder washing is a suitable material to measure in terms of single cell suspension, while the urine appears inadequate because of the low sensitivity for detecting bladder cancer. Second, the role of FCM depends on the different features of superficial, in situ or invasive bladder cancer. In superficial cancers, FCM can detect the presence of cancer, predict intravesical recurrence and evaluate the response of intravesical instillation therapy. In carcinomas in situ, frequent occurrence of polyclonal aneuploid may indicate the aggressive nature of high grade malignancy. Early removal of bladder may be endorsed unless BCG instillation is effective. For invasive cancers, FCM does not seem to be so useful, but FCM relates well to other prognostic parameters.


[Flow cytometric evaluation of DNA ploidy pattern in uterine cancer]. The distribution of DNA ploidy levels and its prognostic significance in cervical cancer (including squamous cell carcinoma and adenocarcinoma) and endometrial cancer is discussed. DNA aneuploidy was observed in most of the cases with either the histological type of cervical cancer and in half of those with endometrial cancer. The DNA ploidy level of the tumor showed a characteristic distribution according to its histological type or grade. Although several investigators have already reported that patients with DNA diploid uterine tumors had a better survival than those with DNA aneuploid uterine tumors, further research is required before a definite conclusion can be attained on the prognostic value of the degree of DNA ploidy measurement in uterine cancer.


[Flow cytometric analysis of DNA ploidy pattern in breast cancer: malignant potential and prognostic implication]. Flow cytometric DNA analysis in solid tumors has developed over the past ten years. In human breast cancer, flow cytometric analysis of DNA ploidy pattern recently reported were reviewed. There are conflicting data regarding the value of DNA ploidy pattern obtained by flow cytometric analysis for determining prognosis for breast cancer patients. Some data seem to indicate that S-phase fraction alone or in combination with DNA ploidy pattern may be more important prognostically than DNA ploidy pattern alone, although the results have not been uniform. These suggests that a consensus of determination for DNA ploidy pattern in breast cancer and further investigations on DNA ploidy pattern are required.


[Separation and measurement of lymphocyte functional subpopulations by two color FCM method]. With the popularity of flow cytometry (FCM) and the availability of a wide variety of monoclonal antibodies to human blood cells, analysis of cell surface antigens is now conducted at many facilities as part of their routine testing. Analysis of cell surface antigens by FCM has contributed greatly to the diagnosis and the determination of therapeutic effects in many diseases. In the present study, for detection of lymphocyte surface antigens by two color analysis and selection of appropriate antibodies are described. In addition, the significance of two color analysis is explained by citing hematological neoplasms and several other clinical cases. It has been recently reported that the immunological system is involved in many disease processes. In the future, more detailed studies will be conducted to elucidate the immunological mechanisms in these diseases.


[Detection of IgG receptor subtype on basophils using two-color FCM]. It was extremely difficult to phenotype basophils because they are most sparse among blood leukocytes. We developed a new method which identified basophils on the basis of negative reactivity with mixed monoclonal antibodies (CD2, CD14, CD16, CD19). Using this method we could detect Fc gamma R II (CDw32) but not Fc gamma R I (CD64) on basophil surface. The method seems useful for the study of clinical allergy and also of the biology of basophil.


[Simultaneous analysis of c-myc protein expression and cell cycle with monoclonal antibody and flow cytometry]. C-myc protein plays an important role in the regulation of cell proliferation and differentiation. In this paper, c-myc protein and DNA were doubly stained and analysed simultaneously with flow cytometry (FCM). At first, three fixatives, ethanol, methanol and paraformaldehyde, were examined using HL-60 cells, among which 50% ethanol was found to be optimal for each staining. After fixation, the cells were stained with a monoclonal antibody against c-myc protein, followed by DNA staining with PI. Simultaneous analysis demonstrated that c-myc protein was constantly expressed through the cell cycle and that the protein amount at the G2 + M phases was 1.5 times higher than that at the G1 phase. Further, the differentiation study with TPA and RA revealed that the growth, S phase and also c-myc protein expression were suppressed during the differentiation.


[The expression of IL-2 receptor subunits on various leukemic cells]. Interleukin-2 receptors are composed of at least two polypeptide chains of alpha (55KD) and beta (75KD). The IL-2R beta chain is an essential component of the functional receptor for signal transduction of IL-2. We previously reported the distribution of IL-2R subunits among peripheral blood mononuclear cells. We here present some data regarding the expression of IL-2R subunits on various hemopoietic malignant cells. Fresh leukemic cells obtained from adult T cell leukemia patients expressed both alpha and beta chains, and leukemic cells derived from some patients with T cell leukemia, B cell leukemia or myeloid leukemia expressed the alpha and/or beta chain of IL-2R. The IL-2R beta chain on these leukemic cells were demonstrated to be functional for cell growth signaling. IL-2R alpha and beta chains should be tumor markers.


[Flow karyotyping and chromosome sorting]. For flow karyotyping and sorting, chromosomes are stained in suspension with appropriate DNA fluorochromes, and passed singly through a beam of laser light. The emitted pulses of fluorescence and scattered light are measured and stored in the form of a histogram displaying the number of pulses versus fluorescence or scattered light intensity. A data processing system was devised to remove signals from cell debris or chromosome fragments. A selective window was set in a dot plot of fluorescence versus scattered light intensities and a program was made to count pulses from the gated area only. As application of chromosome fractionation, flow cytometry has been used for karyotype analysis, gene mapping, and chromosomal DNA library construction.


[Flow cytometric analysis of chromosome aberration on tumor cells]. Chromosomal changes plays an important role in malignant transformation. Generally, the process of karyotype instability with grows aneuploidy occurs in tumor. But it is very difficult to obtained Flow karyotype from tumor cells. There are many technical problem in the analysis of chromosomes aberration in tumor cells. Problem is technical procedure of isolation from metaphase chromosomes. It is important to choice of swelling buffer and treatment times. Such technic affect for Flow karyotype pattern. We try to obtained Flow karyotype from chinese hamster cell and V-79 cells and we reported a recent new techniques of cell preparation and chromosomes suspension.


[Progress in microscopic image analysis and the trend of image cytometry]. Image cytometry has recently developed prominently based on the digital imaging of cell morphology brought about by great advances in microcomputer hardware and software, electronics etc. This method is characterized by the quantitation of both the amount of intracellular bioactive materials and cell morphology, and is expected to further advance many cell analysis techniques involving automated cytology, chromosome image analysis, automated histopathology, viable cell analysis, 3D image construction, etc. The present use is still minimal, and advances in both hardware and software for cyto-histologic recognition are essentially necessary. Perhaps by the end of this century, more sophisticated and faster machines for automated cyto-histologic analysis will appear, replacing many microscopic techniques of presently subjective observation in the biomedical field.


[The advancement of image cytometry in immunohistology]. An image processing system has been developed by the combination of recent electronic and computer technologies. The system may have a potential power for the quantitative analysis on the immunohistology. The image cytometry was applied to quantitative analysis on Ki-67-positive cells in lymphomas of Waldeyer's ring and nasal cavity. High grade lymphomas showed significantly larger number of Ki-67-positive cells than intermediate-grade lymphomas, even analyzed separately by immunophenotypes. A large mean area per Ki-67-positive cell was significantly associated with T-cell phenotype and unfavorable clinical outcome. Thus, Ki-67 immunostaining combined with image cytometry is a novel method for determining a tumor proliferative index which provides useful clinical data of non-Hodgkin's lymphomas.


[Quantitative non-radioactive in situ hybridization for measurement of specific mRNA using image analyzer]. To understand cells, information on the state of gene expression in each cell is essential. We determined the copy number of c-myc mRNA in HL60 cell by first carrying out in situ hybridization using thymine-thymine dimerized anti-sense oligonucleotides probes to c-myc mRNA and the hybridized thymine dimers were localized by the immunoperoxidase method. The density of each cell was measured by an image analyzer and converted to the mRNA copy number in each cell, using dot blot hybridization results, as a reference standard. It was found that the numbers of c-myc mRNA copies varied 0-4,000 when expressed at the unit of cell.


[The grading of cellular and structural atypia in cancer and related lesions using multivariate analysis]. In morphologically diagnosing cancer, pathologists' attention is focussed on the presence or absence of atypia, i.e., the form of cells and tissues deviated from the norm, and if it is present, on its grade. However, due to retarded development of techniques for the objective evaluation of its grade, separation of cancer from premalignant lesions still meets great ambiguity. Particularly, the boundaries of "dysplasia" with cancer and noncancerous lesions are not defined in clear morphological terms and are susceptible to a strong between-observers fluctuation. In this paper we outlined our recent efforts to establish in various organs a statistically most adequate, and therefore reproducible, classification of atypical lesions, resorting to morphometry and multivariate analysis. Two examples were given: atypia of the pancreatic duct epithelia as an object for the study of purely cellular abnormalities, and hepatocellular carcinoma, which required also to quantify the atypia of tissue structures, The classifications thus established proved to be the most adequate, in that, they closely reflected the grade of malignancy in a clinical as well as biological sense, as shown by the clear between-categorier differences in oncogene mutation, DNA ploidy pattern, the patients' prognoses, and so on.


[Application of image cytometry to cytological diagnosis of breast tumors]. Computerized morphological analysis of the cell nuclei was performed on 66 cases (2222 nuclei) of aspirated materials from breast tumors: 25 benign cases (302 nuclei), 41 malignant cases (1420 nuclei). Its diagnostic significance and the correlation between nuclear analysis and clinical staging were studied. Nuclear diagram and intranuclear chromatin distribution pattern were evaluated using computer system named 6400. The malignant tumor had larger, more round-shaped nuclei and less uniform chromatin distribution. Under tnm classification, stage IV tumors had larger, more flattened nuclei and more irregular chromatin distribution pattern than the others. Computerized morphological analysis of breast tumors may give a quantitative aspect to classical diagnostic process. Furthermore, this analytical method provides information on clinical stage of breast cancer.


[Quantitative DNA image cytometry by color image analyzer]. We developed a new method to analyze DNA quantitatively by a color image analyzer. A microscopic image of the nucleus stained with Feulgen's reaction, is taken by a CCD color camera and inputted in a color image analyzer. DNA quantity can be calculated directly from the RGB components of the color image instead of the monochrome image taken through band-pass filter, which is specific for the absorption of Feulgen's stain. It takes about 10 minutes to measure 500 nuclei by this method and its accuracy is almost the same as flow cytometry (CV = 3%). The nuclear color image can also be analyzed morphometrically. This method can select tumor cells in a usual cytologic specimen and measure their DNA contents.


[Confocal laser scanning microscopy for image cytometry]. By using a focused laser beam as the light source of the microscope, to minimized flare, together with a pinhole in front of a photo detector to eliminate out-of-focus data, the confocal laser scanning microscope (CLSM) provides a depth-discriminated fluorescence image with high spatial resolution. The CLSM, therefore, has been widely used as a tool for accurate morphometric and densitometric analyses. We discussed here the biological applications of CLSM for demonstrating the three-dimensional features of the embryonic heart tube and chromosome and for imaging fast dynamic changes of [Ca2+]i in the living heart muscle cell pairs.


[Progress on chromosome image analysis systems]. This is a review of the development of automated systems of chromosome analysis. The history of its development reflects the progress of image analysis systems and that of method of chromosome analysis in man. It can be chronologically summarized as follows. Period I: 1960s-early 1970s when chromosome banding methods were discovered. Period II: early 1970s-early 1980s when the first image analysis system of chromosome was commercialized. Period III: early 1980s-present. Research and development of the automated analysis system of radiation induced chromosome aberrations, conducted under the national project of cross-over research on underlying technology of nuclear energy, Science and Technology Agency, Japan, is briefly reported.


[Advances in molecular parasitology].

[Advances in molecular parasitology]. Recent studies on molecular biology of parasites revealed novel mechanisms for the control of expression of parasitic genes. In trypanosomatid protozoa, unusual characteristics of gene regulation, including polycistronic transcription, trans-RNA splicing and RNA editing have been discovered. This review describes the recent progress of RNA editing in trypanosomes, and trans-splicing in both trypanosomes and nematodes. Studies of the editing of mRNAs in trypanosomatid mitochondria indicate the involvement of small guide RNA molecules, which are encoded in the minicircles and/or maxicircles of kinetoplast DNA. A proposed transesterification model for RNA editing suggests an analogous reaction between RNA editing and RNA splicing. In addition, the development of stable transfection system and success of homologous gene targeting in Trypanosoma and Leishmania are reported.


[Effects of smoking and drinking habits and vitamin A intake on serum concentrations of beta-carotene and retinol]. Serum samples from 341 males aged 10 to 59 years were obtained and stored at -40 degrees C until examined for retinol and beta-carotene concentrations by HPLC, and their relationships to smoking habit, alcohol drinking habit and vitamin A intake were studied. In univariate analysis the serum beta-carotene level was significantly lower in the smokers than in the non-smokers (smokers: 4.6 micrograms/dl, non-smokers: 7.1 micrograms/dl, p less than 0.01) and lower in the drinkers than in the non-drinkers (drinkers: 4.6 micrograms/dl, non-drinkers: 7.3 micrograms/dl, p less than 0.01). The serum retinol level was not different by smoking habit but was higher in the drinkers than in the non-drinkers (drinkers: 80.4 micrograms/dl, non-drinkers: 67.0 micrograms/dl, p less than 0.01). Serum beta-carotene was higher in the group with a greater intake of vitamin A of vegetable origin (6.1 micrograms/dl) than in the group with a smaller intake of it (4.7 micrograms/dl) (p less than 0.01), but serum retinol was not different by the amount of vitamin A intake of animal food origin. To estimate the respective effects and interactions of the above factors on serum beta-carotene and retinol levels by adjusting for the confounding effects of age, serum total cholesterol, HDL-cholesterol and triglyceride, analysis of covariance was performed. For serum beta-carotene, smoking habit (p less than 0.01), drinking habit (p less than 0.01) and the amount of vitamin A intake of vegetable food origin (p less than 0.05) had significant main effects.(ABSTRACT TRUNCATED AT 250 WORDS)


[Improved thermogenesis during acute cold exposure in exercise-trained rats]. The present study was carried out to examine the effects of enhanced cold tolerance induced by exercise training on carbohydrate metabolism during cold exposure. Five-week-old male Wistar rats were divided into three groups. The warm-acclimated control group (WA) was exposed to the environment at 25 degrees C, and the cold-acclimated group (CA) was acclimated to cold by exposure to 5 degrees C. The exercise-trained group (ET) was controlled to run at the rate of 35 meters per minute for 9 weeks on a small driven treadmill. The degree of carbohydrate metabolism due to cold exposure significantly decreased with the drop in environmental temperature in the WA and CA groups, but did not change in the ET group. When glucose utilization was blocked with 2-deoxyglucose administration, oxygen consumption during cold exposure decreased in the WA and ET groups, but did not decrease in the CA group. The liver glycogen content decreased during cold exposure in the WA and ET groups, but there was no change in the CA group. It appeared that the fall of the plasma glucose level after the infusion of insulin was significantly greater in the ET and CA groups than in the WA group. These results indicate that the improved capacity for cold-induced thermogenesis in the ET group may be caused by enhanced carbohydrate metabolism.


[Effects of an essential trace element agent (TE-5) for total parenteral nutrition on the mineral nutrition in rats fed a trace element-deficient diet]. TE-5 is an essential trace element agent containing iron, zinc, copper, manganese and iodine for total parenteral nutrition (TPN). We have already reported that TE-5 improved the reduction of trace element concentrations induced by TPN. However, effects of TE-5 on the changes in biological function relating to trace elements are poorly understood. The present study was designed to clarify the effects of TE-5 on these functions. Rats fed a trace element (iron, zinc, copper, manganese and iodine)-deficient diet for 7 weeks showed reductions in the following parameters: plasma and various tissue concentrations of iron, zinc, copper, manganese and iodine, growth rate, erythrocyte (iron), hemoglobin (iron), hematocrit (iron), mean corpuscular constants (iron), plasma alkaline phosphatase activity (zinc), serum ceruloplasmin concentration (copper), liver pyruvate carboxylase activity (manganese) and serum thyroxine concentration (iodine). On the other hand, when TE-5 (0.008, 0.04 and 0.2ml/kg: x 0.2, x 1 and x 5 the usual clinical dose, respectively) was intravenously administered once a day for 7 weeks under the conditions described above, there was a tendency to prevent the reductions of plasma and various tissue concentrations of iron, zinc and manganese. In addition, TE-5 prevented the reductions of growth rate, iron metabolism functions, plasma alkaline phosphatase activity, serum ceruloplasmin concentration and liver pyruvate carboxylase activity. The present study shows that TE-5 prevents both reductions of trace element contents and trace element-related functions, and suggests that TE-5 is useful for treatment of trace element deficiency in TPN.


[Effect of cadmium on lipid components: relation of cadmium to thyroid hormone and growth hormone]. To clarify the relationship of cadmium (Cd), thyroid hormone (TH) and growth hormone (GH) to lipid components, 4-week-old SD rats were dosed orally with Cd (CdCl2) at a dose of 2.0 mg/kg body weight five times a week, orally with TH at a dose of 2.5 mg/kg body weight five times a week and subcutaneously with GH (somatotropin) at a dose of 1.0 IU/kg body weight three times a week, all for 4 weeks. As lipid components, the serum concentrations of triglycerides, free fatty acids, lipid peroxides and long-chain fatty acids were determined. We have devised a new method for determining the fatty acid composition in the femur using gas chromatography-mass spectrometry and made a simultaneous analysis of fatty acids, from myristic acid (C14:0) to cholesterol. The results of the present study led to the following conclusions. 1. Cd may inhibit lipogenesis by binding with SH of coenzyme A, thereby reducing the serum levels of free fatty acids and lipid peroxides. 2. When TH and Cd were administered in combination, the addition of Cd produced an inhibitory effect on lipid components, although TH given alone stimulated the lipid metabolism. Therefore, Cd and TH may have an interaction in lipid components. 3. When GH and Cd were administered in combination, Cd modulated the action of GH, which enhanced the effect of somatomedin on the lipid metabolism. The inhibitory effect of Cd on somatomedin activity via Zn was suggested. 4. A sex difference was found in the composition of fatty acids in blood. The males had higher proportions of palmitic acid (C16:0) and linoleic acid (C18:2), while the females had a higher proportion of arachidonic acid (C20:4). There was no sex difference in fatty acid composition in the femur. 5. It was confirmed that TH produced a peroxide of dehydrocholesterol, a precursor of vitamin D3, in the diaphysis of the femur in the increased metabolic state.


[The effect of noise on sleep--changes in hypnograms of subjects exposed to repeated truck-passing sound]. The effect of traffic noise on sleep was evaluated in an experimental room using repeated recorded truck-passing sounds. The peak sound level was 55, 60 or 65 dB (A) with frequencies of three times per hour. The background noise of the exposure night was Leq 42 dB (A) and that of the control night was Leq 35 dB (A). The subjects were five students 23 to 24 years old. The sleep stage of each epoch with a 20-second duration was judged visually based on the criteria of Rechtschaffen & Kales and the data of the second night of noise-exposure and the control night were used. The sleep parameters used were total sleep time (TST), each sleep stage, % of sleep stage against TST, sleep efficiencies, sleep latency, REM latency, REM cycle, REM duration, waking from sleep, number of stage shifts, and subjective sleep judged by the OSA sleep inventory. A paired t test was used for the statistical analysis. TST, REM stage in minutes, and % of REM stage against TST of the exposure night were significantly decreased compared with those of the control night. Other objective and subjective parameters showed no changes during noise exposure. Dividing the all-night into halves, the amount and % of each sleep stage were compared between the exposure and control night. There was no change of sleep stage in the former half, but the % and amount of REM sleep in the latter half were significantly decreased as a result of the noise exposure. The decreases of REM sleep and % REM against TST, especially in the latter half of all-night sleep, were caused by the intermittent truck-passing sounds 20 to 30 times per night.


[Peripheral airway obstruction and treatment]. The effect of medication on peripheral airway obstruction was examined in cases of bronchial asthma. Subjects were 1) patients with exercise-induced asthma, 2) an animal model of hyperventilation-induced asthma and 3) patients with chronic asthma. Peripheral airway obstruction was induced in 30 of 51 patients with exercise-induced asthma. Induction of peripheral airway obstruction was protected significantly by procaterol. Cromoglycate was effective in 12 of 17 patients but ipratropium was not effective against induction of peripheral airway obstruction. In the animal model, humid air inhalation, procaterol and ipratropium completely prevented hyperventilation-induced bronchoconstriction, but cromoglycate caused only partial prevention. In cases of chronic asthma with peripheral airway obstruction, beclomethasone inhalation reduced the symptom rating rapidly, but no changes were observed in pulmonary function and threshold of airway responsiveness. Cromoglycate was started in patients with chronic asthma who had been treated with beclomethasone. After cromoglycate administration, therapeutic rating decreased, but increased again after 8 weeks of cromoglycate therapy. Peripheral airway obstruction induced by exercise or hyperventilation could be prevented by adequate premedication, but chronic peripheral airway obstruction was difficult to treat.


[Circadian rhythm in the treatment of asthma]. The circadian rhythm of respiratory function was examined in normal males and asthmatic subjects. In both groups, maximal flow decreased in the early morning. The major factor contributing to the circadian change in V50 was lung elastic recoil pressure in normal males and upstream resistance in asthmatic subjects. In some asthmatic patients, beclomethasone inhaled in the afternoon and at night was more effective than in the morning in increasing both the circadian maximal and minimal peak expiratory flow rates. This indicates the importance of chronotherapy in asthma.


[Features and treatment of immediate and late asthmatic responses]. In atopic asthma, the antigen-induced airway reaction consists of three phases; (1) immediate asthmatic response, LAR, (2) late asthmatic response, LAR, and (3) post-late asthmatic response, pLAR. Election microscopic examination of biopsy specimens of the bronchial mucosa, and examination of desquamated bronchial epithelial cells in sputum after antigen challenge showed that eosinophil infiltration in the bronchial epithelium contributes to airway hyperresponsiveness in asthma. Since LAR and pLAR are considered to be inflammatory responses caused by eosinophil infiltration, suppression of the inflammation by anti-allergic drugs and corticosteroids is effective in the treatment of LAR and pLAR. PAF antagonists and immunosuppressive agents have been investigated for their use in asthma treatment because of their anti-inflammatory effects. In addition to these anti-inflammatory agents, an antagonist of leukotriene C4 and D4 which are potent bronchoconstricting mediators has been shown to suppress both IAR and LAR, suggesting its possible use in asthma treatment.


[Clinical study of synchronous multiple primary cancer involving the lung]. Ten cases of synchronous multiple primary cancer involving the lung were experienced in our hospital during the period from January 1962 to May 1987. The incidence rate was 1.3% of all hospitalized cases of primary lung cancer. The counterpart organs were thyroid gland (4 cases), stomach (2 cases), colorectum (2 cases) and lung (2 cases). Three of four thyroid cancer cases were resected at the same time of pulmonary resection, whereas the stomach and colorectal cancer cases underwent two stage operation. Lung perfusion scintigraphy was very useful in determining the operative method for synchronous multiple primary lung cancer. The 5-year survival rate of all cases of synchronous multiple primary cancer involving the lung was 26.3%, with no hospital death. Surgical treatment is recommended for such cases when both cancers are expected to be resected completely.


[The effects of oxygen and vasodilators on pulmonary hemodynamics and blood gases in chronic lung disease]. The effects of oxygen inhalation, nitroglycerin (NTG) and prostaglandin E1 (PGE1) on pulmonary hemodynamics and blood gases were studied in 15 patients with chronic lung disease (CLD). Cardiac catheterization was performed with Swan-Ganz catheter, and pulmonary hemodynamics and cardiac output were measured. After baseline hemodynamics and blood gas measurements were made, 15 patients inhaled oxygen for 15 minutes and the same measurements were repeated. Twenty minutes later, oxygen inhalation was stopped. Then the 15 patients were divided into two groups; one group was administered sublingual NTG (7 patients) and the other was administered PGE1 by continuous intravenous infusion (8 patients). Pulmonary hemodynamics and blood gas measurements were then performed. Oxygen inhalation significantly reduced mean pulmonary artery pressure (from 22.2 +/- 8.8 to 20.0 +/- 6.4 mmHg; p < 0.01) and heart rate (from 78.1 +/- 12.0 to 75.5 +/- 12.5 beats/min; p < 0.05). With respect to oxygenation, oxygen inhalation significantly increased PaO2 (from 68.6 +/- 10.7 to 113.4 +/- 31.4 mmHg; p < 0.01), PvO2 (from 35.1 +/- 3.7 to 38.0 +/- 3.3 mmHg; p < 0.01). Therefore, oxygen inhalation was thought to be useful in patients with chronic lung disease with pulmonary hypertension. Sublingual administration of NTG significantly reduced mean pulmonary artery pressure (from 24.1 +/- 0.2 to 17.6 +/- 6.8 mmHg; p < 0.01), C.I. (from 2.9 +/- 0.2 to 2.3 +/- 0.2 ml/min/m; p < 0.01), O2-transport (from 589.1 +/- 168.4 to 457.0 +/- 105.8 ml/min; p < 0.01), and had a tendency to decrease PvO2 (from 34.3 +/- 3.0 to 32.1 +/- 1.8 mmHg; p < 0.1).(ABSTRACT TRUNCATED AT 250 WORDS)


[Six cases of primary pulmonary cryptococcosis]. Six patients with asymptomatic primary pulmonary Cryptococcosis are reported. In all of the patients, the disease was detected by annual chest X-ray during mass screening for lung cancer or during follow-up for pulmonary tuberculosis or gastric cancer. The chest X-ray findings consisted of a solitary pulmonary nodule in 4 patients and multiple pulmonary nodules in 2. Only one patient who could not be histologically diagnosed by bronchofiberscopy underwent surgical resection. However, the other 5 patients were histologically diagnosed by transbronchial biopsy with bronchofiberscopy. They were treated with oral antifungal agents, namely flucytosine (5-FC) and/or fluconazole, with marked improvement of chest X-ray findings. These results indicate that transbronchial biopsy with bronchofiberscopy and oral administration of antifungal agents instead of initial surgical resection are useful in the diagnosis and treatment of primary pulmonary cryptococcosis.


[Relation of airway obstruction and respiratory muscle weakness to energy metabolism in pulmonary emphysema]. Resting energy expenditure (REE) was assessed and its relationship to nutritional status, pulmonary function and respiratory muscle function was studied in 25 patients with pulmonary emphysema. The mean value of REE was 1413 +/- 251 Cal and the ratio of REE/REEpred was 1.398 +/- 0.23, suggesting the existence of a hypermetabolic state in these patients. REE/REEpred ratio was inversely correlated with plasma amino acid BCAA/AAA ratio and body muscle mass; and PImax. REE was inversely correlated with FEV1.0%. REE in the patient subgroup with severe hyperinflation (%RV > or = 200) was significantly higher than that in the subgroup with moderate hyperinflation. Malnourished patients showed significantly more severe hyperinflation than well-nourished patients. These findings suggest that augmented REE contributes to malnutrition in patients with emphysema, and that the increase in REE was related to the increase in mechanical work load on the basis of airway obstruction, respiratory muscle weakness and hyperinflation.


[Characteristics of chronic interstitial pneumonia seen in the lung operated for lung cancer]. A total of 480 lungs operated for lung cancer were reviewed in an attempt to clarify the characteristics of idiopathic chronic interstitial pneumonia (UIP), seen in the cancer bearing lungs. UIP was identified in 30 cases (6.3%). The mean patients age was 68 years, and 26 cases were male. Squamous cell carcinoma was the dominant tumor type (17 cases) and the lower lobe was the dominant location (18 cases). All cancers were situated in the periphery of lungs which had UIP, but not necessarily in the center of the fibrosis. According to extent, UIP is divided into two categories, localized and diffuse types. Twenty-seven cases had localized type UIP. UIP can be divided into another three categories, according to its microscopic findings; Intramural (IM), emphysematous (E), and intraluminal (IL) types. Eight cases had IM-type and 22 cases had E-type UIP. Some IM-type UIP cases showed only increased density of the subpleural space on CT, without visible cysts. E-type UIP cases had no decrease of lung capacity, and showed various sized cysts on CT. Some cases showed postoperative worsening of UIP and acute exacerbation. We feel that the existence of localized UIP (especially emphysematous type) is important in the early detection of cancer, and as a cue for careful post-operative management to prevent serious complications.


[Measurement of CA19-9 in bronchial lavage fluid from patients with lung cancer]. Carbohydrate antigen 19-9 (CA19-9) was measured in bronchial lavage fluid in 21 patients with lung cancer and 4 patients with benign lung diseases (2 patients with DPB, 2 patients with BE). Bronchial lavage of the tumor-related bronchus was performed. In normal subjects, levels of CA19-9 in lavage fluid were less than 1000 IU/ml. On the other hand, in 6 patients with lung cancer, levels of CA19-9 were higher than 1000 IU/ml, and in 3 of these cases, CA19-9 levels were higher than 8000 IU/ml. All six three cases were histologically diagnosed as adenocarcinoma. Tumor resected at operation was then stained by antibody recognizing CA19-9. Tumor in cases with high levels of CA19-9 was stained immunohistochemically. These results indicate that measurement of CA19-9 in bronchial lavage fluid of the tumor-related bronchus is a useful auxiliary method in the diagnosis of lung cancer.


[The role of bronchial responsiveness to leukotrienes in antigen-induced immediate and late asthmatic response]. In order to clarify the role of leukotriene in the antigen-induced asthmatic response, we carried out mite-antigen, leukotriene D4 (LTD4) and histamine inhalation tests in 21 mite-sensitive atopic asthma patients, and investigated the relationship between the bronchial responsiveness to mite (PD20-Mite) and that to LTD4 and histamine (PD20-LTD4, PD20-Hist). We then compared PD20-LTD4 and PD20-Hist between the groups with and without late asthmatic response (LAR), and investigated the relationship between the peak percentage fall of FEV1.0 in LAR (magnitude of LAR) and PD20-LTD4 and PD20-Hist. There were significant correlations between PD20-LTD4 and PD20-Mite (r = 0.590, p < 0.01), and between PD20-Hist and PD20-Mite (r = 0.669, p < 0.001). The bronchial responsiveness to LTD4 and histamine was significantly higher in the group with LAR. In addition, PD20-LTD4 showed a significant negative correlation with the magnitude of LAR (r = 0.724, p < 0.001). Furthermore we analyzed the relationship between PD20-LTD and the ratio of the magnitude of LAR to that of IAR (immediate asthmatic response), because there was a significant correlation between the magnitude of IAR and that of LAR (r = 0.842, p < 0.001) in 16 asthmatic patients with dual asthmatic response, and found that the correlation coefficient was also very high (r = -0.836, p < 0.001). These results suggest an important role of leukotriene in the antigen-induced asthmatic response.


[Sputum leukotrienes in chronic airway diseases and bronchial asthma attacks]. To determine the pathogenetic role of leukotrienes in chronic airway diseases, sputum samples from patients with bronchial asthma (BA), diffuse panbronchiolitis (DPB), sinobronchial syndrome (SBS), and bronchiectasis (BE) were examined for the presence of leukotrienes (LT) C4, D4, E4, B4, all apparently derived from airway inflammatory cells. Sputum samples from 21 patients, including 10 with BA attack (5 atopic type, 5 non-atopic type) and 11 with chronic obstructive airway diseases (3 DPB, 6 SBS, and BE), were studied. Patients expectorated sputum directly into test-tubes containing 80% ethanol. Following ethanol extraction and partial purification by a C18 SEP-PAK column, LTs were further purified by high performance liquid chromatography (HPLC). Fractions from HPLC with elution times corresponding to synthetic LTC4, D4, E4, and B4 were used to quantify LTs by radioimmunoassay. Eosinophils and neutrophils in the sputum (0.5 ml) were counted following Papanicolaou staining. Percentages of eosinophils in sputum were higher in BA, while those of neutrophils were higher in DPB. Sputum levels of LTC4 were 1.2 +/- 1.6 in BA, 0.12 +/- 0.11 in SBS, and 0.42 +/- 0.14 ng/ml in DPB. Those of LTD4 were 0.21 +/- 0.27 in BA, 0.9 +/- 0.13 in DPB, and 0.10 +/- 0.07 in SBS. LTE4 levels were 5.06 +/- 3.83 in DPB and 2.66 +/- 4.32 in BA. Levels of LTB4 were 1.36 +/- 1.19 in DPB, 0.28 +/- 0.27 in BA, 0.12 +/- 0.07 in SBS, and 0.04 +/- 0.04 in BE. In asthmatics, peptide leukotriene content in sputum was higher than that in chronic airway disease patients.(ABSTRACT TRUNCATED AT 250 WORDS)


[Correlation of signal intensity of blood flow in the pulmonary artery on MR images and clinical features of two cases of primary pulmonary hypertension]. ECG-gated spin-echo MR images of the chest were obtained in two patients with primary pulmonary hypertension (PPH). In transverse section at the level of the right main pulmonary artery (rPA), flow signals in the rPA were quantitatively evaluated, and the correlations with MR signal intensity of intravascular flow and data of routine clinical examinations and the severity of clinical manifestations were studied. In one case, the signal intensity of flowing blood markedly increased with exacerbation of hypoxemia and other clinical manifestations. However, in the other case with a stable course, the signal intensity of intravascular flow did not change significantly. Increase of flow signal in the rPA can reflect decrease of flow velocity that may be cause by a state of high pulmonary vascular resistance or low cardiac output. Therefore, it is suggested that MRI is a useful modality to evaluate the severity of disturbance of the pulmonary circulation in PPH.


[Two probable cases of adult Williams-Campbell syndrome--advocation of Williams-Campbell-type bronchiectasis]. Williams-Campbell syndrome is a unique type of bronchiectasis which shows ballooning during expiration and collapse during inspiration. Its etiology is thought to be a congenital decrease in bronchial cartilage. We experienced two adult cases of Williams-Campbell syndrome. Since the mucociliary transport system was normal in both cases and neither case had chronic sinusitis, these cases differed from sino-bronchial syndrome. Cases of Williams-Campbell syndrome reported in Japan show characteristic bronchography, but a decrease in bronchial cartilage may or may not be present and its congenital nature is very difficult to demonstrate. We therefore advocate referring to such cases of bronchiectasis with these bronchographic characteristics Williams-Campbell-type bronchiectasis, even if a decrease in bronchial cartilage is not observed.


[Thrombosis-inducing activity (TIA) in plasma of a patient with lung cancer, which disappeared after radiotherapy]. A 58-year-old man had been suffering from persistent right shoulder pain was admitted following detection of a Pancoast tumor in the right lung. On admission, there was elevation of peripheral platelet count, plasma FDP and fibrinogen levels and thrombosis-inducing activity (TIA) was detected in his plasma. After receiving 30 Gy of Liniack, the mass shadow and the shoulder pain disappeared, which was accompanied by the normalization of laboratory data and the disappearance of TIA from plasma. However, plasma TIA became positive again one month later, in association with reappearance of shoulder pain and elevation of FDP and fibrinogen levels, which were considered due to recurrence of the tumor, 30 Gy of Liniack together with thermotherapy was again given. There was no subsequent recurrence of tumor, nor TIA has been detected for more than 3 years.


[Congenital absence of the right main pulmonary artery--a case report]. A 32-year-old man was admitted to hospital for investigation of bloody sputum. Chest roentgenogram showed a decrease in the volume of the right lung, with mediastinal shift to the right. Chest CT examination revealed narrowing of the right main pulmonary artery. Bronchofiberscopic examination was normal. Pulmonary arteriogram demonstrated complete absence of the right pulmonary artery. Cardiac catheterization showed no concomitant cardiac malformation. Therefore, this case was diagnosed as congenital absence of the right main pulmonary artery.


[A case of Hodgkin's disease with endotracheal tumor presenting with severe airflow obstruction]. A 22-year-old woman was admitted to our hospital complaining of productive cough and dyspnea even at rest, and marked cervical lymphadenopathy. Marked stridor and orthopnea were observed, and auscultation of the chest revealed widespread expiratory wheeze which was not relieved by bronchodilators administered intravenously. Chest X-ray and CT scan revealed hilar lymphadenopathy and invasive tumor of the mediastinum. Bronchoscopy demonstrated narrowing of the trachea anteriorly and posteriorly and a submucosal nodular tumor protruding from the right anterior wall, causing approximately 90% occlusion of the lumen of the lower third of the trachea, but distal bronchi were intact. Microscopic findings of inguinal lymph node biopsy specimen revealed mixed cellular lymphoma compatible with Hodgkin's disease. Systemic chemotherapy resulted in relief of symptoms, and two months later, the endotracheal tumor had disappeared bronchoscopically, with slight residual stenosis of the trachea. Before treatment, pulmonary function tests indicated markedly impaired forced volume in 1 second in both expiratory and inspiratory cycles, especially in the latter phase. After remission, however, obstructive ventilatory dysfunction was observed. The cause of prolonged air flow obstruction was thought to be marked infiltration and almost total involvement of the tracheal wall by tumor with a nodular appearance of the lumen. Endotracheal tumor in Hodgkin's disease is rare, and there are few reports on pulmonary function associated with intrathoracic involvement of malignant lymphoma.


[A case of bucillamine-induced interstitial pneumonia]. Bucillamine [N-(2-mercapto-2-methylpropionyl)-L-cysteine] is an anti-rheumatic drug which was developed in Japan. Interstitial pneumonia induced by bucillamine has been reported in two patients with rheumatoid arthritis. In this report, we describe another patient who developed bucillamine-induced interstitial pneumonia. In August 1990, a 53-year-old man was admitted because of fever, dry cough and dyspnea on exertion (DOE). Five months previously, he had noted polyarthritis, and treatment with bucillamine was started in May 1990. His polyarthritis improved about 2.5 months after starting bucillamine, but he developed fever, dry cough, and DOE in August 1990. Chest X-ray on admission showed diffuse acinar and interstitial shadows predominantly in the bilateral upper lung fields. Pulmonary function tests revealed decreased vital capacity and diffusing capacity. Arterial blood gas (ABG) analysis revealed moderate hypoxemia with PaO2 67 Torr. After stopping bucillamine, the fever, dry cough, and DOE improved gradually, and the abnormal shadows on chest X-ray and ABG analysis showed moderate improvements. Bronchoalveolar lavage studies showed that total cell counts and proportion of lymphocytes were increased, and CD4+/CD8+ ratio of T cell subsets was decreased to 0.56. Transbronchial lung biopsy specimen revealed lymphocytic alveolitis and mild interstitial thickening. Lymphocyte stimulation test to bucillamine was negative, but patch test with bucillamine was positive. From the patient's clinical course, laboratory data, and pathologic findings, we concluded that this is a case of bucillamine-induced interstitial pneumonia. After treatment with corticosteroid, his chest X-ray and pulmonary function test showed marked improvements.(ABSTRACT TRUNCATED AT 250 WORDS)


[Two cases of idiopathic pulmonary hemosiderosis: analysis of chest CT findings]. Chest CT findings are reported in two cases of idiopathic pulmonary hemosiderosis. In both cases, CT was performed after remission of an acute exacerbation following corticosteroid therapy. Case 1 was a 17-year-old woman with Down's syndrome. Chest radiograph showed diffuse ground-glass like and reticulonodular shadows, which were predominant in the bilateral lower lung fields. Chest CT showed a diffuse increase of lung filed density, especially in the dorsal zone of both lower lobes. Open lung biopsy revealed hemorrhage and numerous hemosiderin-laden macrophages in the alveoli, and in addition, marked fibrous thickening of the alveolar septa. Case 2 was a 7-year-old girl. Chest radiograph showed diffuse micronodular shadows in both lungs. Chest CT showed diffuse poorly-circumscribed micronodular lesions with uniform distribution, and lung field density was normal except for the right upper lobe with patchy infiltrates. Although lung biopsy was not performed, fibrous thickening of the alveolar septa was presumed to be mild even if present, since pulmonary function and blood gas analysis were within normal limits. In these two cases, lung field density of CT seemed to reflect the degree of diffuse fibrous thickening of the alveolar septa, and it is suggested that CT is valuable in the evaluation of fibrous thickening of the alveolar septa secondary to recurrent pulmonary hemorrhages. Comprehensive review of CT findings of idiopathic pulmonary hemosiderosis was also performed.


[Sudden death due to bilateral pulmonary thromboembolism in a patient with multiple myeloma: an autopsy case report]. We encountered a patient with multiple myeloma who died suddenly, in whom bilateral pulmonary artery thrombosis was found at autopsy. The patient was a 50-year-old woman who had received chemotherapy for multiple myeloma at a local clinic for 4 years, and was transferred to our hospital because of recurrence of multiple myeloma in August 1990. Despite chemotherapy performed after admission, serum globulin level increased and her low back pain worsened, and she was generally restricted to bed. On October 21, she developed sudden dyspnea, became markedly cyanotic, lost consciousness, and then suffered a cardiopulmonary arrest. Autopsy revealed bilateral pulmonary artery thrombosis as well as thrombosis in the pelvic veins. Hyperviscosity due to multiple myeloma and long-term recumbency were the probable causes of pelvic venous thrombosis and subsequent pulmonary artery thrombosis. We report a rare case of bilateral pulmonary artery thrombosis which developed during the course of multiple myeloma and led to sudden death.


[A case of superior vena cava syndrome due to extensive venous thrombosis of an idiopathic nature]. A 37-year-old man was admitted with facial edema and right arm swelling. Venography, computed tomography and magnetic resonance imaging showed massive organizing thrombi in the superior vena cava and bilateral internal jugular as well as subclavian veins, and showed no mass lesions occluding the veins in the mediastinum. Angioscopy demonstrated a white thrombus at the entrance of the right subclavian vein. All results of blood coagulation tests were normal. The patient was diagnosed as having superior vena cava syndrome caused by idiopathic venous thrombosis. Anti-coagulant therapy with intravenous tissue plasminogen activator injection and continuous urokinase and heparin infusion into the thrombi through a catheter were not effective in lysing the thrombi. Collateral circulation gradually developed and his symptoms decreased. We decided to follow this patient on warfarin medication because of the difficulty in removing the thrombi surgically.


[A case of endoscopic laser polypectomy for endobronchial hamartoma]. A 70-year-old man was admitted to our hospital because of left-sided chest pain and dyspnea. Chest X-ray film showed a tumor in the left main bronchus and left pleural effusion. On bronchoscopy, a tumor which moved with respiration was observed in the left main bronchus. We successfully performed endoscopic laser polypectomy. Pathological examination revealed non-chondromatous hamartoma.


[A case of pulmonary alveolar proteinosis and disseminated atypical mycobacteriosis complicating chronic myelogenous leukemia]. A 47-year-old woman with chronic myelogenous leukemia was treated with daily busulfan (total dose approximately 500 mg) from December 1988 to January 1990. The disease thereafter remained stable with no evidence of blastic transformation. In February 1990 she developed productive cough and abnormal acinar lung shadows appeared transiently on her chest X-ray. In October 1990, productive cough and linear and abnormal acinar lung shadows reappeared. Expectorated sputa contained acid-fast bacilli (Gaffky 6, 10). Antituberculous therapy was started, which caused severe liver dysfunction. She was admitted to our hospital for evaluation of abnormal lung shadows. Transbronchial lung biopsy revealed pulmonary alveolar proteinosis with thickening of alveolar septa. The alveolar septal thickening was suspected to be a pathological change following pulmonary alveolar proteinosis. Cultures from sputum, cerebrospinal fluid, and bone marrow aspiration specimens revealed atypical mycobacterium (M. avium complex), and the diagnosis of disseminated atypical mycobacteriosis was established. The pathogenesis of the disseminated atypical mycobacteriosis was considered to be superinfection by mycobacteria.


[A case of anomalous course of the pulmonary vein with a tracheobronchus]. A 28-year-old man was admitted to our hospital for further evaluation of a loop-like abnormal shadow in the middle lower lung field on chest X-ray film. Pulmonary angiograms demonstrated staining of the abnormal shadow during the venous phase. The abnormal vein descended from the right upper lobe to the middle lobe and then tortuously made a loop upward to drain into the left atrium. A part of the right apical vein was stenosed. The right apical segmental bronchus directly branched from the trachea. These observations indicate that the abnormalities may have originated in the prenatal period. Cases of anomalous course of the pulmonary vein are very rare.


The diagnosis and treatment of Lyme disease. Lyme disease has probably been reported for at least 80 years, with the cause unknown until the discovery of the Ixodes ricinus complex tick-borne spirochete B. burgdorferi. The incidence of diagnosed cases appears to be increasing in Kansas and elsewhere. A negative laboratory test does not exclude the possibility of the disease. At present, no accurate laboratory screen for the illness is available. The diagnosis must be considered when acute or chronic clinical manifestations reviewed in this paper appear in patients. Additional clinical criteria and modified CDC guidelines are proposed to help clinicians determine who should be treated. Current treatment recommendations are presented.


[The treatment of joint cicatricial lesions of the esophagus and stomach]. Treatment of 108 patients with concurrent cicatricial stenosis of the esophagus and stomach is analysed. Various operations were performed on the stomach in 70 patients. The patency of the esophagus was restored by forced bougienage and intubation. The late-term results were studied in 48 patients in follow-up periods of 2-13 years. Forty-two patients feel quite well and are engaged in physical work, they live on a common diet. Six have to chew their food thoroughly. Two patients died in the postoperative period.


[Pylorus-preserving gastric resections in gastroduodenal ulcers]. The article analyses the results of pylorus-preserving resections of the stomach in 116 patients among whom 80 had gastric ulcer, 29 ad duodenal ulcer, and 7 had gastroduodenal ulcers. Changes of acidity and motor-evacuation function of the gastric stump were studied according to the method of pylorus-preserving resection. The late-term results of treatment in follow-up periods of 2 to 5 years are shown, they were found to be good in 96.6% of patients treated by operation and poor in 1.7%. It is noted that the function of the gastric stump is better when the vagus innervation of the pylorus is preserved. Mediogastric resection with selective proximal vagotomy in patients suffering from duodenal ulcer with a superacidic stomach led to stable normalization of acid production, which was a reliable measure for the prevention of a recurrent ulcer.


[The use of enteral tube feeding in patients with gastric and duodenal peptic ulcer]. The authors used enteral tube feeding with a multicomponent balanced mixture "Ovolakt" in complex treatment of gastric and duodenal ulcer in 69 patients among which 42 had complicated forms (stenosis, penetration, bleeding) and 27 had no complications. In establishing the diagnosis laboratory tests and radiological and endoscopic examination were conducted. The gastric secretory function was studied by intragastric pH-metry and pH-chromoscopy conducted during enteral feeding and in dynamics after it was ended. In most cases of cicatrization of the gastric and duodenal defects, hypo- and hyperchlorhydria respectively, persisted. Among 42 patients with complicated course of peptic disease, 21 were treated by operation and 19 received nonoperative treatment. The postulcer cicatrix formed 2-3 weeks after enteral feeding was ended in 27 cases of uncomplicated peptic ulcer and in 19 patients with complicated ulcers who were given nonoperative treatment. Complications after operative treatment were not encountered. Patients not treated by operation spent 20-30 days in the hospital.


[Emergency surgery of the esophagus].

[Emergency surgery of the esophagus]. Emergency surgery of the esophagus is undertaken in acute diseases and injuries to the esophagus: among 926 cases of acute pathology of the esophagus 709 were mechanical ruptures of its wall, 380 z (53.6%) of an instrumental character. A rational therapeutic tactics developed at the Institute and basically new methods of treatment allowed the results to be essentially improved and the mortality rate in traumatic purulent mediastinitis to be reduced from 35% to 12%. The total mortality rate in acute surgical pathology in the last 20 years was 9.5%.


[The restoration of the natural intestinal passage after Hartmann's operation in a short stump of the rectum]. Analysis showed restorative-reconstructive operations in patients who underwent Hartmann's operation with preservation of a short rectal stump to be technically difficult and injurious interventions. The interventions must be undertaken only after complete removal of the causes of suppuration of the colostomy in a patient whose condition is satisfactory. The degree of the cicatricial process in the pelvis and the method of the colorectal anastomosis formation essentially influence the results of the restorative-reconstructive operations. The best results were produced in retrorectal pull-through of the colon with the formation of colorectal anastomoses after the type of Duhamel's operation.


[A comparative evaluation of different methods of forming a colonic anastomosis in repeat surgical interventions]. The results of repeated operations on the large intestine, mainly reconstructive operations, are analysed. Surgical interventions with the performance of a triangular anastomosis, including that by a method modified by the authors, were carried out on 50 patients. The operative results, particularly those in the last group of patients, are very promising. The number of complications, such as suppuration of the postoperative wound and incompetence of the anastomosis sutures, reduced from 28.8% to 10%, which allows the method to be recommended for wide application in the clinic.


[The potentials of computed tomography in the treatment of cysts and inflammatory diseases of the internal organs]. Computed tomography (CT) is used more and more often recently in the treatment of some surgical diseases of the internal organs. The subject of discussion in this work are the results of treatment of cysts and pyoinflammatory diseases of the internal organs. The work was conducted on CT IV generation "QUAD 1" of the CMS firm (England). Needles measuring 14-18d in diameter (Charrière scale) were used for aspiration. A total of 23 therapeutic interventions were performed in 19 patients with exudative pleurisy (1 patient), pleuritis (4), pulmonary abscesses (1), abscesses of the liver (1), abscesses of the subhepatic and subdiaphragmatic spaces (2), cysts of the liver (1) and kidneys (1 patient). Among the interventions 21 were aspirations and 2 drainage of the abscess. A mechanical syringe is used lately for more effective evacuation of the contents. The removed contents, ranging in volume from 10.0 to 600.0 ml, was subjected to cytological, biochemical, and bacteriological analysis. A single puncture is sufficient, as a rule, for aspiration of the cyst contents, one-stage aspiration from cysts of both kidneys may be conducted (as was the case in two of our patients). 96% alcohol was used as the sclerosing agent. In cases of small abscesses aspiration may be repeated for removal of the contents, irrigation of the cavity, and administration of antibiotics. In abscesses measuring more than 10 cm drainage is advisable. Control CT examination is conducted once a week. The condition of the cavity is appraised by combining CT with its contrast study. The immediate results were good and satisfactory in 17 (94.7%) patients.(ABSTRACT TRUNCATED AT 250 WORDS)


[Specific immunity in patients with inflammatory diseases of the abdominal cavity]. In most patients with pyoinflammatory diseases of the abdominal cavity caused by Cl. perfringens microbial associations accumulation of agglutinins occurred during the disease. In purulent abdominal processes caused by Cl. perfringens microbial associations a connection exists between the increased antibody level and wound suppuration. The rise of the antibody level was most marked, whatever the character of the bacterial associations, in patients with generalized peritonitis, abdominal abscess, and in young patients (10-40 years of age) and least in those with phlegmonous cholecystitis and in elderly patients (61-70 years of age). Patients with destructive forms of appendicitis and individuals aged 41-60 took an intermediate position according to these values. Antibody production was weaker in patients with 5 species in the microbial association than in monoinfection. A agglutinin titer in the blood of patients beginning with 1:80 and higher should be considered diagnostic.


[Combined operations as a method for increasing the capacity of a surgical clinic]. The work shows the economical effect of concurrent surgery on the abdominal organs and organs of the retroperitoneal space undertaken in 404 patients whose ages ranged from 16 to 81. The time taken by the concurrent operations and the duration of treatment at the hospital were studied. In 63.1% of patients the concurrent operations lasted up to 3 hours. The duration of treatment was least in concurrent appendectomy (14.2 +/- 0.5 days) and longest on oncological patients (40.9 +/- 2.2 days). It is noted that the sum terms of treatment reduce, while in separate performance of the operations they are prolonged. The highest economy of finances is noted in urolithiasis, nephroptosis, acute appendicitis, hernias, and cholelithiasis. The national economic effect in the treatment of 404 patients at the clinic came to 60,471 rubles and 18 kopecks.


[Means for optimizing the conditions for the timely hospitalization of patients with acute surgical diseases of the abdominal cavity organs]. Complex research made it possible to reveal factors influencing timely hospitalization of patients by studying the causes of timely and late hospitalization, medical erudition of the urban and rural population, as well as the organization of emergency medical service in the prehospital stage. New methodical approaches to health education among the population as well as measures for raising the confidence of the sick in doctors of emergency aid service and surgeons are suggested. From the example of urban health service the authors emphasize the importance of the principle of necessary hospitalization of patients for profound examination in all cases of a doubtful diagnosis of an acute surgical disease.


[The mechanisms of the evacuatory function of the gastric stump after a Hofmeister-Finsterer resection]. Attention is drawn to the fact that evacuation from the gastric stump is determined not only by the size of the anastomosis, the manner by which it is formed, whether transversely or longitudinally but also by the functional condition of the gastric stump and the efferent intestinal loop. Motor-evacuation disorders on the part of the stump and efferent intestinal loop precede the development of most postgastric resection syndromes. The authors examined 197 patients who underwent Hofmeister-Finsterer gastrectomy. Five types of evacuation from the gastric stump were distinguished, to which various combinations of motor function of the stump and efferent intestinal loop corresponded. After studying the type of combinations of the motor function of the stomach and efferent intestinal loop, the authors conducted purposeful correction which promoted normalization of the motor function of these organs. For instance, evacuation in portions, which is most efficient in respect of function, was encountered in 42% of patients without correction and in 75.9% of those in whom correction was undertaken. This allowed postoperative functional complications to be reduced from 38% to 12.08% and the adaptation period to be shortened by 2-3 weeks on the average.


[Errors, hazards and complications in the treatment of patients with diffuse suppurative peritonitis]. Errors and complications encountered in the treatment of 208 patients with generalized purulent peritonitis are analysed. The importance of preoperative management of patients is shown, when properly conducted it improves the results of treatment. The author discusses tactical and technical errors which are encountered most frequently during the operation and lead to increase of the mortality rate. Rational postoperative management requires particular attention. The immediate results of treatment are improved when errors, hazards, and complications occurring in the treatment of patients with generalized purulent peritonitis are taken into account.


[The use of methyldioxyline for the prevention of local complications following hemorrhoidectomy]. The article discusses the results of clinico-laboratory study of the application of methyl dioxyline ointment after hemorrhoidectomy with complete restoration of the anal canal mucosa. It is shown that the ointment is a reliable measure for the prevention of postoperative purulent complications, it improves the reparative processes. Its use in the treatment of wounds developing as the result of excision of hemorrhoids shortens the term of in-patient treatment after the operation and the period of general incapacity.


[A method of preparing the large intestine for surgery by using the peroral administration of a polyethylene oxide solution]. A new method for preparing the large intestine for operation is suggested, it is based on oral administration of a solution of polyethylene oxides with chloramphenicol. The preparation possesses a marked hydrophilic activity, which causes inhibition of absorption processes in the lumen of the intestinal tube. This leads to accumulation of the intestinal contents and their quicker passage. Polyethylene oxides also ensure accumulation of antibiotics in the tissues of the intestinal wall. The suggested method was used to prepare the intestine for operation in 93 patients. Comparative evaluation proved its advantages according to the main clinico-laboratory criteria: the method is technically simple, reduces the period of preparation of the intestine, has no negative effect on the homeostasis values, ensures adequate preparation of the large intestine with decrease of the level of its microbial contamination. With the use of the method the number of postoperative purulent complications in interventions on the large intestine reduced.


[The modelling of diffuse peritonitis].

[The modelling of diffuse peritonitis]. Peritonitis is still an urgent problem and its solution is determined to a great measure by purposeful studies in the field of pathogenesis and treatment. The creation of adequate models of peritonitis acquires particular significance. Eighty-four animals (dogs) were used to create the model. It is shown in the work that a standard model of generalized peritonitis can be produced by two injections of a polymicrobial suspension consisting of the most commonly encountered peritoneal microbes E. coli and B. fragilis. The suspension is injected into different levels of the abdominal cavity. With this method of administration of the polymicrobial suspension the model was reproduced in 90% of cases. Thus, the developed model resembles human peritonitis in the clinical picture and the laboratory and morphological values.