Preferential localisation of elastase in cytosol of human myeloid leukemia HL-60 cells.

The subcellular distribution of the elastase in human myeloid leukemia HL-60 cells was studied in comparison with that in normal leukocytes. On differential centrifugation, most of the elastase activity of HL-60 cell lysates was recovered in the 105,000 x g supernatant, while that of human peripheral blood leukocyte lysates was recovered in the 500 x g precipitate (azurophil granule-rich fraction). Moreover, on Percoll density gradient centrifugation, the elastase activity in HL-60 cell extracts was recovered in the lightest fraction with none in the azurophil granule-rich fractions, whereas most of the activity in leukocyte extracts was recovered in the azurophil granule-rich fractions. This subcellular localization of elastase did not change when HL-60 cells differentiated into monocytes and granulocytes by induction with 12-O-tetradecanoyl phorbol-13-acetate and retinoic acid, respectively. Furthermore, on Sephadex G-75 gel filtration, the elastase activity in HL-60 cell extracts was eluted earlier than that in leukocyte extracts. The size estimation indicated that the elastase of HL-60 cells was 36-30 kDa, corresponding to the size of an elastase precursor reported. The relevance of a large form of the elastase in HL-60 cells to its subcellular localization is discussed.     

A repeating unit of the DYZ1 family on the human Y chromosome consists of segments with partial male-specificity

An average-sized human Y chromosome contains about 3,000 copies of the repeating DNA family DYZ1. A major repeating unit of the family, pHY10, has been cloned and an entire 3,564-bp sequence has already been determined by Nakahori et al. (1986). In the present study, pHY10 was divided into six consecutive segments, A to F, which were independently amplified by the PCR technique to see if they were strictly male-specific. pHY10 appears to consist of segments of various male-specificity. The B segment was apparently male-specific; however, the use of additional techniques (Southern-blot analysis or second PCR amplification in combination with the standard PCR) revealed homologous sequences in some females. None of the six segments of pHY10 may be male-specific in a strict sense. Different segments appear to be conserved during evolution to different extents. The 323-bp E segment appears to be the least conserved and to be responsible for the generation of most variations within the DYZ1 family.     

Electrophoretic karyotyping and chromosomal gene mapping of Chlorella

Molecular karyotypes for six strains of four Chlorella species were obtained by using an alternating-field gel electrophoresis system which employs contour-clamped homogenous electric fields (CHEF). The number and migration pattern of the chromosomal DNA molecules varied greatly from strain to strain: for example, nine separated chromosomes of C. ellipsoidea C87 ranged from 2.5 to 6.5 megabase pairs (mbp) in size, whereas 16 chromosomes of C. vulgaris C169 were from 980 kilobase pairs (kbp) to 4.0 mbp. Depending on the chromosome migration patterns, the six strains were classified into two major chromosome-length polymorphism groups. Using hybridization techniques, the genes for alpha-tublin, chlorophyll-a, b-binding proteins, ribosomal RNAs, and the small subunit of ribulose-1, 5-bisphosphate carboxylase/oxygenase (RuBisCO) were mapped on the separated chromosomes of C. vulgaris C169. Since Chlorella chromosomes are small enough to separate and isolate individually by CHEF gel electrophoresis under ordinary conditions, they should serve as excellent materials to study the fundamental molecular structure of plant-type chromosomes.     

Neural coding of speech sound in the telencephalic auditory area of the mynah bird

Summary The responses of neurons in field L in the auditory neostriatum of the mynah bird, Gracula religiosa, were recorded during presentation of intact or manipulated mimic voices. A typical mimic voice konnichiwa elicited responses in most of the neurons. Neurons in the input layer (L2) of field L showed many peaks on peristimulus time histograms while those in other layers (L1 and L3) exhibited only one or two peaks. Several neurons in L1 and L3 responded only to the affricative consonant /t/ in the intact mimic voices. They did not respond to the affricative consonant in the isolated segment or to the one in the playbacked voice in reverse. Forty-five percent of the neurons (33/ 73) decreased in firing rates at the affricative consonant in the isolated segment compared with in the intact voice. Some of these neurons, in which neither the affricative consonant in the isolated segment nor bursts of noise alone elicited responses, exhibited clear phasic responses to /t/ in the case when bursts of noise with particular central frequencies preceded the affricative consonant. The responsiveness of these neurons appears to receive temporal facilitation. These results suggest that these neurons code the temporal relationship of speech sound.Abbreviations HVc hyperstriatum ventrale, pars caudale - TFN temporally facilitated neuron - TSN temporally suppressed neuron     

Mutual interactions between optic lobe circadian pacemakers in the cricket Gryllus bimaculatus

The coupling mechanism between the bilaterally paired optic lobe circadian pacemakers in the cricket Gryllus bimaculatus was investigated by recording locomotor activity, under constant light or constant red light, after the optic nerve was unilaterally severed.

Diagnosis of a case of pulmonary carcinosarcoma by detection of rhabdomyosarcoma cells in sputum

Cytologic examination of sputum samples from an elderly patient revealed the presence of two cell populations: squamous cell carcinoma cells and rhabdomyosarcoma cells. The abnormal squamous cells showed both keratinizing and nonkeratinizing forms while some of the rhabdomyosarcoma cells showed cross striations. Sputum cytology was thus able to suggest a diagnosis of pulmonary carcinosarcoma. Histologically, the tumor was composed mainly of sarcomatous tissue showing various kinds of cells: fusiform or fibrous cells, round anaplastic cells, spindled cells with typical cross striations and myoblastic cells. A partially myxomatous degeneration was present. In addition, squamous cell carcinoma proliferated along the bronchi and formed small invasive cell nests in the sarcomatous tissue. No transition between the two components was noted. Both cellular constituents had metastasized to an interlobar lymph node.     

Immunogenic protein MPB57 from Mycobacterium bovis BCG: molecular cloning, nucleotide sequence and expression

Using a single-probe method, we have cloned the gene for an immunogenic MPB57 protein of Mycobacterium bovis BCG. The nucleotide sequence includes an ORF of 300 base pairs encoding a protein of 99 amino acids with an Mr of 10,818. This cloned gene was expressed in an Escherichia coli expression vector to give a mature protein which reacted with a polyclonal antibody raised against MPB57.     

Structure of the horseradish peroxidase isozyme C genes

We have isolated, cloned and characterized three cDNAs and two genomic DNAs corresponding to the mRNAs and genes for the horseradish (Armoracia rusticana) peroxidase isoenzyme C (HPR C). The amino acid sequence of HRP C1, deduced from the nucleotide sequence of one of the cDNA clone, pSK1, contained the same primary sequence as that of the purified enzyme established by Welinder [FEBS Lett. 72, 19-23 (1976)] with additional sequences at the N and C terminal. All three inserts in the cDNA clones, pSK1, pSK2 and pSK3, coded the same size of peptide (308 amino acid residues) if these are processed in the same way, and the amino acid sequence were homologous to each other by 91-94%. Functional amino acids, including His40, His170, Tyr185 and Arg183 and S-S-bond-forming Cys, were conserved in the three isozymes, but a few N-glycosylation sites were not the same. Two HRP C isoenzyme genomic genes, prxC1 and prxC2, were tandem on the chromosomal DNA and each gene consisted of four exons and three introns. The positions in the exons interrupted by introns were the same in two genes. We observed a putative promoter sequence 5' upstream and a poly(A) signal 3' downstream in both genes. The gene product of prxC1 might be processed with a signal sequence of 30 amino acid residues at the N terminus and a peptide consisting of 15 amino acid residues at the C terminus.     

The effects of ovarian hormones on glucose and fatty acid oxidation during exercise in female ovariectomized rats

The effects of ovarian hormones on glucose and fatty acid oxidation during exercise were investigated in adult female ovariectomized rats. Rats subdivided into 3 groups received intraperitoneal injections of hormones or sesame oil for 8 days. Estrogen (E) treated rats received 17-beta estradiol in daily doses of 2 micrograms. Estrogen and progesterone treated rats (EP) received 17-beta estradiol in daily doses of 2 micrograms and 2 mg, respectively. Control rats (S) received sesame oil alone. After an overnight fast, rats ran at the speed of 25 m.min-1 for 60 min. [U-14C]glucose or [1-14C]palmitate was injected into rats at 5 min of exercise and before 10 min of exercise, respectively. Expired 14CO2 was collected using bottomless chamber on a treadmill belt. No significant differences were found in mean blood glucose, lactate and plasma free fatty acid concentrations after the exercise. Until the end of the exercise 34.7 +/- 2.6 (E, n = 5), 40.8 +/- 2.9 (EP, n = 5) and 43.7 +/- 3.5% (S, n = 6) (mean +/- SE) of 14C which was injected as 14C-glucose was recovered as 14CO2. During 60 min of the exercise 27.5 +/- 1.0 (E, n = 7), 19.8 +/- 2.7 (EP, n = 6) and 25.0 +/- 1.9% (S, n = 6) of 14C which was injected as 14C-palmitate was recovered as 14CO2. A significant difference was found in this rate between E and EP (P less than 0.05). It was concluded that estrogen treatment stimulated fatty acid oxidation compared with the estrogen plus progesterone treatment and tended to inhibit glucose oxidation during prolonged exercise.