Nucleotide and deduced amino acid sequence of bovine adrenal medulla chromogranin B (secretogranin I)

1. A novel 1745-dalton pyroglutamyl peptide (BAM-1745)6 was recently isolated and characterized from bovine adrenal medulla chromaffin granules. Its amino acid sequence was found to be 93% identical to residues 580-593 of human chromogranin B (secretogranin I). 2. Based on this sequence a degenerate oligonucleotide probe was synthesized and used to identify a 2.4-kb bovine adrenal medulla chromogranin B cDNA. 3. The deduced polypeptide is 647 amino acids long and begins with a putative signal sequence of 20 residues as in the human, rat, and mouse proteins. Also conserved in the bovine protein is a tyrosine residue which may be sulfated, two N-terminal cysteines, and many paired basic amino acids which may serve as sites of posttranslational processing. The peptide BAM-1745 is flanked by paired basic amino acids and therefore is most likely a product of posttranslational processing. Bovine chromogranin B is 67, 58, and 58% identical to the human, rat, and mouse chromogranin B proteins, respectively. 4. The carboxyl terminus of bovine chromogranin B, including BAM-1745, was found to be the most conserved region of the polypeptide and may identify it as an important functional domain.     

Coccidioidomycosis in South America. A review of its epidemiology and geographic distribution

Argentina. Twenty-seven human cases and coccidioidin skin-test surveys have located the endemic area of coccidioidomycosis between the 27th and 40th south parallels. Climate is of the arid steppe type in the southern zone, arid hill and prairie in the intermediate zone, and arid hill and prairie plus hot tropical in the northern zone. Temperature ranges from 5° C to 29° C, vegetation is xerophytic and annual rainfall is from 300 to 500 mm. Paraguay. On the basis of human cases and coccidioidin surveys, the endemic area has been delimited between the 19th and 24th south parallels. It was a hot, dry, windy climate with temperature reaching 45° C, an annual rainfall average of 500 mm and xerophytic vegetation. Colombia. On the basis of two human cases and coccidioidin test surveys, an endemic area of low prevalence was confirmed in the northeast between the 10th and 12th north parallels. Altitude in this region is from 2 to 300 meters above sea level, temperature averages about 29° C. Within this region two different areas can be differentiated — one in the north where vegetation is tropical desert brush type and rainfall ranges between 125 and 500 mm; the second in the south with grass and cotton culture and rainfall from 500 to 2000 mm. Venezuela. Thirty-five human cases and nearly 60,000 skin tests made from east to west in the northern part of the country, where the population is concentrated, showed that the endemic area is situated between the 9th and 12th north parallels. This is an arid region with desert soils. Altitude ranges from sea level to 800 meters, annual temperature averages 24° C and rainfall 500 mm in some places, and 29° C and less than 400 mm in others. More than 172 species of plants have been identified in the zone but cacti predominate.C. immitis was isolated from soil collected at a site where a patient had become infected. Bolivia, Peru and Ecuador.Mackinnon studied a patient coming from Bolivia, but he has expressed doubt about the Bolivian origin of the infection because the patient had lived in the Paraguayan Chaco the previous year. More information is necessary to evaluate the human case mentioned in Peru byBinder. Cases reported from Ecuador appear to have been paracoccidioidomycosis and leishmaniasis rather than coccidioidomycosis.Many species of rodents and other wild and domestic animals share with man the possibility of infection in the four countries where the endemic areas have been confirmed.Paper read at the Eighth International Congresses for Tropical Medicine and Malaria, September 1968, Teheran (Iran).     

The behavior of normal peripheral nerve in Tissue culture

Summary A phase contrast and time lapse cinematographic study of normal mouse sciatic nerve cultured in vitro was made. The Rose chamber and chicken plasma clot methods were employed. The growth was characterized by three basic cell types: a spindle-shaped cell with a bulging nucleus, a racket-shaped cell with a short wide fan-shaped process and an opposite filiform process, and a kite-shaped cell with abundant ectoplasm. The spindle-shaped cells exhibited a pulsatile rhythmic activity. The rhythm of contraction varied from two to eighteen minutes. No contractile activity was observed in the case of the racket-shaped cells nor in the kite-shaped cells. The spindle-shaped cells were thought to be Schwann cells, the kite-shaped cells were considered of a fibroblastic nature, whereas no source could be found for the racket-shaped cells, although the perineurium was considered as a possible origin. The cultures were maintained up to 80 days, but at no time were phagocytes, observed. With the methods employed no transformation of cells from one type to another took place, and the Schwann cells did not transform themselves into phagocytes.This work was supported in part by grant P-405A from the American Cancer Society and by NIH grant NB-06391-02.     

Isolation of Histoplasma capsulatum from an oil bird (steatornis caripensis) cave in Venezuela

Summary 1.Histoplasma capsulatum was recovered from two of fifteen soil specimens collected in a Venezuelan cave. This cave, located near Caripe in the state of Monagas, harbors a large colony of the oil bird,Steatornis caripensis.2. This finding confirms the previous discovery in Tingo Maria, Peru, of the association ofH. capsulatum with oil birds.3. The relationship ofH. capsulatum with animal habitats is discussed along with the public health importance of this association.

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Zusammenfassung 1.Histoplasma capsulatum ist in zwei Bodenproben aus fünfzehn, die einer Höhle in Venezuela entnommen wurden, gefunden worden. Diese Höhle, die sich in der Nähe von Caripe, im Staate Monagas befindet, herbergt eine grosse Kolonie von Fettvögeln,Steatornis caripensis.2. Dieser Befund bestätigt die Vergesellschaftung vonH. capsulatum mit den Fettvögeln, wie es vorher bereits in Tingo Maria, Peru, entdeckt worden ist.3. Die Beziehung vonH. capsulatum zum tierischen Habitat wird diskutiert unter Betonung der Wichtigkeit dieser Vergesellschaftung vom Standpunkte des öffentlichen Gesundheitswesens.

     

Characterization of SKM1, a Saccharomyces cerevisiae gene encoding a novel Ste20/PAK-like protein kinase

Ste20/PAK serine/threonine protein kinases have been suggested as playing essential roles in cell signalling and morphogenesis as potential targets of Cdc42 and Rac GTPases. We have isolated and characterized the Saccharomyces cerevisiae SKM1 gene, which codes for a novel member of this family of protein kinases. The amino acid sequence analysis of Skm1p revealed the presence of a PH domain and a putative p21-binding domain near its amino terminus, suggesting its involvement in cellular signalling or cytoskeletal functions. However, deletion of SKM1 produced no detectable phenotype under standard laboratory conditions. Moreover, disruption of each of the two other S. cerevisiae Ste20/PAK-like kinase-encoding genes, STE20 and CLA4 , in skm1 backgrounds, showed that Skm1p is not redundant with Ste20p or Cla4p. Interestingly, overexpression of SKM1 led to morphological alterations, indicating a possible role for this protein in morphogenetic control. Furthermore, overproduction of Skm1p lacking its N-terminus caused growth arrest. This effect was also seen when similarly truncated versions of Ste20p or Cla4p were overexpressed. We further observed that overproduction of this C-terminal fragment of Skm1p complements the mating defect of a ste20 mutant strain. These results suggest that the N-terminal domains of S. cerevisiae Ste20/PAK-like protein kinases share a negative regulatory function and play a role in substrate specificity.     

New records of large branchiopods (Branchiopoda: Anostraca,Notostraca, and Spinicaudata) in Mexico

This paper reports new distribution records of large branchiopods for Mexico following a three year survey of the Baja California peninsula. The occurance of the anostracans Thamnocephalus mexicanus (Linder, 1941) and T. platyurus (Packard, 1877), the notostracan Lepidurus lemmoni (Holmes, 1894), and the spinicaudatans Eulimnadia cylindrova (Belk, 1989) and E. texana (Packard,1871), all represent the first records for the peninsula. An undescribed species of the anostracan genus Streptocephalus is recorded from the state of Baja California (Norte). The occurrence of the notostracan genus Triops and four other anostracan species on the peninsula is also confirmed. The conchostracan Cyclestheria hislopi (Baird, 1859) is recorded from the state of Quintana Roo. The collections of Lepidurus and Cyclestheria are the first records for México. These records increase the number of species of large branchiopods reported fromMéxico to 36: 20 Anostraca, 3 Notostraca, 11 Spinicaudata, and 2Laevicaudata. This revised version was published online in August 2006 with corrections to the Cover Date.     

On the taxonomic status of a Dugesia species from Buenos Aires (Platyhelminthes: Turbellaria: Tricladida)

The taxonomic and distributional statuses of the freshwater planarians of South America are imperfectly understood, and a comprehensive review is sorely needed. The species known from the vicinity of Buenos Aires, Argentina, was first described by Böhmig (1902) under the name Planaria dubia Borelli, 1895. Subsequent authors (Kenk, 1930, 1974; Cazzaniga & Curino, 1987) have cast doubt on this assignation. Some have assigned it to Dugesia anceps Kenk; Hyman (1959) was inclined to regard the Buenos Aires planarian as a geographical variant of Dugesia sanchezi Hyman. A comparison of D. sanchezi from the mountainous region of central Chile with specimens from Buenos Aires confirms that the Argentinian material is a distinct species.The differences between the two species are subtle. The Argentinian specimens are distinctive in their possession of infranucleate epithelia of the atria, penis, and bursal canal, as well as in their penial structure. In both species, the epithelia of the atria and bursal canal are glandular. Those of D. sanchezi are nucleate and their secretions are cyanophilous. In the Argentinian specimens, these same epithelia are infranucleate; but while the secretion of the bursal canal is indeed cyanophilous, that of the atrium is erythrophilous. The presence, in these same specimens, of cyanophilous glands opening into the penial vesicles, and of erythrophilous glands whose secretions contribute to a viscous plug in the ejaculatory duct, suggests a difference in the physiology of copulation.It is apparent that the Argentinian specimens are distinct from D. sanchezi Hyman, and probably they represent a new species, one to be described in detail elsewhere. In any case, Hyman's (1959) suggestion that D. sanchezi occurs near Buenos Aires is mistaken; so also is Cazzaniga's & Curino's (1987) identification of specimens from Buenos Aires as D. anceps.     

Pre-column derivatization of free bile acids for high-performance liquid chromatographic and gas chromatographic—mass spectrometric analysis

Pentachlorophenyl (PCP) esters of five free bile acids (FBA) were obtained by reacting the FBA and Kovacs' complex (KC) in a 1:8 molar ratio in acetone at 65°C, and were purified by column chromatography on silica gel. The esters were crystallized from benzene—hexane, derivatized as trimethylsilyl ethers for gas chromatography on a DB-1 capillary column and for gas chromatography—mass spectrometry with a DB-5 column, and mass spectrometry (MS) in the electron-impact (EI) positive-ion mode at 70 eV. The reaction is specific for FBA even in the presence of glycine and taurine conjugates of bile acids. The PCP esters were treated with benzylamine in chloroform or methanol to produce N-benzyl derivatives of FBA. The N-benzylamides were separated by high-performance liquid chromatography (HPLC) on a 4-μm Nova-Pak C₁₈ column, studied by thermospray—LC—MS, and in the direct insertion probe—EI positive-ion mode.     

Effects of growth hormone on diaphragmatic recovery from malnutrition.

A 25% weight loss was induced in adult Fisher 344 rats by nutritional deprivation. Subsequently, normal feeding was resumed. Refed animals were divided into three groups and received recombinant human growth hormone (rhGH) injections during 5 wk of refeeding, saline injections during 5 wk of refeeding, or 9 wk of refeeding without injections. The effects of nutritional deprivation and the various refeeding protocols on the cross-sectional areas (CSA) of each of the four types of myofibers [typed immunohistochemically with antibodies against four myosin heavy chain (MHC) isoforms known to be present in the rat diaphragm] were determined. Malnutrition decreased the CSA of myofibers containing MHC2X, MHC2B, and MHC2A (i.e., fast myofibers), with the greatest effect on muscle mass being due to the atrophy of fibers containing MHC2X. Fibers containing MHC beta/slow failed to undergo malnutrition-induced atrophy. Whereas refeeding for 5 wk in the absence of rhGH allowed the recovery of CSA of fibers containing MHC2A, fibers containing MHC2B and MHC2X remained smaller than fibers of similar type in control animals. In contrast, 5 wk of refeeding supplemented with rhGH returned all fiber CSAs to control values. Even when refeeding alone was extended to 9 wk to allow for weight stabilization, the CSA of the fibers containing MHC2B and MHC2X remained smaller than similar fibers in control muscle. Serum insulin-like growth factor, a marker of malnutrition (R. Reeves and J. Elders, J. Nutr. 109: 613-620, 1979), was significantly decreased after nutritional deprivation and returned to normal after 5 wk of refeeding and GH supplementation.     

Heavy metal composition of polysomal fractions following cadmium challenge

Endogeneous levels of zinc and copper were found to be 1.2±0.1×10⁻² and 0.3±0.1×10⁻² μg/A260 unit, respectively, in polysomal fractions from control animals; cadmium, however, was undetectable. In experimental animals (injected with cadmium) zinc, copper, and cadmium were found in polysomal fractions isolated by two different methods. One hour after a cadmium injection there was a rise in both the zinc and copper content of the polysomal fractions, which then declined steadily to below control levels by 16 h. Neither zinc nor cadmium were dialyzable from these fractions by a TRIS buffer; however, addition of 0.01M EDTA to the buffer resulted in removal of 75% of the zinc and all of the detectable cadmium. The addition of cadmium (CdCl₂) to control supernatants (adjusted to the cadmium concentration present in supernatants 6 h after in vivo exposure) resulted in metal binding to polysomal fractions in levels comparable to those observed after in vivo exposures to the metal. When cadmium was added in the form of cadmium thionein, a smaller fraction of the metal was isolated with the polysomal fraction. Cadmium bound to polysomal fractions in vivo (24 h after exposure) was sensitive to release by protease digestion, but insensitive to release by ribonuclease digestion.