New developments in vertebrate cytotaxonomy IX Chromosome numbers in the order Insectivora (Mammalia)

A reference list of chromosome numbers of about 110 species in the order insectivora is presented. Cytotaxonomic data are known for six families: Solenodontidae, Tenrecidae (incl. Potamogalidae), Erinaceidae, Soricidae, Talpidae and Macroscelididae (only the Chrysochloridae remain unstudied). The list provides, wherever possible, the diploid chromosome number(s), Nombre Fondamental, and references; sometimes information on geography, synonymy, as well as short comments are added.This is the ninth in a series of articles, by different authors, to appear in this journal under the editorship of Professor Chiarelli, supplementing the data published in Cytotaxonomy and Vertebrate Evolution (A. B. Chiarelli & E. Capanna, eds, Academic Press, London & New York, 1973).-Eds.     

New developments in vertebrate cytotaxonomy VI. Cytotaxonomy and evolution of lower chordates

A correlation between evolutionary trends and karyotype has been recognized in tunicates. In the ambit of lower chordates and related species, Hemichordata, Cephalochordata and, in the Tunicata, some ascidians seem to maintain a primitive organization of chromosomes: haploid numbers around 20, absence of sex chromosomes, and rod-shaped bivalents. Within the class Ascidiacea, at the karyological level Clavelina depadiformis seems to be very specialized and the family Perophoridae (suborder Phlebobranchiata) very primitive. Thaliacea and Larvacea appear specialized in comparison with the primitive ascidians.This is the sixth in a series of articles, by different authors, to appear in this journal under the editorship of Professor Chiarelli, supplementing the data published in Cytotaxonomy and Vertebrate Evolution (A. B. Chiarelli & E. Capanna, eds, Academic Press, London & New York)-Eds.     

New developments in vertebrate cytotaxonomy VIII. A current list of references on avian karyology

A list of 256 references dealing with avian karyology is presented. This list is updated to April 1983 and covers 587 bird species, 385 non-passerines and 202 passerines. It includes 37 non-passerines and 29 passerines that were only studied in the period from 1902 onwards to the early 1950s using less sophisticated techniques (mostly on sectioned material of gonadal tissues). The remaining 348 non-passerines and 173 passerines were studied with more advanced techniques using colchicine-treated material of feather-pulp, embryonic tissue, bone-marrow, leucocyte or tissue culture. The species studied to date belong to 25 of the 26 avian orders and to 96 of the 160 avian families.This is the eighth in a series of articles, by different authors, to appear in this journal under the editorship of Professor Chiarelli, supplementing the data published in Cytotaxonomy and Vertebrate Evolution (A. B. Chiarelli & E. Capanna, eds. Academic Press, London & New York, 1973)-Eds.     

Polytene chromosomes of Simulium craigi (Diptera: Simuliidae)

Simulium craigi Adler and Currie is a polymorphic species based on polytene chromosome banding patterns in the long arm of chromosome III (IIIL). Three cytotypes are described based on the predominant IIIL sequences. These correspond to three broad geographic areas: cytotype CC from Pennsylvania; cytotype AF from Ontario and Manitoba; and cytotype ACF/BCF from New Hampshire. In the absence of sympatric populations, these cytotype differences are best explained by clinal variation within a single species. The relationship of S. craigi to other described members of the S. vernum group is discussed.     

Interactions of protein kinase CK2 subunits

Several approaches have been used to study the interactions of the subunits of protein kinase CK2. The inactive mutant of CK2 that has Asp 156 mutated to Ala (CK2A156) is able to bind the CK2 subunit and to compete effectively in this binding with wild-type subunits and . The interaction between CK2A156 and CK2 was also demonstrated by transfection of epitope-tagged cDNA constructs into COS-7 cells. Immunoprecipitation of epitope-tagged CK2A156 coprecipitated the subunit and vice-versa. The assay of the CK2 activity of the extracts obtained from cells transiently transfected with these different subunits yielded some surprising results: The CK2 specific phosphorylating activity of these cells transfected with the inactive CK2A156 was considerably higher than the control cells transfected with vectors alone. Assays of the immunoprecipitated CK2A156 expressed in these cells, however, demonstrated that the mutant was indeed inactive. It can be concluded that transfection of the inactive CK2A156 affects the endogenous activity of CK2. Transfection experiments with CK2 and subunits and CK2A156 were also used to confirm the interaction of CK2 with the general CDK inhibitor p21WAF1/CIP1 co-transfected into these cells. Finally a search in the SwissProt databank for proteins with properties similar to those derived from the amino acid composition of CK2 indicated that CK2 is related to protein phosphatase 2A and to other phosphatases as well as to a subunit of some ion-transport ATPases.     

Combined use of 13C chemical shift and 1Hα−13Cα heteronuclear NOE data in monitoring a protein NMR structure refinement

Summary A large portion of the ¹³C resonance assignments for murine epidermal growth factor (mEGF) at pH 3.1 and 28°C has been determined at natural isotope abundance. Sequence-specific ¹³C assignments are reported for 100% of the assignable C, 96% of the C, 86% of the aromatic and 70% of the remaining peripheral aliphatic resonances of mEGF. A good correlation was observed between experimental and back-calculated C chemical shifts for regions of regular -sheet structure. These assignments also provide the basis for interpreting ¹H–¹³C heteronuclear NOE (HNOE) values in mEGF at natural isotope abundance. Some of the backbone polypeptide segments with high internal mobility, indicated by these ¹H–¹³C HNOE measurements, correlate with locations of residues involved in the putative mEGF-receptor binding site. Using four families of mEGF structures obtained over the last few years, we demonstrate that standard deviations between experimental and back-calculated C values can be used to monitor the refinement of this protein's structure, particularly for -sheet regions. Improved agreement between calculated and observed values of C is correlated with other measures of structure quality, including lowered values of residual constraint violations and more negative values of conformational energy. These results support the view that experimental conformation-dependent chemical shifts, C, can provide a reliable source of information for monitoring the process of protein structure refinement and are potentially useful restraints for driving the refinement.Abbreviations HSQC heteronuclear single-quantum coherence spectroscopy - PFG pulsed-field gradient - TOCSY ¹H-¹H total correlation spectroscopy - EGF epidermal growth factor - mEGF murine EGF - hEGF human EGF - hTGF human type- transforming growth factor - DIPSI spm-locking pulse sequence - NOE nuclear Overhauser effect - HNOE heteronuclear Overhauser effect     

A new type of inversion of a human Y chromosome

Summary Using chromosome banding techniques, a phenotypically normal male was found to have an abnormal banding pattern of the Y chromosome. By the constitutive heterochromatin staining method, a darkly stained band was located on the short arm and the proximal region of the long arm. The quinacrine staining method also showed a similar abnormal banding pattern: a brightly fluorescing band was seen on the short arm and the proximal region of the long arm. By the conventional Giemsa staining method, however, no specific morphological abnormality was detected in the aberrant Y. On detailed karyotype analyses no recognizable abnormality of banding patterns of any other chromosome was found aside from the abnormal Y. The abnormality was determined to be a complex inversion of the Y chromosome, which is described as 46,X,inv(Y)(pterp11::q11q12::cen::q12qter).     

A comparative study of karyotypes of muntjacs by chromosome painting

We have used a combination of chromosome sorting, degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR), chromosome painting and digital image capturing and processing techniques for comparative chromosome analysis of members of the genus Muntiacus. Chromosome-specific paints from a female Indian muntjac were hybridised to the metaphase chromosomes of the Gongshan, Black, and Chinese muntjac by both single and three colour chromosome painting. Karyotypes and idiograms for the Indian, Gongshan, Black and Chinese muntjac were constructed, based on enhanced 4, 6-diamidino-2-phenylindole (DAPI) banding patterns. The hybridisation signal for each paint was assigned to specific bands or chromosomes for all of the above muntjac species. The interspecific chromosomal homology was demonstrated by the use of both enhanced DAPI banding and comparative chromosome painting. These results provide direct molecular cytogenetic evidence for the tandem fusion theory of the chromosome evolution of muntjac species.Deceased     

Interaction of cytochrome c L with methanol dehydrogenase from Methylophaga marina 42:

The reaction of methanol dehydrogenase with cytochrome c _L from Methylophaga marina and the reactions of the non-physiological substrates, Wurster's blue and ascorbic acid, with both proteins were studied as a function of temperature (4–32 °C), pressure (1–2000 bar) and ionic strength using the optical high pressure stopped-flow method. The thermodynamic parameters H, S and V were determined for all reactions where electron transfers are involved. These data allowed the determination of the Maxwell relationships which proved the internal thermodynamic consistency of the system under study. A conformational change on the cytochrome c _L level was deduced from both breaks in the Arrhenius plots and the variation of the V with temperature.Abbreviations MOPS 4-morpholinepropanesulfonic acid - CHES 2-(cyclohexylamino)ethanesulfonic acid - MDH methanol dehydrogenase - EDTA ethylenedinitrilotetraacetic acid disodium salt - BTB bromothymol blue (3,3-dibromothymolsulfoneph-thalein) - PQQ 2,7,9-tricarboxy-lH-pyrrolo-[2,3f]quinoline-4,5-dione - cytochrome c _HH mammalian horse heart cytochrome c     

Over-expression of the gene (bglBC1) from Bacillus circulans encoding an endo-β-(1→ 3),(1→ 4)-glucanase useful for the preparation of oligosaccharides from barley β-glucan

An endo--(13),(14)-glucanase gene (bglBC1) from Bacillus circulans ATCC21367 was modified by substituting its native promoter with a strong promoter, BJ27X, to increase expression of the gene when cloned into B. subtilis RM125 and B. megaterium ATCC14945. A 771-bp endo--(13),(14)-glucanase open reading frame was inserted into a new shuttle plasmid, pBLC771, by ligating the ORF and pBE1, the latter of which contained the strong promoter, BJ27X. B. subtilis, transformed with the recombinant plasmid pBLC771, produced an extracellular endo--(13),(14)-glucanase that was 130 times (7176 mU ml^–1) more active than that of the gene donor cells (55 mU ml^–1), while the enzyme from the transformed B. megaterium was 7 times (378 mU ml^–1) more active than that of the gene donor cells. M _r of the enzyme was 28 kDa, with proteolytic processing of the enzyme being observed only in B. subtilis cells. The major products of water-soluble -glucan hydrolyzed by over-produced endo--(13),(14)-glucanase were tri- and tetra-oligosaccharides which can be developed as useful products such as anti-hypercholesterolemic, anti-hypertriglyceridemic, and anti-hyperglycemic agents.     

Measurement of residual dipolar couplings from 1Hα to 13Cα and 15N using a simple HNCA-based experiment

Novel NMR pulse schemes for simultaneous measurement of ¹ D _CHand ² D _NHresidual dipolar couplings in proteins is presented. We show that ² D _NHcoupling can be very useful for protein structure determination. The ² D _NHcoupling can be measured from ¹⁵N dimension with good accuracy on a slowly relaxing TROSY resonance, utilizing HNCA-TROSY-based experiments, which concomitantly supply large ¹ D _CHcoupling. The dynamic range of ² D _NHcoupling is comparable to ¹ D _NC coupling, but instead, it also serves non-redundant information on the course of protein backbone, thanks to rotational degree of freedom with respect to peptide bond. The HNCA-TROSY-based experiments are optimal for measuring residual dipolar couplings at high magnetic fields owing to absence of rapid transverse relaxation of carbonyl carbon. The reliability of the proposed approach was tested on ¹⁵N/¹³C human ubiquitin. A very good correlation with ubiquitin solution as well as crystal structure, for both ¹ D _CHand ² D _NHcouplings, was obtained.     

Alteration by centric fission of the diploid chromosome number in Vicia faba L.

An individual with a diploid chromosome complement of 2n=14 instead of 2n=12 is deseribed for the broad bean Vicia faba L. This karyotypic deviation resulted from centromere splitting of the wildtype metacentric satellite chromosome pair as demonstrated by Giemsa banding pattern. It is the first case of centric fission observed in this species. Our data support the hypothetic mechanism of Robertsonian rearrangements suggested by Holmquist and Dancis (Genetica 52–53: 151–163, 1980).     

Recherche D'enzymes intracellulaires dans le genre schizosaccharomyces. Implications systématiques

In the genus Schizosaccharomyces intracellular osidases and nitrite and nitrate reductases are revealed; particularly all the species possessing invertase, -glucosidase and -galactosidase. These characters underline the homogeneity of the genus. On the basis of osidases, nitrite and nitrate reductases results, 2 groups can be distinguished in this genus.     

Monosomy 21 in spontaneous abortus

Summary Chromosome analysis of the amnion from a missed abortus revealed a 45,XX,G-karyotype. Giemsa banding pattern technique identified the chromosome involved in monosomy as No. 21.

---

Zusammenfassung Die Chromosomenanalyse des Amnions eines verhaltenen Abortus ergab einen 45,XX,G-Karyotyp. Dank der Anwendung der Giemsa banding pattern-Technik konnte festgestellt werden, daß die Monosomie das Chromosom Nr. 21 betrifft.

---

---

This study was supported by the Ford Foundation Population Program, Grant No. 640-0411 B, and by the World Health Organisation.     

Carvedilol prevents cardiac hypertrophy and overexpression of hypoxia-inducible factor-1α and vascular endothelial growth factor in pressure-overloaded rat heart

Summary The use of -blockers has emerged as a beneficial treatment for cardiac hypertrophy. Hypoxia-inducible factor-1 (HIF-1) is tightly regulated in the ventricular myocardium. However, the expression of HIF-1 in cardiac hypertrophy due to pressure overload and after treatment with -blocker is little known. To evaluate the effect of carvedilol on both myocardial HIF-1 expression and cardiac hypertrophy, infra-renal aortic banding was performed for 4 weeks in adult Sprague-Dawley rats to induce cardiac hypertrophy. Carvedilol at 50 mg/kg body weight per day after surgery was given. Heart weight and the ratio of heart weight and body weight increased significantly after aortic banding for 4 weeks in the absence of drug treatment. Mean arterial pressure increased from 80 ± 9 mmHg in the sham group to 94 ±5 mmHg (p < 0.001) in the banding group. Echocardiography showed concentric hypertrophy after aortic banding. Mean arterial pressure decreased after treatment with carvedilol. The increased wall thickness and heart weight was reversed to normal by carvedilol. Western blot showed that HIF-1, vascular endothelial growth factor (VEGF) and brain natriuretic peptide (BNP) proteins were up-regulated and nerve growth factor- (NGF-) down-regulated in the banding group. Treatment with valsartan, doxazosin, or N-acetylcysteine did not significantly affect HIF-1 and VEGF proteins expression in the banding groups. Real-time polymerase chain reaction showed that mRNA of HIF-1, VEGF and BNP increased and mRNA of NGF- decreased in the banding group. Treatment with carvedilol reversed both protein and mRNA of HIF-1, VEGF, BNP, and NGF- to the baseline values. Increased immunohistochemical labeling of HIF-1, VEGF, and BNP in the ventricular myocardium was observed in the banding group and carvedilol again normalized the labeling. In conclusion, HIF-1, VEGF, and BNP mRNA and protein expression were up-regulated, while NGF- mRNA and protein was downregulated in the rat model of pressure-overloaded cardiac hypertrophy. Treatment with carvedilol is associated with a reversal of abnormal regulation of HIF-1,VEGF, BNP, and NGF- in the hypertrophic myocardium.     

A few distinct ‘molecular sandwiches’ are basis for structural and functional similarities of subspecies of interferon α and of families of growth-promoting hormones

Molecular sandwiches composed of two aromatic amino acids separated by a hydrophilic one were found on eleven subspecies of human interferon , on murine interferon 2, and human interferon 1. In addition, another type of the sandwiches was found on several species of interferon. This confirms and extends the observations concerning the similarities between some interferons and several classical hormones. Furthermore, we are presenting evidence that a distinct type of the molecular sandwiches: Tyr-Cys...Cys and/or Cys...Cys-Cys...Cys, that participate in formation of disulfide bonds, is a characteristic marker of most, if not all of the growth-promoting hormones including growth factors. The sandwiches appear to be important for receptor binding.     

Fine structure and function of ocelli and sagittocysts of acoel flatworms

We studied the taxonomy and distribution of the Conchostraca in Yugoslavia. A diverse fauna occurs in the country. However, the systematics of this group is still incomplete and has many problems. There is considerable variability of morphological characters. By a comparative-morphological analysis of the populations found, we re-evaluated the taxonomic significance of a series of structural characters, and offer comments on the results.In Yugoslavia four families were found: Cyzicidae Stebbing, 1910; Limnadiidae Burmeister, 1843; Imnadiidae Botnariuc & Orghidan, 1941 and Leptestheriidae Daday, 1923, with a present total of 8 species and 4 subspecies. Within the genus Imnadia Hertzog, 1935, three new species were found: Imnadia cristata Marinek, 1972; Imnadia banatica Marinek & Valvajter, 1982 and Imnadia panonica Marinek & Petrov, 1984. Within the genus Eoleptestheria Daday, 1923, the species Eoleptestheria spinosa Marinek, 1978, with four distinct subspecies was discovered.     

Identification of a 1B/1R wheat-rye chromosome translocation

Summary The common wheat selection 79-4045 was identified as a wheat-rye 1B/1R chromosome translocation line, by means of C-banding patterns and test cross with Chinese Spring double-ditelosomic line. The translocation chromosome consisted of the long arm of wheat chromosome 1B, including its centromere, and the short arm of rye chromosome 1R or tis portion.     

Intergeneric somatic hybridization of sexually incompatible parents: Citrus sinensis and Atalantia ceylanica

Protoplast fusion using polyethylene glycol (PEG) was conducted to combine Citrus sinensis (L.) Osbeck cv. Hamlin sweet orange protoplasts, isolated from nucellus-derived embryogenic callus with Atalantia ceylanica (Arn.) Oliv, leaf protoplasts. Five plants regenerated from independent fusion events following protoplast culture were identified as intergeneric allotetraploid somatic hybrids of Hamlin sweet orange and A. ceylanica, and confirmed by isozyme analysis and chromosome number determination in root tip cells (2n=4x=36). Two different types of leaf morphology were observed among the hybrids (normal and narrow), although no differences in chromosome number nor isozyme banding patterns were observed. This is the first report of the production of hybrid plants between these sexually incompatible genera.Florida Agricultural Experiment Station Journal Series No. R-03069.     

Chromosome number 18

Summary 4 cases of ? chromosome abnormality, referred to this department in the past 3 months, have shown an aberrant complement of chromosome 18. Chromosome 18 has been identified using a modification of Seabright's (1971) method of banding human chromosomes.

---

Zusammenfassung Vier Fälle mit fraglicher Chromosomen-Anomalie, die während der vergangenen 3 Monate an diese Abteilung überwiesen wurden, zeigten ein abnormes Chromosom Nr. 18. Chromosom 18 wurde mit Hilfe einer Modifikation von Seabright (1971) für die Darstellung von Bandenmustern identifiziert.

---

---

Supported in part by United Sheffield Hospitals Research Grant Code 309.