Constitutive heterochromatin patterns of G-group chromosomes in Down's syndrome

Summary 2 male patients with G-trisomic Down's syndrome, were studied for constitutive heterochromatin pattern by the technique of Arrighi and Hsu (1971). 2 G-chromosomes were found to have heterochromatin distributed around the centromeric areas and the remaining 3 were free of such localized heterochromatin blocks; the extra G-chromosome belonging to the non-localized heterochromatin members.

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Zusammenfassung 2 männliche Patienten mit G-Trisomie (Down-Syndrom) sind im Hinblick auf das konstitutive Heterochromatinmuster mit Hilfe der Technik von Arrighi u. Hsu (1971) untersucht worden. Es fanden sich 2 G-Chromosomen, die eine Heterochromatinverteilung um die Zentromerregion zeigen, während die restlichen 3 frei von derartig lokalisierten Heterochromatinblocks sind; das überzählige G-Chromosom gehört zu den letztgenannten.

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Supported in part by the Robert A. Welch Foundation, The National Foundation —March of Dimes and USPHS Grant No. FR-05425.     

Zwei subterminale Heterochromatinregionen bei einer seltenen Form einer 21/21-Translokation

Zusammenfassung Bei einem 31/2jährigen Kind mit leichter Form eines Down-Syndroms wurde ein dem Chromosom D ähnliches Translokationschromosom gefunden, das nach Atebrin-Färbung als 21/21-Translokation identifiziert werden konnte. Es fehlt ein G₂₁-Chromosom. Da das Translokationschromosom an beiden Enden Satelliten trägt, wurde zur Aufklärung des Entstehungsmechanismus die Heterochromatinfärbung nach Arrighi u. Hsu (1971) durchgeführt. Die Färbung ergab 2 subterminale Heterochromatinregionen an diesem Chromosom. Unter diesem Gesichtspunkt werden die möglichen Entstehungsmechanismen diskutiert.

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Two subterminal heterochromatin regions in a rare form of 21/21 translocation

Summary A translocation resembling a chromosome D but with satellites at both ends in most metaphases was found in a child with a mild form of Down's syndrome. This chromosome was identified as a 21/21 translocation by atebrin staining. One G₂₁-chromosome was missing.To examine the origin and nature of this chromosome further, heterochromatin staining (Arrighi and Hsu, 1971) was performed. 2 subterminal heterochromatin regions were found in the translocation chromosome.On the basis of this finding various possible origins of this chromosome are discussed.

     

Studies of multipolar mitoses in euploid tissue cultures

Cultures of kidney epithelium and fibroblasts of 39 specimens of Microtus agrestis were investigated. In all 77 cultures multipolar mitoses were found. They were studied in living state and after pulse labelling with ³H-thymidine. The ploidy of the multipolar mitoses and of their daughter nuclei was determined by measuring the relative Feulgen-DNA content and by counting the predominantly constitutive heterochromatic sex chromosomes. Constitutive heterochromatin was demonstrated by late replication, retarded separation of the chromatids in anaphase, heteropycnosis and by the Giemsa technique of Arrighi and Hsu (1971). The latter stained also the spindle apparatus of mitoses.—In living cells, transformation of multipolar mitoses into bipolar mitoses was observed. The chromosomes of multipolar mitoses are separated into complete genomes; the daughter nuclei can be haploid, diploid, triploid or tetraploid. The chromosomes of haploid and triploid metaphases were studied with the Giemsa banding technique. The banding pattern shows an exact monosomy and trisomy, respectively, for each chromosome. Haploid nuclei are likely to be viable only in multinucleate cells, whereas triploid cells behave like diploid cells during the S period and the mitosis.Dedicated to Prof. Dr. K. Goerttler on the occasion of his 75th birthday.Supported by the Bundesministerium für Bildung und Wissenschaft of the Federal Republic of Germany.     

Study of constitutive heterochromatin with a new and simplified fluorescence staining technique

A new fluorochrome that preferentially binds itself to the centromeric or the constitutive heterochromatin is described. This stain allows an easy assay, through fluorescence, of the repetitive DNA or bands, supposedly composed of constitutive heterochromatin, in insectivores, rodents and man, without following the in situ hybridization of Pardue & Gall (1970) or the DNA denaturation-renaturation processes of Arrighi & Hsu (1971). The staining patterns with this derivative of a Benzimidazole compound (Hoechst 33258) are induced in the chromosomes without incubation or pretreatment with SSC and are identical to those produced by other techniques. This stain may eventually contribute to elucidating the hitherto unknown molecular mechanisms involved in the relationship between the repetitious DNA sequences and the banding patterns, and to interpreting the mechanisms responsible for the chromosomal rearrangements and aberrations involving the peri-and non-pericentric regions of the chromosomes.Paper read by P.K.S. at the NATO Advanced Study Institute on Comparative Biology of Primates Turin 7–19 June 1972. Supported by the Alexander von Humboldt-Stiftung, Bonn-Bad Godesberg.     

On the Fertility and Survival of Deep Frozen Boar Spermatozoa Thawed in Skim Milk

Satisfactory conception rates of deep frozen boar spermatozoa were obtained, with insemination by way of the cervix, after thawing the deep frozen spermatozoa in boar seminal plasma, both in preliminary trials (Crabo & Einarsson 1971, Crabo et al. 1972 b) and in a large field trial (Einarsson et al. 1972). Fertility with pellet frozen boar spermatozoa, thawed without dilution, was reported by Graham et al. (1971 a, b) and Pursel & Johnson (1971).     

A Field Outbreak Caused by Bovine Respiratory Syncytial Virus

Bovine respiratory syncytial virus (BRSV) caused a large epizootic of acute respiratory disease in Japan in 1968—69 (Inaba et al. 1970, Inaba et al. 1972). A much smaller outbreak occurred in Switzerland (Paccaud & Jacquier 1970). In Belgium the virus has been isolated from an outbreak of respiratory disease (Wellemans et al. 1970). BRSV has later been proved an important causal agent of respiratory disorders in the same country (Wellemans & Leiinen 1975). In England and USA the virus has caused and been isolated from outbreaks of acute respiratory disease in calves (Jacobs & Edington 1971, Rosenquist 1974, Smith et al. 1974). In Denmark BRSV has sporadically been isolated from pneumonic calf lungs (Bitsch et al. 1976).     

Giemsa banding patterns in chronic lymphocytic leukaemia.

The banding patterns of chromosomes from 20 patients with chronic lymphocytic leukaemia (C.L.L.) have been analyzed. 97 of 100 metaphases examined had a normal banding pattern. The 3 remaining metaphases, all from one patient had bands similar to those seen after aging. It is concluded that the chromosomes in C.L.L. have normal banding patterns. The majority of cytogenetic studies in chronic lymphocytic leukaemia have reported normal chromosomes (Fitzgerald and Adams 1965; Oppenheim et al., 1965; Lawler et al., 1968). An inherited abnormality of G group chromosome (No. 22) has been reported in a family, three members of whom developed C.L.L. (Fitzgerald and Hamer, 1969), but further investigations of cases of familial leukaemia failed to reveal a similar abnormality (Fitzgerald et. al., 1966). The development of new techniques which allow the positive identification of individual chromosomes (Caspersson et al., 1969; Dutrillaux and Lejeune, 1971; Sumner et al., 1971; Seabright, 1971), has revolutionised human cutogenetics and revealed additional information regarding chromosome abnormalities and leukaemia (Rowley, 1973; Lobb et al., 1972; Milligan and Garson, 1974). The purpose of this investigation was to determine whether the chromosomes in C.L.L. have normal banding patterns.     

Banding in Human Chromosomes treated with Trypsin

THE differential staining properties of the Giemsa stain were first observed by Pardue and Gall¹. They were studying in situ hybridization between mouse satellite DNA and mouse chromosomes and observed that following certain pretreatment the centromeric regions of mouse chromosomes were more densely stained by Giemsa stain than other regions. The darkly stained regions were considered to consist of constitutive heterochromatin. Similar observations were later made on human chromosomes by Arrighi and Hsu² and Gagné et al.³. Through modifications of the original methods used in the DNA hybridization work, techniques have been developed which make each chromosome identifiable^4–6.     

Pattern of repetitive DNA of the chromosomes of Microtus agrestis

The distribution of repetitive DNA in the chromosomes of Microtus agrestis was studied with the method for demonstrating constitutive heterochromatin given by Yunis et al. (1971) and the reassociation technique described by Schnedl (1971). All autosomes can be individually recognized by means of the position of their bands. The euchromatic segment of the X1 chromosome shows the same banding pattern as the corresponding segment of X2 which consists of facultative heterochromatin. The short arms of the Y chromosome are not deeply stained with either method and therefore do not contain noticeable amounts of repetitive DNA. The relative distances between the bands remain constant during chromosome contraction in mitosis.     

Preliminary data on the chromosomes of Echinostoma caproni Richard, 1964 (Trematoda: Echinostomatidae)

The karyotype of Echinostoma caproni Richard, 1964 is studied. There are eleven pairs of chromosomes (2n=22). Two pairs are submetacentric (Nos. 4 and 5). C-banding methods revealed a large block of heterochromatin in No. 3 and two blocks in No. 5. E. cinetorchis (see Terasaki et al., 1982) and E. barbosai (see Mutafova & Kanev, 1983) also have 22 chromosomes, but the metacentric or submetacentric chromosomes are Nos. 8 and 11 for the former and No. 10 for the latter. ac]19860425     

A Cytological Study of Six Populations of Disporum cantoniense (Liliaceae)

Disporum cantoniense (Lour.) Merr. is widely distributed in the area from the Himalayas to Indonesia, via south China, Indo-China and Taiwan, especially in the various parts of Yunnan Province. In this paper, the karyotype variation of six populations of the apecies from southeastern, middle and northwestern part of Yunnan are studied. The result shows that the chromosome number of all the populations are 2n= 14. The species was reported to have 2n= 16, 30 (Hasegawa 1932, Mehra and Pathamia 1960, Kurosawa 1966, 1971, Tang et al. 1984) and 2n= 14 (Kurosawa 1971, Mehra and Sachdeva 1976a). 2n= 14, 2n= 16 and 2n=32 were observed in the material from Taiwan (Chuang, et al. 1962, Chao, 1963, Hsu, 1971, 1972, Chang, 1974). Based on the cytological study of D. megalanthum Wang et Tang and seven other species in this genus reported by other authors, Hong and Zhu (1990) consider that the basic number of this genus is x= 8, because species with 2n= 16 was more than those with 2n= 14, despite some number variation of chromosomes in this genus. Based on the results of the present paper, we consider that x= 7 might be one of the basic numbers of this genus. In the karyotypes studied here, the relative chromosome lengths and the ratio of the longest/the shortest chromosomes of the six populations are rather approximate. Moreover, all the karyotypes belong to Stebbins’3B type. However, karyotype variation was detected in these populations. The homologues of the 2nd, 3rd and 6th pair of chromosomes are different from each other, the numbers and popsition of satellites are found very different, among the populations except for the Lijiang population, the 1st, 2nd, 3rd or 4th pair of all the populations exhibited heterozygosity. Although all the karyotypes belong to Stebbins ‘3B type, the homologues were more regular in the Lijiang population than in the other populations, and the most irregular in the Wenshan population, because it hadfour pairs of heterozygous chromosomes.     

Induction of Swine Dysentery with a Pure Culture of Treponema Hyodysenteriae in Vitamin E and Selenium Deficient Pigs

Several successful attempts have been made to induce swine dysentery in pigs using pure cultures of Treponema hyodysen-teriae (Taylor & Alexander 1971, Harris et al. 1972, Akkermans & Pomper 1973, Hughes et al. 1975). In these studies, either conventional or specific-pathogen-free pigs were used. In the present study, 2 approximately 8 weeks old conventional pigs (Nos. 1 and 2) were purchased and fed the same basic ration as used by Teige et al. (1977). In addition, 10 % cod liver oil was incorporated in the diet at each feeding. After a feeding period of 25 days rectal swabs were applied and examined for the presence of spirochaetes. The pigs were then fed a 3 days old primary and pure culture of T. hyodysenteriae on TSA-S400 medium (Songer et al. 1976). The culture originated from the colon of a pig with swine dysentery (Pig No. 4, Teige et al. 1977). Each pig received the agar contents of 5 petri dishes which were mixed with the food.     

Infectious Pancreatic Necrosis First Isolation of Virus from Fish in Norway

Infectious pancreatic necrosis (IPN) is a disease of salmonid fishes. It has been reported in many countries throughout the world (M’Gonigle 1940, Wood et al. 1955, Besse & Kinkelin 1965, Vestergård Jørgensen & Bregnballe 1969, Sano 1971, Ball et al 1971, Ljungberg & Vestergård Jørgensen 1972, Schlotfeldt et al. 1975). Outbreaks of the disease can cause serious losses in populations of hatchery reared salmonids, the mortality rate varying between 10 and 90 % (Vestergård Jørgensen & Kehlet 1971). There are at least four different serotypes of the virus distinguished by neutralization tests (Wolf et al. 1968). Twenty-five isolates of IPN virus in Denmark proved to represent only two serotypes (Sp and Ab) (Vestergård Jørgensen & Kehlet). The present paper reports the first isolation of IPN virus from the stock at a fish farm in Norway.     

Distribution of constitutive heterochromatin in mammalian chromosomes

Using a special staining technique, a survey of the chromosomes of many mammalian species showed that constitutive heterochromatin is present in all cases and that the heterochromatin pattern appears to be specific and consistent or each chromosome and each taxon. Usually heavy heterochromatin is found in the centromeric areas, but terminal heterochromatin is not uncommon. Occasionally interstitial heterochromatin bands occur. In some species, such as the Syrian hamster and Peromyscus, many chromosome arms are completely heterochromatic.Supported in part by Research Grant GB-13661 from the National Science Foundation.     

The mitotic configuration of Chaetomium globosum

The somatic nuclear division ofChaetomium globosum was studied utilizing acetocarmin and aceto-orcein techniques. Nuclear division in hyphae of this species was found to be mitotic, but diversity in morphology and division configuration was noted. Identifiable chromosomes, the metaphase plate, and the chromosome bridge were commonly seen.A combination of extremely small nuclei, difficulties in staining, multinucleate conditions, and protoplasmic streaming in hyphae presented difficulties for these studies. Contradictory views are held on the structure of the nucleus, presence of the centriolar body, and other karyological features as described byFinley (1970)Rabinow &Bakerspigel (1965). Nuclear division structural details in a few other fungal species such as the centriolar body, spindle apparatus, and nuclear membrane disassociation have been examined by electron microscopy (Motta, 1969;Ichida &Fuller, 1968;Namboodiri &Lowry, 1967).The present study and the previous report onC. globosum (Hsu, Yu &Volz, 1972) presents comparative data for a NASA Appollo 16 MEED Mycology investigation now in progress.     

Evidence for a highly differentiated sex chromosome heteromorphism in the salamander Necturus maculosus (Rafinesque)

The mitotic chromosomes of the neotenic (sensu Gould, 1977, and Alberch et al., 1979) salamander Necturus maculosus (Rafinesque) have been examined using a C-band technique to demonstrate the distribution of heterochromatin. The C-banded mitotic chromosomes provide evidence of a highly differentiated XY male/XX female sex chromosome heteromorphism, in which the X and Y chromosomes differ greatly in size and morphology, and in the amount and distribution of C-band heterochromatin. The X chromosome represents one of the largest biarmed chromosomes in the karyotype and is indistinguishable from similar sized autosomes on the basis of C-band heterochromatin. The Y chromosome, on the other hand, is diminutive, morphologically distinct from all other chromosomes of the karyotype, and is composed almost entirely of C-band heterochromatin. The discovery of an X/Y chromosome heteromorphism in this species is consistent with the observation by King (1912) of a heteromorphic spermatogenic bivalent. Karyological and phylogenetic implications are discussed.     

The taxonomy of the family Paramphistomidae Fischoeder, 1901 with special reference to the morphology of species occurring in ruminants. II. Revision of the genus Paramphistomum Fischoeder, 1901

Summary The genus Paramphistomum Fischoeder, 1901 is redefined and restricted and only the following species are retained and considered valid: P. cervi (Zeder, 1790) (type species); P. liorchis Fischoeder, 1901; P. gracile Fischoeder, 1901 P. epiclitum Fischoeder, 1904; P. gotoi Fukui, 1922, P. ichikawai Fukui, 1922; P. leydeni Näsmark, 1937 and P. hiberniae Willmott, 1950. These are redescribed and illustrated. A new species, Paramphistomum cephalophi is described and illustrated from the black-fronted duiker (Cephalophus nigrifrons) in Rwanda. It differs from the rest of the species in the genus by the presence of an anterior sphincter in the pharynx and the characteristic posterior notch of the acetabular rim. Scanning electron photomicrographs of the tegumental surfaces of the species in the genus are provided. Cotylophoron indicum Stiles & Goldberger, 1910 (=Paramphistomum thapari Price & McIntosh, 1953), C. madrasense Gupta, 1958, C. chauhani Gupta & Gupta, 1972, Paramphistomum indicum Stiles & Goldberger, 1910 (in part), P. malayi Lee & Lowe, 1971 and Srivastavaia indica Singh, 1970 are considered synonyms of Paramphistomum epiclitum Fischoeder, 1904. Paramphistomum indicum Stiles & Goldberger, 1910 (in part) and P. bombayiensis Gupta & Verma in Gupta & Nakhasi, 1977 are regarded as synonyms of Paramphistomum gracile Fischoeder, 1901. P. scotiae Willmott, 1950, P. julimarinorum Velázquez-Maldonado, 1976, P. nicabrasilorum Velázquez- Maldonado, 1976, P. procapri Wang, 1979 and Cotylophoron skrjabini Mitskevich, 1958 are considered synonyms of Paramphistomum leydeni Näsmark, 1937. Cotylophoron vigisi Davydova, 1963 is considered synonymous with Paramphistomum ichikawai Fukui, 1922. Paramphistomum birmense Railliet, 1924, P. microon Railliet, 1924, P. chinensis Hsu, 1935 and P. pseudocuonum Wang, 1979 are regarded as species inquirendae.The genera Liorchis Velichko, 1966 and Srivastavaia Singh, 1970 are synonymized with Paramphistomum Fischoeder, 1901.A key to the species of the genus is provided.Part of a thesis approved by the University of London for the award of the Ph.D. degree.Part of a thesis approved by the University of London for the award of the Ph.D. degree.     

Mus and Peromyscus chromosome homology established by FISH with three mouse paint probes

Fluorescence-labeled DNA probes constructed from three whole house mouse (Mus domesticus) chromosomes were hybridized to metaphase spreads from deer mouse (Peromyscus maniculatus) to identify homologies between the species. Mus Chr 7 probe hybridized strongly to the ad-centromeric two-thirds of Peromyscus Chr 1q. Most of Mus 3 probe hybridized principally to two disjunct segments of Peromyscus Chr 3. Mus Chr 9 probe hybridized entirely to the whole Peromyscus Chr 7. Three Peromyscus linkage groups were assigned to chromosomes, based on linkage homology with Mus. The data also are useful in interpretation of chromosomal evolutionary history in myomorphic rodents. Received: 1 December 1998 / Accepted: 17 February 1999     

Facilitated diffusion: The problem of boundary conditions

Summary The model equations suggested by Wyman (1966) to explain Wittenberg's (1966) experiments on oxygen diffusion facilitated by haemoglobin have been studied by various authors. Kreuzer and Hoofd (1970) use a semi-analytical and numerical approach; Kutchaiet al. (1970) use a purely numerical approach; and Murray (1971) solved the equations analytically. Although the results they obtain are in good agreement with experiment, Kreuzer and Hoofd (1970) and Kutchaiet al. (1970) on the one hand and Murray (1971) and Murray and Wyman (1971) on the other use fundamentally different boundary conditions. This paper reconsiders the problem and proves that these different boundary conditions are equivalent for practically all situations of biological interest. The conclusion is that the simple algebraic result of Murray (1971) suffices for most experimental situations. In the extreme situations where his procedure is not applicable, which are distinguished in the text, the numerical scheme of Kutchaiet al. (1970) is recommended.P. J. M. would like to thank the Science Research Council for their financial support.     

Freeze-resistant Trichinella (Trichinella nativa) Established on the Scandinavian Peninsula

Until the 1970’s, Trichinella spiralis (Owen 1835) was considered the only species within the genus Trichinella. Then T. pseudospiralis (Garkavi 1972) was classified as a separate species on the basis of morphological and biological features. The remaining morphologically homogenous “T. spiralis-group” has been split into 4 different species (or subspecies) on the basis of their biological and biochemical characteristics; T. nativa (Britov & Boev 1972), T. nelsoni (Britov & Boev 1972), T. spiralis sensu stricto and T. britovi (Pozio et al. 1992).