bluuebunny/crossref_metadata_2025 huggingface dataset:
Auditory Evoked Response Test Strategies to Reduce Cost and Increase Efficiency: The Postauricular Muscle Response Revisited
Authors: Patuzzi Robert B. , Thomson Susmita M.
Published: April 2003

We describe a number of techniques in auditory evoked response (AER) testing for hearing loss which should decrease its cost and increase its efficiency, making its use in infant hearing screening more viable. We demonstrate the use of bit-stream averaging of the electrical signals from the head as a cheap alternative to analogue averaging and show that the average waveforms obtained are similar with both techniques. We demonstrate how the postauricular muscle response (PAMR) can be potentiated by lateral rotation of the eyes and argue that uncontrolled eye movements in previous studies have led to an unfounded belief that the PAMR is not stable. When eye rotation is used to potentiate the PAMR, the response becomes very stable and so large in most subjects that it is clearly visible in the raw traces. We also demonstrate that when the PAMR is potentiated by eye rotation, stable PAMR waveforms can be reliably obtained with tone bursts with frequencies up to and above 8 kHz and with sound levels within 30 dB of the subjective detection threshold. As a result the PAMR can be used to rapidly determine an objective audiogram in most subjects within minutes. Finally, we demonstrate a correlation technique for detecting the PAMR without waveform averaging and the need for an expensive computer. We are sure that a combination of these techniques can be used to increase the efficiency of AER screening for infant deafness and lower its cost dramatically.
Viral Hepatitis Infection Should Be Considered for Evaluating Uremic Pruritus in Continuous Ambulatory Peritoneal Dialysis Patients
Authors: Hung Kuan-Yu , Shyu Ren-Shi , Tsai Tun-Jun , Chen Wan-Yu
Published: April 2003

Determining the possible association of viral hepatitis infection and degree of pruritus is the primary concern of this study. Ninety-six adequately dialyzed CAPD patients (47 male and 49 female) and 526 normal controls (266 male and 260 female) were enrolled. Blood hemoglobin, ferritin, electrolytes, calcium, phosphate, albumin, urea, creatinine, aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase, and bilirubin were analyzed by routine methods. Serum HBsAg was examined, using a radioimmunoassay method and the anti-HCV, an enzyme immunoassay method. All cases were interviewed with a standardized questionnaire. The highest possible pruritus score (PS) was 22. The prevalences of HBsAg(+) and anti-HCV(+) were 14.6% and 17.7%, respectively. The mean PS in all 96 CAPD patients was 11.6 (range 7–22). The mean PS were 11.8 ± 0.6 and 12.5 ± 1.0 for patients infected with HBV and HCV, respectively. Both were significantly higher than that (10 ± 0.9) of patients without hepatitis infection. AST and ALT were significantly higher in patients infected with viral hepatitis than those without. The other biochemical parameters were not significant. Thirty-seven (38.5%) of our 96 patients had mild pruritus (PS &IE; 7) and 11 (15.9%) had severe pruritus (PS ≧15). Of the 83.9% (26/31) patients with viral hepatitis, the grades of skin itching were moderate to severe; whereas those of the patients without viral hepatitis, 53.6% (37/69) belonged to the group of moderate to severe pruritus (p = 0.003, χ2 test with Yates’ correction). The authors recommended screening of viral hepatitis infection to be undertaken for uremic patients with unexplained skin itching.
Is Post-Dialysis Urea Rebound Significant with Long Slow Hemodialysis?
Authors: Jean G. , Chazot C. , Charra B. , Terrat J.C. , Vanel T. , Calemard E. , Laurent G.
Published: April 2003

Background: According to previous studies, postdialysis urea rebound (PDUR) is achieved within 30–90 min, leading to an overestimation of Kt/V of between 15 and 40% in 3- to 5-hour dialysis. The purpose of the study was to assess the impact of PDUR on the urea reduction ratio (URR), Kt/V and normal protein catabolic rate (nPCR) with long 8-hour slow hemodialysis. Methods: This study was performed in 18 patients (13 males/5 females), 62.5 ± 11.7 years of age, hemodialyzed for 3–265 months. Initial nephropathies were: 3 diabetes; 2 polycystic kidney disease; 3 interstitial nephritis; 2 nephrosclerosis; 3 chronic glomerulonephritis, and 5 undetermined. Residual renal function was negligible. The dialysis sessions were performed using 1- to 1.8-m2 cellulosic dialyzers during 8 h, 3 times a week. Blood flow was 220 ml/min, dialysate flow 500 ml/min, acetate or bicarbonate buffer was used. Serial measurements of the urea concentration were obtained before dialysis, immediately after dialysis (low flow at t = 0), and at 5, 10, 20, 30, 40, 60, 90 and 120 min, and before the next session. The low-flow method was used to evaluate the access recirculation, second-generation Daugirdas formulas for Kt/V, and Watson formulas for total body water volume estimation. The difference between the expected urea generation (UG) and urea measured after dialysis (global PDUR) defines net PDUR (n-PDUR). Results: The n-PDUR usually became stable after 58 ± 25 (30–90) min. Its mean value was 17 ± 10% of the 30-second low-flow postdialysis urea (3.9 ± 2 mmol/l). This small postdialysis urea value and the importance of UG in comparison with shorter dialysis justify the use of n-PDUR. Ignoring n-PDUR would lead to a significant 4% overestimation (p < 0.001) of the URR (79 ± 7 vs. 76 ± 8%), 12% of Kt/V (1.9 ± 0.4 to 1.7 ± 0.38) and 4% of the nPCR (1.1 ± 0.3 to 1.05 ± 0.3). n-PDUR correlated negatively with postdialysis urea (r = 0.45 p = 0.05), positively with URR (r = 0.31 p = 0.01) and Kt/V (r = 0.3 p = 0.03) but not with K, and negatively with the urea distribution volume (r = 0.33 p = 0.05). Mean total recirculation, ultrafiltration rate, predialysis urea levels and urea clearance did not correlate with n-PDUR. Conclusion: We found a significant PDUR in long-slow hemodialysis after a mean of 1 h after dialysis. This PDUR has a less important impact upon dialysis delivery estimation than short 3- to 5-hour hemodialysis, especially for the lower Kt/V or URR ranges. This is explained by the low-flux, high-efficiency, and long-term dialysis. Its inter-individual variability incites us to calculate PDUR on an individual basis.
Effect of Plasma Exchange on Serum Tissue Inhibitor of Metalloproteinase 1 and Cytokine Concentrations in Patients with Fulminant Hepatitis
Authors: Nakamura Tsukasa , Ushiyama Chifuyu , Suzuki Shingo , Shimada Noriaki , Ebihara Isao , Suzaki Masataka , Takahashi Toshimasa , Koide Hikaru
Published: April 2003

Aims: The present study assessed whether the serum concentrations of tissue inhibitor of metalloproteinase 1 (TIMP-1) and cytokines are altered in patients with fulminant hepatitis and whether plasma exchange affects these concentrations. Methods: Fifteen patients with fulminant hepatitis, 14 patients with severe acute hepatitis, and 20 healthy controls were included in this study. The serum levels of tumor necrosis factor alpha (TNF-α), interleukin 1β (IL-1β), interleukin 6 (IL-6), transforming growth factor beta (TGF-β), and TIMP-1 were determined in all patients upon hospital admission and before and after a single course of plasma exchange in the patients with fulminant hepatitis. Results: Ten out of the 15 patients with fulminant hepatitis and all patients with severe acute hepatitis survived. Serum TNF-α, IL-6, TGF-β, and TIMP-1 levels in patients with fulminant hepatitis were significantly higher than the levels in patients with severe acute hepatitis (p < 0.01). IL-1β was not detectable in either group. Plasma exchange reduced the increased serum concentrations of TNF-α, IL-6, TGF-β, and TIMP-1 in patients with fulminant hepatitis (p < 0.01). Conclusions: These data suggest that increased serum levels of TIMP-1 and cytokines may reflect severe hepatic inflammation and that plasma exchange is an effective therapy to reduce these levels.
Double-Chamber On-Line Hemodiafiltration: A Novel Technique with Intra-Treatment Monitoring of Dialysate Ultrafilter Integrity
Authors: Pizzarelli F. , Tetta C. , Cerrai T. , Maggiore Q.
Published: April 2003

On-line hemodiafiltration is a technique that relies on the re-injection of pyrogen-free substitution fluid obtained by cold filtration of dialysate. Therefore, safety of this treatment modality depends on the quality of dialysate and, mainly, on the integrity of the ultrafilter(s) employed. Double-chamber on-line hemodiafiltration is a new technique where re-infusion takes place inside the dialyser by means of dialysate backfiltration. The peculiar geometry of the dialyser allows intra-treatment assessment of its fibre integrity. In this paper, we tested feasibility and safety of this new modality of on-line treatment. The extracorporeal blood and infusate pressure values resulted well inside the safety range. Blood urea clearances and β2 removal were consistent with the figures usually found in standard hemodiafiltration. Whole blood production of cytokines was similar when blood was exposed to saline or infusate, both values being comparable to the spontaneous whole blood cytokine release. The on-line dialyser fibre integrity check showed a great sensitivity even for minimal dialyser damage. We conclude that double-chamber on-line hemodiafiltration is a feasible and safe procedure. Our preliminary results encourage the undertaking of multicentre, prospective, randomised studies.
Artificial Cell Biotechnology for Medical Applications
Authors: Chang Thomas Ming Swi
Published: April 2003

Artificial cells are prepared in the laboratory for medical and biotechnological applications. The earliest routine clinical use of artificial cells is in the form of coated activated charcoal for hemoperfusion. Implantation of encapsulated cells are being studied for the treatment of diabetes, liver failure and the use of encapsulated genetically engineered cells for gene therapy. We recently found that daily orally administered artificial cells containing a genetically engineered microorganism can lower the elevated urea level in uremic rats to normal levels and increase the survival of the animal. Furthermore, this can remove potassium, phosphate, uric acid and other waste metabolites from uremic plasma. Blood substitutes based on modified hemoglobin are already in phase-III clinical trials in patients with as much as 20 units infused into each patient during trauma surgery. Artificial cells containing enzymes are being developed for clinical trials in hereditary enzyme deficiency diseases and other diseases. Artificial cells are also being investigated for drug delivery and other uses in biotechnology, chemical engineering and medicine.
Increase in Blood Volume during Dialysis without Ultrafiltration
Authors: Nette Robert W. , Akçahüseyin Emin , Krepel Harmen P. , Weimar Willem , Zietse Robert
Published: April 2003

Combined dialysis and ultrafiltration leads to more frequent episodes of hypotension than isolated ultrafiltration. It has been suggested that decreased plasma volume preservation could be responsible for this phenomenon. The present study evaluates the effects of diffusive dialysis on the changes in relative blood volume (RBV). Six stable hemodialysis patients, without the need of ultrafiltration, were studied during 10 sessions of diffusive dialysis (bicarbonate) lasting 4 h. RBV was monitored continuously by measurement of hematocrit. During the 1st and 2nd h RBV increased by 2.4 ± 1.4 and 2.5 ± 0.8% respectively, returning to baseline levels at the end of dialysis. No changes in blood pressure or heart rate were noted. We conclude that during diffusive dialysis without ultrafiltration RBV is increased. A decrease in vascular resistance, or changes in regional blood distribution could explain these findings.
Evolution by polyploidy in Amphibia: new insights
Authors: Beçak M.L. , Beçak W.
Published: April 2003

Polyploidy is an important mechanism of evolution in lower vertebrates, resulting in gene duplication and loci duplication evolving to diploidization. In polyploid anurans DNA is increased, but RNA and protein synthesis is kept at the same levels as in their diploid counterparts. Recent cytogenetic findings in Odontophrynus americanus show: (1) amphiplasty and asynchrony of the cell division cycle in 4n and (2) intra- and interindividual alterations of chromosome 4 morphology in 4n and 2n through breaks at secondary constrictions and at/or around the centromeres, followed by the appearance of changes of centromeres position, dicentrics, bisatellited chromosomes, precocious sister-chromatid segregation, and loss or unequal segregation of the chromosomes. This genome instability suggests the presence of chromosomal aberrations with chromodomain alterations at the centromeres affecting the mechanisms ensuring normal segregation of the chromatids in anaphase of mitosis and meiosis. Cell division asynchrony and chromosome abnormalities, novel findings in polyploids, are discussed respectively in function of gene activity and putative translocations producing polymorphisms in nucleolus organizers and secondary constrictions.
Tetranucleotide short tandem repeat polymorphisms and their possible mode of origin
Authors: Bender K. , Beller G. , Lautsch S.
Published: April 2003

An analysis of three autosomal, one X-chromosomal, and one Y-chromosomal tetranucleotide short tandem repeat loci in a large southern German population sample (including family studies) and in several nonhuman primate species revealed remarkable similarities in structure of already known and some newly detected alleles and in allele frequency distributions. These similarities, which are briefly discussed, can best be explained by transspecific evolution, followed by gene-conversion-like mutational events.
The Y* rearrangement in mice: new insights into a perplexing PAR
Authors: Burgoyne P.S. , Mahadevaiah S.K. , Perry J. , Palmer S.J. , Ashworth A.
Published: April 2003

In essence, the Y* rearrangement in the mouse is a Y chromosome that has been hijacked by a non-Y centromere attached distal to the pseudoautosomal region (PAR). All the Y-unique material is thought to be unaltered, but the recombinatory behaviour of the Y* with the X during male meioisis led to the conclusion that part of the PAR is inverted. In the course of a cross set up to introduce the X-linked mutation Patchy fur (Paf) into XY* males, the Y* chromosome was found to carry the wild type allele of Paf. Paf maps close to the X PAR boundary, so we hypothesised that the inverted region of the Y* PAR originated from an X chromosome that provided not only an inverted copy of proximal PAR, but also an X PAR boundary together with some adjacent X-unique material that included the Paf locus. This hypothesis was validated by Southern analysis using an X PAR boundary probe to show that Y* has an X PAR boundary. Thus the Y* PAR has resulted from an end to end fusion of an X and a Y PAR. Furthermore, it was shown that in conjuction with this PAR-PAR fusion, there has been deletion of both copies of the distally located pseudoautosomal gene Steroid sulfatase (Sts).
Gene dosage in the evolution and function of mammalian sex chromosomes
Authors: Graves J.A.M. , Disteche C.M. , Toder R.
Published: April 2003

Ohno’s early suggestions about the origin of sex chromosomes and the consequences of alterations of dosage of X and Y genes have provided an important framework for understanding sex chromosome organization, function and evolution. Here we review evidence that heteromorphic sex chromosomes evolved from an autosomal pair, and that one of the consequences of X–Y differentiation is the evolution of dosage compensation by X inactivation and upregulation of the active X, which in turn, has selected for a highly conserved X chromosome.
Did the first adaptive immunity evolve in the gut of ancient jawed fish?
Authors: Matsunaga T.
Published: April 2003

Gut-associated immune tissue, as represented by lymphocytes and lymphoid cell aggregates, occurs in all jawed vertebrates. Considering the lymphopoietic function of the cryptopatches in the mouse intestine, the lymphoid aggregates in the gastrointestinal lamina propria of birds and cold-blooded vertebrates seem to serve the same function, thus providing an explanation for their conservation during a period of over 400 million years. Some γδ+ intraepithelial T cells are likely to develop and mature in gut-associated immune tissues. Intraepithelial γδ+ T cells may therefore have represented the first step in the evolution of adaptive immunity, reinforcing the gastrointestinal defense against microbial invasion as a result of increasing traumatization by injury and infection due to jaw development in host fish.
Y;16 translocation breakpoint associated with a partial Turner phenotype identifies a foamy virus insertion
Authors: Schmidt Drury, S. , Erickson R.P. , Glover T.W.
Published: April 2003

We have previously identified YACs containing the Y breakpoint in a male patient with Turner-like hydrops in the newborn period and a Y;16 translocation (Erickson et al., 1995). We have now subcloned these YACs and built a lambda contig across the Y breakpoint. A subclone near the Y;16 breakpoint of our patient shows great evolutionary conservation by hybridization to a zoo blot. We have sequenced the region responsible for the hybridization and found a foamy (spuma) virus sequence. These viruses are known to be xenotropic and a very similar cross-species hybridization (using the same commercial zoo blot) was recently reported with a human foamy virus clone (Cordonnier et al., 1995). Thus, we have identified a foamy virus insertion near the Y heterochromatin boundary.
Detailed genetic mapping around a putative prostate-specific membrane antigen locus on human chromosome 11p11.2
Authors: Maraj B.H. , Leek J.P. , Karayi M. , Ali M. , Lench N.J. , Markham A.F.
Published: April 2003

Physical mapping of the human prostate-specific membrane antigen gene (FOLH) has not been straightforward. Previously, localisations of this gene to either 11p11.2 or 11q14 have been described. This raised the possibility of the presence of more than one related FOLH gene in man. We now present detailed characterisation of the region around a FOLH gene in a 500-kb non-chimaeric YAC clone, putatively assigned to 11p11.2. This clone contains two known microsatellites, D11S1326 and D11S1357 which have been previously mapped unequivocally to the 11p11.2 region at 62.5 cM. This data strongly supports the original 11p11.2 localisation of FOLH. The YAC clone also has a putative EST at each of its ends and these have thus been physically positioned on this chromosome. However, the two regions previously highlighted at 11p11.2 and 11q14 are apparently extensively duplicated, as evidenced by the appearance of dual FISH signals with a number of different genomic probes selected across this 500-kb 11p11.2 region.
In situ hybridization of five loci to cattle chromosome 1
Authors: Schmutz S.M. , Moker J.S. , Berryere T.G.
Published: April 2003

The genes for pituitary-specific transcription factor (PIT1), propionyl coenzyme A carboxylase, β-polypeptide (PCCB), transferrin (TF), trichohyalin (THH), and involucrin (IVL) were mapped to cattle chromosome 1 (BTA 1) by isotopic in situ hybridization. Two of the loci were mapped from cattle PCR products and three from human ATCC probes. PIT1 localized to segment 1q2; PCCB to 1q3; and TF, THH, and IVL to 1q4. These localizations agree with the homology previously shown between BTA 1 and human chromosome 3 (HSA 3). Some homology with HSA 1 has been established with the mapping of THH and IVL to BTA 1q4.
Physical and linkage mapping of the gene for the α3 chain of type IX collagen, COL9A3, to human chromosome 20q13.3
Authors: Tiller G.E. , Warman M.L. , Gong Y. , Knoll J.H.M. , Mayne R. , Brewton R.G.
Published: April 2003

Type IX collagen is a minor cartilage component which associates with mixed fibrils of types II/XI collagen. We have determined the precise physical and genetic locations for the gene encoding the α3 chain of type IX collagen, COL9A3. Utilizing fluorescence in situ hybridization, radiation hybrid mapping, and multipoint linkage analysis, we have mapped COL9A3 to human chromosome 20q13.3, 13 cM telomeric to D20S173.
Assignment of the mouse and cow CXC chemokine genes
Authors: Modi W.S. , Amarante M.R.V. , Hanson M. , Womack J.E. , Chidambaram A.
Published: April 2003

Gene specific PCR primers were constructed for five mouse and three bovine CXC chemokine genes. The mouse genes were assigned using SSCP analyses of the Jackson BSS backcross panel to two groups on chromosome 5. One group containing Gro1 and Mip2 cosegregated with reference markers Alb1 and Btc, and was positioned 2.2 cM proximal to a group comprising Ifi10, Mig, and Scyb5. The bovine genes IL8, GRO1, and GRO3, mapped using bovine × hamster somatic cell hybrids, were all found to be located on chromosome 6. The locations of these genes in these two animal species are consistent with the positions in humans (4q13→q21), and previous syntenic relationships among these three mammals.
Comparative mapping of two adjacent regions of MMU19 with their human counterpart on HSA11q13
Authors: Fernandes M. , Lespinasse F. , Rotomondo F. , Poirier C. , Guenet, J.-L. , Gaudray P. , Carle G.F.
Published: April 2003

High resolution physical maps of two adjacent regions of MMU19 were constructed in order to establish a comparative map between the pericentromeric region of MMU19 and its human counterpart on HSA11q13. These two physical maps span 2.5 and 0.5 megabases on MMU19. Long range restriction analysis and YAC contigs have been built, five genes were located on MMU19 and eight new STSs were generated. The 0.5-Mb map which has been positioned close to the centromere of MMU19, based on dual-color FISH experiments and genetic data, includes eight genes (Type I markers), three microsatellites (Type II markers) and five new STSs. The 2.5-Mb map is located more telomeric and contains seven genes, four microsatellites and four new STSs. Gene order and physical distances appear to be similar in human and in mouse in this 2.5-Mb region. Strikingly, the 0.5-Mb region has a similar size in human but gene order is shuffled. The overall comparative map shows that these two regions are inverted on MMU19 when compared with HSA11q13.
Expressed copies of the MN7 (D15F37) gene family map close to the common deletion breakpoints in the Prader-Willi/Angelman syndromes
Authors: Buiting K. , Groß S. , Ji Y. , Senger G. , Nicholls R.D. , Horsthemke B.
Published: April 2003

Approximately 70% of patients with Prader-Willi syndrome or Angelman syndrome have a similar sized de novo deletion of 3–4 Mb in the proximal region of 15q. The distal breakpoints appear to cluster between the P gene (OCA2) and D15S24, whereas two deletion breakpoint clusters have been identified on the proximal side (one centromeric to D15S541 and one between D15S541 and D15S9). Based on the identification of a gene family in 15q11→q13 (MN7, D15F37), we have previously proposed that the presence of multiple copies of this sequence may be related to the instability of this region. Using fluorescence in situ hybridization and YAC mapping, we have found that at least one D15F37 locus is centromeric to D15S9 and at least two are between OCA2 and D15S24. As determined by cDNA cloning and sequence analysis, each of the individual loci is expressed. The close proximity of the D15F37 loci and the deletion breakpoints suggests that the common deletions arise by unequal crossover events at or near these loci.
Genetic mapping of mouse centromere protein (Incenp and Cenpe) genes
Authors: Fowler K.J. , Saffery R. , Kile B.T. , Irvine D.V. , Hudson D.F. , Trowell H.E. , Choo K.H.A.
Published: April 2003

Inner centromere protein (INCENP) and centromere protein E (CENPE) are two functionally important proteins of the higher eukaryotic centromere. Using a mouse Incenp genomic DNA and a mouse Cenpe cDNA to analyze recombinant inbred mouse sets, as well as interspecific backcross panels, we have mapped these genes to the proximal regions of mouse Chromosomes 19 and 6, respectively. Comparison of Cenpe and human CENPE, which maps to chromosome region 4q24→q25, has further identified a new region of homology between the two species.
Localization of FCGR1 encoding Fcγ receptor class I in primates: molecular evidence for two pericentric inversions during the evolution of human chromosome 1
Authors: Maresco D.L. , Blue L.E. , Culley L.L. , Kimberly R.P. , Anderson C.L. , Theil K.S.
Published: April 2003

The human high-affinity receptor for immunoglobulin G, FcγRI (FCGR1), is encoded by a family of three genes that share over 95% sequence homology. Curiously, the three genes in this recently duplicated gene family flank the centromere of human chromosome 1, with FCGR1B located at 1p12 and both FCGR1A and FCGR1C located at 1q21. We have previously speculated that a pericentric inversion could account for the separation of the genes in the FCGR1 family and explain their current chromosomal location. Here we present evidence, obtained through fluorescence in situ hybridization analysis, that in the rhesus monkey (Macaca mulatta) and baboon (Papio papio) FCGR1 is located adjacent to the centromere on the chromosomal arm with greatest homology to human 1p, whereas in the chimpanzee (Pan troglodytes) it is located adjacent to the centromere on the chromosomal arm with greatest homology to human 1q. The separation of the FCGR1 gene family in humans suggests that the location of a second pericentric inversion, known to distinguish the human from the chimpanzee chromosome 1, is within the FCGR1 gene family. This finding refines the assignment of homology between the human and chimpanzee chromosomes 1.
A baboon (Papio hamadryas) with an isochromosome for the long arm of the X
Authors: Moore C.M. , Leland M.M. , Brzyski R.G. , McKeand J. , Witte S.M. , Rogers J.
Published: April 2003

A 5.5-yr-old female baboon was evaluated for sexual immaturity. She was small for her age and had normal external female genitalia. However, she lacked cyclical perineal turgescence and displayed atypical coloration of the perineal skin. Laparoscopy revealed a small uterus and absence of both ovaries. Cytogenetic analysis revealed a 42,X,i(X)(q10) karyotype. DNA analysis using loci DXS1683, which maps to Xp22.1, and DXS297, which maps to Xq27.3, was consistent with inheritance of the normal X chromosome from the dam and formation of the isochromosome Xq from the paternal X.
Fine mapping of 12 previously unassigned EST clones to individual YACs in the familial Alzheimer’s disease (FAD) region of chromosome 14q24.3
Authors: Ardley H.C. , Markham A.F. , Robinson P.A.
Published: April 2003

Abstract. We have constructed an approximately 4-Mb YAC contig spanning the microsatellite markers D14S1028–D14S61 on chromosome 14q24.3 using oligonucleotide primers to microsatellite markers, ESTs and the genes PSEN1, FOS, TGFB3 and UBE2L1. This contig consists of 44 ICI YACs and 5 megaCEPH YACs. The data reported here should aid the final construction of an integrated map of chromosome 14 and allow the identification and accurate mapping of additional novel ESTs within this region.
Assignment of Ptprn2, the gene encoding receptor-type protein tyrosine phosphatase IA-2β, a major autoantigen in insulin-dependent diabetes mellitus, to mouse chromosome region 12F
Authors: van den Maagdenberg A.M.J.M. , Schepens J.T.G. , Schepens M.T.M. , Pepers B. , Wieringa B. , Geurts van Kessel A.Geurts , Hendriks W.J.A.J.
Published: April 2003

The receptor-type protein tyrosine phosphatase IA-2β gene (mouse gene symbol Ptprn2) encodes a major autoantigen in insulin-dependent diabetes mellitus. We physically mapped Ptprn2 by fluorescence in situ hybridization to band F of mouse chromosome 12, a region that lacks diabetes susceptibility loci. The mapping confirms the proposed synteny of mouse 12F with band q36 of human chromosome 7.
A versatile image analysis approach for simultaneous chromosome identification and localization of FISH probes
Authors: Christian A. , McNiel E. , Robinson J. , Drabek R. , LaRue S. , Waldren C. , Bedford J.
Published: April 2003

Modern cytogenetic techniques, such as comparative genomic hybridization (CGH) and the multi-color fluorescence in situ hybridization (FISH) techniques of multiplex fluorescence in situ hybridization (M-FISH) and spectral karyotyping (SKY), require a coordinated banding analysis to maximize their usefulness. All of the methods currently used, including Giemsa (G-) banding, Alu banding, and 4′,6-diamidino-2-phenyl-indole (DAPI) banding, have serious drawbacks. A simple and effective method to band chromosomes concurrently with FISH is needed. To address this problem, we stained chromosomes with DAPI and chromomycin A3, and then used an image analysis program to generate banding by dividing the image taken with a DAPI excitation filter by the image taken with a chromomycin A3 excitation filter. The result was a metaphase spread in which the chromosomes possessed a banding pattern characteristic of R-banding. The image analysis program was then used to generate linescans of pixel intensity versus relative position along the length of chromosomes that were banded using this technique, which we have called D/C R-banding. Each chromosome in a genome was represented by a characteristic scan profile, which was unaffected by FISH signals. Reference linescans were prepared by karyotyping D/C R-banded chromosomes for a given species, and then drawing lines along the length of the known chromosomes. The linescans were combined into a spreadsheet database, which was linked by dynamic data exchange to the image analysis program and normalized for length and intensity. The linescan of an unknown chromosome was then transferred to the spreadsheet, where it was normalized for length and intensity and overlaid on the linescans of each chromosome in the genome. Unknown chromosomes were identified by comparison of their graphs with graphs in the standardized reference genome. We have used this approach to create reference linescan karyotypes of several species, and to identify chromosomes on which FISH was performed.
Refined chromosomal localization of the putative tumor suppressor gene TP73
Authors: Lo Cunsolo C. , Casciano I. , Banelli B. , Tonini G.P. , Romani M.
Published: April 2003

We report the refined chromosomal localization of the putative tumor suppressor gene TP73 (alias p73) within the genomic region between the anonymous loci D1Z2 and D1S47. The region measures less than 6 Mb and covers a genetic distance of 16 cM. The present mapping considerably restricts the previous cytogenetic localization of TP73.
Sperm segregation analysis of a complex chromosome rearrangement, 2;22;11, by whole chromosome painting
Authors: Cifuentes P. , Navarro J. , Míguez L. , Egozcue J. , Benet J.
Published: April 2003

Using the human sperm–hamster oocyte fusion technique and whole chromosome painting, we studied sperm chromosome segregation in a male heterozygous for a complex chromosome rearrangement, 46,XY,–2,+der(2)t(2;11)(q13; q23),–11,+der(11)t(11;22)(q23;q11.2),–22,+der(22)t(2;22)(q13; q11.2). A total of 208 sperm complements were analyzed. The frequency of sperm carrying a normal or a balanced complement was 13.5% (9.62% and 3.85%, respectively). The frequency of unbalanced sperm was 86.5% (64.9% from 3:3 segregation, including 30 different types; 20.7% from 4:2 segregation, including 21 different types; and 0.96% from 5:1 segregation, including 2 different types). The sex ratio, determined in 134 sperm complements, did not differ from the expected 1:1 ratio. The results obtained in this study are compatible with the formation, during the synaptic process, of a complex hexavalent figure involving chromosomes 2, 11, and 22. The behavior and segregation of this complex figure would explain the high frequency (86.5%) of unbalanced complements observed in this carrier.
Fluorescence in situ hybridization analysis of the replication properties of the myotonic dystrophy protein kinase (DMPK) gene region
Authors: Rajcan-Separovic E. , Barcelo J.M. , Korneluk R.G.
Published: April 2003

Abstract. Myotonic dystrophy (DM) is caused by an expansion of a CTG repeat sequence in the 3′ noncoding region of a protein kinase gene (DMPK) at 19q13.3. We used in situ hybridization to analyse the replication timing of the genomic region containing DMPK in fibroblasts and myoblasts from controls and myotonic dystrophy patients. In this method the relative proportion of singlet to doublet hybridization signals is used to infer the relative time of replication of specific loci or regions. Our results show that in cells from normal individuals approximately 65% of signals appear as doublets, indicating early replication. In DM patients with a number of CTG repeats ranging from about 600–1800 we observed a significant increase of singlet-doublets compared to the background level. These results suggest the existence of replication alternations and/or structural differences between the normal and mutant alleles induced by the presence of the DM mutation.
A satellite DNA of the Sparidae family (Pisces, Perciformes) associated with telomeric sequences
Authors: Garrido-Ramos M.A. , de la Herrán R. , Ruiz Rejón C. , Ruiz Rejón M.
Published: April 2003

This paper reports on the isolation and localization of the subtelomeric DraI satellite DNA in the Sparidae family. Gene cloning determined that the DraI satellite DNA is present in only 3 species (Pagrus pagrus, P. auriga, and Pagellus erythrinus) of the 10 Sparidae species analyzed. The results were confirmed by PCR amplification. This satellite DNA is located in a subtelomeric position in all 48 acrocentric chromosomes of these species. However, interstitial loci are also observed. Sequence analysis of monomers of this repetitive family indicates that the satellite DNA is associated with telomeric sequences, (TTAGGG)n, in at least one species, P. erythrinus. This is the first direct demonstration of the existence of the consensus telomere sequences of vertebrates in fish. Likewise, this report also demonstrates that the ends of fish chromosomes have a structure similar to those of most eukaryote chromosomes, viz., telomere sequences and subtelomeric sequences associated by a boundary in which both types of sequences are interspersed. The recent origin of the DraI satellite DNA and its use as a phylogenetic marker is discussed.
Description and physical localization of the bovine survival of motor neuron gene (SMN)
Authors: Pietrowski D. , Goldammer T. , Meinert S. , Schwerin M. , Förster M.
Published: April 2003

Proximal spinal muscular atrophy (SMA) is an autosomal recessive disease in humans and other mammals, characterized by degeneration of anterior horn cells of the spinal cord. In humans, the survival of motor neuron gene (SMN) has been recognized as the SMA-determining gene and has been mapped to 5q13. In cattle, SMA is a recurrent, inherited disease that plays an important economic role in breeding programs of Brown Swiss stock. Now we have identified the full- length cDNA sequence of the bovine SMN gene. Molecular analysis and characterization of the sequence documents 85% identity to its human counterpart and three evolutionarily conserved domains in different species. Physical mapping data reveals that bovine SMN is localized to chromosome region 20q12→q13, supporting the conserved synteny of this chromosomal region between humans and cattle.
Organization and expression of rat Tspy
Authors: Dechend F. , Schubert S. , Nanda I. , Vogel T. , Schmid M. , Schmidtke J.
Published: April 2003

We have isolated both a full-length rat Tspy cDNA from testicular mRNA by rapid amplification of cDNA ends (RACE) and RT-PCR and a full-length rat Tspy gene from genomic DNA by PCR. In contrast to the mouse, where Tspy is present in a single copy and is apparently functionless, and to man and cattle, where TSPY is organized in a moderately repetitive cluster, the rat Tspy locus apparently consists of one complete functional and one truncated, probably nonfunctional, copy, coherently localized on Yp, as revealed by FISH analysis.
Analysis of the monomeric alphoid sequences in the pericentromeric region of human chromosome 7
Authors: de la Puente A. , Velasco E. , Pérez Jurado L.A. , Hernández-Chico C. , van de Rijke F.M. , Scherer S.W. , Raap A.K. , Cruces J.
Published: April 2003

To further define the structure of the pericentromeric region of human chromosome 7, we have identified and characterized a YAC clone (YAC 311.H5) containing the D7S1480 locus, which maps to the short arm near the centromere of this chromosome, by linkage in CEPH families and radiation hybrid analysis. This YAC contains two new blocks of alphoid DNA (named Z5 and Z6). Both Z5 and Z6 show monomeric structures and a lack of higher-order repeats, and, therefore, belong to suprachromosomal family type 4 (M1). The orientation of the two blocks and the physical distances over the region were defined by pulsed-field gel electrophoresis (PFGE) and fluorescence in situ hybridization on chromatin fibers (FiberFISH). A YAC contig spanning the centromeric region has been developed by STS content.
Fine mapping of distal 1p loci reveals TP73 at D1S468
Authors: Perri P. , Praml C. , Savelyeva L. , Pillmann A. , Schwab M.
Published: April 2003

In the present study we establish a FISH fine-map of 1p36.3 loci. This region is frequently altered in different types of human tumors suggesting the existence of cancer-related genes. Identification of cosmids carrying both D1S468 and TP73 sequences leads to the assignment of TP73 to the most frequently deleted locus in colon and breast cancer and integrates this gene in human genetic maps. Localization of other distal loci was determined as follows: distal–CDC2L1–D1Z2–D1S94–TP73/D1S468–D1S1615–proximal. D1S1615, earlier reported as a telomeric sequence, is considerably more proximal than previously thought.
Synteny mapping of five human chromosome 7 genes on bovine chromosomes 4 and 21
Authors: Antoniou E. , Womack J.E. , Grosz M.D.
Published: April 2003

Five genes on human chromosome 7 (HSA 7) were assigned to bovine chromosome 21 (BTA 21) and 4 (BTA 4) using a bovine-rodent somatic hybrid cell panel. These five genes were alpha-I subunit of adenylate cyclase-inhibiting G-protein (GNAI1), alpha/beta preprotachykinin (TAC1), reelin (RELN), c-AMP dependant protein kinase type II beta regulatory chain (PRKAR2B) and apolipoprotein A1 regulatory protein 1 (TFCOUP2). Four genes mapped to BTA 4 (GNAI1, TAC1, RELN, PRKAR2B) while one gene mapped to BTA 21 (TFCOUP2). This study confirms the synteny conservation between HSA 7 and BTA 4, finely maps the breakpoints of conserved synteny on HSA 7 and defines a new synteny conservation between HSA 7 and BTA 21.
Genomic organization of the human complex I 13-kDa subunit gene NDUFA5
Authors: Tensing K. , Pata I. , Wittig I. , Wehnert M. , Metspalu A.
Published: April 2003

We have characterized the human gene NDUFA5 encoding a 13-kDa subunit of the mitochondrial respiratory chain complex I (NADH: ubiquinone oxidoreductase). The gene contains 5 exons and 4 introns, and spans 14 kb of genomic DNA. In the untranscribed region we observed potential transcription factor binding sites. We determined a single nucleotide variant (C/T) at –318, and its frequency in the German population. The functional gene was localised by FISH to 7q31 and by radiation hybrid panel near marker D7S648 in YAC 883_a_2.
Cytogenetic assignment of 53 microsatellites from the USDA-MARC porcine genetic map
Authors: Lopez-Corrales N.L. , Beattie C.W. , Rohrer G.A.
Published: April 2003

This study provides 53 new fluorescent in situ hybridization cytogenetic assignments for microsatellite markers linked on the swine genetic map. Forty microsatellites are physically assigned for the first time. The chromosomal locations of eight markers were either confirmed or refined, while five loci were assigned to locations different from those given in previous reports. Markers were selected to provide physical anchors based on their presumed proximity to centromeres or telomeres and at approximately 30 cM intervals across the genetic map. The number of physical anchors for pig (SSC) chromosomes 8, 15, and 18 linkage groups was significantly improved. Centromeric regions were localized to areas less than 10 cM for SSC 1, 2, 3, 6, 7, 8, and 9. Although the recombination rate was generally higher across small biarmed chromosomes and lowest for large acrocentric chromosomes, two regions with particularly low (1q2.1→q2.9 and 13q2.3→q4.1) and three regions with extremely high (5p1.5→p1.2, 6p1.4→p1.3, and 12p1.5→p1.4) rates of recombination were detected. These assignments represent an overall 10% increase in the number of physically assigned markers in Sus scrofa and more than a 20% increase in the number of Type II loci assigned to the pig cytogenetic map.
Assignment of human teratocarcinoma derived growth factor (TDGF) sequences to chromosomes 2q37, 3q22, 6p25 and 19q13.1
Authors: Scognamiglio B. , Baldassarre G. , Cassano C. , Tucci M. , Montuori N. , Dono R. , Lembo G. , Barra A. , Lago C.T. , Viglietto G. , Rocchi M. , Persico M.G.
Published: April 2003

The human teratocarcinoma derived growth factor 1 (TDGF1) gene maps on chromosome (Chr) 3p21.3. One pseudogene (TDGF3) maps on Chr Xq21→q22. We now report the nucleotide sequence and chromosome location of three additional TDGF pseudogenes. The three new sequences (TDGF2, TDGF4 and TDGF5) are truncated at the 5′ end and have accumulated several point mutations, deletions and insertions. TDGF2, TDGF4 and TDGF6 map on Chrs 2q37, 6p25 and 3q22, respectively. Finally, Southern blot analysis of DNA from normal individuals shows a highly variable restriction pattern of the TDGF sequences.
The application of AFLP fingerprinting to construct a YAC contig containing ADH2 and MTP on sheep chromosome 6
Authors: Lumsden J.M. , Lord E.A. , Cato S.A. , Richardson T.E. , van Stijn T.C. , Broom M.F. , Patel K. , Montgomery G.W.
Published: April 2003

The low density of genetic markers on livestock maps limits progress in positional cloning projects. We demonstrate a strategy of combining comparative mapping with AFLP fingerprinting to develop physical maps in a defined region of the sheep genome. Sequence tagged sites for alcohol dehydrogenase 2 (ADH2) and microsomal triglyceride transfer protein (MTP) were developed and used to screen a sheep yeast artificial chromosome (YAC) library. Nine YACs were identified containing the microsatellite marker BM1329 and either ADH2 or MTP. Additional markers in the region were not available, and AFLP analysis was developed to identify sheep-specific bands within the YACs to determine their degree of overlap. Fourteen bands common to more than one YAC were analysed and provided the markers necessary to develop a YAC contig containing the three STS markers. One YAC (yac260B5) containing all three markers (ADH2, MTP, and BM1329) was mapped to sheep chromosome 6q1.6→q1.8 by FISH analysis.
Genomic organization and chromosome location of the rat CD3ζ locus (Cd3z)
Authors: Matsuura A. , Kinebuchi M. , Itoh Y. , Kasai K. , Yamada K. , Yoshida M.C. , Horie M. , Kikuchi K. , Sato N.
Published: April 2003

The structure and pattern of expression of the CD3ζ chain have apparently been conserved throughout mammalian evolution. The organization of the rat CD3ζ locus was determined by genomic cloning and nucleotide sequencing. Most of the rat CD3ζ coding region was similar to mouse and human CD3ζ sequences. Whereas the 3′ region involving alternative splicing was relatively well conserved at the nucleotide level, the deduced amino acid sequences were different in rats, mice and humans due to the presence of deletion and insertion mutations. Alternative splicing products of the CD3ζ locus, such as CD3η, CD3θ and CD3τ, which have been reported for mice, were not expressed by rat T cells. By using fluorescence in situ hybridization, we have localized rat CD3ζ to chromosome 13q22→q23.
The novel contiguous gene syndrome of myotubular myopathy (MTM1), male hypogenitalism and deletion in Xq28:report of the first familial case
Authors: Bartsch O. , Kreß W. , Wagner A. , Seemanova E.
Published: April 2003

Hu et al. (1996) and Laporte et al. (1997) recently proposed a novel contiguous gene syndrome of myotubular myopathy, abnormal male genital development and deletion in Xq28. We studied a family where two male infants, both deceased, had myotubular myopathy and intersexual genitalia. Using FISH we detected in the mother a hemizygous deletion including the myotubularin gene MTM1 and F18 (a gene of yet unknown function). DNA studies with STR-markers (short tandem repeats) within and flanking the deleted segment confirmed the deletion in the family and were used for prenatal diagnosis. Our findings confirm the existence of this novel contiguous gene syndrome and support that the deletion of the F18 gene, or a neighboring gene, may cause ambiguous genitalia or severe hypospadias in males. The mother had low muscle power and marked menstrual irregularities which may indicate that she is a manifesting carrier and that the deletion may include a gene (F18 or other) for gonadal function in females.
Cloning, structural organization and chromosomal localization of the mouse Aquaporin-8 water channel gene (Aqp8)
Authors: Calamita G. , Spalluto C. , Mazzone A. , Rocchi M. , Svelto M.
Published: April 2003

The gene encoding the mouse Aquaporin-8 water channel protein (Aqp8) was cloned and its genomic structure was defined. Aqp8 consists of six exons with boundaries at amino acids 1–4, 5–87, 88–129, 130–201, 202–246 and 247–261 which partially correspond to those of other known aquaporin genes. All splice sites conform to the GT–AG rule except the first one which is GT–GG. Primer extension and RNase protection analyses using mouse liver RNA demonstrated three initiation transcription sites located 385, 156 and 146 bp upstream from the translational start codon. No defined TATA box was found in the 5′-flanking region where numerous CAAT motifs and one GATA box were identified. Fluorescence in situ hybridization localized the Aqp8 locus to mouse chromosome 7F3. The 7F region is syntenic with human chromosomes 11, 16 and 10. These results (i) reveal marked structural distinction between the Aqp8 gene and the other known mammalian aquaporin genes, (ii) may now permit the molecular characterization of Aqp8 expression and (iii) represent a fundamental step for the construction of a target vector to generate transgenic Aqp8 knockout mice.
Chromosome duplications and deletions and their mechanisms of origin
Authors: Tharapel A.T. , Michaelis R.C. , Velagaleti G.V.N. , Laundon C.H. , Martens P.R. , Buchanan P.D. , Teague K.E. , Tharapel S.A. , Wilroy Jr. R.S.
Published: April 2003

Duplications and deletions of the same gene loci or chromosome regions are known to produce different clinical manifestations and are significant factors in human morbidity and mortality. Extensive cytogenetic and molecular cytogenetic studies with cosmid and YAC probes in two patients with unique mosaicism for reciprocal duplication-deletion allowed us to further understand the origin of these abnormalities. The first patient’s mosaic karyotype was 46,XX,inv dup(11) (q23q13)/46,XX,del(11)(q13q23). The second patient had a 46,XY,dup(7)(p11.2p13)/46,XY,del(7)(p11.2p13)/46,XY karyotype. Fluorescence in situ hybridization studies on the first patient placed the two breakpoints near the folate-sensitive fragile sites FRA11A and FRA11B. The presence of repeated sequences responsible for these fragile sites may have been involved in the patient’s duplication-deletion. Our investigation leads us to conclude that, in addition to known mechanisms (such as unequal crossovers between homologs, unequal sister chromatid exchanges, excision of intrachromatid loops, and meiotic recombination within a single chromatid), duplication-deletion can also arise by the formation of an overlying loop followed by an uneven crossover at the level of the DNA strand.