Search

The Chinese soft-shelled turtle (Pelodiscus sinensis, Trionychidae, Testudines) has ZZ/ZW-type micro-sex chromosomes where the 18S-28S ribosomal RNA genes (18S-28S rDNA) are located. The W chromosome is morphologically differentiated from the Z chromosome by partial deletion and amplification of 18S-28S rDNA and W-specific repetitive sequences. We recently found a functional gene (TOP3B) mapped on the P. sinensis Z chromosome, which is located on chicken (Gallus gallus, GGA) chromosome 15. Then we cloned turtle homologues of 4 other GGA15-linked genes (GIT2, NF2, SBNO1, SF3A1) and localized them to P. sinensis chromosomes. The 4 genes all mapped on the Z chromosome, and 2 of them (SBNO1, SF3A1) were also localized to the W chromosome. Our mapping data suggest that at least one large inversion occurred between GGA15 and the P. sinensis Z chromosome, and that there are homologous regions in the distal portions of both the short and long arms between the Z and W chromosomes. W chromosomal differentiation in P. sinensis probably proceeded by the deletion of the proximal chromosomal region followed by 18S-28S rDNA amplification, after a paracentric inversion occurred at the breakpoints between the distal region of 18S-28S rDNA and the proximal region of SBNO1 on the Z chromosome. Copyright © 2009 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Cancer vaccines represent a promising new strategy for immunotherapy against cancer, but their effects are insufficiently understood. The effect of heat treatment against mouse colon adenocarcinoma cell line (colon-26), and combined effects of heat treatment and immunizing host animals with heat treated colon-26 cell extracts were investigated. Heat treatment of colon-26 cells induced heat-shock protein 70 (HSP70), but not other HSP. Immunization of BALB/cJ mice with heat treated colon-26 cell extract, which was enriched in HSP70, elicited antitumour immunity against subcutaneously injected colon-26 cells. Furthermore, combination therapy of heat treatment and immunization with heat treated colon-26 cell extract significantly reduced tumour volumes compared with heat treatment alone. Similar immunization enhanced the cytotoxic activity of mouse splenic lymphocytes against untreated and heat treated colon-26 cells in an in vitro assay, as well as against heat treated allogenic mouse lymphoma cell line (YAC1). These findings suggest possible usefulness of heat treated cancer cell extract as a cancer vaccine, especially if given in combination with hyperthermia. [ABSTRACT FROM AUTHOR]

1. Major biliary conjugates of the male Wistar rat dosed orally with 2,4-dinitrotoluene (2,4-DNT) or 2,6-dinitrotoluene(2,6-DNT) were examined by hplc using potassium 2,4- dinitrobenzyl glucuronide (potassium 2,4-DNB- G), potassium 2,6-dinitrobenzyl glucuronide (potassium 2,6-DNB- G), pyridinium 2,4-dinitrobenzyl sulphate (pyridinium 2,4- DNB- S) and pyridinium2,6-dinitrobenzylsulphate (pyridinium2,6-DNB- S) as authentic compounds. Other metabolites were also examined by hplc. In addition, metabolites formed by incubation of potassium 2,4-DNB- G and potassium 2,6-DNB- G with rat intestinal microflora under nitrogen were examined by hplc. 2. Conjugates detected directly from bile following administration of 2,4-DNT and 2,6-DNT were 2,4-DNB- G and 2,6-DNB- G, which accounted for 35 0 and 51 5 % of the administered dose respectively. No peaks corresponding to pyridinium 2,4-DNB- S and pyridinium 2,6-DNB- S were detected in bile samples. 3. 2-Amino- 4-nitrotoluene,4-amino- 2-nitrotoluene,2,4-diaminotolueneand 4-acetylamino- 2-nitrobenzoic acid (0 02-0 12 % of the dose excreted in 24 h), in addition to the known metabolites 2,4-dinitrobenzyl alcohol (2,4-DNB), 2,4-dinitrobenzalde hyde and 2,4-dinitrobenzoic acid (0 09-0 14 %), were detected in ether extracts of bile of rat given 2,4-DNT. 2,6-Dinitrobenzylalcohol (2,6-DNB), 2-amino- 6-nitrotolueneand 2,6-dinitrobenzaldehyde(0 02-0 03 %), which are known metabolites, were detected in ether extracts of bile from rat given 2,6-DNT. 4. Potassium 2,4-DNB- G was transformed by the anaerobic incubationof rat intestinal microflora into 2,4-DNB, 4-amino- 2-nitrobenzyl alcohol and 2-amino- 4-nitrobenzyl alcohol. Potassium 2,6-DNB- G was transformed into 2,6-DNB and 2-amino- 6-nitrobenzyl alcohol by the anaerobic incubation. Time-course studies showed that 2,4-DNB, 4-amino- 2-nitrobenzyl alcohol, 2-amino- 4-nitrobenzyl alcohol and 2,6-DNB, 2-amino- 6- nitrobenzyl alcohol peaked at 30, 75, 120 and 10, 50 min respectively. 5. These results, together with previous findings, show that 2,4-dinitrobenzalde hyde and 2,6-dinitrobenzalde hyde, which are potent mutagens, are formed either by the hepatic metabolism of 2,4-DNB and 2,6-DNB formed by the intestinal metabolism of 2,4-DNBG and 2,6-DNB- G excreted in bile or by the direct hepatic metabolism of 2,4-DNT and 2,6-DNT. [ABSTRACT FROM AUTHOR]

Neutrophils play an essential role in innate immune responses to bacterial and fungal infections, and loss of neutrophil function can increase the risk of acquiring lethal infections in clinical settings. Here, we show that engineered neutrophil-primed progenitors derived from human induced pluripotent stem cells can produce functional neutrophil-like cells at a clinically applicable scale that can act rapidly in vivo against lethal bacterial infections. Using 5 different mouse models, we systematically demonstrated that these neutrophil-like cells migrate to sites of inflammation and infection and increase survival against bacterial infection. In addition, we found that these human neutrophil-like cells can recruit murine immune cells. This system potentially provides a straight-forward solution for patients with neutrophil deficiency: an off-the-shelf neutrophil transfusion. This platform should facilitate the administration of human neutrophils for a broad spectrum of physiological and pathological conditions., (© 2021 by The American Society of Hematology.)

Since the Fukushima Dai-ichi Nuclear Power Plant accident, radiation effects on nonhuman biota in the contaminated areas have been a major concern. Here, we analyzed the frequencies of chromosomal aberrations (translocations and dicentrics) in the splenic lymphocytes of large Japanese field mice (Apodemus speciosus) inhabiting Fukushima Prefecture. A. speciosus chromosomes 1, 2, and 5 were flow-sorted in order to develop A. speciosus chromosome-specific painting probes, and FISH (fluorescence in situ hybridization) was performed using these painting probes to detect the translocations and dicentrics. The average frequency of the translocations and dicentrics per cell in the heavily contaminated area was significantly higher than the frequencies in the case of the noncontaminated control area and the slightly and moderately contaminated areas, and this aberration frequency in individual mice tended to roughly increase with the estimated dose rates and accumulated doses. In all four sampling areas, the proportion of aberrations occurring in chromosome 2 was approximately >3 times higher than that in chromosomes 1 and 5, which suggests that A. speciosus chromosome 2 harbors a fragile site that is highly sensitive to chromosome breaks induced by cellular stress such as DNA replication. The elevated frequency of chromosomal aberrations in A. speciosus potentially resulting from the presence of a fragile site in chromosome 2 might make it challenging to observe the mild effect of chronic low-dose-rate irradiation on the induction of chromosomal aberrations in A. speciosus inhabiting the contaminated areas of Fukushima.

Both ionizing radiation (IR) and psychological stress (PS) cause detrimental effects on humans. A recent study showed that chronic restraint-induced PS (CRIPS) diminished the functions of Trp53 and enhanced radiocarcinogenesis in Trp53-heterozygous (Trp53 +/- ) mice. These findings had a marked impact on the academic field as well as the general public, particularly among residents living in areas radioactively contaminated by nuclear accidents. In an attempt to elucidate the modifying effects of CRIPS on radiation-induced health consequences in Trp53 wild-type (Trp53 +/+ ) animals, investigations involving multidisciplinary analyses were performed. We herein demonstrated that CRIPS induced changes in the frequency of IR-induced chromosomal aberrations (CAs) in splenocytes. Five-week-old male Trp53 +/+ C57BL/6J mice were restrained for 6h per day for 28 consecutive days, and total body irradiation (TBI) at a dose of 4Gy was performed on the 8th day. Metaphase chromosome spreads prepared from splenocytes at the end of the 28-day restraint regimen were painted with fluorescence in situ hybridization (FISH) probes for chromosomes 1, 2, and 3. The results obtained showed that CRIPS alone did not induce CAs, while TBI caused significant increases in CAs, mostly translocations. Translocations appeared at a lower frequency in mice exposed to TBI plus CRIPS than in those exposed to TBI alone. No significant differences were observed in the frequencies of the other types of CAs (insertions, dicentrics, and fragments) visualized with FISH between these experimental groups (TBI+CRIPS vs. TBI). These results suggest that CRIPS does not appear to synergize with the clastogenicity of IR., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Following the Fukushima Dai-ichi Nuclear Power Plant accident, radiation effects on nonhuman biota in the contaminated areas have been a great concern. The induction of chromosomal aberrations in splenic lymphocytes of small Japanese field mice (Apodemus argenteus) and house mice (Mus musculus) inhabiting Fukushima Prefecture was investigated. In mice inhabiting the slightly contaminated area, the average frequency of dicentric chromosomes was similar to that seen in mice inhabiting a noncontaminated control area. In contrast, mice inhabiting the moderately and heavily contaminated areas showed a significant increase in the average frequencies of dicentric chromosomes. Total absorbed dose rate was estimated to be approximately 1 mGy d(-1) and 3 mGy d(-1) in the moderately and heavily contaminated areas, respectively. Chromosomal aberrations tended to roughly increase with dose rate. Although theoretically, the frequency of chromosomal aberrations was considered proportional to the absorbed dose, chromosomal aberrations in old mice (estimated median age 300 days) did not increase with radiation dose at the same rate as that observed in young mice (estimated median age 105 days).

Reptiles have a wide diversity of sex-determining mechanisms and types of sex chromosomes. Turtles exhibit temperature-dependent sex determination and genotypic sex determination, with male heterogametic (XX/XY) and female heterogametic (ZZ/ZW) sex chromosomes. Identification of sex chromosomes in many turtle species and their comparative genomic analysis are of great significance to understand the evolutionary processes of sex determination and sex chromosome differentiation in Testudines. The Mexican giant musk turtle (Staurotypus triporcatus, Kinosternidae, Testudines) and the giant musk turtle (Staurotypus salvinii) have heteromorphic XY sex chromosomes with a low degree of morphological differentiation; however, their origin and linkage group are still unknown. Cross-species chromosome painting with chromosome-specific DNA from Chinese soft-shelled turtle (Pelodiscus sinensis) revealed that the X and Y chromosomes of S. triporcatus have homology with P. sinensis chromosome 6, which corresponds to the chicken Z chromosome. We cloned cDNA fragments of S. triporcatus homologs of 16 chicken Z-linked genes and mapped them to S. triporcatus and S. salvinii chromosomes using fluorescence in situ hybridization. Sixteen genes were localized to the X and Y long arms in the same order in both species. The orders were also almost the same as those of the ostrich (Struthio camelus) Z chromosome, which retains the primitive state of the avian ancestral Z chromosome. These results strongly suggest that the X and Y chromosomes of Staurotypus turtles are at a very early stage of sex chromosome differentiation, and that these chromosomes and the avian ZW chromosomes share the same origin. Nonetheless, the turtles and birds acquired different systems of heterogametic sex determination during their evolution.

The black marsh turtle (Siebenrockiella crassicollis) has morphologically differentiated X and Y sex chromosomes. To elucidate the origin and evolutionary process of S. crassicollis X and Y chromosomes, we performed cross-species chromosome painting with chromosome-specific DNA from Chinese soft-shelled turtle (Pelodiscus sinensis) and chromosome mapping of the sex-linked genes of S. crassicollis using FISH. The X and Y chromosomes of S. crassicollis were hybridized with DNA probe of P. sinensis chromosome 5, which is homologous to chicken chromosome 5. S. crassicollis homologues of 14 chicken chromosome 5-linked genes were all localized to the X long arm, whereas two genes were mapped to the Y short arm and the other 12 genes were localized to the Y long arm in the same order as the X chromosome. This result suggests that extensive linkage homology has been retained between chicken chromosome 5 and S. crassicollis X and Y chromosomes and that S. crassicollis X and Y chromosomes are at an early stage of sex chromosome differentiation. Comparison of the locations of two site-specific repetitive DNA sequences on the X and Y chromosomes demonstrated that the centromere shift was the result of centromere repositioning, not of pericentric inversion.

Crocodilians have several unique karyotypic features, such as small diploid chromosome numbers (30-42) and the absence of dot-shaped microchromosomes. Of the extant crocodilian species, the Siamese crocodile (Crocodylus siamensis) has no more than 2n = 30, comprising mostly bi-armed chromosomes with large centromeric heterochromatin blocks. To investigate the molecular structures of C-heterochromatin and genomic compartmentalization in the karyotype, characterized by the disappearance of tiny microchromosomes and reduced chromosome number, we performed molecular cloning of centromeric repetitive sequences and chromosome mapping of the 18S-28S rDNA and telomeric (TTAGGG)( n ) sequences. The centromeric heterochromatin was composed mainly of two repetitive sequence families whose characteristics were quite different. Two types of GC-rich CSI-HindIII family sequences, the 305 bp CSI-HindIII-S (G+C content, 61.3%) and 424 bp CSI-HindIII-M (63.1%), were localized to the intensely PI-stained centric regions of all chromosomes, except for chromosome 2 with PI-negative heterochromatin. The 94 bp CSI-DraI (G+C content, 48.9%) was tandem-arrayed satellite DNA and localized to chromosome 2 and four pairs of small-sized chromosomes. The chromosomal size-dependent genomic compartmentalization that is supposedly unique to the Archosauromorpha was probably lost in the crocodilian lineage with the disappearance of microchromosomes followed by the homogenization of centromeric repetitive sequences between chromosomes, except for chromosome 2.

1. Conjugates of 2,4-dinitrobenzyl alcohol (2,4-DNB) and 2,6-dinitrobenzyl alcohol (2,6-DNB), which were major urinary metabolites of the male Wistar rat dosed orally with 2,4-dinitrotoluene (2,4-DNT) or 2,6-dinitrotoluene (2,6-DNT), were examined by hplc using potassium 2,4-dinitrobenzyl glucuronide (2,4-DNB-G), potassium 2,6-dinitrobenzyl glucuronide (2,6-DNB-G), pyridinium 2,4-dinitrobenzyl sulphate (2,4-DNB-S), and pyridinium 2,6-dinitrobenzyl sulphate (2,6-DNB-S) as authentic compounds. Other metabolites were also examined by hplc. 2. Conjugates detected from urine following administration of 2,4-DNT and 2,6-DNT were 2,4-DNB-G and 2,6-DNB-G, which accounted for about 10.7 and 17.4% of the administered dose respectively. No peaks corresponding to pyridinium 2,4-DNB-S and pyridinium 2,6-DNB-S were detected in urine samples. 3. 2-Amino-4-nitrobenzoic acid (0.71%), 4-amino-2-nitrobenzoic acid (0.52%) and 4-acetylamino-2-nitrobenzoic acid (3.9%), in addition to known metabolites 4-amino-2-nitrotoluene (0.04%), 2,4-DNB (0.25%), 2,4-dinitrobenzoic acid (6.9%) and 4-acetylamino-2-aminobenzoic acid (3.4%), were detected in ether extracts of urine of rat given 2,4-DNT. 2,6-Dinitrobenzoic acid (0.17%) and two known metabolites, 2-amino-6-nitrotoluene (0.44%) and 2,6-DNB (0.53%), were detected in ether extracts of urine of rat given 2,6-DNT.

Background: The prognosis of pancreatic adenocarcinoma still remains poor because of the lack of reliable diagnostic tests for early stages of the disease. Monoclonal antibody 17-1A (MoAb 17-1A) has been studied extensively, and the antigen recognized by MoAb 17-1A is expressed by adenocarcinomas of the pancreas and stomach, as well as other normal and malignant epithelial tissues. The potential of MoAb 17-1A was investigated for its ability to detect pancreatic carcinomas. The use of MoAb 17-1A in treatment also was studied., Methods: Immunoreactivity of MoAb 17-1A with human pancreatic carcinoma cell line HuP-T4 was examined histochemically by the avidin-biotinylated enzyme complex method. MoAb 17-1A was labeled with 125I by the Iodogen method and 111In using either diethylenetriaminepentaacetic anhydride (cDTPA) or 1-(p-benzyldiazonium) diethylenetriaminepentaacetic acid (aDTPA). After injection in nude mice bearing HuP-T4 xenografts, the biodistribution of 111In- and 125I-labeled MoAb 17-1A was examined at various time points., Results: Positive staining of MoAb 17-1A was noted for HuP-T4 cells. A statistically significant (P < 0.01) greater tumor uptake was observed at 3 days after intravenous injection of 125I-labeled MoAb 17-1A when compared with 125I-labeled nonspecific immunoglobulin G. 125I- and 111In-labeled MoAb 17-1A was concentrated in HuP-T4 carcinoma 1.9-4.8 times higher than in the spleen, heart, liver, and pancreas., Conclusions: MoAb 17-1A was found to bind selectively to human pancreatic carcinoma HuP-T4. Tumor exhibited higher uptake of radiolabeled MoAb 17-1A compared with adjacent normal tissues. These results suggest that MoAb 17-1A may be applicable to the radioimmunodetection and radioimmunotherapy of pancreatic adenocarcinomas.