Characterization of a deletion in the Hsp70 cluster in the bovine reference genome.

The 70 kilodalton heat shock proteins (Hsp70) are highly conserved molecular chaperones which have a crucial role in the stress response of the cell. In mammals, the Hsp70 proteins are encoded by a cluster of three genes: HSPA1A, HSPA1B and HSPA1L. In bovines, this cluster is located on chromosome 23 downstream of the major histocompatibility complex (BoLA). We detected inconsistencies in the location of markers on the Hsp70 genes reported in the literature that pointed to a potential deletion in the bovine reference genome UMD 3.1.1. An in silico analysis of the bovine genomic region of the Hsp70 cluster, using available information from public databases, confirmed the existence of a deletion of 11.1-kb spanning the HSPA1B gene and the intergenic region between HSPA1B and HSPA1A. Although we originally considered this an assembly error, it is most likely a particular condition of L1 Dominette 01449, the cow sequenced in the Bovine Genome Project. Moreover, we suggest a new classification of bovine Hsp70 sequences reported in NCBI and a reassignment of the location of SNPs from dbSNP that map to the deletion on BTA23. We also compared the location of selected transcription factor binding sites on the promoters of HSPA1A and HSPA1B. The results generated in the present work could be helpful to refine the reference genome of an important livestock species and also to understand the role and the regulation of the bovine Hsp70 genes.

Mapping and characterization of new EST-derived microsatellites for potato (Solanum tuberosum L.).

Microsatellites, or simple sequence repeats (SSRs) are very useful molecular markers for a number of plant species. They are commonly used in cultivar identification, plant variety protection, as anchor markers in genetic mapping, and in marker-assisted breeding. Early development of SSRs was hampered by the high cost of library screening and clone sequencing. Currently, large public SSR datasets exist for many crop species, but the number of publicly available, mapped SSRs for potato is relatively low (approximately 100). We have utilized a database mining approach to identify SSR-containing sequences in The Institute For Genomic Research Potato Gene Index database (http://www.tigr.org), focusing on sequences with size polymorphisms present in this dataset. Ninety-four primer pairs flanking SSR sequences were synthesized and used to amplify potato DNA. This study rendered 61 useful SSRs that were located in pre-existing genetic maps, fingerprinted in a set of 30 cultivars from South America, North America, and Europe or a combination thereof. The high proportion of success (65%) of expressed sequence tag-derived SSRs obtained in this work validates the use of transcribed sequences as a source of markers. These markers will be useful for genetic mapping, taxonomic studies, marker-assisted selection, and cultivar identification.

Variability among inbred lines and RFLP mapping of sunflower isozymes

Eight isozyme systems were used in this study: acid phosphatase (ACP), alcohol dehydrogenase (ADH), esterase (EST), glutamate dehydrogenase (GDH), malate dehydrogenase (MDH), phosphoglucoisomerase (PGI), 6-phosphogluconate dehydrogenase (PGD), and phosphoglucomutase (PGM). The polymorphism of these enzyme systems was studied in 25 elite inbred lines. A total of 19 loci were identified, but only eight of them were polymorphic in the germplasm tested. The polymorphic index for the eight informative markers ranged from 0.08 to 0.57, with a mean value of 0.36. Five isozyme loci were mapped in F2:3 populations with existing RFLP data. Est-1, Gdh-2 and Pgi-2 were mapped to linkage groups 3, 14 and 9, respectively. As in previous reports, an ACP locus and a PGD locus were found to be linked, both located in linkage group 2 of the public sunflower map

Blood vitamin and choline concentrations in healthy domestic cats, dogs, and horses.

Blood concentrations of thiamin, biotin, nicotinates, pantothenates, folates, riboflavin, vitamins A, B6, B12, C, E, beta-carotene and choline were analyzed in healthy animals (23 horses, 25 dogs, and 29 cats). B-Complex vitamins and choline also were analyzed in the liver of the dogs and cats. Vitamin concentrations in the blood and livers of dogs were similar; however, blood vitamin A and beta-carotene concentrations were lower in the cat than in the dog. Horses had a higher B12 blood concentration than did the dogs and cats. These data can be useful for detecting overt and hidden vitamin deficits in these species due to various conditions.

Improved response of a murine fibrosarcoma (Meth-A) to interstitial radiation when combined with hyperthermia.

Removable Iridium-192 implants provided a dose of 10 Gy/day or 41.5 cGy/hr at 0.5 cm from the center of 1.0 cm diameter tumors. The total radiation (x) alone was 20, 40 or 60 Gy, representing 2, 4, or 6 days of continuous radiation. The doses used for the combined treatments at elevated temperatures were 10, 20 or 30 Gy. The local tumor hyperthermia (LTH) treatment (43.6 for 35 min, water bath) was administered once for each 10 Gy of dose. The combined radiation + LTH was clearly superior to that achieved with radiation or LTH alone and yielded Thermal Enhancement Ratios (TER) of 3.4-3.9. Local tumor control rate was 67% and 89% for the 20 and 30 Gy x + LTH groups, respectively. A comparison of the results obtained in this study with those of earlier studies on the same tumor system indicate that the effects of Iridium-192 alone on this rapidly proliferating tumor were comparable to similar total doses of fractionated external beam radiation. Iridium + LTH produced a tumor response comparable to that achieved with external fractionated radiation + LTH. Combined treatment effect of elevated temperature appears to be less dose rate dependent in the range of 40 cGy/hr to 100 cGy/min and more dependent upon total dose accumulation.

Vitamin content of some normal human brain segments.

Nicotinates, pantothenates, riboflavin, vitamins B6 and B12' free (acetyl) and total (free and bound) choline, biopterin, thiamin, biotin, methylated and nonmethylated folates in frontal, temporal, precentral, postcentral, and occipital cortex, thalamus, cerebellum, pons, basal ganglia, and substantia nigra were estimated. Nicotinates are significantly more concentrated in basal ganglia and thalamus than pons. Nonmethylated folate content is not significantly varied in brain segments; the pons contains more methylated folate. Riboflavin content is higher in the basal ganglia and temporal cortex than frontal cortex. Biotin is concentrated in pons and basal ganglia. Thiamin concentration is less in the postcentral cortex than the thalamus and substantia nigra. Biopterin is significantly higher in substantia nigra and basal ganglia than the other brain segments. Total choline content is high in substantia nigra, pons, and thalamus; free (acetyl) choline is significantly elevated in basal ganglia. B12 content is less concentrated in the cortex segments. B6 is highly concentrated in the basal ganglia. Pantothenate content is elevated in pons when compared to the various cortex segments and cerebellum.

Recovery from aspermia induced by low-dose radiation in seminoma patients.

Gonadal dosimetry and spermatogenic activity was monitored in patients given radiation therapy (RT) after unilateral orchiectomy for seminoma. The RT given was, with minor variations, 3200 rad in 16 fractions in four weeks to the para-aortic and ipsilateral pelvic inguinal lymphatics in order to include the orchiectomy scar. The incidental amount of radiation to the remaining testicle averaged 78.4 +/- 7.4 rad and ranged from 32-178 rad as determined by thermoluminescent dosimetry. Induction of aspermia was documented in ten out of 14 patients who received over 65 rad to the gonad. At lower doses, aspermia may not have occurred or was of short duration. Recovery of sperm in the semen occurred in 12 patients within 30-80 weeks after start of treatment. The data suggest that the time of recovery may be dose dependent within the range of 19-148 rad. During the period of recovery, patients with oligospermic semen may be fertile and should be so advised.

Role of placenta in maternal-fetal vitamin transfer in humans.

Vitamins B12, B6, biotin, folate, thiamine, riboflavin, pantothenate, and nicotinate were determined in maternal and fetal blood and placental tissue of normovitaminemic and hypovitaminemic mothers who disclaimed supplemental vitamin intake during pregnancy. No biotin or pantothenate deficits were observed in the gravidas. Hypovitaminemic mothers transferred less B12, folate, and B6 to the fetus and placenta than normovitaminemic mothers. Vitamins given by mouth increased maternal fetal, and placental levels of folate, but B6 increased only in maternal blood and the placenta; biotin and pantothenate increased only in fetal blood. Except for riboflavin, nicotinate, and pantothenate, the intramuscular administration of vitamins increased the levels of other vitamins in maternal and fetal blood and placental tissue. Results suggest that the placenta stores vitamins and the tissue vitamin receptors must be saturated before adequate transfer of vitamins to the fetus occurs.

Elevated vitamin levels in colon adenocarcinoma as compared with metastatic liver adenocarcinoma from colon primary and normal adjacent tissue.

Twenty-four samples of colon adenocarcinoma removed at surgery and autopsy together with adjacent uninvaded normal colon from the same subjects were analyzed for vitamin B12 and B6, biopterin, nicotinate, riboflavin, pantothenate, thiamin, biotin, and folates. Nine specimens of metastatic liver adenocarcinoma from colon primary together with adjacent uninvaded normal liver were also analyzed for these same vitamins. Primary colon adenocarcinoma contains significantly (P less than 0.001) more of the above vitamins than normal colon; 1.8- to 3.5-fold higher concentrations of vitamins were found in this tumor. In contrast, vitamin B12 levels were almost two-fold lower. Unlike colon tumor, metastatic liver adenocarcinoma from colon primary contained from 1.2- to 28-fold lower vitamin concentration than normal liver tissue. The present findings suggest that those types of primary tumors with conspicuously high vitamin content needed for the enhanced growth and catalysis of tumor metabolism may be arrested with antivitamins targeted at metabolic sites other than those involved with nucleic acid synthesis.

Concrete and verbal reinforcement in creative thinking of disadvantaged cildren.

The effects of verbal and concrete reinforcement on creative thinking were investigated in disadvantaged Israeli seventh-graders (N = 90) in a baseline-treatment design. When compared to a no-incentive condition, both concrete (giving a piece of candy for each response) and verbal reinforcement (giving verbal praise to each response) raised the level of ideational fluency on the abbreviated Wallach and Kogan creativity battery by 114% and 61%, respectively. The high correlations between baseline and incentive creativity scores were interpreted as supporting either cognition or motivation but not exclusively the former as the source of individual differences in creative thinking among these disadvantaged children.

The measurement of skin blood flow in peripheral vascular disease by epicutaneous application of Xenon133.

Radioactive xenon133 applied epicutaneously was used to study the skin blood flow below the knee in sixteen normal subjects, in eight patients with peripheral vascular disease not requiring amputation, and in a blind study of twenty-nine patients requiring amputation. Following these twenty-nine amputations, the flow rates were made known and correlated with the rates of healing. When the flow rates were above 1.5 milliliters per minute per 100 grams of tissue (skin), the wounds generally healed. In thirteen other patients, Syme or below-the-knee amputations were performed on the basis of the flow rates, and all of the wounds healed. The test, therefore, is now used routinely prior to amputation for peripheral vascular disease as an adjunct to clinical judgment in the determination of the level of amputation.

Aspermia and recovery of spermatogenesis in cancer patients following incidental gonadal irradiation during treatment: a progress report.

The 11 patients in this study who received incidental gonadal irradiation from clinical therapy all became aspermic within 8-34 weeks following treatment. The estimated gonadal dose was 118-228 rads. Five of these patients have shown recovery of spermatogenesis; 3 of these have reached fertilizable concentrations. Recovery of semen sperm was noted at 44-77 weeks following treatment. The other 2 patients, after 110-176 weeks, have sub-fertile concentrations of semen sperm. Five patients have not been followed for a sufficient length of time to permit recovery to be noted.