Phytoremediation of trichloroethylene and dichlorodiphenyltrichloroethane-polluted water using transgenic Sesbania grandiflora and Arabidopsis thaliana plants harboring rabbit cytochrome p450 2E1

Sesbania grandiflora (L.) pers (Fabaceae) and Arabidopsis thaliana (L.) (Brassicaceae) were genetically engineered to constitutively express the rabbit cytochrome p450 2E1 enzyme aiming at increasing their activity toward trichloroethylene (TCE) and dichlorodiphenyltrichloroethane (DDT) removal Successful generation of Sesbania and Arabidopsis transgenic plants was verified using p450 2E1 specific PCR and confirmed by western blot analysis. Gas chromatography (GC) analysis revealed that small cuttings of Sesbania and third generation (F3) Arabidopsis transgenic plants exposed to TCE and DDT in small hydroponics' vessels accumulated more TCE and DDT compared to plants transformed with the empty vector. Furthermore, both transgenic plants were more effective in breaking down TCE and DDT with a 2-fold increase in TCE metabolism. Two independent Arabidopsis lines showed that DDT was metabolized about 4-fold higher than that detected in non transformed plants. Similarly, S. grandiflora cuttings removed 51 to 90% of the added DDT compared with only 3% removal in controls transformed with the null vector. Notably, stability of rabbit cytochrome p450 2E1 was confirmed using third generation Arabidopsis plants that displayed higher potential for the removal of two important pollutants, TCE and DDT compared with the controls.     

Ionic composition of endolymph and perilymph in the inner ear of the oyster toadfish, Opsanus tau

The concentrations of free Na+, K+, Ca(+, and Cl(-)in endolymph and perilymph from the inner ear of the oyster toadfish, Opsanus tau, were measured in vivo using double-barreled ion-selective electrodes. Perilymph concentrations were similar to those measured in other species, while endolymph concentrations were similar to those measured previously in elasmobranch fish, though significantly different from concentrations reported in mammals. Perilymph concentrations (mean +/- std. dev.) were as follows: Na+, 129 mmol l(-1) +/- 20; K+, 4.96 mmol l(-1) +/- 2.67; Ca2+, 1.83 mmol l(-1) +/- 0.27; and Cl(-), 171 mmol l(-1) +/- 20. Saccular endolymph concentrations were Na+, 166 mmol l(-1) +/- 22; K+, 51.4 mmol l(-1) +/- 16.7; Ca2+, 2.88 mmol l(-1) +/- 0.27; and Cl(-), 170 mmol l(-1) +/- 12; and semicircular canal (utricular vestibule) endolymph concentrations were Na+, 122 mmol l(-1) +/- 15; K+, 47.7 mmol l(-1) +/- 13.2; Ca2+, 1.78 mmol l(-1) +/- 0.48; Cl(-), 176 mmol l(-1) +/- 27. The relatively high concentrations of Ca2+ and Na+ in the endolymph may have significant implications for the physiological function of the mechanoelectrical transduction channels in the vestibular hair cells of fish compared to those of their mammalian counterparts.     

Characteristics of ovarian follicular dynamics throughout the estrous cycle of Egyptian buffaloes

Data of 56 normal and 9 abnormal estrous cycles were collected from 9 Egyptian buffaloes (Bublus bublis) to describe the follicular growth wave pattern. Heat was checked twice daily while, ovaries were scanned daily to monitor the patterns of follicular waves. Day of ovulation was determined when the largest follicle was replaced by corpus haemorrhgicum (CH). Number of waves/cycle, day of emergence of the follicular wave, characteristics of the dominant follicle and corpus luteum (CL) growth features were monitored. Buffaloes displayed mainly two types of follicular waves; two (46.4%) and three (53.6%). In cycles of three wave pattern, time of emergence of the 1st wave post-heat was longer (P < 0.05) and number of recruited follicles/wave were larger (P < 0.05) compared to the corresponding values of the two wave pattern. Number of recruited follicles in early follicular waves (1st or the 2nd) had larger number (P < 0.05) compared to the subsequent ones. Follicles that reached ovulation in both types of estrous cycle had shorter life-span (P < 0.05) than the previous ones. Life-span of CH, growing and regressed CL were 3.6 ± 0.6, 11.2 ± 0.8 and 4.4 ± 0.5 days, respectively with no difference in both types of follicular wave. Three types of ovarian disorders were observed. Follicular waves and CL growth features showed unique pattern for each individual. These results demonstrate that buffaloes display two main types of follicular waves with dominance of three wave type.     

Complex vertebral malformation in Holstein cows in Japan and its inheritance to crossbred F1 generation

Complex vertebral malformation (CVM) is an autosomal recessive inherited defect in the Holstein breed. It causes intra-uterine mortality leading to repeat breeding and involuntary culling of cows. This study was carried out to show the prevalence of CVM carriers among Holstein cows in five dairy farms in Hiroshima Prefecture, South Western Japan and whether the defect could be inherited to crossbred F1 generation, when Japanese Black semen was used to inseminate a Holstein cow. Two hundreds Holstein cows were used in the study. Blood samples were collected from all cows in a heparinized tubes and genomic DNA was extracted with a commercial kit. Apart of the bovine solute carrier family 35 member 3 (SLC35A3) gene around the suspected mutation (G to T) was amplified with an allele specific polymerase chain reaction (AS-PCR). For DNA sequencing, PCR products of 522 bp were obtained from the genomic DNA of the cows. Out of 200 examined cows, 26 cows (13.0%) were found to be CVM carriers. Wild type cows showed amplification with the G-specific primer pair but not with the T-specific primer pair, while carrier ones showed amplification of both the G- and T-specific primers, exhibited two DNA bands of 395 bp. Based on the clinical history of the cows, lowered reproductive performance was noticed in carrier ones. Out of four crossbred F1 calves (Holstein x Japanese Black), two were CVM carrier. Crossbred F1 could inherit the CVM defect when Japanese Black semen used to inseminate a carrier Holstein cow. In conclusion, the study reports the occurrence of CVM among Holstein cows in Hiroshima Prefecture, Japan; moreover, it describes the first cases of CVM in crossbred F1 generation.     

The role of bacteria on heavy-metal extraction and uptake by plants growing on multi-metal-contaminated soils

Four bacterial isolates were examined for their ability to increase the availability of water soluble Cu, Cr, Pb and Zn in soils and for their effect on metals uptake by Zea mays and Sorghum bicolor. Random Amplified Polymorphic DNA (RAPD) analysis was used to show that the bacterial cultures were genetically diverse. Bacterial isolates S3, S28, S22 and S29 had 16S rRNA gene sequences that were most similar to Bacillus subtilis, Bacillus pumilus, Pseudomonas pseudoalcaligenes and Brevibacterium halotolerans based on 100% similarity in their 16S rDNA gene sequence, respectively. Filtrate liquid media that had supported B. pumilus and B. subtilis growth significantly increased Cr and Cu extraction from soil polluted with tannery effluent and from Cu-rich soil, respectively, compared to axenic media. The highest concentrations of Pb (0.2 g kg⁻¹), Zn (4 g kg⁻¹) and Cu (2 g kg⁻¹) were accumulated in shoots of Z. mays grown on Cu-rich soil inoculated with Br. halotolerans. The highest concentration of Cr (5 g kg⁻¹) was accumulated in S. bicolor roots grown in tannery-effluent-polluted soil inoculated with a mixed inoculum of bacterial strains. These results show that bacteria play an important role in increasing metal availability in soil, thus enhancing Cr, Pb, Zn and Cu accumulation by Z. mays and S. bicolor.     

Increasing myocardial contraction and blood pressure in C57BL/6 mice during early postnatal development

Knowledge of the developmental changes of cardiovascular parameters in the genetic background of a mouse strain is important for understanding phenotypic changes in transgenic or knockout mouse models for heart disease. We studied arterial blood pressure and myocardial contractility in mice of the common background strain C57BL/6, aged 21 days [postnatal day 21 (P21)] to 580 days. Heart rate increased during maturation from 396 beats/min at P21 to 551 beats/min at postnatal day 50 (P50), and mean arterial blood pressure increased in parallel from 86 to 110 mmHg and remained constant afterward. Echocardiographically determined left ventricular myocardial wall dimensions (R = 0.79, P < 0.0001) and left ventricular mass calculated using the area-length algorithm correlated strongly with histomorphometrical measurements (R = 0.93, P < 0.001). Sarcomere shortening records from isolated ventricular myocytes used as a measure for myocardial contractility revealed a negative shortening-frequency relation under a pacing frequency of 2 Hz and a positive relation above 2 Hz. Shortening amplitudes recorded from P21 myocytes were smaller, and the shortening-frequency relation was less steep than in adult myocytes. A stimulation pause was followed by a negative "staircase" at pacing frequency of < or =6 Hz and a positive staircase at > or =6 Hz. P21 myocytes developed positive staircases at 8 and 10 Hz, and adult myocytes also developed them at 6 Hz. Blood pressure increase during maturation until P50 may originate from increasing single cardiomyocyte contractility.