Expression patterns of the BRCA1 splicing variants in canine normal tissues and mammary gland tumors

Human BRCA1 is familial breast cancer susceptibility gene. Recently, decreased BRCA1 mRNA and protein expression has been identified in sporadic breast tumors. In the reported human BRCA1 splicing variants, delta11b lacks the majority of exon11 and is suspected to have a distinct function in normal tissues. The splicing variants display a variety of expression pattern in breast cancer samples. Although mammary gland tumor is important disease in dog, there are few reports for BRCA1 in the canine tumors. In this study, we examined the relative amounts of BRCA1 splicing variants mRNA in canine normal and mammary tumor samples by RT-PCR to investigate whether there is the altered expression of variant mRNAs in the canine tumor as reported in human. The exon11b-defecting RT-PCR products were observed in all the normal tissues examined and the nucleotide sequence was quite similar to that of human BRCA1 delta11b. In some tumor samples, we did not detect the products targeted for exon10-13 and exon14-15, while these products were observed in all the normal samples examined. Especially, the relative amounts of the exon11-defecting products were remarkably decreased in most of the tumors (11/16).     

Jack mackerel <Emphasis Type="Italic">Trachurus japonicus</Emphasis> juveniles use jellyfish for predator avoidance and as a prey collector

ABSTRACT:   Juveniles of carangid fishes including jack mackerel Trachurus japonicus are known to associate with jellyfishes. The function of this association behavior was studied through rearing experiments and underwater visual observations. Association behavior of jack mackerel with moon jellyfish in experimental tanks was more frequent in the presence compared to the absence of predators (chub mackerel Scomber japonicus ). In the experimental tanks, the presence of jellyfish, however, did not mitigate predation by these predators. Although jack mackerel did not feed on the jellyfish itself, they frequently fed on the captured prey ( Artemia nauplii) whilst in the gut cavity of the jellyfish. Underwater observations of giant jellyfish Nemopilema nomurai off Kyoto and Fukui prefectures revealed that approximately 30% of these jellyfish were accompanied by jack mackerel juveniles with body sizes ranging 10–45 mm standard length (SL). Considering that jack mackerel juveniles found in subtidal rocky reefs ranged 40–120 mm SL, we considered that jack mackerel from 10 to 45 mm SL associate with jellyfish as a hiding place as well as a food collector, until they find a suitable reef habitat when they attain approximately 40 mm SL.     

Production and characterization of monoclonal antibodies to leukocytes and serum immunoglobulin in Japanese flounder <Emphasis Type="Italic">Paralichthys olivaceus</Emphasis>

Two monoclonal antibodies (MAbs: JFW1 and JFW10) were produced against peripheral blood leukocytes (PBL) in Japanese flounder. Additionally, MAbs against flounder immunoglobulin (Ig; JFW20 and JFW21) were generated for the surface marker of Ig⁺ leukocytes using purified serum Ig as an antigen. MAb JFW1 recognized the surface marker of granulocytes and monocytes and MAb JFW10 specifically bound to the surface antigen of thrombocytes. Flow cytometric analysis of PBL incubated with JFW1, JFW10, JFW20 and JFW21 revealed that 2.5–7.4, 23.7–50.1, 25.2–26.1 and 5.2–8.3% of all leukocytes were positive for these markers. Analysis of head kidney leukocytes (HKL) showed that JFW1, JFW10, JFW20 and JFW21 bound to 30.5–36.3, 1.9–2.8, 6.4–8.3 and 1.9–3.0% of all leukocytes, respectively. Western blot analysis after SDS-PAGE showed that JFW10 recognizes a protein of 115 kDa from lysed PBL. JFW20 recognized the 70 and 74 kDa proteins of the heavy chain of Ig from serum. No band was observed for either JFW1 or JFW21. These antibodies will be useful for the identification and isolation of Japanese flounder leukocyte subpopulations and will facilitate immunological studies of flounder.     

Molecular characterization of kisspeptin 2 dodecapeptide in sixteen species of Scombridae

Fisheries Science - Kisspeptin is an endogenous ligand that modulates gonadotropin-releasing hormone (GnRH) secretion and plays an essential role in the reproduction of mammals. In fish, two...     

Age and growth of the lizardfish <Emphasis Type="Italic">Saurida elongata</Emphasis> from the Tsushima/Korea Strait

We studied the age and growth of the lizardfish Saurida elongata using ground thin sections of otoliths (sagittae) from specimens collected in the Tsushima/Korea Strait between May 1999 and June 2001. A total of 695 individuals with fork lengths (FL) ranging from 189 to 478 mm were examined. The frequency of translucent zone occurrence at the outer margin of the otoliths indicated that translucent zones formed once a year between November and February. Most of the males examined were estimated to be 2–7 years old and the females, 3–9 years old. The maximum estimated age of a male specimen was 10 years and that of a female, 11 years. The estimated von Bertalanffy growth curves were FL _t  = 451{1 − exp[−0.172(t + 2.50)]} and FL _t  = 515{1 − exp[−0.151(t + 2.47)]} for males and females, respectively. At all ages, the FL at a specific age for females was greater than that for males, suggesting that females of this species grow faster than males.     

Milk production,nutrient digestibility and nitrogen balance in lactating cows fed total mixed ration silages containing steam‐flaked brown rice as substitute for steam‐flaked corn,and wet food by‐products

The objective of this study was to evaluate the effect of substituting brown rice grain for corn grain in total mixed ration (TMR) silage containing food by‐products on the milk production, whole‐tract nutrient digestibility and nitrogen balance in dairy cows. Six multiparous Holstein cows were used in a crossover design with two dietary treatments: a diet containing 30.9% steam‐flaked corn (corn TMR) or 30.9% steam‐flaked brown rice (rice TMR) with wet soybean curd residue and wet soy sauce cake. Dietary treatment did not affect the dry matter intake, milk yield and compositions in dairy cows. The dry matter and starch digestibility were higher, and the neutral detergent fiber digestibility was lower for rice TMR than for corn TMR. The urinary nitrogen (N) excretion as a proportion of the N intake was lower for rice TMR than for corn TMR with no dietary effect on N secretion in milk and fecal N excretion. These results indicated that the replacement of corn with brown rice in TMR silage relatively reduced urinary N loss without adverse effects on feed intake and milk production, when food by‐products such as soybean curd residue were included in the TMR silage as dietary crude protein sources.     

Acute lipoprivation suppresses pulsatile luteinizing hormone secretion without affecting food intake in female rats

The present study examined the effect of acute lipoprivation on pulsatile luteinizing hormone (LH) secretion in both normal-fat diet, ad libitum-fed and fasted female rats. To produce an acute lipoprivic condition, mercaptoacetate (MA), an inhibitor of fatty acid oxidation, was administered intraperitoneally to ad libitum-fed or 24-h fasted ovariectomized (OVX) rats with or without an estradiol (E2) implant, that produces a negative feedback effect on LH pulses. The steroid treatment was performed to determine the effect of estrogen on lipoprivic changes in LH release, because estrogen enhances fasting- or glucoprivation-induced suppression of LH pulses. Pulsatile LH secretion was suppressed by MA administration in a dose-dependent manner in the ad libitum-fed OVX and OVX+E2 rats. LH pulses were more severely suppressed in the 24-h-fasted OVX and OVX+E2 rats compared to the ad libitum-fed rats. Estrogen slightly enhanced lipoprivic suppression but the effect was not significant. In the present study, increased plasma glucose and free-fatty acid concentrations may indicate a blockade of fatty acid metabolism by the MA treatment, but food intake was not affected by any of the MA doses. Acute vagotomy did not block lipoprivic suppression of LH pulses. Thus, the present study indicates that lipid metabolism is important for maintenance of normal reproductive function even in rats fed a normal-fat diet and lipoprivation may be more critical in fasted animals that probably rely more heavily on fatty acid oxidation to maintain appropriate metabolic fuel levels. In addition, failure of blockade of lipoprivic LH inhibition by vagotomy suggests that lipoprivic information resulting in LH suppression is not transmitted to the brain via the vagus nerve.     

Collateral uterine blood flow in Holstein cows during the third trimester of pregnancy

Blood flow to the gravid and nongravid uterine horns of four multiparous Holstein cows (mean +/- SD, BW=641.8 +/- 95.4 kg; age=4.8 +/- 1.2 years; parity=3.0 +/- 1.2) was measured on days 225, 248, and 266 of gestation. Surgery was conducted on day 214.5 +/- 4.0 of gestation through the flank of the standing cows. Transit-time ultrasonic flow probes (diameter 12 or 14 mm) were fitted surgically around the uterine arteries of each cow. Surgery was completed within two hours of anesthesia, and the animals recovered rapidly following surgery. Uterine blood flow (UBF, l/min) was recorded at 10 sec intervals for approximately 23.5 hours; these values were averaged to determine UBF. The mean gravid UBF was significantly (P<0.05) greater than the nongravid UBF in this study. The range of the gravid and nongravid UBFs varied from 3.61 to 14.05 and 0.72 to 6.54 l/min, respectively. There were no changes (P>0.1) in the mean gravid and nongravid UBFs from day 225 to 266 of gestation.     

Time course of final oocyte maturation and ovulation in chub mackerel <Emphasis Type="Italic">Scomber japonicus</Emphasis> induced by hCG and GnRHa

Abstract:   The question of whether the ovulation and spawning time in chub mackerel Scomber japonicus is entrained by a circadian rhythm was raised by our previous experiments. Further questions were also raised about whether the time course of human chorionic gonadotropin (hCG)-induced final oocyte maturation (FOM) and ovulation reflected the natural time course induced by endogeneous pituitary gonadotropin (GtH). To address these questions, hCG and gonadotropin-releasing hormone analog (GnRHa) were administered at two 'opposite' times, 14:00 and 02:00 hours, and the time courses of FOM and ovulation were compared. When hCG was injected, ovulation occurred 33 h post-injection in both groups, regardless of the timing of the hCG injection. The timing of ovulation in chub mackerel depends on the timing of hCG injection, but apparently not on circadian rhythms. When GnRHa was injected, ovulation began at 36 h post-injection of GnRHa, regardless of the timing of injection. These results indicate that the time course of FOM and ovulation in the chub mackerel followed a similar pattern whether stimulated by hCG injection or spontaneous luteinizing hormone (LH) surge because GnRHa induces the secretion of endogenous GtH (primarily LH) from the fish pituitary. Thus, it is concluded that the time course of hCG-induced FOM and ovulation in chub mackerel follows the natural time course induced by endogenous pituitary LH.