Relationship between α-melanocyte-stimulating hormone- and neuropeptide Y-containing neurons in the goldfish hypothalamus

Intracerebroventricular (ICV) injection of α-melanocyte-stimulating hormone (α-MSH) inhibits, whereas ICV injection of neuropeptide Y (NPY) stimulates food intake in the goldfish. However, there is little information about the functional relationship between α-MSH-induced anorexigenic and NPY-induced orexigenic actions in the goldfish. In this study we examined the relationship between α-MSH- and NPY-containing neurons in the goldfish hypothalamus to investigate whether these α-MSH- and NPY-containing neurons have direct mutual inputs. α-MSH- and NPY-like immunoreactivities were distributed throughout the brain, especially in the diencephalon. In particular, α-MSH-containing nerve fibers or endings lay in close apposition to NPY-containing neurons in a specific region of the hypothalamus, the nucleus posterioris periventricularis (NPPv). NPY-containing nerve fibers or endings also lay in close apposition to α-MSH-containing neurons specifically in the interior part of the nucleus lateralis tuberis (NLTi). We also investigated the effect of ICV injection of melanocortin 4 receptor agonist (melanotan II) at 100 pmol/g body weight (BW), which is enough to suppress food intake, or NPY at 10 pmol/g BW, which is enough to enhance food intake, on expression levels of mRNA for NPY or proopiomelanocortin (POMC) in the hypothalamus. ICV injection of melanotan II and NPY induced a significant decrease in the expression levels for NPY and POMC mRNA, respectively. These observations suggest that α-MSH- and NPY-containing neurons share direct mutual inputs in the NPPv and the NLTi of the hypothalamus, and that α-MSH and NPY functionally interact or exhibit mutual inhibition to regulate feeding behavior in the goldfish.     

Regenerative responses after mild heart injuries for cardiomyocyte proliferation in zebrafish

Background: The zebrafish heart regenerates after various severe injuries. Common processes of heart regeneration are cardiomyocyte proliferation, activation of epicardial tissue, and neovascularization. In order to further characterize heart regeneration processes, we introduced milder injuries and compared responses to those induced by ventricular apex resection, a widely used injury method. We used scratching of the ventricular surface and puncturing of the ventricle with a fine tungsten needle as injury‐inducing techniques. Results: Scratching the ventricular surface induced subtle cardiomyocyte proliferation and responses of the epicardium. Endothelial cell accumulation was limited to the surface of the heart. Ventricular puncture induced cardiomyocyte proliferation, endocardial and epicardial activation, and neo‐vascularization, similar to the resection method. However, the degree of the responses was milder, correlating with milder injury. Sham operation induced epicardial aldh1a2 expression but not tbx18 and WT1. Conclusions: Puncturing the ventricle induces responses equivalent to resection at milder degrees in a shorter time frame and can be used as a simple injury model. Scratching the ventricle did not induce heart regeneration and can be used for studying wound responses to epicardium. Developmental Dynamics 243:1477–1486, 2014. © 2014 Wiley Periodicals, Inc.     

Association analysis of toll-like receptor 4 polymorphisms in Japanese primary biliary cirrhosis

Primary biliary cirrhosis (PBC) is characterized by portal inflammation and immune-mediated destruction of intrahepatic bile ducts that often result in liver failure. Toll-like receptor (TLR) 4 recognizes lipopolysaccharides of Gram-negative bacteria. Infectious agents have been suspected to play a crucial role in PBC pathogenesis since TLR4 expression was found in bile duct epithelial cells and periportal hepatocytes in liver tissues of PBC. To assess the potential contribution of TLR4 SNPs to the development of this disease, we genotyped five SNPs in TLR4 in 261 PBC patients and 359 controls using a TaqMan assay. No significant positive associations with either PBC susceptibility or progression were uncovered. These results indicate that TLR4 polymorphisms do not play a prominent role in the development of PBC in Japanese patients.     

Use of puncture force measurement to investigate the conditions of blood vessel needle insertion

Central venous catheterization involves venous puncture and catheter insertion for transfusions. Quantitative conditions that facilitate insertion of the needle, such as the angle and velocity of insertion, have not been clarified. We previously developed a robotic system for guiding the needle along a specified puncture path with high precision and are currently implementing a hardware design for a robotic system to assist in blood vessel puncture. In this study, we proposed the insertion angle and velocity for stopping the needle in a blood vessel, assuming that a robotic system such as ours is used during the procedure. We inserted a needle into a segment of porcine jugular vein and obtained the puncture reaction force. Evaluation indices were the magnitude of the sudden decrease in reaction force at the point at which the needle advances and the length of time that the needle is present within the vein. Results indicated that the conditions under which it was easiest to stop the needle inside the vein were an insertion angle range of 10–20 and an insertion velocity of 3 mm/s.     

Genetic evidence for Dnmt3a‐dependent imprinting during oocyte growth obtained by conditional knockout with Zp3‐Cre and complete exclusion of Dnmt3b by chimera formation

In the male and female germ‐lines of mice, both of the two de novo DNA methyltransferases Dnmt3a and Dnmt3b are expressed. By the conditional knockout experiments using the Tnap‐Cre gene, we previously showed that deletion of Dnmt3a in primordial germ cells disrupts paternal and maternal imprinting, however, Dnmt3b mutants did not show any defect. Here, we have knocked out Dnmt3a after birth in growing oocytes by using the Zp3‐Cre gene and obtained genetic evidence that de novo methylation by Dnmt3a during the oocyte growth stage is indispensable for maternal imprinting. We also carried out DNA methylation analysis in the mutant oocytes and embryos and found that hypomethylation of imprinted genes in Dnmt3a‐deficient oocytes was directly inherited to the embryos, but repetitive elements were re‐methylated during development. Furthermore, we show that Dnmt3b‐deficient cells can contribute to the male and female germ‐lines in chimeric mice and can produce normal progeny, establishing that Dnmt3b is dispensable for mouse gametogenesis and imprinting. Finally, Dnmt3‐related protein Dnmt3L is not only essential for methylation of imprinted genes but also enhances de novo methylation of repetitive elements in growing oocytes.     

Huntingtin-associated protein 1 (HAP1) interacts with androgen receptor (AR) and suppresses SBMA-mutant-AR-induced apoptosis

Huntingtin-associated protein 1 (HAP1), an interactor of huntingtin, has been known as an essential component of the stigmoid body (STB) and recently reported to play a protective role against neurodegeneration in Huntington's disease (HD). In the present study, subcellular association between HAP1 and androgen receptor (AR) with a long polyglutamine tract (polyQ) derived from spinal-and-bulbar-muscular-atrophy (SBMA) was examined using HEp-2 cells cotransfected with HAP1 and/or normal ARQ25, SBMA-mutant ARQ65 or deletion-mutant AR cDNAs. The results provided the first clear evidence that HAP1 interacts with AR through its ligand-binding domain in a polyQ-length-dependent manner and forms prominent inclusions sequestering polyQ-AR, and that addition of dihydrotestosterone reduces the association strength of HAP1 with ARQ25 more dramatically than that with ARQ65. Furthermore, SBMA-mutant-ARQ65-induced apoptosis was suppressed by cotransfection with HAP1. Our findings strongly suggest that HAP1/STB is relevant to polyQ-length-dependent modification on subcellular AR functions and critically involved in pathogenesis of not only HD but also SBMA as an important intrinsic neuroprotectant determining the threshold for cellular vulnerability to apoptosis. Taking together with previous reports that HAP1/STB is selectively expressed in the brain regions spared from degenerative targets in HD and SBMA, the current study might explain the region-specific occurrence of neurodegeneration in both diseases, shedding light on common aspects of their molecular pathological mechanism and yet-to-be-uncovered diagnostic or therapeutic applications for HD and SBMA patients.     

Surgical margin status and recurrence pattern in invasive vulvar Paget’s disease: A Japanese Gynecologic Oncology Group study

ObjectiveTo examine the association between surgical margin status and recurrence pattern in invasive vulvar Paget’s disease.MethodsThis is a preplanned secondary analysis of a previously organized nationwide retrospective study in Japan (JGOG-1075S). Women with stage I-IV invasive vulvar Paget’s disease who received surgical treatment from 2001-2010 were examined (n=139). Multivariable analysis was performed to assess local-recurrence, distant-recurrence, and all-cause mortality based on surgical margin status.ResultsThe median age was 70 years. The majority had stage I disease (61.2%), and the median tumor size was 5.0cm. Nodal metastasis was observed in 15.1%. Simple vulvectomy (46.0%) was the most common surgery type followed by radical vulvectomy (28.1%). More than half received vulvar reconstructive surgery (59.0%). Positive surgical margin was observed in 35.3%, and close margin <1cm was observed in 29.5%. Vulvectomy type was not associated with surgical margin status (P=0.424). The median follow-up was 5.8 years. Positive surgical margin was associated with increased local-recurrence (5-year cumulative rates for positive versus negative margin: 35.8% versus 15.0%, P=0.010) but not distant-recurrence (18.3% versus 16.0%, P=0.567). Positive surgical margin was also associated with increased all-cause mortality (5-year overall survival rates for positive versus negative margin: 72.6% versus 88.2%, P=0.032). In multivariable analysis, positive surgical margin remained an independent factor associated with increased risk of local-recurrence (hazard ratio 2.80, 95% confidence interval 1.18-6.63) and all-cause mortality (hazard ratio 2.87, 95% confidence interval 1.20-6.83).ConclusionPositive surgical margin appears to be common in invasive vulvar Paget’s disease that is associated with increased local-recurrence and all-cause mortality risks. Role of alternative surgical technique or adjuvant therapy merits further investigation to improve local disease control.     

Live donor renal transplants using kidneys with arteriographic evidence of mild renovascular disease

The objective of the study was to determine whether using a living donor kidney with arteriographic evidence of renovascular disease affects the outcome of the recipient's transplanted graft. Twenty-eight patients who had unilateral renovascular abnormalities on conventional angiography underwent donor nephrectomy of the ipsilateral kidney. Results in the recipients who received these kidneys were reviewed. Comparison was made to a control group of living donor renal transplant recipients who were matched for donor, recipient age. Graft survival, patient survival, serum creatinine for all 28 recipients were evaluated. All 28 donors underwent a successful donor nephrectomy. Recipient graft survival was 96% at 1 yr, 92% at 3 yr, 71% at 5 yr, 62% at 10 yr. Mean serum creatinine values at 1 month, 1 yr, 3 yr were 1.5, 1.5, 1.6, respectively. Similar results were observed in the control group. We conclude that renal transplantation utilizing kidneys with arteriographic evidence of renovascular disease results in excellent short, long-term renal allograft survival.     

Ammonium acid urate urinary stone caused by a low-caloric diet: a case report

A 32-year-old woman complained of right back pain and pyuria. The plain radiograph (KUB) and drip infusion pyelography (DIP) demonstrated a right renal stone and hydronephrosis. The stone was successfully treated using extracorporeal shock wave lithotripsy. Infrared spectrophotometry revealed that the stone was composed of pure ammonium acid urate. The patient had a 3-year history of excessive anorexia. The low-caloric diet was considered to have caused the disease.