Reconstitution of (1→3)-ß-D-glucans measurement system using recombinant Limulus polyphemus Factor G.

Horseshoe crab Factor G is a heterodimeric serine protease zymogen that is activated by (1→3)-ß-D-glucans (BDG) from fungal cell walls. This reaction is used in diagnostic agents for deep-seated mycosis. At present, functional analysis using Factor G from Tachypleus tridentatus has been performed, and genetic information has been published, but reconstitution using recombinant proteins has not yet been achieved. In this study, we cloned the genes for Factor G α and ß from Limulus polyphemus; two gene sequences were obtained for Factor G α and seven for ß. The obtained L. polyphemus Factor G α was used to specifically remove BDG from the culture medium for eliminating the activator BDG. The optimal combination for each sequence was examined with BDG removal medium, and a combination was found that featured BDG-dependent activity. These results indicate that a BDG assay system using recombinant Factor G is feasible in reconstitution. This research will support future reagent development that does not require natural horseshoe crab resources. KEY POINTS: • Cloned novel Factor G α subunit and ß subunit genes from L. polyphemus • Proposed a method of removing BDG without reducing culture medium performance • Identified combination of recombinant α and ß subunits for BDG-dependent activation.

Effects of acupuncture on autonomic nervous function and prefrontal cortex activity.

Acupuncture is helpful in treating various diseases, including autonomic nervous system (ANS) dysfunction caused by mental stress. On the other hand, the frontal lobe is suggested to play an important role in stress responses by modulating the ANS. The aim of the study was to evaluate the effects of acupuncture on ANS and frontal lobe activities. We investigated 18 normal adults. We measured the activity of prefrontal cortex (PFC) caused by real acupuncture (WHO-LI4) and sham acupuncture, employing optical topography. To evaluate ANS function, we analyzed heart rate variability (HRV). Analysis of HRV revealed a decrease of the LF/HF ratio, and an increase of the HF power by real acupuncture, indicating a shift to parasympathetic dominancy. Acupuncture also caused cerebral blood oxygenation changes in both directions, that is, an increase and/or a decrease of oxyhemoglobin (Oxy-Hb) in the bilateral PFC. However, the Oxy-Hb change was not correlated with HRV parameters in the majority of cases. One of the possible explanations of the poor correlations might be that the PFC activity induced by acupuncture is not closely linked with ANS function.

Preferable correlation to blastocyst development and pregnancy rates with a new embryo grading system specific for day 3 embryos.

PURPOSE: To investigate the cleavage stage embryo quality by the correlation between the morphological features and blastocyst development rate to develop a new embryo grading system. METHODS: A retrospective analysis, including 216 cycles of cleavage stage embryo transfer and 251 cycles of blastocyst transfer. The correlation with blastocyst development of the embryo cleavage stage, fragmentation and uniformity of blastomeres was evaluated. RESULTS: There were significant differences in the blastocyst development rate between > or =7 cells and < or=6 cells (68.8% vs. 30.7%), <50% fragmentation and > or =50% fragmentation (51.9% vs. 25.7%), and evenly sized blastomeres and unevenly sized blastomeres (48.7% vs. 30.1%) on day 3. The new grading system defined by these 3 parameters showed a preferable correlation to the pregnancy rate. CONCLUSIONS: The new grading system specific for day 3 embryos is useful for the selection of good quality embryos and may improve the pregnancy rate.

Metastatic renal cell carcinoma of the gallbladder.

A 73-year-old man presented for further evaluation of a gallbladder polyp, 5 years after left radical nephrectomy for renal cell carcinoma (RCC). Computed tomography revealed a 2-cm enhancing pedunculated tumour within the gallbladder. Open simple cholecystectomy was performed and the tumour was histologically confirmed as a metastasis of the RCC to the gallbladder. The patient is alive and disease-free, 2 years after cholecystectomy. Of the reported 23 patients with metastatic RCC of the gallbladder, nine patients were reported to be cancer-free with the longest follow-up interval of 6 years after cholecystectomy. Although metastasis of RCC is a rare differential diagnosis of gallbladder tumours, simple cholecystectomy may offer a chance of long-term survival for patients with RCC.

Popliteal arterial aneurysm associated with Klippel-Trénaunay syndrome: case report and literature review.

A 35-year-old woman with Klippel-Trénaunay syndrome developed a popliteal artery aneurysm in the affected extremity. The aneurysm was successfully treated by aneurysmectomy and bypass grafting with autologous saphenous vein. In Klippel-Trénaunay syndrome, angiodysplasia in the venous system is common. However, reports of an arterial aneurysm in a patient with Klippel-Trénaunay syndrome are extremely rare.

Pretreatment for ovarian endometrial cyst before in vitro fertilization.

Assisted reproductive technology is a widely accepted treatment for infertile women with endometriosis. The presence of an ovarian endometrial cyst reduces the quality of oocytes, while surgical resection of endometrioma may reduce the ovarian reserve for ovarian stimulation by exogenous gonadotropins. To determine what pretreatment should be performed for ovarian endometrial cyst before IVF-ET, we analyzed IVF outcomes with or without pretreatment in patients with endometrioma. Infertile women with endometrioma who underwent IVF-ET were divided into 3 groups, including patients who had received laparotomy or laparoscopy, patients for whom the endometrioma content had been aspirated and treated with or without alcohol fixation, and patients who did not undergo pretreatment. The number of retrieved oocytes, rate of mature oocytes, and fertilization rate were compared among groups. The results showed that pretreatment for endometrioma reduces the number of retrieved oocytes. Although oocyte quality as a rate of mature oocytes was not affected by the presence of an ovarian endometrial cyst, the fertilization rate was improved by cyst aspiration. We propose that surgical pretreatment is not necessary for ovarian endometrial cyst before IVF-ET, but cyst aspiration may be beneficial after several failed attempts of IVF.