Transient azoospermia following rosuvastatin medication for hypercholesterolemia.

The authors report a case of transient azoospermia following hydroxymethylglutaryl-coenzyme A reductase (HMGCR) inhibitor rosuvastatin medication for hypercholesterolemia. While a primary infertile couple with oligoasthenospermia was preparing for an in vitro fertilization program, the male partner had been diagnosed with hypercholesterolemia in a medical check-up and prescribed four-week oral administration of rosuvastatin. No motile spermatozoa were found in the ejaculated semen and urine on the day of follicular aspiration. Azoospermia was confirmed by reexamination in weeks 3 and 7. Spermatozoa appeared in the ejaculated semen in two weeks of drug withdrawal. In week 16, the sperm count and motility increased to the level where intracytoplasmic sperm injection was available.

Do combined psychological stress examinations predict pregnancy outcome in an assisted reproductive technology program?

PURPOSE OF INVESTIGATION: To investigate prospectively if the pregnancy outcome in infertile women undergoing assisted reproductive technology (ART) is predictable by a combination of psychological stress examinations on the day of embryo/blastocyst transfer. MATERIALS AND METHODS: From April 2012 to May 2012, 114 women aged 42 years old or less underwent transfer of morphologically-good embryo/blastocyst(s) in the present in vitro fertilization (IVF) center. Immediately before the transfer, salivary secretion was obtained and frozen. α-amylase and cortisol concentrations were quantified using biochemical assays. In addition, patients were asked to answer General Health Questionnaire 28 (GHQ28) and Zung's Self Rating Depression Scale (SDS) following transfer. The results were compared between the pregnant group and non-pregnant group. RESULTS: There were no significant differences in the age of the infertile couples between the pregnant group and non-pregnant group as well as body mass index of the infertile women. The GHQ28 and SDS scores were similar between the two groups, as were the salivary α-amylase and cortisol concentrations. CONCLUSION: This prospective study failed to demonstrate the predictivity of the pregnancy outcome by psychological stress examinations in infertile women in an ART program, even though these tests were used in combination.

Diclofenac suppository pretreatment in prevention of vasovagal reflex-associated complications for infertile women undergoing local endometrial injury.

PURPOSE OF INVESTIGATION: To assess the effects of the diclofenac suppository pretreatment in prevention of vasovagal reflex-associated complications for infertile women undergoing local endometrial injury (LEI). MATERIALS AND METHODS: Eighty-six infertile outpatients with repeated implantation failure following transfer of morphologically good embryos and/or blastocysts underwent single curettage LEI to improve the pregnancy outcome in the subsequent embryo/blastocyst transfer cycle. Of them, 35 patients chose diclofenac suppository administration prior to LEI, whereas 51 patients did not. The occurrence of palpitations, bradycardia, hypotension, presyncope, and requirement of bed rest was compared between the two groups. RESULTS: There were no significant differences in the demographics between the two groups. The prevalence of presyncope and requirement of bed rest was significantly lower in the diclofenac suppository group than in the control group. The pregnancy outcome was similar between the two groups. CONCLUSION: The diclofenac suppository administration is a low-cost effective method to reduce the risk of the vasovagal reflex-associated complications in infertile women undergoing LEI.

Vaginal preparation with povidone iodine disinfection and saline douching as a safe and effective method in prevention of oocyte pickup-associated pelvic inflammation without spoiling the reproductive outcome: evidence from a large cohort study.

PURPOSE OF INVESTIGATION: The aim of this study was to investigate if vaginal preparation procedure affects the occurrence of oocyte pickup-associated pelvic inflammation (OPU-PI) and the reproductive outcome in an in vitro fertilization (IVF) program. MATERIALS AND METHODS: The occurrence of OPU-PI and the reproductive outcome were compared between 956 infertile patients undergoing vaginal preparation with saline douching alone versus 1,216 infertile patients undergoing a combination ofpovidone iodine disinfection and subsequent saline douching in an IVF program. RESULTS: OPU-PI occurred in four patients (0.042%) in the saline douching alone group, whereas there were no cases in the combination group (p = 0.016). There were no significant differences in the rate of fertilization, morphologically good embryo acquisition, clinical and ongoing pregnancy between the two groups (p > 0.23). CONCLUSIONS: This large cohort study demonstrated that a combination of vaginal povidone iodine disinfection and subsequent saline douching is more effective procedure than saline douching alone to prevent OPU-PI, without spoiling the oocyte quality.

Dimensional accuracy of optical bite registration in single and multiple unit restorations.

The dimensional accuracy of optical bite registration in the CEREC system was compared to that of the conventional physical method in vitro using a bite registration material. Maxillary and mandibular full-arch dentate epoxy models mounted on an articulator were used to measure the interarch distance and the angles created by the occlusal planes. The preparations for a single restoration on the maxillary first molar or for multiple restorations on the maxillary posterior quadrant were made on the model. Optical impression and bite registration data were collected to construct virtual models using computer-aided design software. A silicone material was used for the physical method, and the dimensional accuracy was measured by means of the coordinate measuring machine. The discrepancy relative to the baseline before preparation was analyzed in each registration record. For the single restoration, the optical method created a mean discrepancy of 243.2 µm relative to baseline at the prepared tooth, which was insignificantly but slightly lower than the mean discrepancy of 311.1 µm obtained with the physical method. The mean rotational deviation in the horizontal plane was significantly lower for the optical method. For the multiple preparations, the optical method showed significantly larger discrepancy on the right molar and on the left premolar and molar sites. In the frontal view, the optical method created significantly larger rotational deviation than the physical method. The result indicates that the optical bite registration was effective in terms of dimensional accuracy for single posterior restorations.

Successful living related kidney transplantation across an anti-donor HLA antibody.

In preconditioning highly sensitized kidney transplant candidates, renal allograft outcomes have been better when the serum titer for class I anti-HLA donor-specific antibody (DSA) is low in the recipient at the time of transplantation. However, the ideal level to which the titer should be lowered is still controversial. We report a primary living related kidney transplant in a 34-year-old highly sensitized woman (pretransplant panel-reactive antibody=70%) with end-stage renal disease secondary to chronic glomerulonephritis. We sought to desensitize by lowering the DSA titer specifically to 1:4 pretransplant. A standard complement-dependent cytotoxicity cross-match with her donor (sister) was repeatedly negative, although she was positive for class I antibody against her mismatched HLA antigen (A*2402) at a titer up to 1:16 by the single-antigen flowbead assay. The target DSA titer of 1:4 before transplant was achieved by 12 sessions of plasmapheresis (PP) over 7 weeks, plus two intravenous immune globulin infusions (IVIG) (500 mg/kg/infusion). The patient outcome was excellent. Neither IVIG nor PP was needed posttransplant. The serum creatinine ranged between 0.5 mg/dL and 1.2 mg/dL, and no rejection episode was documented at 28 weeks posttransplant. Therefore, we encourage the use of IVIG and PP to lower the DSA titer to at least 1:4 before kidney transplantation in highly sensitized patients. Large prospective trials are needed to establish a consensus for pretransplant risk assignment and to evaluate the need for desensitization.

Average P/F ratio during donor management for finding out the potential donor of lung procurement.

We investigated the potential donors from cases without lung procurement in spite of considering procurement as the donor. We reviewed 207 cases considered for lung procurement. Donors were divided into 2 groups according to whether or not their lungs were harvested: 50 did and 157 did not. We investigated their age, gender, donor management periods, blood pressure, heart rate, P/F ratio, PCO2, HCO3, positive end-expiratory pressure (PEEP), respiratory rate, abnormal findings in the chest X-ray, blood chemistries, sputum culture, and intravenous administration of steroid. Univariable or multivariable logistic regression analysis was used to predict lung harvesting by various factors. Univariable logistic regression analysis revealed maximum heart rate (HR) and respiratory rate (RR), maximum, minimum, and average P/F ratio, and abnormal findings in the chest X-ray, to be predictors to perform transplant. The maximum HR and RR of the non-harvest group are higher than those of the harvest group. Multivariable logistic regression analysis revealed average P/F ratio only to be a predictor to transplant. We divided the non-harvest group into 2 groups according to whether or not their P/F ratio was >300: 55 did (P/F 300 group) and 102 did not (P/F 299 group). Between P/F 299 group and the harvest group, univariable logistic regression analysis revealed maximum HR, maximum and minimum RR, maximum PEEP, maximum, minimum, and average P/F ratio, and abnormal findings in the chest X-ray to be predictors to transplant. Maximum HR, PEEP, and maximum and minimum RR of the P/F 299 group are higher than those of the harvest group. Maximum, minimum, and average P/F ratio of the P/F 299 group are lower than those of the harvest group. The average PCO2 of P/F 299 group is higher than that of the harvest group. The rate of abnormal findings in the chest X-ray of the P/F 299 group is higher than that of the harvest group. Multivariable logistic regression analysis revealed average P/F ratio only to be a predictor to transplant. There were no predictors between the P/F 300 group and the harvest group in both univariable and multivariable logistic regression analyses. Furthermore, there were no different factors between the P/F 300 group and the harvest group. Our study showed that 35.5% of the non-harvest group had a P/F ratio of >300 and had no difference compared with the harvest group. Our data suggest that potential donors existed in the non-harvest group and increased the number of lung procurement to 27.3% from 13.0% of all donors.

Renal biopsy donor group: the influence of glomerulosclerosis on transplant outcomes.

The UNOS donor population was examined from 1999 to 2002, and approximately 25% of the over 23,000 donors were biopsied (Bx). There was a significant trend (P < .001) of older donors, cardiovascular accident, and hypertension in the Bx group versus the non-Bx group. The percent GS was directly correlated (P < .001) to graft survival, delayed graft function, and primary nonfunction. Cox regression showed significant relative risk (RR) for >10% GS, hypertension, donors over the age of 50, and African American recipients. RR in donors with >10% GS could be ameliorated (P < .001) by choosing donors with <5 HLA-A, -B, or -DR mismatches (MM), or recipients who were nonsensitized, and/or first transplant. Risk should be managed in donors by choosing appropriate recipients and high-risk immunosuppresion protocols.

Multiplexed analysis of polymorphisms in the HLA gene complex using bead array chips.

A novel custom bead array technology is introduced, and it is applied to multiplexed analysis of highly polymorphic human leukocyte antigen (HLA) genetic loci. Our technology combines the construction of probe libraries on color-encoded microparticles (beads) with semiconductor chip processing to produce custom-designed high-density bead array chips in large quantities. Using this novel assay format, two modes of parallel molecular typing of the HLA complex were implemented, namely direct hybridization, illustrated here for class II HLA-DR, and a novel format of on-chip polymerase-mediated primer elongation, illustrated here for class I HLA-A, HLA-B, and HLA-DR using patient and reference cell-line DNA samples. Hybridization-mediated multiplexed analysis of polymorphism method was validated with 142 samples, resulting in 100% concordance with sequence-specific oligonucleotide typing results and a concomitant average of 40% less allele ambiguity. In addition, elongation and hybridization reactions were combined to identify multiple polymorphisms on the same phase of DNA for allele identification.

Clinical, electrophysiological, and serological overlap between Miller Fisher syndrome and acute sensory ataxic neuropathy.

We report a patient with severe sensory ataxia, areflexia, and ophthalmoplegia with preservation of limb muscle strength. Electrophysiological examinations revealed peripheral sensory nerve involvement. A serological examination showed the elevation of IgG antibodies to various b-series gangliosides as well as GT1a. These indicated that this case is an overlap between acute sensory ataxic neuropathy and Miller Fisher syndrome. Autoantibody is implicated as potential pathogenic agents in some cases of acute sensory ataxic neuropathy.

[Effect of different concentrations of tranilast on posterior capsule opacification].

PURPOSE: To evaluate the effect of different concentrations(0.5, 1.0, 2.0%) of tranilast eyedrops in preventing fibrous posterior capsule opacification (PCO) in rabbits. METHODS: Experimental phacoemulsification procedures and in-the-bag placement of intraocular lens(IOL) implant were performed. An anterior eye segment analysis system(EAS-1000, Nidek Co, Ltd) was used to evaluate the degree of PCO during 5 weeks after surgery. RESULTS: The development of PCO was significantly suppressed in the eyes treated with tranilast eyedrops from 1 week after surgery. The mean PCO density after treatment with 0.5% tranilast was 59.1 +/- 9.3 (mean deviation +/- standard error) and 57.1 +/- 8.2, while for 1.0% tranilast it was 40.1 +/- 6.8 and 46.6 +/- 8.4, and for 2.0% tranilast it was 38.5 +/- 6.0 and 37.5 +/- 5.6 (computer compatible tape, CCT) at 1 week and 5 weeks, respectively. In the control group, the mean PCO density was 89.3 +/- 10.4 and 137.4 +/- 32.8 at 1 week and 5 weeks, respectively(p < 0.05). However, no significant statistical difference was observed in any of the findings for the 3 different tranilast concentrations. CONCLUSION: These results suggest that the 0.5% tranilast eyedrops, which are already available on the market, are sufficiently effective for inhibiting PCO.

Differentiation of responses to allergenic and irritant compounds in mouse popliteal lymph node assay.

The mouse popliteal lymph node assay (PLNA) has been proposed as an immunotoxicological test to predict allergenic chemicals. However, PLN response is also observed in association with non-specific activation induced by some irritants. We, therefore, examined the kinetics of the PLN cellularity indices of primary and secondary responses. Flow cytometric analysis was used to measure the proportions of T and B cells in PLN. Male ICR mice were subcutaneously injected with TNBS (an allergenic compound) or SDS (an irritant compound) in the left hind footpad, and with vehicle in the right one. On day 28 after first injection, mice were injected with 1/10 or 1/100 dose of the initial injection. On day 7 after first injection of TNBS, primary response reached maximal PLN cellularity index (16.5). On day 2 after second injection, secondary response reached maximal PLN cellularity index (13.1). A marked increase in proportion of B cells was observed in the PLN. On the other hand, after first injection of SDS, primary response reached maximal PLN cellularity index (2.8) on day 10, but neither secondary response nor increase in the B cell proportion were observed. These results suggest that the detection of secondary response and the flow cytometric analysis are useful in differentiating responses to allergenic and irritant compounds in the PLNA.

Gene delivery to human chondrocytes by an adeno associated virus vector.

OBJECTIVE: To investigate the efficiency of gene transduction to human chondrocytes using an adeno associated virus (AAV) vector. METHODS: We transduced green fluorescent protein (GFP) gene using AAV vector to primary human chondrocytes as well as human cartilage organ cultures, in which chondrocytes are surrounded by extracellular matrix. Expression of GFP gene was analyzed at various time points after transduction by fluorescence microscopy and immunohistochemistry. RESULTS: In primary chondrocytes, the percentages of GFP positive cells were 15.9% or 16.0% on Day 1 and 95.0% or 93.7% on Day 7 after gene transduction. In cartilage organ cultures, gene delivery was observed in cells located not only in the superficial layer but also in the deep layer within the cartilage tissue. Up to 45.3+/-7.4% or 46.0+/-3.9% of chondrocytes expressed GFP for at least 28 days. CONCLUSION: AAV vector could be useful for direct gene delivery to chondrocytes in situ.

Type 1 diabetes mellitus caused by treatment with interferon-beta.

A 57-year-old man was referred to our outpatient clinic after interferon-beta (IFN-beta) treatment for 7 weeks. While IFN-beta therapy was continued in our outpatient clinic, his blood glucose level increased gradually, and he was admitted to our hospital for hyperglycemia. The patient was prescribed a 1,600-kcal diet and intensive insulin therapy was performed. GAD antibody became positive 15 months after the start of IFN therapy, and disappeared 27 months after the start of IFN therapy. Insulin secretion was depleted and the patient had HLA-DR4, B54, and DRB1*0405. This appears to be a case of type 1 diabetes mellitus induced by administration of IFN-beta alone.

[Poison elimination methods in acute poisoning].

This paper presents current ideas about the application of poison elimination methods in acute poisoning. The methods are divided into two groups: poison elimination before absorption; and enhancement of elimination after drug absorption. The former include washes, emesis, gastric lavage, and administration of activated charcoal and cathartics, while the latter include forced diuresis and extracorporeal toxin elimination such as hemodialysis or hemoperfusion. The indications, contraindications, techniques, and complications, associated with these elimination methods, except for extracorporeal toxin elimination, are discussed. In the treatment of poisoning, early and comprehensive poison elimination is important.

Effects of tranilast on cultured rabbit corneal keratocytes and corneal haze after photorefractive keratectomy.

PURPOSE: In vitro and in vivo studies were performed to elucidate the effects of tranilast on cellular proliferation and collagen synthesis. METHODS: Subculturing was carried out using keratocytes from rabbits that underwent photorefractive keratectomy (PRK) and developed corneal haze, and keratocytes from normal rabbit cornea. RESULTS: Tranilast suppressed proliferation in cultured keratocytes from the corneal haze region at doses of 30 and 300 micromol/L and collagen synthesis at doses of 3, 30, and 300 micromol/L. Normal corneal cultures showed suppression of keratocyte proliferation and collagen synthesis only at a high dose of tranilast (300 micromol/L). Betamethasone suppressed proliferation of keratocytes in both haze and normal cornea at a dose of 10 micromol/L, as well as collagen synthesis at respective doses of 1 and 10 micromol/L. Diclofenac sodium suppressed collagen synthesis of keratocytes in haze cornea at a high dose of 100 micromol/L, and in keratocytes in normal cornea, at doses of 10 and 100 micromol/L. In an in vivo study, either 0.5% tranilast, 0.1% betamethasone phosphate eye drops, or a tranilast base solution (control) was instilled four times daily to rabbits that had undergone PRK. Weekly evaluation of the inhibitory effect of these drugs on the development of haze was performed 2 weeks after surgery. Tranilast suppressed haze 6-13 weeks after PRK, but betamethasone phosphate showed no effect. CONCLUSION: These results indicate that tranilast is potentially effective for inhibiting the corneal haze that occurs after PRK.

Effect of tranilast eyedrops in preventing posterior capsule opacification: preliminary report.

PURPOSE: To evaluate the effect of tranilast eyedrops in preventing fibrous opacification of the posterior lens capsule after cataract extraction and intraocular lens (IOL) implantation. SETTING: The Second Department of Ophthalmology, Toho University School of Medicine, Tokyo, and Shohzankai Medical Foundation, Miyake Eye Hospital, Nagoya, Japan. METHODS: This study comprised eyes having continuous curvilinear capsulorhexis and phacoemulsification/aspiration followed by implantation of a posterior chamber IOL in the capsular bag. In this prospective, randomized, controlled, and double-masked trial, tranilast 0.5% (Rizaben) eyedrops (15 eyes) or its placebo eyedrops (20 eyes) were given 4 times a day for 3 months after surgery. An anterior eye segment analysis system (EAS 1000, Nidek Co., Ltd.) was used to evaluate the degree of fibrous posterior capsule opacification (PCO) 1 week and 1 and 3 months after surgery. RESULTS: The mean PCO density in the tranilast group was 17.1 cct +/- 4.6 (SD), 20.0 +/- 3.6 cct, and 23.0 +/- 7.7 cct (cct = computer compatible tape) at 1 week and 1 and 3 months, respectively. In the control group, it was 18.2 +/- 5.3, 30.2 +/- 7.8, and 38.4 +/- 8.0 cct, respectively. There was a significant difference in the 1 and 3 month findings between the 2 groups (P < .001). CONCLUSION: Tranilast was effective in preventing fibrous PCO at an early postoperative stage. The possible mechanisms of its effect may be prevention of collagen synthesis by minimizing transforming growth factor type beta released during lens epithelial cell metaplasia.

Isolation of recombinant adeno-associated virus vector-cellular DNA junctions from mouse liver.

Recombinant adeno-associated virus (rAAV) vectors allow for sustained expression of transgene products from mouse liver following a single portal vein administration. Here a rAAV vector expressing human coagulation factor F.IX (hF.IX), AAV-EF1alpha-F.IX (hF.IX expression was controlled by the human elongation factor 1alpha [EF1alpha] enhancer-promoter) was injected into mice via the portal vein or tail vein, or directly into the liver parenchyma, and the forms of rAAV vector DNA extracted from the liver were analyzed. Southern blot analyses suggested that rAAV vector integrated into the host genome, forming mainly head-to-tail concatemers with occasional deletions of the inverted terminal repeats (ITRs) and their flanking sequences. To further confirm vector integration, we developed a shuttle vector system and isolated and sequenced rAAV vector-cellular DNA junctions from transduced mouse livers. Analysis of 18 junctions revealed various rearrangements, including ITR deletions and amplifications of the vector and cellular DNA sequences. The breakpoints of the vector were mostly located within the ITRs, and cellular DNA sequences were recombined with the vector genome in a nonhomologous manner. Two rAAV-targeted DNA sequences were identified as the mouse rRNA gene and the alpha1 collagen gene. These observations serve as direct evidence of rAAV integration into the host genome of mouse liver and allow us to begin to elucidate the mechanisms involved in rAAV integration into tissues in vivo.