A pragmatic RCT of conventional versus increased concentration sucrose in freezing and thawing solutions for human embryos.

BACKGROUND: Intact frozen-thawed embryos have a greater potential than damaged embryos to establish successful pregnancies. This study aimed to determine whether elevated concentrations of sucrose during freezing would increase the proportion of patients with ≥ 50% of embryos intact after thawing (primary outcome), and improve clinical outcome. METHODS: In a two arm, parallel group, pragmatic trial, IVF/ICSI couples were randomized prospectively to have their supernumerary embryos frozen in a medium containing 0.1 M sucrose (control; n = 99) or 0.3 M sucrose (intervention; n = 102). RESULTS: More control (74/99) than intervention (63/102) couples had at least one embryo thawed (P = 0.07). Significantly more (P = 0.005) intervention (53/63) than control (45/74) couples had ≥ 50% of embryos intact. Freezing in a medium containing 0.3 M sucrose increased by 3.4-fold [95% confidence interval (CI) (1.45, 7.82)] the likelihood of a couple having ≥ 50% of their embryos intact. In the fresh cycle, live birth rate per transfer was similar in the control (35/95) and intervention (36/93) groups (P = 0.91). More control (19/63) than intervention (9/59) couples had a live birth after frozen embryo transfer (P = 0.08). When fresh and frozen cycles were combined, fewer intervention (n = 102) than control (n = 99) couples had at least one live birth (42 versus 53%). The difference in cumulative live birth rate was not significant [hazard ratio = 0.75, 95% CI (0.49, 1.13); P = 0.17]. CONCLUSIONS: Increasing the concentration of sucrose in the freezing medium improves embryo survival, but this is not reflected by increased cumulative birth rates. CLINICAL TRIALS REGISTRATION NUMBER: ISRCTN93314892.

Monitoring of Tribolium castaneum (Coleoptera: Tenebrionidae) in Rice Mills using Pheromone-Baited Traps.

The red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae), is a common pest in rice mills. With limited information in the literature addressing T. castaneum in rice processing facilities, we examined the spatial and temporal distribution of T. castaneum inside and outside of three commercial rice milling facilities and one rice packaging plant from June 2012 to August 2014 using pheromone-baited dome traps. Each mill had very different population trends with fewer numbers collected in rough rice storage areas. T. castaneum were more commonly collected in processing areas. Beetle infestation at all the mills was evaluated using the threshold of mean beetle capture of 2.5 beetles per trap per 2 wk period. Trap captures were below threshold for all but one facility. Temperatures inside were ~1°C warmer than outside temperatures, with these temperature differences more noticeable during cool months (October-March). Higher numbers of T. castaneum were captured in 2012 in comparison to 2013, with higher beetle numbers observed during warmer (April-September) than cooler months. With variation in trap capture of T. castaneum occurring among all facilities, this study illustrates that having a monitoring program designed for each facility is important to help managers decide when and where to apply pest management tactics. The use of pheromone traps could provide information to mill managers to find locations within a mill that are most vulnerable to infestation by T. castaneum, and to assist with the timing of control interventions.

Lentiviral transduction of the murine lung provides efficient pseudotype and developmental stage-dependent cell-specific transgene expression.

Gene transfer for cystic fibrosis (CF) airway disease has been hampered by the lung's innate refractivity to pathogen infection. We hypothesized that early intervention with an integrating gene transfer vector capable of transducing the lung via the lumen may be a successful therapeutic approach. An HIV-based lentiviral vector pseudotyped with the baculovirus gp64 envelope was applied to the fetal, neonatal or adult airways. Fetal intra-amniotic administration resulted in transduction of approximately 14% of airway epithelial cells, including both ciliated and non-ciliated epithelia of the upper, mid and lower airways; there was negligible alveolar or nasal transduction. Following neonatal intra-nasal administration we observed significant transduction of the airway epithelium (approximately 11%), although mainly in the distal lung, and substantial alveolar transduction. This expression was still detectable at 1 year after application. In the adult, the majority of transduction was restricted to the alveoli. In contrast, vesicular stomatitis virus glycoprotein pseudotyped virus transduced only alveoli after adult and neonatal application and no transduction was observed after fetal administration. Repeat administration did not increase transduction levels of the conducting airway epithelia. These data demonstrate that application at early developmental stages in conjunction with an appropriately pseudotyped virus provides efficient, high-level transgene expression in the murine lung. This may provide a modality for treatment for lung disease in CF.

Quercetin attenuates lactate production and extracellular matrix secretion in keratoconus.

Keratoconus(KC) is an ecstatic corneal disease leading to corneal-thinning and the formation of a cone-like cornea. Elevated lactate levels, increased oxidative stress, and myofibroblast formation have all been previously reported. In the current study, we assess the role of Quercetin on collagen secretion and myofibroblast formation in KC in vitro. Human corneal fibroblasts(HCFs) and human keratoconus cells(HKCs) were treated with a stable Vitamin C derivative and cultured for 4 weeks, stimulating formation of a self-assembled extracellular matrix. All samples were analyzed using Western blots and targeted tandem mass spectrometry. Our data showed that Quercetin significantly down regulates myofibroblast differentiation and fibrotic markers, such as α-smooth muscle actin (α-SMA) and Collagen III (Col III), in both HCFs and HKCs. Collagen III secretion was reduced 80% in both HCFs and HKCs following Quercetin treatment. Furthermore, Quercetin reduced lactate production by HKCs to normal HCF levels. Quercetin down regulated TGF-ßR2 and TGF-ß2 expression in HKCs suggesting a significant link to the TGF-ß pathway. These results assert that Quercetin is a key regulator of fibrotic markers and ECM assembly by modulating cellular metabolism and TGF-ß signaling. Our study suggests that Quercetin is a potential therapeutic for treatment of corneal dystrophies, such as KC.

An RGD-oligolysine peptide: a prototype construct for integrin-mediated gene delivery.

We have synthesized a linear, bifunctional peptide that comprises an integrin-targeting domain containing an arginine-glycine-aspartic acid tripeptide motif and a DNA-binding moiety consisting of a short stretch of 16 lysine residues. This peptide can form distinctive, condensed complexes with DNA and is capable of mediating its delivery and expression in a variety of mammalian cells in culture. Internalization is mediated by cell surface integrin receptors via a mechanism that is known to be phagocytic. We have analyzed the relationship between DNA and peptide and have investigated the conditions suitable for optimal gene delivery. The formation of condensed peptide DNA complexes leads to resistance to nuclease degradation. The level of reporter gene expression obtained is dependent on the peptide-to-DNA ratio and is enhanced in the presence of the endosomal buffer chloroquine, polyethyleneimine, and deactivated adenovirus during gene delivery. Under optimal conditions the levels of reporter gene expression obtained approach or even exceed those obtained with DNA delivered with the commercial liposome Lipofectamine. The ability to produce an efficient gene delivery system using small, easily modified, and well-defined constructs that have no constraint of particle size demonstrates the advantages of integrin-targeting peptides for gene transfer.

Efficient, all-solid-state, Raman laser in the yellow, orange and red.

We report efficient operation of a KGd(WO(4))(2) Raman laser pumped by a small, 1 W, 532 nm laser module. By changing the output coupler and Raman crystal orientation, more than 8 wavelengths in the yellow-to-red spectral region were generated including 555 nm, 559 nm, 579 nm, 589 nm, 606 nm, 622 nm, 636 nm and 658 nm, ie., the first 4 Stokes orders on the two orthogonal high-gain Raman shifts of KGd(WO(4))(2). We have also demonstrated spectrally pure output (typically >90% pure) for selected Stokes order with output power up to 400 mW. High slope efficiency (up to 68%) and high beam quality (M(2)~1.5) of Stokes output are obtained even at the highest pump power.

Natural transformation of Streptococcus crista.

Over the years Streptococcus gordonii (sanguis) Challis has become the workhorse of genetic manipulations for the sanguis group of oral streptococci. This is because strain Challis was shown in early studies to be highly naturally competent for transformation. However, Challis is not usually the most appropriate strain to use in studies which focus on oral microbial adherence. We report that other members of the newly reorganized sanguis group, particularly within the species S. crista, display reasonable transformation frequencies, with both plasmid and chromosomal DNA, if transformed at the appropriate time during the growth curve. The ability to transform S. crista may be especially important for genetic studies of biological properties that appear to be limited to these specific streptococcal strains.

Limited efficacy of preoperative computed tomographic scanning for the evaluation of lymph node metastasis in patients before radical prostatectomy.

OBJECTIVES: To evaluate the efficacy of preoperative computed tomographic (CT) scanning in patients with presumed localized prostatectomy prior to radical retropubic prostatectomy. METHODS: A retrospective study of 173 consecutive patients believed to be candidates for radical retropubic prostatectomy who underwent preoperative CT scanning regardless of preoperative prostate-specific antigen (PSA) value, clinical stage, or Gleason grade was undertaken. All patients underwent radical retropubic prostatectomy with bilateral pelvic lymph node dissection or aspiration needle biopsy of abnormal nodes on CT scanning. RESULTS: One hundred sixty-five of 173 patients (95.4%) were believed to have normal CT scans preoperatively. Of these 165 patients, 156 (94.5%) were found to have negative lymph nodes confirmed histologically at the time of lymphadenectomy. Nine patients (5.5%) were found to have lymph node metastases confirmed histologically, despite a negative CT scan. Computed tomographic scanning understaged 9 of 12 (75%) patients with proven metastases. Incidental abdominal pathology of clinical significance was documented in 4 patients (2.3%), including 2 with renal cell cancers, 1 with colon cancer, and 1 with a large (8 cm) abdominal aortic aneurysm. Prostate-specific antigen levels in patients with metastatic lymph nodes ranged from 0.7 to 130 ng/mL (Hybritech Tandem assay), with a mean level of 42 ng/mL. Although 9 of 33 patients (27.3%) with PSA levels greater than 25 ng/mL had node metastases, only 3 of these 33 patients (9.1%) were correctly diagnosed by CT scanning. CONCLUSIONS: Although additional numbers of patients with high PSA levels need to be evaluated, we could not find any justification for routine preoperative CT scanning in patients with a PSA of less than 25 ng/mL. These results suggest that significant savings can be realized by abandoning the practice of routine CT scanning for lymph node metastasis in all patients with newly diagnosed prostate cancer.

7 alpha-Alkyltestosterone derivatives: synthesis and activity as androgens and as aromatase inhibitors.

The 7 alpha-ethyl,propyl,butyl,3'-t-butoxypropyl,allyl,3'-hydroxypropyl 17-acetate, and 3'-chloropropyl 17-acetate derivatives of testosterone and the 7 alpha-3'-t-butoxypropyl, 3'-hydroxypropyl,3'-acetoxypropyl,3'-bromoacetoxypropyl, 3'-chloropropyl, and 2'-oxo-3'-bromopropyl derivatives of 4-androstene-3,17-dione were synthesized. The testosterone derivatives were found to lose androgenic and anabolic activity rapidly as the size of the group at the 7 position increased. Many of the compounds were tested as inhibitors of aromatase. The 17-keto compounds were more active than the corresponding alcohols and the enzyme was found to tolerate at least the bulk of a hydroxypropyl group at the C-7 alpha position.

Semantic activation of noun concepts in context.

A modified Stoop procedure was used to examine the role that context plays in guiding semantic access of unambiguous nouns in sentence contexts. The sentences either emphasized a high- or a low-dominant property of a noun that was the last word in the sentence or were control sentences. Each sentence was followed by the relevant high- or low-dominant property either immediately or after a 300-or 600-ms delay. There was significant color-naming interference (relative to control) for high-dominant properties regardless of biasing context in the immediate and delayed conditions. There was also significant color-naming interference for low-dominant properties in the immediate condition regardless of context. However, in the delayed conditions, the low-dominant properties led to color-naming interference only when preceded by sentence contexts biasing interpretation toward the low-dominant property. It was concluded that high-dominant properties function as core, or invariant, aspects of meaning and that initial semantic access is context independent.

Methylprednisolone in spinal cord compression.

In acute nonsurvival studies, eight anesthetized lambs were subjected to cord compression at T13 by means of an epidural balloon distended to a pressure of 200 mm Hg for 40 minutes. Subsequent to withdrawal of the balloon, each animal received 30 mg/kg of methylprednisolone succinate in an intravenous bolus followed by a continuous infusion of 10 mg/kg/hr for the duration of the experiment. Spinal cord blood flow (SCBF) and spinal evoked potential (SEP) determinations were obtained sequentially prior to, during, and at 1/2, 1 1/2, and 2 1/2 hours following compression. In spite of the absence of ischemia following compression, SEPs failed to recover. Methylprednisolone had no apparent effect on blood flow or on the recovery of SEPs when compared with results in ten control animals that received saline alone.

Nicardipine after spinal cord compression in the lamb.

To study the effects of the calcium channel blocker nicardipine on spinal cord blood flow and spinal evoked potentials, the following study was undertaken. After cord compression, which was productive of paraparesis, nicardipine was administered intravenously in 10 anesthetized lambs. Ten control animals were subjected to compression but received saline instead. Nicardipine produced a significant decrease in mean arterial pressure when compared to the control group. Thirty minutes after compression, spinal cord blood flow also was lower in the nicardipine group compared with controls. Spinal evoked potentials did not recover after compression in either group.

Relative rates of absorption of fluid and protein from the peritoneal cavity in cats.

The relative rates of fluid and protein absorption from the peritoneal cavity of anesthetized cats were measured over 6 hours at an intraperitoneal pressure of 15 mm Hg and with intraperitoneal protein concentrations from 1-8 g%. The fractional absorption rates of fluid and protein did not change significantly over the 6 hours and were not significantly different from each other within each one hour period. In addition both fractional absorption rates were unaffected by the protein concentration of the fluid within the peritoneal cavity. Although the absolute rate of absorption is greatly increased by elevation of the intraperitoneal pressure, these data indicate that the process remains iso-oncotic as would be expected for lymphatic rather than transcapillary absorption.

Cytomegalovirus prostatitis. Case report and review of the literature.

We report the case of a patient undergoing chemotherapy for multiple myeloma discovered to have cytomegalovirus prostatitis. The findings of a hypoechoic prostatic lesion on ultrasound and a slightly elevated prostatic specific antigen of 4.6 ng/ml prompted a prostate biopsy. Cytopathologic examination and immunohistochemical staining demonstrated cytomegalovirus within the prostate. This virus is a common pathogen in the immunosuppressed patient, but its presence in the male genital tract is relatively rare. No previous reports of biopsy-proven cytomegalovirus prostatitis appear in the literature. The relationship of cytomegalovirus to the prostate is discussed in detail.

Sex effects on free recall of transforming orally and visually presented nouns into printed, picture, or imagery form.

Two experiments using male and female college students studied the effects on free recall of transforming auditory and visual stimuli by imagination or penciling. Experiment 1 (N = 152) presented nouns either auditorily by a tape recorder or visually as printed words, and Ss either drew them as pictures or printed them as words. Significantly more items were recalled as a result of drawing pictures during the presentation of stimuli (p less than .001). Female Ss recalled significantly more items than male Ss (p less than .01). There was no significant difference found between auditory or visual presentation. The procedure in Experiment 2 (N = 112) was similar, except that Ss transformed the nouns by imagination or penciling. Results were similar to those of Experiment 1. There was no significant difference between imagery and penciling.

Time estimation: effects of attentional focus and a comparison of interval conditions.

Time estimation was studied as a function of the subject's attention to the stimulus interval and the stimulus condition of the standard interval. 111 subjects from lower level psychology classes were randomly assigned to a interval condition. Time estimation of a standard interval was significantly lower when sujects attended to the stimulus material than when attending to the interval of passing time. A significant difference was not obtained among the three intervals.