Vibratome Sections Of Difficult Tissues

After 1 hour of aldehyde fixation, 4 to 20 hours of soaking in 2% BSA (bovine serum albumin) solution and another 17 hours in the same fixative, the vibratome will produce smooth, even sections from agar embedded guinea pig thyroid, skeletal muscle or larynx with the same ease as it will from soft, homogeneous tissues such as liver or spleen.     

Highly parallel and short-acting amplification with locus-specific primers to detect single nucleotide polymorphisms by the DigiTag2 assay

The DigiTag2 assay enables analysis of a set of 96 SNPs using Kapa 2GFast HotStart DNA polymerase with a new protocol that has a total running time of about 7 hours, which is 6 hours shorter than the previous protocol. Quality parameters (conversion rate, call rate, reproducibility and concordance) were at the same levels as when genotype calls were acquired using the previous protocol. Multiplex PCR with 192 pairs of locus-specific primers was available for target preparation in the DigiTag2 assay without the optimization of reaction conditions, and quality parameters had the same levels as those acquired with 96-plex PCR. The locus-specific primers were able to achieve sufficient (concentration of target amplicon ≥5 nM) and specific (concentration of unexpected amplicons <2 nM) amplification within 2 hours, were also able to achieve detectable amplifications even when working in a 96-plex or 192-plex form. The improved DigiTag2 assay will be an efficient platform for screening an intermediate number of SNPs (tens to hundreds of sites) in the replication analysis after genome-wide association study. Moreover, highly parallel and short-acting amplification with locus-specific primers may thus facilitate widespread application to other PCR-based assays.     

Isolierung von Auxinen aus dem Chromatin regenerierender Erbsenkeimlinge

Summary Etiolated pea epicotyls were treated with IAA, NAA or 2,4-D for 48 hours. From the chromatin (purified by gel-chromatography) of the treated seedlings growth substances could be isolated which significantly promote growth of Avena coleoptiles. The isolated growth substances showed the same RF as the applied auxins. When the seedlings were treated for 48 hours either with auxin or with water only the extract from the auxin treated plants promoted Avena growth significantly. The ether extract from the auxin treated seedlings still had growth promoting effect when the seedlings were treated 24 hours with auxin and then 24 hours with water to remove most of the free auxin in the cells and in the cell walls.     

The different roles of serum and calcium in the control of proliferation of BALB/c 3T3 mouse cells.

Proliferatively inactive BALB/c 3T3 mouse cells in dense cultures initiate a growth-division cycle upon exposure to fresh calf serum in a low-calcium (0.01 mM) medium. If these calcium-deprived cells are not supplied with calcium sometime during the first 10 hours after serum stimulation, they will rapidly return to a proliferatively inactive state without initiating DNA synthesis. The prereplicative development of such stimulated calcium-deprived cells appears to stop at an advanced stage, because addition of calcium as late as 10 hours after serum exposure rapidly initiates DNA synthesis, and enables the culture's DNA-synthetic activity subsequently to reach its peak value at the same time as in control cultures.     

Interference of MHV3 virus on ascites tumor growth.

In this experimental study the Authors have evaluated the interference of MHV3 hepatitis virus on Ascites Tumor's growth, using 4 groups of mice, the first one considered as control, the others inoculated with the virus 48 hours before, at the same time and 48 hours after tumor's inoculation. In each group labeling index, mitotic index and ascites volume were assessed a week after tumor's inoculation. The results seem to reveal an inhibition of tumor's growth in the second group. For this reason on the base of these encouraging observation, the Authors will continue researches to further confirm these preliminary results.     

Rapid dissemination of SIV follows multisite entry after rectal inoculation

Receptive ano-rectal intercourse is a major cause of HIV infection in men having sex with men and in heterosexuals. Current knowledge of the mechanisms of entry and dissemination during HIV rectal transmission is scarce and does not allow the development of preventive strategies. We investigated the early steps of rectal infection in rhesus macaques inoculated with the pathogenic isolate SIVmac251 and necropsied four hours to nine days later. All macaques were positive for SIV. Control macaques inoculated with heat-inactivated virus were consistently negative for SIV. SIV DNA was detected in the rectum as early as four hours post infection by nested PCR for gag in many laser-microdissected samples of lymphoid aggregates and lamina propria but never in follicle-associated epithelium. Scarce SIV antigen positive cells were observed by immunohistofluorescence in the rectum, among intraepithelial and lamina propria cells as well as in clusters in lymphoid aggregates, four hours post infection and onwards. These cells were T cells and non-T cells that were not epithelial cells, CD68(+) macrophages, DC-SIGN(+) cells or fascin(+) dendritic cells. DC-SIGN(+) cells carried infectious virus. Detection of Env singly spliced mRNA in the mucosa by nested RT-PCR indicated ongoing viral replication. Strikingly, four hours post infection colic lymph nodes were also infected in all macaques as either SIV DNA or infectious virus was recovered. Rapid SIV entry and dissemination is consistent with trans-epithelial transport. Virions appear to cross the follicle-associated epithelium, and also the digestive epithelium. Viral replication could however be more efficient in lymphoid aggregates. The initial sequence of events differs from both vaginal and oral infections, which implies that prevention strategies for rectal transmission will have to be specific. Microbicides will need to protect both digestive and follicle-associated epithelia. Vaccines will need to induce immunity in lymph nodes as well as in the rectum.     

Women in pediatrics: the experience in Quebec.

OBJECTIVES: To compare the practice patterns of female pediatricians in Quebec with those of their male counterparts and to identify specific factors influencing these practice patterns. DESIGN: Matched cohort questionnaire survey. SETTING: Primary, secondary and tertiary care pediatric practices in Quebec. PARTICIPANTS: All 146 female pediatricians and 133 of the 298 male pediatricians, matched for age as well as type and site of practice; 119 (82%) of the female and 115 (86%) of the male pediatricians responded. MAIN OUTCOME MEASURES: Demographic and family data as well as detailed information about the practice profile. RESULTS: The two groups were comparable regarding demographic data, professional work and patient care. Compared with the male respondents, the female pediatricians were younger and saw more outpatients. The mean number of hours worked per week, excluding on-call duty, was 40.5 (standard deviation [SD] 12.4) for the women and 48.9 (SD 12.0) for the men (p < 0.001). The female pediatricians were more likely than their male counterparts to have spouses who were also physicians (40%) or in another profession (45%). The female pediatricians without children worked significantly fewer hours than the male pediatricians with or without children (p < 0.001). Children (p = 0.006), but not the number of children (p = 0.452), had a significant effect on the number of hours worked by the female pediatricians. CONCLUSION: The duality of the role of female physicians as mothers and professional caregivers must be considered during workload evaluations. If the same style of practice and the increase in the proportion of female pediatricians continue, about 20% more pediatricians will be needed in 10 years to accomplish the same workload.     

Circadian activity rhythms in the crayfish Paranephrops zealandicus (Crustacea)

The diel activity rhythm shown by Paranephrops is usually unimodal with most activity occurring at night. In constant darkness, locomotor activity is expressed for up to 60 days as a free-running circadian rhythm. The period varies from 18 to 38 hours following a seasonal cycle (summer mean period 24 hours, winter mean 30 hours). No adaptive significance can be attached to either seasonal or individual variations in period and it is suggested that such variations are a consequence of the way in which the clock mechanism operates, the period being linked to the overall activity level following Aschoff's ( 1960 ) "Circadian Rule". Activity levels will inevitably vary individually and seasonally.     

The combined action of gamma irradiation and hyperthermia on the structure of the cell population in the Chinese hamster]

A change in the structure of FAF-28 Chinese hamster cell population occurred during 24 h following gamma-irradiation or hyperthermia heating, or the effect of both factors was studied by flow cytofluorometry. With radiation delivered immediately after heating the distribution of cells among cycle phases was nearly the same as with hyperthermia alone: the share of cells at the S-phase was invariable during the first 4-6 h, then it slowly diminished; at G1 it slowly decreased and at G2 increased. When irradiation preceded heating the pattern of cell redistribution during the first hours was the same as that with radiation alone: the "wave" of transition from G1 to S phase was the same, but shorter in amplitude and longer in time; then cells were accumulated at G2+M and remained there for 24 h. Thus, of the two factors applied, the first was the major one in changing the cell population structure during the first hours after treatment. In 24 h the result was the same, that is, the considerable accumulation of cells at G2+M.     

Phenylboronic acid selectively inhibits human prostate and breast cancer cell migration and decreases viability

We compared the in vitro effect of boric acid (BA) versus phenylboronic acid (PBA) on the migration of prostate and breast cancer cell lines and non-tumorigenic cells from the same tissues. Treatment at 24 hours with BA (≤ 500 µM) did not inhibit chemotaxis on fibronectin in any cell line. However, treatment over the same time course with concentrations of PBA as low as 1 µM significantly inhibited cancer cell migration without effecting non-tumorigenic cell lines. The compounds did not affect cell adhesion or viability at 24 hours but did alter morphology; both decreased cancer cell viability at 8 days. These results suggest that PBA is more potent than BA in targeting the metastatic and proliferative properties of cancer cells.     

Tissue and Organ Specificity of Eimeria tenella (Railliet and Lucet, 1891) Fantham, 1909 in Cecectomized Chickens*†

SYNOPSIS. The ceca of 2-week-old chicks were surgically removed. One week post-operation each cecectomized bird was given 2 × 10⁶ sporulated Eimeria tenella oocysts per os. Birds from the same hatch, with intact ceca, served as controls and were infected the same time as the cecectomized birds. However, in order to reduce mortality, control birds were each given 1 × 10⁴ sporulated E. tenella oocysts per os. Four cecectomized and 5 control birds were killed 12, 24, 48, 72, 96, 120, and 144 hours after inoculation. Tissues from the small and large intestine of each bird proximal to the cecal junction were removed, processed by the dioxan method, and studied microscopically for developmental stages of the parasite. Developmental stages of the parasite were observed in all sections from the large intestine of cecectomized chickens. Initial sporozoite penetration and subsequent development of the parasite in this location was similar to that observed in the cecal mucosa of non-cecectomized chickens. No parasites were observed in sections of the small intestine of cecectomized birds 12 or 24 hours after inoculation, and findings after 48 and 72 hours were inconsistent. However, numerous parasites were observed in sections 96, 120, and 144 hours post-inoculation. In contrast, endogenous stages of the parasite were not seen in tissue sections of the small and large intestine of birds with intact ceca until 120 hours after inoculation. Numerous young gametocytes were then observed in sections from all birds. Similarly, mature gametocytes were observed in all fixed sections 144 hours after inoculation. No evidence was found that would indicate whether or not infection in the small and large intestine of birds with intact ceca or the small intestine of cecectomized birds was initiated by sporozoites or merozoites, nor was evidence found to suggest that development of any stage of the parasite was suppressed in these organs.     

Feulgen Hydrolysis with Perchloric Acid

In tissue fixed with Carnoy's acetic alcohol (1:3), the hydrochloric acid hydrolysis performed as part of the Feulgen reaction is optimal for only a very short period of time. When 10% perchloric acid is used as the hydrolytic agent, the same color maximum is obtained, and the optimal hydrolysis time at 25°C. extends from 12 hours to 24 hours. During this time the intensity of color does not change. The events which take place during the period of suboptimal hydrolysis are the same as diose which take place during the corresponding period of hydrochloric acid hydrolysis. Part of the decrease in ultraviolet extinction of nuclei during the first 12 hours is due to the splitting off of purine bases from the desoxyribose nucleic acid. This is consistent with the increase in the amount of Feulgen dye bound by nuclei during this period of time. Between 12 hours and 24 hours no ultraviolet absorbing material is lost from nuclei, which is consistent with the fact that during this time the Feulgen color produced remains at a maximum.     

Replacement of L-T4 suppository in MMI treated rabbits

L-T4 suppositories containing 50, 100 or 200 micrograms of L-T4 were given to rabbits treated with MMI. The serum T4 gradually increased within 3.5 and 6.5 hours following the use of 100 and 200 micrograms of L-T4 suppositories, respectively, without an acute increase in serum T3. The increase in serum T4 continued up to 48 hours. The area under the curve (AUC) for serum T4 was evidently dose-dependent. It is concluded from these results that the T4 suppository will be useful as a replacement therapy for patients with hypothyroidism.     

Inexpensive Multiplexed Library Preparation for Megabase-Sized Genomes

Whole-genome sequencing has become an indispensible tool of modern biology. However, the cost of sample preparation relative to the cost of sequencing remains high, especially for small genomes where the former is dominant. Here we present a protocol for rapid and inexpensive preparation of hundreds of multiplexed genomic libraries for Illumina sequencing. By carrying out the Nextera tagmentation reaction in small volumes, replacing costly reagents with cheaper equivalents, and omitting unnecessary steps, we achieve a cost of library preparation of $8 per sample, approximately 6 times cheaper than the standard Nextera XT protocol. Furthermore, our procedure takes less than 5 hours for 96 samples. Several hundred samples can then be pooled on the same HiSeq lane via custom barcodes. Our method will be useful for re-sequencing of microbial or viral genomes, including those from evolution experiments, genetic screens, and environmental samples, as well as for other sequencing applications including large amplicon, open chromosome, artificial chromosomes, and RNA sequencing.     

Aprepitant versus ondansetron in preoperative triple-therapy treatment of nausea and vomiting in neurosurgery patients: study protocol for a randomized controlled trial

ABSTRACT: BACKGROUND: The incidence of postoperative nausea and vomiting is 50% to 80% after neurosurgery. The common prophylactic treatment for postoperative nausea and vomiting is a triple therapy of droperidol (Inapsine), promethazine (Phenergan) and dexamethasone (Decadron). Newer, more effectives methods of prophylaxis are being investigated. We designed this prospective, double-blind, single center study to compare the efficacy of ondansetron (Zofran) to a neurokinin-1 antagonist, aprepitant (Emend), as a substitute for droperidol, in the prophylactic treatment of postoperative nausea and vomiting after neurosurgery. METHODS: After obtaining institutional review board approval, One hundred-seventy-six patients, 18-85 years of age with ASA I to III, who did not receive anti-emetics 24 hours before surgery and are expected to undergo general anesthesia for neurosurgery lasting longer than two hours were included in this study. After meeting the inclusion and exclusion criteria and providing written informed consent, patients will be randomly assigned in a 1:1 ratio to one of two treatment groups: aprepitant or ondansetron. Because ondansetron is given intravenously and aprepitant orally, patients will be given an oral or intravenous placebo to maintain the double blind. Patients will receive aprepitant 40 mg PO/placebo within 2 hours prior to induction. At induction, a combination of intravenous dexamethasone 10 mg, promethazine 25 mg and ondansetron 4 mg/placebo will be given. The primary outcome measures are the episodes and severity of nausea and vomiting; administration of rescue antiemetic; and opioid consumption for 120 hours postoperatively. Standard safety assessments will include adverse event reports, physical and laboratory data, awakening time and duration of recovery from anesthesia. Logistic regression will be used to test the efficacy of aprepitant compared to ondansetron with demographic characteristics as potential covariates in the model. For the number of rescue therapy treatments used during the postoperative period, a Wilcoxon rank sum test will be performed. DISCUSSION: The results of this comparative study will potentially identify an improved treatment regimen that will decrease the incidence and severity of postoperative nausea and vomiting in patients undergoing neurosurgery. This will serve to enhance patient recovery and overall satisfaction of neurosurgical patients in the immediate postoperative period. Registered at The Ohio State University Biomedical Sciences Institutional Review Board: Protocol Number: 2007H0053 KEYWORDS: aprepitant, postoperative nausea and vomiting, craniotomy, ondansetron. Word Count: 347.     

Parental employment,family routines and childhood obesity

Using the Early Childhood Longitudinal Survey-Kindergarten Class of 1998–1999 (ECLS-K) data from kindergarten through eighth grade, this paper investigate the relationships among maternal employment, family routines and obesity. More hours worked by the mother tend to be negatively related to positive routines like eating meals as a family or at regular times, or having family rules about hours of television watched. Many of these same routines are significantly related to the probability of being obese, implying that family routines may be a mechanism by which maternal employment intensity affects children's obesity. However, inclusion of family routines in the obesity regression does not appreciably change the estimated effect of maternal employment hours. Thus, the commonly estimated deleterious effect of maternal employment on children's obesity cannot be explained by family routines, leaving the exact mechanisms an open question for further exploration.     

Sequence of prolactin effects on phospholipid synthesis in mouse mammary gland explants

In cultured mouse mammary gland explants derived from 12-14 day pregnant mice, the effect of prolactin (PRL) on the rate of incorporation of several precursors into neutral lipids and phospholipids was determined. Employing [14C]-acetate as a substrate, PRL stimulates its incorporation into a) neutral lipids by 4-6 hours, b) phosphatidyl choline (PC) and phosphatidyl inositol-phosphatidyl serine (PI-PS) by 1-2 hours, and c) phosphatidyl ethanolamine (PE) by 2-4 hours. Using [3H]-glycerol as a substrate, the temporal response to PRL for its incorporation into the neutral lipids was the same as that for [14C]-acetate, however, PRL did not enhance the rate of [3H]-glycerol incorporation into the phospholipids at any time through 16 hours. PRL similarly had no effect on the rates of [3H]-choline, [3H]-serine, [3H]-ethanolamine, or [32P]O4 incorporation into the phospholipids at hormone exposure periods of 8 hours or more. And finally, PRL had no effect on the rates of [3H]-arachidonate or [14C]-linoleate incorporation into neutral lipids or phospholipids at culture periods up to 18 hours. These data suggest that the early effect of PRL on [14C]-acetate incorporation into the phospholipids is due to either the insertion of newly synthesized fatty acids and/or the extension of fatty acids contained in the phospholipids.     

Reversibility of chilling injury to corn seedlings

Seedlings of corn (Zea mays) were tested for recovery from chilling injury incurred at 0.3 ± 0.3 C. At 0.3 C visual leaf injury appeared in 36 hours, whereas stem and root injuries appeared later. Appearance of leaf injury was preceded by a rise in O₂ uptake and a lessened effect of 2,4-dinitrophenol on O₂ uptake by leaf segments and was accompanied by increased ion leakage from the leaves. These effects were reversible, in that transfer of seedlings to 21 C after 36 hours at 0.3 C produced a return of O₂ uptake, 2,4-dinitrophenol stimulation, and ion leakage to the levels of unchilled leaves, as well as a disappearance of leaf symptoms, within 72 hours. For most seedlings, transfer to 21 C after 48 to 60 hours at 0.3 C reversed the chilling effects on O₂ uptake, 2,4-dinitrophenol stimulation, and injury symptoms but not on ion leakage within 108 hours. However, some seedlings collapsed during 48 to 60 hours of chilling, and these never recovered. Transfer to 21 C after 72 hours at 0.3 C did not produce recovery from any symptom of chilling injury examined, and these seedlings soon died. No growth occurred at 0.3 C, but growth began soon after transfer to 21 C. Seedlings chilled 24 or 36 hours grew at reduced rates during the first 72 hours at 21 C, but within 96 hours at 21 C were growing at the same rate as nonchilled seedlings. These results demonstrate considerable capacity of growing plants to recover from short chilling treatments even though significant physiological changes occurred at low temperatures.     

Tumor cell biotransformation products of prostaglandin A1 with growth inhibitory activity

The growth inhibitory effect and the fate of prostaglandin A1 (10(-6) M) were followed in cultures of rat B104 neuroblastoma and C6 glioma cells. More than 40% and 85% of the drug were neither recognized by a prostaglandin A1 antiserum nor extracted from the acidified medium with ethyl acetate, after 6 h and 24 h-incubation, respectively. When the supernatant of cells cultured in the presence of prostaglandin A1 during 24 hours was transferred to other cells and used as culture medium, the same growth inhibitory effect as with prostaglandin A1 was observed even when no prostaglandin A1 was added. After extensive purification and reverse phase HPLC of supernatant, four peaks more polar than prostaglandin A1 were shown; two of them were still active as growth inhibitors. This biotransformation was not observed with normal cells like L 929 or chick embryo fibroblasts, for which prostaglandin A1 had no inhibitory effect. The identification of these metabolites will allow the study of the structure-activity relationship.     

Analysis of the myogenic lineage in chick embryos. IV. Effects of conditioned medium

Immunocytochemical analysis of small myogenic clones was used to compare the effects of fresh medium (FM) and conditioned medium (CM) on muscle differentiation. In order to compare the same population of cells, clones were initiated in FM and then switched to either new FM or to CM. Clones were fixed at 12-hour intervals up to 76 hours, then assayed for the presence of post-mitotic myoblasts by immunoperoxidase staining for muscle myosin heavy chain (MHC) or M-creatine kinase (MCK). In both media, myogenic cells occurred predominantly in homogeneous positive clones (all cells (MHC +/MCK +) which contained 2" cells. At 76 hours, the percentages of 1-, 2-, and 4-cell positive clones did not differ statistically in the two conditions; however, the percentages of 8- and 16-cell positive clones were significantly reduced in CM, and the percentages of small negative clones were concomitantly increased. We conclude from these data that CM affects myogenesis by slowing progression through a predetermined lineage rather than by changing the number of mitoses an individual cell will undergo before terminally differentiating. These results further support the idea that progress through the myogenic lineage is mediated by cell divisions.