Detection of HIV and HCV RNA in semen from Brazilian coinfected men using multiplex PCR before and after semen washing

INTRODUCTION: Prolonged survival of patients under HAART has resulted in new demands for assisted reproductive technologies. HIV serodiscordant couples wish to make use of assisted reproduction techniques in order to avoid viral transmission to the partner or to the newborn. It is therefore essential to test the effectiveness of techniques aimed at reducing HIV and HCV loads in infected semen using molecular biology tests. METHODS: After seminal analysis, semen samples from 20 coinfected patients were submitted to cell fractioning and isolation of motile spermatozoa by density gradient centrifugation and swim-up. HIV and HCV RNA detection tests were performed with RNA obtained from sperm, seminal plasma and total semen. RESULTS: In pre-washing semen, HIV RNA was detected in 100% of total semen samples, whereas HCV RNA was concomitantly amplified in only one specimen. Neither HIV nor HCV were detected either in the swim-up or in the post-washing semen fractions. CONCLUSIONS: Reduction of HIV and/or HCV shedding in semen by density gradient centrifugation followed by swim-up is an efficient method. These findings lead us to believe that, although semen is rarely found to contain HCV, semen processing is highly beneficial for HIV/HCV coinfected individuals.

Sperm tail flexibility test: a simple test for selecting viable spermatozoa for intracytoplasmic sperm injection from semen samples without motile spermatozoa

PURPOSE: The objective was to describe the results of the injection of immotile spermatozoa with flexible tails when only immotile spermatozoa are present in the semen sample. METHODS: A retrospective study was conducted to analyze the procedure results for 10 couples who participated in our intracytoplasmic sperm injection program. The sperm tail was considered flexible when it moved up and down independently of the head movement, and it was considered inflexible when the movement occurred together (tail plus head). The fertilization and pregnancy rate were analyzed. RESULTS: The normal fertilization rate (presence of 2 pronuclei) was 30.3% (40/132), and the abnormal fertilization rate (presence of less than or more than 2 pronuclei) was 6.81% (9/132). A total of 52 embryos were obtained with 9 transfer procedures performed (pregnancy rate: 11.12%). CONCLUSIONS: The sperm tail flexibility test (STFT) is an easy and cost-effective way for selecting viable immotile spermatozoa and can be used as an alternative method for determining the viability of spermatozoa. This test seems to be a simple and risk-free method when compared to the swelling test.

Congelación de semen en porcinos: 4. Estudio trianual de la calidad del semen congelado en sistemas monodosis mediante la evaluación por pruebas de laboratorio y de fertilidad

La congelación del semen porcino se ha caracterizado por ser una técnica que no ha logrado alcanzar altos índices de fertilidad, hecho que ha dificultado su aplicación práctica a terreno y parte de su progreso genético. El problema particular radica en que la calidad espermática, por factores poco conocidos, disminuye considerablemente al momento de la descongelación. Como consecuencia, las tasas de fecundidad, fertilidad y prolificidad son inferiores a las obtenidas mediante monta natural o inseminación artificial con semen fresco refrigerado. Pese a esta dificultad, la fertilidad es adecuada para casos de importación, exportación, introducción de nuevas líneas de sangre y uso de semen de machos con alto mérito genético. (...) Entre los factores más importantes que afectan la fertilidad del semen congelado se citan el efecto individual de los machos, de mala congelabilidad, el efecto raza, el momento óptimo para inseminar a la hembra en celo, la estación del año y el tiempo de almacenaje del semen, entre otros. (...) Objetivos Generales: El objetivo de este trabajo es desarrollar un programa anual destinado, en una primera etapa, a mejorar la técnica de laboratorio con equipamiento de mayor desarrollo tecnológico y utilizando nuevos envases; en una segunda etapa a estudiar el efecto raza y el efecto individual, con un pool de semen de diferentes machos; y en una tercera etapa, consistente en aplicar el mejor semen criopreservado, en el momento biológico óptimo de la hembra en celo, con el fin de aumentar la fertilidad y prolificidad. Objetivos Específicos: 1. Primer Año Mejorar las técnicas de congelación utilizando nuevos equipos y estudiar la calidad del semen porcino congelado en micropajuelas (Mp) y bolsas plásticas (Bp) evaluando la capacidad fecundante por pruebas de laboratorio y pruebas de fertilidad en animales vivos. 2. Segundo Año Estudiar la calidad del semen porcino congelado en Mp y/o Bp utilizando un pool de semen de tres machos de razas diferentes y evaluar su capacidad fecundante por pruebas de fertilidad a campo. 3. Tercer Año Estudiar la calidad del semen porcino congelado en Mp y/o Bp, utilizando un pool de semen de tres machos diferentes e inseminándolo en el momento biológico óptimo del celo.

Social freezing in Switzerland and worldwide--a blessing for women today?

This is a critical review of the medical, ethical, judicial and financial aspects of the so called "social freezing", the cryopreservation of a woman's oocytes for non-medical purposes. The possibility of storing the eggs of fertile women in order to prevent age-related fertility decline is being widely promoted by fertility centres and the lay press throughout the world. Research data has shown that social freezing should ideally be performed on women around 25 years of age in order to increase their chances of a future pregnancy. In reality, it is mostly performed after the age of 35. Unfortunately, social freezing is in general not a solution for the underlying societal problems to fit in with professionally active women and having children. It only delays the existing problems. Furthermore, it creates a lot of potential new problems. A great deal more should be undertaken to offer real solutions to the underlying societal problems which are in part: pre-school education, care in the event of childhood illness, and the many weeks of school holidays, acceptance of professionally active women having children, and more job offers with a workload <100%.). Furthermore, society should be informed about the decreasing chances of pregnancy with increasing maternal (and paternal) age as well as the increasing risks of miscarriage and obstetric/neonatal complications. Detailed information for woman considering social freezing is crucial. Every doctor, proposing social freezing to his patients, should be up to date with all these details. Follow-up studies on the outcome of these children are needed.

Freezing of 4He and its liquid-solid interface from density functional theory

We show that, at high densities, fully variational solutions of solidlike types can be obtained from a density functional formalism originally designed for liquid 4He . Motivated by this finding, we propose an extension of the method that accurately describes the solid phase and the freezing transition of liquid 4He at zero temperature. The density profile of the interface between liquid and the (0001) surface of the 4He crystal is also investigated, and its surface energy evaluated. The interfacial tension is found to be in semiquantitative agreement with experiments and with other microscopic calculations. This opens the possibility to use unbiased density functional (DF) methods to study highly nonhomogeneous systems, like 4He interacting with strongly attractive impurities and/or substrates, or the nucleation of the solid phase in the metastable liquid.

Development and Demonstration of a Freezing Drizzle Algorithm for Roadway Environmental Sensing Systems, Final Report, October 2012

The primary goal of this project is to demonstrate the accuracy and utility of a freezing drizzle algorithm that can be implemented on roadway environmental sensing systems (ESSs). The types of problems related to the occurrence of freezing precipitation range from simple traffic delays to major accidents that involve fatalities. Freezing drizzle can also lead to economic impacts in communities with lost work hours, vehicular damage, and downed power lines. There are means for transportation agencies to perform preventive and reactive treatments to roadways, but freezing drizzle can be difficult to forecast accurately or even detect as weather radar and surface observation networks poorly observe these conditions. The detection of freezing precipitation is problematic and requires special instrumentation and analysis. The Federal Aviation Administration (FAA) development of aircraft anti-icing and deicing technologies has led to the development of a freezing drizzle algorithm that utilizes air temperature data and a specialized sensor capable of detecting ice accretion. However, at present, roadway ESSs are not capable of reporting freezing drizzle. This study investigates the use of the methods developed for the FAA and the National Weather Service (NWS) within a roadway environment to detect the occurrence of freezing drizzle using a combination of icing detection equipment and available ESS sensors. The work performed in this study incorporated the algorithm developed initially and further modified for work with the FAA for aircraft icing. The freezing drizzle algorithm developed for the FAA was applied using data from standard roadway ESSs. The work performed in this study lays the foundation for addressing the central question of interest to winter maintenance professionals as to whether it is possible to use roadside freezing precipitation detection (e.g., icing detection) sensors to determine the occurrence of pavement icing during freezing precipitation events and the rates at which this occurs.

Freezing and low temperature photoinhibition tolerance in cultivated potato and potato hybrids

Selostus: Perunan ja perunahybridien jäätymisen ja fotoinhibition kestävyys

Subgrade Insulation to Prevent Soil Freezing, HR-87, 1965

The basic purpose of this study was to determine if an expanded polystyrene insulating board could prevent subgrade freezing and thereby reduce frost heave. The insulating board was placed between a nine inch P. C. concrete slab and a frost-susceptible subgrade. In one section at the test site, selected backfill material was placed under the pavement. The P. C. pavement was later covered by asphalt surfacing. Thermocouples were installed for obtaining temperature recordings at various locations in the surfacing, concrete slab, subgrade and shoulders. This report contains graphs and illustrations showing temperature distributions for two years, as well as profile elevations and the results of moisture tests.

Investigation into Freezing-Thawing Durability of Low-Permeability Concrete with and without Air Entraining Agent, 2009

The aim of the present study is to investigate the effect of low-permeability concrete, made with reduced water‐to‐binder ratios (w/b) and/or supplementary cementitious materials (SCMs), on the need for air entrainment to achieve freezing‐thawing (F‐T) durability. In the present study, concrete mixes were made with different types of cement (Types I and IP), with or without fly ash replacement (15%), with different water‐to‐binder ratios (w/b =0.25, 0.35, 0.45 and 0.55), and with or without air entraining agent (AEA). All concrete mixtures were controlled to have a similar slump by using different dosages of superplasticizer. The rapid chloride permeability and F-T durability of the concrete samples were determined according to ASTM C1202 and ASTM C666A, respectively. The air void structure of the concrete was studied using the Air Void Analyzer, RapidAir, and porosity tests (ASTM C642). In addition, the general concrete properties, such as slump, air content, unit weight, and 28‐day compressive strength, were evaluated. The results indicate that all concrete mixes with proper air entrainment (ASTM C231 air content ≥ 6%) showed good F‐T resistance (durability factor ≥85%). All concrete mixes without AEA showed poor F‐T resistance (durability factor < 40%), except for one mix that had very low permeability and high strength. This was the concrete made with Type IP cement and with a w/b of 0.25, which had a permeability of 520 coulombs and a compressive strength of 12,760 psi (88 MPa). There were clear relationships between the F‐T durability and hardened concrete properties of non–air entrained concrete. However, such relationships did not exist in concrete with AEA. For concrete with AEA, good F‐T durability was associated with an air void spacing factor ≤ 0.28 mm (by AVA) or ≤ 0.22 mm (by RapidAir).

Freezing Point Depressions of Dilute Solutions of Alkali Metal Chlorides and Bromides

Freezing point depressions (¿Tf) of dilute solutions of several alkali metal chlorides and bromides were calculated by means of the best activity coefficient equations. In the calculations, Hückel, Hamer and Pitzer equationswere used for activity coefficients. The experimental ¿Tf values available in the literature for dilute LiCl, NaCl and KBr solutions can be predicted within experimental error by the Hückel equations used. The experimental ¿Tf values for dilute LiCl and KBr solutions can also be accurately calculated by corresponding Pitzer equations and those for dilute NaCl solutions by the Hamer equation for this salt. Neither Hamer nor Pitzer equations predict accurately the freezing points reported in the literature for LiBr and NaBr solutions. The ¿Tf values available for dilute solutions of RbCl, CsCl or CsBr are not known at the moment accurately because the existing data for these solutions are not precise. The freezing point depressions are tabulated in the present study for LiCl, NaCl and KBr solutions at several rounded molalities. The ¿Tf values in this table can be highly recommended. The activity coefficient equations used in the calculation of these values have been tested with almost allhigh-precision electrochemical data measured at 298.15 K.

Asset freezing, corporate political resources and the Tullock paradox

In 1967, Gordon Tullock asked why firms do not spend more on campaign contributions, despite the large rents that could be generated from political activities. We suggest in this paper that part of the puzzle could come from the fact that one important type of political activity has been neglected by the literature which focuses on campaign contributions or political connections. We call this neglected activity "asset freezing": situations in which firms delay lay-offs or invest in specific technologies to support local politicians' re-election objectives. In doing so, firms bear a potentially significant cost as they do not use a portion of their economic assets in the most efficient or productive way. The purpose of this paper is to provide a first theoretical exploration of this phenomenon. Building on the literature on corporate political resources, we argue that a firm's economic assets can be evaluated based on their degree of "political freezability," which depends on the flexibility of their use and on their value for policy-makers. We then develop a simple model in which financial contributions and freezing assets are alternative options for a firm willing to lawfully influence public policy-making, and derive some of our initial hypotheses more formally.

Toxoplasma gondii in semen of experimentally infected swine

Eight reproductive boars were divided into three groups and inoculated with Toxoplasma gondii [GI (n=3) 1.5x10(4) oocysts strain P; GII (n=3) 1.0x10(6) tachyzoites strain RH; and GIII (n=2) non-inoculated control]. Clinical, hematological, parasitemia and serological tests and studies of the parasite in the semen through bioassay and PCR, and in reproductive organs (Bioassay and immunohistochemical analyses) were conducted to evaluate the toxoplasmic infection. Blood and semen were collected on day -2, -1, 1, 3, 5, 7, 9, 11, 14 and weekly up to 84 days post-inoculation (DPI). No clinical or hematimetric alteration was observed in the boars. Parasitemia was detected in one boar inoculated with oocysts at the 7th DPI and in another boar infected with tachyzoites (GII) at the 3rd and 49th DPI. Serological tests revealed antibodies against T. gondii in animals inoculated with oocysts or tachyzoites at the 7th DPI with dilutions of 1:256 and 1:64, which reached peaks of 1:4096 at day 11 and 9, respectively. The bioassays revealed the presence of the parasite in semen samples of a boar inoculated with oocysts (GI) at 3, 49 and 56 DPI and from two boars infected with tachyzoites (GII), one animal at 5 and two animals at 49 days DPI. Mice inoculated with semen from the control group (GIII) remained serologically negative. PCR analysis showed T. gondii DNA in the semen of Boar 1 and Boar 3 inoculated with tachyzoites and oocysts, respectively. The immuno-histochemical tests showed T. gondii in the reproductive organs of Boar 1 and Boar 2, inoculated with tachyzoites and oocysts, respectively. These findings suggest the possible occurrence of venereal transmission of T. gondii in swine.