Wright's Solution Research Articles

DNA integrity of canine spermatozoa during chill storage assessed by the sperm chromatin dispersion test using bright-field or fluorescence microscopy

The objective of this study was to evaluate the effect of chill storage on canine sperm DNA fragmentation assessed by the sperm chromatin dispersion test using bright-field microscopy with Wright solution (sDF-B) or fluorescence microscopy with propidium iodide (sDF-F). The relationship and agreement between the results obtained with both staining methods were analyzed. The values of DNA fragmentation indexes (sDF-F and sDF-B) were compared at each time of chill storage (0, 24, 48, 72, and 96 hours). Additionally, the sperm DNA fragmentation rate (slope) was compared between the methods during chill storage. Good agreement and no significant differences between values obtained with both staining procedures were observed. Finally, the effect of chill storage for up to 96 hours was assessed on sperm motility parameters and DNA fragmentation indexes. Significant differences were found after 48 hours of chill storage, obtaining greater values of fragmented DNA. Progressive sperm motility was lower just after 96 hours of chill storage, and no effect was found in total sperm motility. In conclusion, the Sperm-Halomax kit, developed for canine semen and based on the sperm chromatin dispersion test, can be used accurately under bright-field or fluorescence microscopy to assess the sperm DNA integrity of canine semen during chill storage. The sperm DNA fragmentation index increased after 48 hours of chill storage, thereby detecting sperm damage earlier than other routine sperm parameters, such as sperm motility.

Characterization of the OSF-band structure in n-type Cz-Si using photoluminescence-imaging and visual inspection

Oxygen induced stacking faults (OSFs) are mainly seen in oxygen rich wafers from the seed end of Cz-silicon crystals. In wafers this ring shaped OSF-region delineates a border between two defect regions; usually silicon self-interstitials dominate outside and vacancies inside this ring. High temperature treatment (>800°C) leads to oxygen precipitation in the border region. These precipitates act as nucleation sites for stacking faults. The standard procedure for characterizing the OSF-rings is to expose an oxidized sample to a preferential etchant, e.g. the highly toxic Wright solution. In this work photoluminescence-imaging is compared to preferential etching and visual inspection. Vertical samples from an n-type Cz-crystal containing an OSF-boundary are studied before and after wet oxidation. High resolution PL-images, which allow for a close inspection of the OSF-area, reveal a complex band-structure of this border region.

Characterization of Oxidation-Induced Stacking Fault Rings in Cz Silicon: Photoluminescence Imaging and Visual Inspection After Wright etch

Oxidation-induced stacking fault rings in polished Cz silicon samples before and after thermal wet oxidation are investigated by use of photoluminescence imaging. Currently the standard procedure for OSF ring detection is to expose the samples to a toxic preferential etchant, e.g. a Wright solution, after a thermal oxidation. This solution primarily attacks the regions with stacking faults, allowing detection by visual inspection. Samples from the seed end of p-type Cz silicon ingots with resistivities of approximately 1Ohm-cm were measured by PL imaging before and after a thermal oxidation process. Subsequently, Wright-etching was performed on the oxidized samples to expose stacking faults. The lifetime variations in the PL images were correlated with the location of the rings in the preferentially etched surfaces, and good agreement was found. The results show that for this crystal pulling process, even the PL images of unpassivated polished samples can be used to detect the OSF ring location. The thermal oxidation at 1100°C enhanced the contrast between the OSF ring and the rest of the sample in the PL image.

313 ASSESSMENT OF SPERM DNA FRAGMENTATION IN CANINE EJACULATES USING THE Sperm-Halomax® KIT: PRELIMINARY RESULTS

Recently, a new procedure for the analysis of sperm DNA fragmentation has been developed for humans and different mammalian species, using a commercial kit based on the sperm chromatin dispersion (SCD) test; however, a descriptive study in canine semen has not been performed. The aim of this work was to assess the sperm DNA fragmentation in canine ejaculates using the SCD test and 2 different staining techniques. For this purpose, ejaculates were collectedby digital manipulation from4 healthy dogs of different breeds (1 German Pointer, 2 Spanish Greyhounds, and 1 Crossbreed). After collection, the sperm-rich fraction of the ejaculates from 3 dogs were pooled each time (n = 4) and then extended in Dulbecco’s phosphate buffered saline. All the pooled semen samples presented physiological values concerning routine semen parameters (motility, morphology, and sperm concentration). The sperm DNA fragmentation was assessed using the Sperm-Halomax® commercial kit specifically developed for canine semen (Halotech DNA SL, Madrid, Spain). Two semen aliquots of the diluted pooled semen samples were processed on each pre-treated slide provided in the kit following the manufacturer’s instructions. The last step was the staining technique. We stained each slide with 2 different staining procedures. The first half of the slide was stained with propidium iodide (Sigma-Aldrich, St. Louis, MO, USA) mixed in a proportion 1 : 1 with an antifading solution. The second half of the slide was stained for 15 min in Wright solution (1.01383.0500, Merck, Whitehouse Station, NJ, USA) 1 :1 in Phosphate Buffer pH 6.88 (1.07294.1000, Merck). The stained slides were observed using fluorescence and light microscopy, respectively. Five hundred sperm per slide were counted. Spermatozoa with fragmented DNA showed a large and spotty halo of chromatin dispersion. Unfragmented sperm only showed a small and compact halo. Statistical analyses were performed using the Statistical Package for Social Science version 12.0 (SPSS Inc., Chicago, IL, USA). The sperm DNA fragmentation index was compared between Wright and fluorescence staining methods by ANOVA. Results were expressed as mean ± standard error of the mean. The first report of the sperm DNA fragmentation index in canine ejaculates was 2.26 ± 0.53% for Wright staining and 1.99 ± 0.10% for fluorescence technique. No differences were found between staining procedures. In conclusion, it was possible to assess the sperm DNA fragmentation of canine ejaculates using 2 different staining procedures, expecting that continuous research could be useful in defining the role of DNA fragmentation SCD test in canine semen evaluation and cryopreservation.

Treatment of K562 cells with 1,25-dihydroxyvitamin D3 induces distinct alterations in the expression of apoptosis-related genes BCL2, BAX, BCLXL, and p21

Apoptosis, or programmed cell death, is a very important phenomenon in cytotoxicity induced by anticancer treatment. 1α,25-Dihydroxyvitamin D(3) (1,25-(OH)(2)D(3)), the active metabolite of vitamin D, inhibits the growth of multiple types of cancer cells including breast, colon, and prostate cancer cell lines. We studied alterations in the mRNA expression levels of BCL2, BAX, CYC, BCL-XL, and VDR genes in the K562 chronic myeloid leukemia cell line in response to treatment with 1,25-(OH)(2)D(3). Morphological observation of K562 cells was evaluated by the staining with Wright's solution. Cell percentage at different phases of the cell cycle was measured, and apoptosis was measured by flow cytometry. The expression levels of the apoptosis-related genes were analyzed by real-time reverse transcription polymerase chain reaction. We found that treatment with 1,25-(OH)(2)D(3) down-regulates BCL2 and BCL-XL mRNA expressions, as well as up-regulates expressions of BAX and p21 mRNA. The expression pattern of CYC and VDR genes were not influenced. However, K562 cells treated with 1,25-(OH)(2)D(3) caused an arrest of cell cycle progression in G1 phase resulting in a decreased number of cells in the S phase, complemented by an accumulation of cells in the G0-G1 phases. Our data show the modulatory effects of 1,25-(OH)(2)D(3) treatment in apoptosis-related genes in K562 cells.

Identificación morfológica de Ehrlichia sp. en las plaquetas de pacientes con infección por virus de la inmunodeficiencia humana, en Venezuela

To determine the frequency of infection by bacteria of the genus Ehrlichia in a population of patients infected with the human immunodeficiency virus (HIV). As is widely recognized, HIV infection is usually followed by the acquired immunodeficiency syndrome (AIDS) after a variable period during which the individual can develop different types of opportunistic infections whose severity depends on his/her immune status. Human ehrlichiosis is a newly-recognized tick-borne infectious disease caused by Ehrlichia spp., which are obligate intracellular, Gram-negative bacteria showing tropism for blood cells (leukocytes and platelets). In Venezuela, the disease was first recognized in humans in 1994, when intra-cytoplasmic microcolonies (morula) were found in human platelets. We collected peripheral blood samples from 87 HIV-positive persons that were received at the Immunology Laboratory of the Oncology and Hematology Institute in Caracas, Venezuela, one of the country's referral centers for cases of HIV infection. Slides prepared from buffy coat that had been stained with Wright's solution and sealed with resin for preservation were examined retrospectively under the light microscope. Ehrlichia spp. were found in the platelets of 12 of the 87 (13.8%) patients whose slides were examined. The presence of Ehrlichia sp. in the platelets of HIV-positive patients suggests that ehrlichiosis should be among the differential diagnoses of opportunistic infection in this group of patients.

Ultrastructural Characteristics of Alginate-Induced Reddish Purple Stained Cells (ARPC) in Mouse Peritoneal Cavity

He, W., Fujiki, K. and Fujihara, N. 2001. Ultrastructural characteristics of alginate-induced reddish purple stained cells (ARPC) in mouse peritoneal cavity. J. Appl. Anim. Res., 20: 15–26. To characterize microscopic features of the alginate-induced reddish purple-stained cells (ARPC) using electron microscope, one ml of sodium alginate solution (10 mg/ml) was injected into mice intraperitoneally. Mice were killed after the injection at various time intervals to collect the peritoneal fluid. The cells contained in the fluid were collected by centrifugation and subjected to cytological examinations. As a result, electron microscopy revealed that the vacuole-like organelles (VLO) contained very small amount of electron-dense substance. The substance was stained reddish purple with Wright solution. ELISA using anti-alginate monoclonal antibody and uronic acid determination indicated that the substance is not phagocytosed sodium alginate but ARPC-derived cellular materials. It is concluded that the ARPC is shown to be obviously cellular organelles.

N + P Junction Leakage Current Caused by Oxygen Precipitation Defects and Its Temperature Dependence

We investigated the relation between the generation leakage current and oxygen precipitation defects (OPD) and the temperature dependence of the generation leakage current and the diffusion leakage current for junctions fabricated on czochralski (CZ)‐Si wafers with different thermal histories. The generation and diffusion leakage current can be calculated from the area current component of the junction leakage current. To separate the area current component from the total leakage current gate‐controlled junction diodes with a wide range of perimeter‐to‐area ratios were investigated. The OPD (plate‐like oxide precipitates and punch‐out dislocations) were created inside CZ‐Si wafers during both the prefabrication heat‐treatments and the junction fabrication process itself. After the electrical measurements, the size and shape of the OPD under the junction regions were investigated by transmission electron microscopy observations. The density of the oxide precipitates was estimated for the surface and cleaved wafer planes from the optical observations of the samples etched in a Wright solution. The generation leakage current at room temperature was strongly affected by the density and size of the oxide precipitates. The activation energy of the generation leakage current for the sample with the highest OPD density was calculated as 0.58 eV, with the dominant generation centers positioned at the oxide precipitate/silicon interface. © 1999 The Electrochemical Society. All rights reserved.

Cytologic Diagnosis of Chlamydia in Cervicovaginal Secretions

To carry out a prospective study on 1,569 females to evaluate the diagnostic utility of adding buffered Wright solution to Papanicolaou stain for observing cytoplasmic inclusions in cervicovaginal cytology. Group A had multivacuolated cells and group B, granules (not in a vacuole) in the cytoplasm of low-intermediate epithelial or parabasal cells. There were 7 patients in group A and 16 in group B. Two duplicate plates of cervicovaginal secretions were obtained from each patient before and after treatment; one was stained with Papanicolaou stain and the other with our variant. The trial of therapy consisted of doxycycline (effectiveness for statistical analysis = 0.9) in oral doses of 100 mg every 12 hours for 7 days; 7 days after the beginning of treatment, check samples were obtained. In group A, six patients had cytoplasmic inclusions with the variant stain, and two had cytoplasmic inclusions with Papanicolaou stain. The binomial test revealed that probably six of the seven patients had infections with Chlamydia (P = .372) and that in 100% of cases, the variant stain showed cytoplasmic inclusions, while Papanicolaou stain was observed in 33% of cases. These findings were morphologically and statistically proven (P = .124) on control slides with posttreatment absence of multivacuolated cells. In relation to the selection criteria for B group, the intracytoplasmic granules found in parabasal and low intermediate cells had no relation to Chlamydia. The study demonstrated the superiority of our variant of Papanicolaou stain for cervicovaginal Chlamydia diagnosis.

A case report of an overseas-traveler's diarrhea probably caused by Chilomastix mesnili infection.

Many active flagellates were detected in stool samples of a 22-year-old Japanese male who traveled China and India. Microscopic observation showed that the organisms stained with Wright's solution were pyriform or rounded in shape, measuring 11.7-17.5 pm in length and 7.9-f2.2 pm in width. The posterior end was pointed. Three free anterior flagella were located in the front end. A spherical nucleus was situated near the anterior pole. Based on these morphological features we identified the organisms as trophozoites of Chilomastix mesnili, a common intestinal protozoon. Although we examined viral, bacterial and parasitic infections other than C.mesnili, no pathogen was found. From these results obtained, it was considered to be highly probable that C. mesnili infection was the cause of the diarrhea.

Dummett's Critique of Wright's Attempt to Resuscitate Frege

Michael Dummett mounts, in Frege: Philosophy of Mathematics, a concerted attack on the attempt, led by Crispin Wright, to salvage defensible versions of Frege's platonism and logicism in which Frege's criterion of numerical identity plays a leading role. I discern four main strands in this attack—that Wright's solution to the Caesar problem fails; that explaining number words contextually cannot justify treating them as enjoying robust reference; that Wright has no effective counter to ontological reductionism; and that the attempt is vitiated by the unavoidable impredicativity of its leading principle—and argue that none of them succeeds.

Electrophysiological properties of three types of granulocytes in circulating blood of the newt.

1. The electrophysiological properties of three subtypes of granulocytes obtained from the circulating blood of the newt, Triturus pyrrhogaster, were studied using the whole-cell variation of the patch-electrode voltage-clamp technique. 2. Neutrophils were identified by their multilobulated nucleus in the cytoplasm. Basophils and eosinophils, having characteristic granular structures in their cells, could definitely be distinguished from other leucocytes. The reliability of cellular identification using phase-contrast microscopy was confirmed by fixing and staining the granulocytes with Wright's solution. 3. In neutrophils under current-clamp conditions, a hyperpolarization-induced conductance increase was observed. With depolarization, however, no changes in regenerative potential were detected. When voltage clamped in standard saline (containing 96 mM-NaCl), hyperpolarizing voltage pulses to a potential more negative than -90 mV evoked slowly decaying inward currents. 4. The hyperpolarization-evoked membrane currents in neutrophils were identified as anomalous rectifying K+ currents, since (1) externally applied Cs+ (0.1 or 1 mM) or Ba2+ (1 mM) produced suppressive effects on the currents, (2) replacement of external Na+ with choline ions eliminated the decay of macroscopically observed currents, and (3) both the amplitude and kinetic properties of the currents were strongly dependent on membrane potential as well as on external K+ concentration; the activation of the conductance depended on the electrochemical force for K+ rather than on membrane potential alone. The magnitude of steady-state conductance was roughly proportional to the square root of the external K+ concentration. 5. In basophils and eosinophils, no major time- or voltage-dependent increase in conductance was detected at voltages between +20 and -130 mV. However, under current-clamp conditions, spontaneous fluctuation of zero-current potentials was clearly apparent, presumably due to the activities of some ion channels generating a small amount of current flux through the membranes of these cells. 6. It was concluded that the three subtypes of granulocytes in the newt differ considerably not only in appearance and structure but also in the electrical properties of their membranes. The anomalous rectifying K+ channels in neutrophils may serve to determine the resting potential of the cell at K+ equilibrium potential. The closure of the channels at depolarization might facilitate the maintenance of depolarization triggered by stimuli accompanying current influx.

The probability of war in then-crises problem: Modeling new alternatives to Wright's solution

In hisStudy of War, Q. Wright considered a model for the probability of warP “during a period ofn crises”, and proposed the equationP=1−(1−p) n , wherep is the probability of war escalating at each individual crisis. This probability measure was formally derived recently by Cioffi-Revilla (1987), using the general theory of political reliability and an interpretation of the “n-crises problem” as a branching process. Two new, alternate solutions are presented here, one using D. Bernoulli's St. Petersburg Paradox as an analogue, the other based on the logic of conditional probabilities. Analysis shows that, while Wright's solution is robust with regard to the general overall behavior ofp andn, some significant qualitative and quantitative differences emerge from the alternative solutions. In particular,P converges to 1 only in a special case (Wright's) and not generally.

Automated staining of bone marrow and peripheral blood by a modified Wright's technic.

A Wright's stain provided for the automatic staining of routine peripheral blood smears is recommended for use in the Hematrak instrument. This stain at a pH range of 6.5-7.15 shows a dull and bluish coloration of erythrocytes, a finding not readily acceptable for visual microscopy of cell morphology or for photomicroscopy. In order to achieve a desirable erythrocyte color, but still retain the staining intensity of leukocyte chromatin, a two-stage staining method was devised. This involves the use of a mixture of Eosin y and a buffered Wright's solution to secondarily stain Wright-stained smears. This modification of Wright's technic offers a stain of high quality for both bone marrow and peripheral blood films with the advantage of little or no day-to-day variability. In comparison to the frequently advocated dual staining methods, it is cheap and easy to automate. The total staining time for peripheral blood films is about 12 minutes and about 20 minutes for bone marrow smears.

Query: On the Estimation of Season Total Number of Different Households Utilizing a Park

Robson and Wright (1977) recently provided an answer to a query on the problem of estimating the number of different households visiting a park during a season. Robson and Wright's solution used only the names of households in attendance for a sample of days. Robson and Wright illustrated their method with a sample of 3 days selected from a population of 6 days. We outline an alternative procedure for estimating the number of distinct park users. The basis for the procedure is a study of Iowa park users conducted by the Statistical Laboratory of Iowa State University for the Iowa Conservation Commission in 1962. In the Iowa study, a stratified unequal probability sample of parks, days. times of day and exit gates was selected. Twenty-eight parks with an estimated attendance of about 4,500,000 visitor days (number of visitors on a day summed over days of the season) were included in the sample. Only a fraction of people exiting from the selected parks on the selected days was sampled. For purposes of our present discussion, only the fact that a random sample of days was selected for the sample parks need concern us. At the end of the season a subsample of the visitors identified in the original sample of days was selected. Individuals in this sample were asked to enumerate their park visits for the entire season. One of the reasons for the end of season study was to obtain a subsample of those who did not respond to a questionnaire distributed to the original sample. Although it was not an objective in the Iowa study, we demonstrate how such a subsample can be used to estimate the number of distinct visitors. We first consider a very simple design. Let a simple random sample of days be selected. The persons visiting the park on those days are contacted at the end of the season and asked to report their total number of visits to the park. Let tij = total number of visits during the season for individual i visiting the park on day j. The probability of individual i being observed given that one day is chosen at random is N1tij, where N is the total number of days in the season. Suppose that day j is randomly selected and M1 different individuals visited the park on dayj. Then Mj Kj= E t N is an unbiased estimator for K, where K is the total number of different individuals visiting the park during the entire season. An unbiased estimator of K based on a sample of n days is then

A COMPARATIVE CYTOSPECTRAL STUDY ON VERTEBRATE LEUCOCYTES STAINED BY THE WRIGHT METHOD

A new method was developed for cytospectral characterization of leucocytes after staining with Wright solution. The spectral types of various vertebrate leucocytes were compared by this method. To characterize vertebrate leucocytes their absorption maximum (A max) and relative absorption (RA) spectra (A/A max, %) were recorded. From the values of A max and the absorptions at 600nm (RA 600) and 460nm (RA 460), the RA spectra were divided into 7 types (I, II-a, II-b, III-A, III-a, III-b, IV). The leucocyte types of individual species could be classified into certain types, but these types differed in different species. However, even in different species, most thrombocytes were classified as type III-A, mammalian eosinophils as type III-b and those of lower vertebrates as type IV. In general the spectral types changed progressively from higher to lower animal species. The blood of lower vertebrates seemed to contain immature cells judging from the RA spectra of these cells. From our data this spectral classification seems promising for comparison of vertebrate leucocytes and should be useful in their cytoanalysis.

Studies on the energy metabolism of erythrocytes during artificial heart pumping.

It was said that hemolysis could take place in the artificial heart pumping, so that some changes in the red blood cells would be expected as a cause of hemolysis. These changes were caused by the trauma to the erythrocytes. In order to elucidate the energy metabolism of the red blood cells, 4 calves, which survived for 408, 150, 64, and 41 hours after the total replacement by the artificial hearts, were subjected to the analysis of the erythrocyte's adenine nucleotides (adenosine-tri-phosphate; ATP, adenosine-di-phosphate; ADP, and adenosine-mono-phosphate; AMP).Analysis of adenine nucleotides were performed by the enzymatic method. Peripheral blood smear was made and stained with Wright's solution. % crenocytes was obtained by this preparation.It was demonstrated that ATP was decreasing and ADP was increasing toward the end of pumping. ATP: ADP ratio went down. AMP showed fluctuation. Close relationship between ATP level and crenocytes was observed.It was considered that glycolytic activities in the red blood cells during the artificial heart pumping were much influenced by its mechanical and physical trauma, so that their ATP contents went down and the crenocytes increased in number.This led to the intravascular and extravascular hemolysis in the artificial hearts pumping.

Increased Blood Platelet Clumping in Thromboembolic Disease

Ordinary routine differential blood films were made on glass slides, stained with Wright's solution and observed under the oil immersion lens. The presence of blood platelet clumps and the number of platelets per clump were noted and recorded. Clumping was considered normal if the blood film did not reveal more than 10 platelets to the clump (Fig. 1), as increased or abnormal if there were more than 10 platelets to the clump (Fig. 2). Clumps containing more than 15 platelets not infrequently presented a syncytial arrangement with an almost transparent hyaline matrix in which platelets of varying sizes and shapes were embedded (Fig. 3). In 100 presumably normal control subjects (Table 1) without detectable clinical disease, clumping was normal in 92 per cent and designated as increased or abnormal in only 8 per cent. Of 100 patients with various types of thromboembolic disease, clumping was of normal type in 19 per cent and designated as increased in 81 per cent.

Internal Drainage of Acute Ears, and Abortion of Acute Mastoiditis by the Use of Sir A. E Wright's Solution

Internal Drainage of Acute Ears, and Abortion of Acute Mastoiditis by the Use of Sir A. E Wright's Solution

Internal drainage of acute ears, and abortion of acute mastoiditis by the use of sir almroth wright's solution.

Internal drainage of acute ears, and abortion of acute mastoiditis by the use of sir almroth wright's solution.