Adaptability and stability of genotypes of sweet sorghum by GGEBiplot and Toler methods.

Sweet sorghum has considerable potential for ethanol and energy production. The crop is adaptable and can be grown under a wide range of cultivation conditions in marginal areas; however, studies of phenotypic stability are lacking under tropical conditions. Various methods can be used to assess the stability of the crop. Some of these methods generate the same basic information, whereas others provide additional information on genotype x environment (G x E) interactions and/or a description of the genotypes and environments. In this study, we evaluated the complementarity of two methods, GGEBiplot and Toler, with the aim of achieving more detailed information on G x E interactions and their implications for selection of sweet sorghum genotypes. We used data from 25 sorghum genotypes grown in different environments and evaluated the following traits: flowering (FLOW), green mass yield (GMY), total soluble solids (TSS), and tons of Brix per hectare (TBH). Significant G x E interactions were found for all traits. The most stable genotypes identified with the GGEBiplot method were CMSXS643 for FLOW, CMSXS644 and CMSXS647 for GMY, CMSXS646 and CMSXS637 for TSS, and BRS511 and CMSXSS647 for TBH. Especially for TBH, the genotype BRS511 was classified as doubly desirable by the Toler method; however, unlike the result of the GGEBiplot method, the genotype CMSXS647 was also found to be doubly undesirable. The two analytical methods were complementary and enabled a more reliable identification of adapted and stable genotypes.

Relationship of inflammatory markers and pain in patients with head and neck cancer prior to anticancer therapy.

Pain is a common symptom in patients with cancer, including those with head and neck cancer (HNC). While studies suggest an association between chronic inflammation and pain, levels of inflammatory cytokines, such as C-reactive protein (CRP) and tumor necrosis factor-alpha (TNF-α), have not been correlated with pain in HNC patients who are not currently undergoing anticancer treatment. The purpose of this study was to examine the relationship between these inflammatory markers and perceived pain in HNC patients prior to anticancer therapy. The study group consisted of 127 HNC patients and 9 healthy controls. Pain was assessed using the Brief Pain Inventory (BPI), and serum levels of CRP and TNF-α were determined using the particle-enhanced turbidimetric immunoassay (PETIA) and ELISA techniques, respectively. Patients experiencing pain had significantly higher levels of CRP (P<0.01) and TNF-α (P<0.05) compared with controls and with patients reporting no pain. There were significantly positive associations between pain, CRP level, and tumor stage. This is the first study to report a positive association between perceived pain and CRP in HNC patients at the time of diagnosis. The current findings suggest important associations between pain and inflammatory processes in HNC patients, with potential implications for future treatment strategies.

Relationship of inflammatory markers and pain in patients with head and neck cancer prior to anticancer therapy

Pain is a common symptom in patients with cancer, including those with head and neck cancer (HNC). While studies suggest an association between chronic inflammation and pain, levels of inflammatory cytokines, such as C-reactive protein (CRP) and tumor necrosis factor-alpha (TNF-α), have not been correlated with pain in HNC patients who are not currently undergoing anticancer treatment. The purpose of this study was to examine the relationship between these inflammatory markers and perceived pain in HNC patients prior to anticancer therapy. The study group consisted of 127 HNC patients and 9 healthy controls. Pain was assessed using the Brief Pain Inventory (BPI), and serum levels of CRP and TNF-α were determined using the particle-enhanced turbidimetric immunoassay (PETIA) and ELISA techniques, respectively. Patients experiencing pain had significantly higher levels of CRP (P<0.01) and TNF-α (P<0.05) compared with controls and with patients reporting no pain. There were significantly positive associations between pain, CRP level, and tumor stage. This is the first study to report a positive association between perceived pain and CRP in HNC patients at the time of diagnosis. The current findings suggest important associations between pain and inflammatory processes in HNC patients, with potential implications for future treatment strategies.

Pregnancy rates and corpus luteum-related factors affecting pregnancy establishment in bovine recipients synchronized for fixed-time embryo transfer.

The objective was to investigate the influence of corpora lutea physical and functional characteristics on pregnancy rates in bovine recipients synchronized for fixed-time embryo transfer (FTET). Crossbred (Bos taurus taurus x Bos taurus indicus) nonlactating cows and heifers (n=259) were treated with the following protocol: 2mg estradiol benzoate (EB) plus an intravaginal progesterone device (CIDR 1.9g progesterone; Day 0); 400 IU equine chorionic gonadotropin (eCG; Day 5); prostaglandin F(2alpha) (PGF(2alpha)) and CIDR withdrawal (Day 8); and 1mg EB (Day 9). Ovarian ultrasonography and blood sample collections were performed on Day 17. Of the 259 cattle initially treated, 197 (76.1%) were suitable recipients; they received a single, fresh, quality grade 1 or 2 in vivo-derived (n=90) or in vitro-produced (n=87) embryo on Day 17. Pregnancy rates (23 d after embryo transfer) were higher for in vivo-derived embryos than for in vitro-produced embryos (58.8% vs. 31.0%, respectively; P<0.001). Mean (+/-SD) plasma progesterone (P(4)) concentration was higher in cattle that became pregnant than that in nonpregnant cattle (5.2+/-5.0 vs. 3.8+/-2.4 ng/mL; P=0.02). Mean pixel values (71.8+/-1.3 vs. 71.2+/-1.1) and pixel heterogeneity (14.8+/-0.3 vs. 14.5+/-0.5) were similar between pregnant and nonpregnant recipients (P>0.10). No significant relationship was detected between pregnancy outcome and plasma P(4), corpus luteum area, or corpus luteum echotexture. Embryo type, however, affected the odds of pregnancy. In conclusion, corpus luteum-related traits were poor predictors of pregnancy in recipients. The type of embryo, however, was a major factor affecting pregnancy outcome.

Avaliação de diferentes técnicas para coloração de sêmen de peixes / Different techniques to evaluate to stain fish semen / Evaluación de técnicas para colorear sémen de peces

A análise da morfologia dos espermatozóides do sêmen colhido de 30 pacus (Piaractus mesopotamicus) consistiu de três soluções fixadoras: formol salina tamponada (FS), citrato sódio diidratado a 3% (CS) e água destilado (Ag) e para corar as células espermáticas três corantes: Vermelho Congo (VC), Rosa Bengala (RB) e Williams-modificado (WM). As combinações de corantes e soluções fixadoras utilizadas foram: RB X FS, WM X FS, VC X FS, VC X Ag, WM X Ag, WM X CS e VC X CS. Na avaliação dos esfregaços levaram-se em consideração as alterações anatômicas, a eficiência do corante, a aglutinação e a sujidade. Quando o sêmen foi diluído em CS e Ag, os espermatozóides sofreram alterações anatômicas e aglutinação, inviabilizando a análise. O FS não gerou alteração anatômica e nem aglutinação dos espermatozóides, sendo possível avaliar a eficiência do corante. Ao analisar a combinação RB X FS verificou-se a melhor média estimada de 9,97 pontos (P<0,05), com baixa sujidade e boa eficiência do corante. A combinação WM X FS resultou na média estimada de 7,35 pontos e a do VC X FS de 6,83 pontos que foram valores menores (P<0,05) em comparação aos esfregaços da combinação RB X FS. De acordo com os resultados obtidos neste estudo, a combinação do RB X FS apresentou os melhores resultados, sendo recomendado preparar esfregaços para analisar a morfologia dos espermatozóides de pacus (Piaractus mesopotamicus).

The analyses of spermatozoa morphology from semen obtained from pacus (Piaractus mesopotamicus) consisted of three fi xers solution: formal-saline buffered (FB), 3 % sodium citrate diihydrated (SC) and distilled water (DW) and to stain the spermatic cells were used three dyeing: Congo red (CR), Bengal rose (BR) and Williams modifi ed (WM). Combinations of dyeing and fi xers solution were: BR x BF, WM x FB, CR x FB, CR x DW, WM x DW, WM x SC and CR x SC. In the rubbing brush stained evaluation was considered spermatozoa anatomic alterations, dyeing effi ciency, agglutination and dirtiness. For semen diluted in SC and DW was observed spermatozoa anatomic alterations and agglutination on the rubbing brush, becoming impossible any analyses. When semen was diluted in FB to prepare rubbing brush was not observed spermatozoa anatomic alterations and agglutination, becoming possible to evaluate dyeing effi ciency. In analyze of BR x FB combination was verifi ed the best estimate average of 9.97 points (P<0.05), showing reduced dirtiness and good effi ciency of dyeing. When combination was WM x FB the estimate average was 7.35 points and CR x FB was 6.83 points which were lower (P<0.05) than the average verifi ed in the rubbing brush stained with BR x FB. According to the results observed in this study, the best combination to evaluate spermatozoa morphology from pacus semen was rubbing brush stained with BR x FB combination.

El análisis de la morfologia de los espermatozoides del semen colectado de 30 pacus (Piaractus mesopotamicus) consistio en tres soluciones fi jadoras: Formol salina tamponada (FS), citrato sodio bihidratado al 3% (CS) y agua destilada (Ag) y para colorear las células espermáticas tres colorantes: Rojo Congo (VC), Rosa Bengala (RB) y Williams ­ modifi cado (WM). Las combinaciones de colorantes y soluciones fi jadoras utilizadas fueron: RB x FS, WB x FS, VC x FS, VC x Ag, WM x Ag, WM x CS y VC x CS. En la evaluación de los frotis se tuvo en cuenta las alteraciones anatómicas, la efi ciencia del colorante, la aglutinacion y la suciedad. Cuando el semen fue diluído en CS y Ag, los espermatozoides sufrieron alteraciones anatómicas y aglutinación, inviabilizando el análisis. El FS no generó alteracion anatómica ni aglutinación de losespermatozoides, siendo posible evaluar la efi ciencia del colorante. Al analizar la combinación RB x FS se verifi có la mejor média estimada de 9,97 puntos (P〈0,05) con baja suciedad y buena efi ciencia del colorante. La combinación WM x FS resultó en el promedio estimado de 7,35 puntos y la de VC x FS de 6,83 puntos que fueron valores menores (P〈0,05) en comparación a los frotis de la combinación RB x FS. De acuerdo con los resultados obtenidos en este estudio, la combinación de RB x FS presentó los mejores resultados, siendo recomendado para preparar frotis para analizar la morfologia de los espermatozoides de pacus (Piaractus mesopotamicus).

In vitro cleavage of hepatitis C virus polyprotein substrates by purified recombinant NS3 protease.

The non-structural protein NS3 of hepatitis C virus has been expressed in bacteria as a polyhistidine fusion protein which can be produced in a soluble form and easily purified by affinity chromatography. Using an in vitro transcription and translation system we have been able to demonstrate that this protein can proteolytically process substrate molecules derived from the non-structural region of the polyprotein. Using this assay system we have been able to optimize basic biochemical characteristics of the purified enzyme. Parallel experiments show that the full-length NS3 protein also possesses ATPase activity, indicating the bifunctional nature of the protein. In contrast, purified NS3 in which the predicted catalytic serine has been mutated loses protease but retains ATPase activity.

Analysis of NS3-mediated processing of the hepatitis C virus non-structural region in vitro.

The protease activity of the hepatitis C virus (HCV) NS3 protein has been investigated using transient expression methods in mammalian cells, as well as in vitro transcription/translation systems. We confirmed that expression of the NS3-5 polyprotein in rabbit reticulocyte lysates results in efficient cis processing at the NS3/NS4 junction. However, processing at the other predicted sites of NS3-mediated cleavage varied markedly in efficiency, the site most susceptible being that between NS5A and NS5B. Time-course analysis of the proteolytic processing of the HCV non-structural precursor showed that the cis cleavage between NS3 and NS4 occurred extremely rapidly. However, efficient cleavage at this position was dependent on the prior removal of the NS2 protein. Furthermore, the presence of uncleaved NS2 sequences on the enzyme severely impeded NS3-mediated proteolysis at downstream sites in the polyprotein. This suggests therefore that efficient cleavage at the NS2/NS3 junction is a pivotal event in HCV replication. During the course of this study a proteolytically inactive mutant of NS3 was characterized carrying a previously unreported amino acid substitution near the proposed active site of the enzyme. Molecular modelling suggested that the amino acid present at this position may influence the conformation of the active site of the enzyme. Recently a number of reports have described a second protease activity, located in the NS2/NS3 region, which is responsible for cleavage at the NS2/NS3 junction. We have identified an isolate of HCV, obtained from a U.K. patient, which has a virtually inactive NS2/NS3 protease. The possible implications of this observation are discussed.

A cassette vector for the construction of antigen chimaeras of poliovirus.

A cassette vector has been constructed which allows the rapid and extensive modification of one of the neutralizing antigenic sites of the Sabin 1 poliovirus vaccine strain, P1/LSc 2ab. Unique restriction endonuclease sites flanking antigenic site 1 have been engineered into a full-length infectious Sabin 1 cDNA clone with minimal alteration to the coding sequence. This facilitates replacement of this region by oligonucleotides encoding foreign amino acid sequences. Our results indicate that this region is highly flexible in terms of the number and sequence of amino acids which can be accommodated.