The potential of mRNA expression evaluation in predicting HER2 positivity in gastroesophageal cancer.

Gastroesophageal cancer (GEC) is an aggressive disease characterized by a high frequency of metastasis and poor overall survival rates. GEC presents HER2 overexpression in 5 to 25% of tumors eligible for HER2-targeted therapy. HER2 evaluation requires protein levels and copy number alteration analyses by immunohistochemistry (IHC) and in situ hybridization (FISH or SISH), respectively. These are semiquantitative methodologies that need an expert and well-trained pathologist. Therefore, the use of new surrogate methods for HER2 evaluation in cancer, such as gene expression analysis, might improve GEC HER2 classification. We evaluated HER2 positivity in GEC through conventional IHC and SISH analyses and investigated the potential application of HER2 mRNA expression by quantitative PCR to categorize GEC samples as HER2-positive or HER2-negative. Among 270 GEC samples, 10.9% were HER2-positive by IHC and SISH analyses. HER2 mRNA was overexpressed in HER2-positive GEC samples and presented high accuracy in distinguishing those tumors from HER2-negative GEC. Nevertheless, HER2 mRNA analysis was not capable of classifying HER2-equivocal GEC samples into HER2-positive or -negative according to SISH data. Quantitative PCR analysis showed HER2 overexpression in HER2-positive GEC samples. Nevertheless, HER2 mRNA analysis failed to classify HER2-equivocal GEC according to SISH data.

Renal amyloidosis: a new time for a complete diagnosis.

Amyloidoses are a group of disorders in which soluble proteins aggregate and deposit extracellularly in tissues as insoluble fibrils, causing organ dysfunction. Clinical management depends on the subtype of the protein deposited and the affected organs. Systemic amyloidosis may stem from anomalous proteins, such as immunoglobulin light chains or serum amyloid proteins in chronic inflammation or may arise from hereditary disorders. Hereditary amyloidosis consists of a group of rare conditions that do not respond to chemotherapy, hence the identification of the amyloid subtype is essential for diagnosis, prognosis, and treatment. The kidney is the organ most frequently involved in systemic amyloidosis. Renal amyloidosis is characterized by acellular pathologic Congo red-positive deposition of amyloid fibrils in glomeruli, vessels, and/or interstitium. This disease manifests with heavy proteinuria, nephrotic syndrome, and progression to end-stage kidney failure. In some situations, it is not possible to identify the amyloid subtype using immunodetection methods, so the diagnosis remains indeterminate. In cases where hereditary amyloidosis is suspected or cannot be excluded, genetic testing should be considered. Of note, laser microdissection/mass spectrometry is currently the gold standard for accurate diagnosis of amyloidosis, especially in inconclusive cases. This article reviews the clinical manifestations and the current diagnostic landscape of renal amyloidosis.

First report and biological characteristics of Heterorhabditis amazonensis in the state of Paraná, Brazil.

Entomopathogenic nematodes (EPNs) from Heterorhabditidae and Steinernematidae families are extensively used to control insect pests. In Brazil, however, relatively few studies have identified and characterized these entomopathogens. The objective of this study was to identify and characterize an EPN isolate obtained from soil samples collected in the state of Paraná, Brazil. An isolate (UEL 08) of Heterorhabditis was detected in a soil sample collected from a pasture area cultivated with Brachiaria grass in Londrina, state of Paraná, Brazil (23°34'311''S, 050°58'298''W), using the insect-baiting technique with Galleria mellonella larvae as hosts. The nematode was identified through morphometric studies and molecular analyses based on amplification of the rDNA ITS region. Although we identified certain morphometric differences compared with the original description, the molecular data indicated that the ITS sequence obtained for the UEL 08 isolate is identical to the reference sequence of H. amazonensis (DQ665222) and presented 100% similarity. Thus, the findings of our morphological and molecular studies confirmed that the isolated nematode is H. amazonensis, which is the first time this species has been registered in Paraná. Study of the biological characteristics of H. amazonensis (UEL 08) revealed that the isolate has two distinct life cycles - one short (216 h) and the other long (288 h) - and produces two generations in both cycles. We observed that H. amazonensis (UEL 8) was pathogenic and virulent to the three evaluated hosts, although with different virulence against these hosts. The larvae of G. mellonella and Alphitobius diaperinus were more susceptible than adult Dichelops (Diacereus) melacanthus, with 100%, 85%, and 46% mortality, respectively. Furthermore, an in vivo production assay revealed a mean daily yield of 3.4 × 103 infective juveniles/g host larvae.

Renal amyloidosis: a new time for a complete diagnosis

Amyloidoses are a group of disorders in which soluble proteins aggregate and deposit extracellularly in tissues as insoluble fibrils, causing organ dysfunction. Clinical management depends on the subtype of the protein deposited and the affected organs. Systemic amyloidosis may stem from anomalous proteins, such as immunoglobulin light chains or serum amyloid proteins in chronic inflammation or may arise from hereditary disorders. Hereditary amyloidosis consists of a group of rare conditions that do not respond to chemotherapy, hence the identification of the amyloid subtype is essential for diagnosis, prognosis, and treatment. The kidney is the organ most frequently involved in systemic amyloidosis. Renal amyloidosis is characterized by acellular pathologic Congo red-positive deposition of amyloid fibrils in glomeruli, vessels, and/or interstitium. This disease manifests with heavy proteinuria, nephrotic syndrome, and progression to end-stage kidney failure. In some situations, it is not possible to identify the amyloid subtype using immunodetection methods, so the diagnosis remains indeterminate. In cases where hereditary amyloidosis is suspected or cannot be excluded, genetic testing should be considered. Of note, laser microdissection/mass spectrometry is currently the gold standard for accurate diagnosis of amyloidosis, especially in inconclusive cases. This article reviews the clinical manifestations and the current diagnostic landscape of renal amyloidosis.

The potential of mRNA expression evaluation in predicting HER2 positivity in gastroesophageal cancer

Gastroesophageal cancer (GEC) is an aggressive disease characterized by a high frequency of metastasis and poor overall survival rates. GEC presents HER2 overexpression in 5 to 25% of tumors eligible for HER2-targeted therapy. HER2 evaluation requires protein levels and copy number alteration analyses by immunohistochemistry (IHC) and in situ hybridization (FISH or SISH), respectively. These are semiquantitative methodologies that need an expert and well-trained pathologist. Therefore, the use of new surrogate methods for HER2 evaluation in cancer, such as gene expression analysis, might improve GEC HER2 classification. We evaluated HER2 positivity in GEC through conventional IHC and SISH analyses and investigated the potential application of HER2 mRNA expression by quantitative PCR to categorize GEC samples as HER2-positive or HER2-negative. Among 270 GEC samples, 10.9% were HER2-positive by IHC and SISH analyses. HER2 mRNA was overexpressed in HER2-positive GEC samples and presented high accuracy in distinguishing those tumors from HER2-negative GEC. Nevertheless, HER2 mRNA analysis was not capable of classifying HER2-equivocal GEC samples into HER2-positive or -negative according to SISH data. Quantitative PCR analysis showed HER2 overexpression in HER2-positive GEC samples. Nevertheless, HER2 mRNA analysis failed to classify HER2-equivocal GEC according to SISH data.

Topologically driven linear magnetoresistance in helimagnetic FeP.

The helimagnet FeP is part of a family of binary pnictide materials with the MnP-type structure, which share a nonsymmorphic crystal symmetry that preserves generic band structure characteristics through changes in elemental composition. It shows many similarities, including in its magnetic order, to isostructural CrAs and MnP, two compounds that are driven to superconductivity under applied pressure. Here we present a series of high magnetic field experiments on high-quality single crystals of FeP, showing that the resistance not only increases without saturation by up to several hundred times its zero-field value by 35 T, but that it also exhibits an anomalously linear field dependence over the entire range when the field is aligned precisely along the crystallographic c-axis. A close comparison of quantum oscillation frequencies to electronic structure calculations links this orientation to a semi-Dirac point in the band structure, which disperses linearly in a single direction in the plane perpendicular to field, a symmetry-protected feature of this entire material family. We show that the two striking features of magnetoresistance-large amplitude and linear field dependence-arise separately in this system, with the latter likely due to a combination of ordered magnetism and topological band structure.

Comparison between ultrasonography and X-ray as evaluation methods of central venous catheter positioning and their complications in pediatrics.

PURPOSE: This study evaluates the capacity of ultrasonography as a diagnostic method to confirm the proper positioning of central venous catheter (CVC) when compared to the current gold standard, chest radiography (CR). METHODS: A prospective study was performed including children from 0 to 14 incomplete years, who underwent CVC placement between March and May 2018 at a teaching hospital in Brazil. A four-chamber view of the heart was performed with ultrasound during a rapid injection of saline solution to identify hyperechoic images and confirm the central position of the catheter. After that, a CR was performed. The diagnostic quality of ultrasound was evaluated based on accuracy, sensitivity, specificity, positive and negative predictive values. RESULTS: A total of 21 patients were analyzed. The mean age was 3.95 ± 4.01 years. The preferred puncture site was the right internal jugular vein (71.4%). Ultrasound accuracy to detect CVC positioning was 81%. Sensitivity, specificity and positive and negative predictive values were 33%, 100%, 100% and 79%, respectively. CONCLUSION: Ultrasound is a reliable method for detection of CVC positioning. Even so, with the four-chamber cardiac view, this method is unable to identify catheters inside heart chambers, therefore, needing to confirm the positioning with CR.

Use of Low Temperature Storage to Preserve Host and Parasitoid to Improve the Rearing of Telenomus podisi (Hymenoptera: Platygastridae) on Euschistus heros (Hemiptera: Pentatomidae) Eggs.

Successful parasitoid rearing is crucial for augmentative biological control. A low temperature preservation protocol allowing the availability of host and parasitoid year-round was evaluated in this study in four bioassays: (1) host eggs [Euschistus heros (Fabricius)] stored at - 196, - 80, and - 20°C for up to 70 days prior to exposure to Telenomus podisi Ashmead parasitism; (2) Euschistus heros eggs removed from storage at - 196°C after 70 days and kept at 5°C for up to 9 days prior to exposure to T. podisi parasitism; (3) Telenomus podisi adult emergence of insects stored as pupae at 5°C; and (4) fitness of adults of T. podisi stored at 5°C. Higher parasitism was observed in parasitoids reared on E. heros eggs stored at - 196 and - 80°C. Host eggs removed from - 196°C and stored at 5°C for up to 6 days did not impact T. podisi parasitism and development. Storage of T. podisi pupae for more than 7 days negatively affected parasitoid biology. Storing T. podisi adults at 5°C for up to 6 days does not alter the biological parameters of the parasitoid. Thus, parasitoids can be stored as pupae or adults as well as its host E. heros eggs. Our findings can be applied to improve the feasibility of year-round insect production.

Use of prostaglandin F2α as ovulatory stimulus for synchronizing dairy cattle.

The aim of this study was to evaluate if prostaglandin F2α (PGF) can be used to induce ovulation in a GnRH-progesterone based protocol. In Experiment 1 crossbred dairy cows (n=32) were synchronized with a progesterone-GnRH based protocol for seven days, where the luteolytic dose of 150µg PGF was given 24h prior progesterone device removal (CIDR). On Day 8 cows were separated into two groups to receive: 1) 2mL of Saline (Control Group, n=15) or 2) 150µg of PGF (PGF Group, n=17). Ovulation rate was higher in the PGF than Control group (100% vs 53.3%, P=0.001, Odds ratio=30.88). The percentage of cows that ovulated synchronously tended to be higher in the PGF than Control group (P=0.1, Odds ratio=9.6). Experiment 2 was performed in a cross-over (3×3) design. Crossbred dairy cows (n=25) received a CIDR for seven days and GnRH on Day 0. Seven days later 150µg of PGF was given and the progesterone device was removed, and 24h later cows were distributed into three groups to receive: 1) 2mL of Saline (Control Group, n=25), 2) 150µg of PGF (PGF Group, n=25) or 3) 1mg of ECP (ECP Group, n=23). Diameter of ovulatory follicle was larger in the PGF and Control than ECP Group (P=0.002, Effect size>4.0). Synchronized ovulation rate (between 72 and 96h after CIDR removal) tended to be higher in PGF group in Control group (P=0.1, Odds ratio=0.35). Results suggest that PGF is equally efficient to ECP to induce synchronized ovulation in dairy cows subjected to progesterone-GnRH based protocols.

Action of natural phytosanitary products on Bacillus thuringiensis subsp. kurstaki S-1905.

The objective of this study was to evaluate the effects of natural phytosanitary products (NPs) on spores and crystals of Bacillus thuringiensis subsp. kurstaki S-1905 (Btk S-1905). For the spore assay, NPs and bacteria were applied in combination and individually. For the combined application, Btk S-1905 + NP mixtures were inoculated on nutrient agar (NA), and for the separate applications, the NPs were spread on NA plates, which were later inoculated with the pathogen. The number of colony-forming units (CFU) per milliliter was quantified after 18 h of incubation. For the crystal protein degradation assay, the Btk S-1905 + NP mixtures were added to the diet of Anticarsia gemmatalis (Lepidoptera: Erebidae), and mortality was evaluated at the following time points: 12, 24, 48, and 72 h. Scanning electron microscopy and agarose gel electrophoresis were carried out. Biogermex and Ecolife® reduced the CFU ml-1 in both combined and separate applications. Biogermex, Ecolife®, and Planta Clean were antagonistic to the action of bacterial toxins, and no product affected the morphology or resulted in the degradation of the crystal proteins. The remaining products evaluated did not reduce the CFU ml-1 and had additive effect when combined with the crystal toxin.

Potential Side Effects of the Entomopathogenic Fungus Metarhizium anisopliae on the Egg Parasitoid Trichogramma pretiosum (Hymenoptera: Trichogrammatidae) Under Controlled Conditions.

The purpose was to evaluate the side effects of strains Metarhizium anisopliae (Metsch.) Sorokin sensu lato Unioeste 43 and M. anisopliae sensu stricto ESALQ 1641 on Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae) under controlled conditions. A free-choice test for parasitism was performed, with the confinement of T. pretiosum females mated with cards (1 × 5 cm) containing age-standardized Anagasta kuehniella Zeller eggs, either sprayed with a fungal strain (109 conidia/ml) or 0.01% v/v Tween 80 (control). For the no-choice tests, T. pretiosum females mated were confined with cards sprayed with fungal strains before or after parasitism, and cards with fungal applications at different times. The number of parasitized eggs, percentage of emergence, longevity, egg-to-adult period, sex ratio, total and confirmed mortality by the fungus, and longevity of females that parasitized previously sprayed eggs, were assessed. Histological analysis of immature phases was also performed. The fungus was repellent to T. pretiosum in the free-choice test, while in the no-choice test, fungal applications before and after parasitism did not affect the number of eggs parasitized by T. pretiosum or the sex ratio of emerging adults. However, both strains affected adult emergence rates, the egg-to-adult period, and longevity. Overall, both M. anisopliae strains had minor effects on these biological parameters of T. pretiosum under controlled conditions. Hyphae were not detected in histological observations of immature stages of the parasitoid.

Comparison between two estradiol-progesterone based protocols for timed artificial insemination in blocks in lactating Nelore cows.

The aim of this study was to compare the use of artificial insemination in time blocks (Artificial Insemination Blocks, AIB) using an 8 and 9 d estradiol-progesterone based protocol. In this experiment, lactating Nelore cows (n=253) were subjected to two estradiol-progesterone based TAI protocols. On the morning of Day 10 (8d group, n=124) or Day 11 (9d group, n=129), cows were examined by ultrasonography to evaluate the diameter of the preovulatory follicle and were inseminated once at one of the following time points, according to the diameter of the pre-ovulatory follicle (POF): Block 0 (POF≥15mm, TAI 0h after conventional TAI), Block 1 (POF 13.0-14.9mm, TAI 6h later), Block 2 (POF 10.1-12.9mm, TAI 24h later), and Block 3 (POF≤10.0mm, TAI 30h later). The pregnancy per AI (P/AI) did not differ between 8d and 9d groups (P>0.05). Considering only multiparous cows, however, P/AI tended to be greater in the 8d (64.1%) than in the 9d group (49.3%; P=0.08). Cows from the 9d group tended to have a larger POF than cows from the 8d group (P=0.07). In conclusion, these results provide evidence that there is no difference between 8d or 9d protocols when using the AIB technique. Use of the 8d estradiol-progesterone based protocol, however, tended to increase pregnancy in multiparous cows.

Selection and Characterization of Bacillus thuringiensis (Berliner) (Eubacteriales: Bacillaceae) Strains for Ecdytolopha aurantiana (Lima) (Lepidoptera: Tortricidae) Control.

The citrus fruit borer, Ecdytolopha aurantiana (Lima, 1927) (Lepidoptera: Tortricidae), is responsible for major losses to the citrus industry because it causes rot and drop of fruits. The current study aimed to select and characterize Bacillus thuringiensis (Berliner, 1911) strains toxic to E. aurantiana. For this purpose, 47 B. thuringiensis strains were evaluated in selective bioassays using first instar larvae of E. aurantiana. The lethal concentration (LC50) of the most toxic strains was estimated, and the strains were characterized by morphological, biochemical, and molecular methods. Of the 47 strains tested, 10 caused mortality above 85% and showed mean lethal concentrations between 1.05E+7 and 1.54E+8 spores mL-1. The lowest LC50 values were obtained for the HD-1 standard strain and the BR145, BR83, BR52, and BR09 strains. The protein profile showed the presence of Cry proteins of 60, 65, 70, 80, and 130 kDa. The molecular characterization showed the presence of cry1, cry2, cry3, and cry11 genes. The morphological analysis identified three different crystalline inclusions: bipyramidal, round, and cuboidal. The cry1 and cry2 genes were the most frequent among the B. thuringiensis strains evaluated and encode Cry proteins toxic to insects of the order Lepidoptera, which agree with the toxicity results obtained by the selective bioassays against E. aurantiana. The results showed four different B. thuringiensis strains toxic to E. aurantiana at the same level as the HD-1 standard strain, and these strains have biotechnological potential for E. aurantiana control through the production of transgenic plants or the formulation of biopesticides.

Production of cellulosic ethanol from sugarcane bagasse by steam explosion: Effect of extractives content, acid catalysis and different fermentation technologies.

The production of cellulosic ethanol was carried out using samples of native (NCB) and ethanol-extracted (EECB) sugarcane bagasse. Autohydrolysis (AH) exhibited the best glucose recovery from both samples, compared to the use of both H3PO4 and H2SO4 catalysis at the same pretreatment time and temperature. All water-insoluble steam-exploded materials (SEB-WI) resulted in high glucose yields by enzymatic hydrolysis. SHF (separate hydrolysis and fermentation) gave ethanol yields higher than those obtained by SSF (simultaneous hydrolysis and fermentation) and pSSF (pre-hydrolysis followed by SSF). For instance, AH gave 25, 18 and 16 g L(-1) of ethanol by SHF, SSF and pSSF, respectively. However, when the total processing time was taken into account, pSSF provided the best overall ethanol volumetric productivity of 0.58 g L(-1) h(-1). Also, the removal of ethanol-extractable materials from cane bagasse had no influence on the cellulosic ethanol production of SEB-WI, regardless of the fermentation strategy used for conversion.

Timed artificial insemination in blocks: A new alternative to improve fertility in lactating beef cows.

The aim of this study was to evaluate whether changing the interval from CIDR removal to timed artificial insemination (TAI) according to the diameter of the preovulatory follicle (POF) would improve pregnancy per AI in cows. In Study 1, a retrospective analysis of TAI experiments (n=96 cows) was performed to characterize the time of ovulation according to the diameter of the dominant follicle. It was observed that cows with a larger POF had ovulations earlier than cows with smaller POF, according to the equation: y=0.72x(2)-26.74x+264.54 (R(2)=0.63; P<0.001). In Study 2, lactating Nelore cows (n=412) were subjected to an EB-CIDR based TAI protocol. On the morning of Day 10 (time of TAI), cows were randomized into Control (n=209) and Block (n=203) groups; (1) Cows in the Control Group were TAI 48 h after CIDR removal (08:00 am on Day 10), and; (2) Cows in the block group were inseminated once at one of the following time points, according to the diameter of the POF on Day 10: B0 (POF≥15mm, TAI 0 h after convetional TAI), B1 (POF 13-14.9 mm, TAI 6h later), B2 (POF 10.1-12.9 mm, TAI 24h later) and B3 (POF≤10mm, TAI 30 h later). The cows of the Block Group had greater pregnancy rates per AI than the Control Group (129/203, 63.5% when compared with 102/209, 48.8%, respectively; P<0.01). In conclusion, results of the present study demonstrate that adjusting the timing of TAI according to the diameter of the POF can be an effective practice for improving fertility of cows in TAI protocols.

Selectivity of Organic Products to Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae).

The selectivity of various entomopathogens and one insecticide (chlorpyrifos = positive control) to Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae) was evaluated in the laboratory, using the protocol established by the Working Group on "Pesticides and Beneficial Organisms" of the IOBC. The evaluated parameters were parasitism (%), adult emergence (%), and product repellency to the parasitoid when sprayed on host eggs prior to parasitism (free-choice and no-choice tests). Most of the studied entomopathogens (Bacillus thuringiensis var. kurstaki, Bacillus thuringiensis var. aizawai, Beauveria bassiana, Metarhizium anisopliae, and Trichoderma harzianum) had no effects on biological parameters and were classified as harmless to T. pretiosum. Emergence of parasitoids (progeny viability) was reduced, but remained above 90%, when host eggs were sprayed with Baculovirus anticarsia prior to parasitism in the free-choice test, and B. anticarsia was therefore considered harmless. Chlorpyrifos (positive control) caused high adult parasitoid mortality in all bioassays. While T. pretiosum and the tested entomopathogens may be used simultaneously in integrated pest management programs, the use of chlorpyrifos should be avoided.

Propiedades biomecánicas de la cornea con corvis ST y ocular response analyser después de diferentes procedimientos quirúrgicos en pacientes con queratocono / Corneal biomechanical properties with corvis ST and ocular response analyser in eyes with keratoconus which were submitted to different surgical procedures

Introducción. Las opciones quirúrgicas disponibles para el tratamiento del queratocono (QC) se han ampliado y mejorado. Nuestro objetivo es comparar las propiedades biomecánicas de la córnea con dos instrumentos de uso clínico en ojos con QC sin cirugía o intervenidos con diferentes tipos de cirugía corneal (cross-linking (CXL), segmentos intra-corneales (ICSR) y queratoplastia penetrante(QP)). Material e Métodos. Se analizaron 91 ojos de paciente con KC no operados, 22 ojos con ICSR, 16 ojos con QP y 6 ojos con CXL. Se analizó el espesor corneal central (ECC), presión intra-ocular y los parámetros obtenidos con ORA (clásicos, y análisis de señal) y Corvis. Resultados. Tanto los valores de presión intra-ocular (PIO) obtenidos directamente con ORA y Corvis como los valores de PIO corregidos por el ORA (PIOcc) fueron significativamente diferentes entre los distintos grupos. Hasta 9 de los parámetros resultantes del análisis de señal de medida del ORA (p1area, p2area, h2, dive2, path1, aplhf, p1area1, p2area1 y w11) mostraron diferencias estadísticamente significativas entre los grupos. En cuanto al Corvis, A1 Time, A2 Time y Peak Distance fueron significativamente diferentes entre los grupos considerados. Conclusiones. Las nuevas tecnologías de evaluación y análisis de datos de la respuesta biomecánica de la cornea permiten cuantificar y diferenciar el comportamiento biomecánico tras diferentes tipos de cirugía para la estabilización, regularización o substitución corneal en pacientes con queratocono (AU)

Background. Surgical options for the treatment of keratoconus (KC) have been expanded and improved. Our goal is to evaluate the changes in biomechanical properties of the cornea measured with two instruments for clinical use in KC eyes not operated or which undergoing different surgical procedures (cross-linking (CXL), intra-corneal ring segment (ICSR) and penetrating keratoplasty (QP)). Material and methods. The population consisted of 91 eyes with KC not operated, 22 eyes that underwent ICRS implant, 16 eyes that were submitted to PK and 6 eyes that underwent CXL. We analyzed corneal thickness, intraocular pressure and the parameters obtained with ORA (classic and signal analysis) and Corvis. Results. Both the values of intraocular pressure (IOP) obtained directly from ORA and Corvis, as IOP values corrected by the ORA (IOPcc) were significantly different between groups. For 9 of the parameters resulting from ORA analysis of signal (p1area, p2area, h2, Dive2, path1, aplhf, p1area1, p2area1 and w11) showed statistically significant differences between groups. Regarding Corvis, A1 Time, A2 Time and Peak Distance were significantly different between the groups. Conclusions. The new technology assessment and analysis of the biomechanical response of the cornea to quantify and differentiate the biomechanical behavior after different types of surgery for stabilization, regularization or corneal replacement in patients with keratoconus (AU)

Increasing storage capability of pacu (Piaractus mesopotamicus) embryos by chilling: development of a useful methodology for hatcheries management.

Cryopreservation of fish gametes has been studied extensively in the last few decades, but the successful cryopreservation of fish embryos remains elusive. However, recent studies using short-term chilling techniques have shown that it is possible to store embryos at low temperatures with no significant loss in viability. Information on cryopreservation of Neotropical freshwater fish embryos has so far been very limited in the literature. In the present study, chilling protocols for storage of pacu embryos at -8°C for up to 24 h were studied using different concentrations of sucrose in methanol. Embryos tolerated the subzero temperature for up to 6 h with no adverse effects (P > 0.05). After 12 h chilling, hatching rate of 64.0 +/- 3.5 percent was recorded. Low temperature storage of pacu embryos by chilling is detailed here for the first time. Further studies are needed to extend the storage time and to improve the hatching rate.

Duplication of the inferior vena cava: case report and a literaturereview of anatomical variation

Anatomical variation in the inferior vena cava can result in misdiagnosis, making a better understanding of suchvariations crucial. Here we report the case of a 29 year-old male, victim of multiple trauma, who in the courseof treatment presented with a pulmonary thromboembolism confirmed by tomography. Given the gravityof the situation and the need for additional surgeries, a decision was made to implant an inferior vena cavafilter. During phlebography, prior to implantation of the filter, the duplication of the vena cava was detectedand classified as a complete duplication. A review of the literature revealed various anatomical descriptions ofduplicated inferior vena cava, the most common of which were incomplete cases showing greater variationin venous contion. All in vivo anatomical descriptions were done via phlebography, demonstrating the valueof this test for the diagnosis of anatomical variation in the abdominal veins. While duplication of the inferiorvena cava was not the cause of the venous thrombosis in our patient, a detailed phlebography test was neededto both identify the anatomical variation and facilitate the placement of the filter to prevent a new pulmonarythromboembolism.

Genetic Variability of Beauveria bassiana and a DNA Marker for Environmental Monitoring of a Highly Virulent Isolate Against Cosmopolites sordidus.

The banana weevil Cosmopolites sordidus (Germar) is one of a number of pests that attack banana crops. The use of the entomopathogenic fungus Beauveria bassiana as a biological control agent for this pest may contribute towards reducing the application of chemical insecticides on banana crops. In this study, the genetic variability of a collection of Brazilian isolates of B. bassiana was evaluated. Samples were obtained from various geographic regions of Brazil, and from different hosts of the Curculionidae family. Based on the DNA fingerprints generated by RAPD and AFLP, we found that 92 and 88 % of the loci were polymorphic, respectively. The B. bassiana isolates were attributed to two genotypic clusters based on the RAPD data, and to three genotypic clusters, when analyzed with AFLP. The nucleotide sequences of nuclear ribosomal DNA intergenic spacers confirmed that all isolates are in fact B. bassiana. Analysis of molecular variance showed that variability among the isolates was not correlated with geographic origin or hosts. A RAPD-specific marker for isolate CG 1024, which is highly virulent to C. sordidus, was cloned and sequenced. Based on the sequences obtained, specific PCR primers BbasCG1024F (5'-TGC GGC TGA GGA GGA CT-3') and BbasCG1024R (5'-TGC GGC TGA GTG TAG AAC-3') were designed for detecting and monitoring this isolate in the field.