Identification of prognostic factors for survival in patients with metastatic gastric adenocarcinoma in a Mexican population.

INTRODUCTION AND AIMS: Gastric adenocarcinoma is among the high-ranking tumors, with respect to frequency and mortality, worldwide. The inflammatory process and immune system activity are associated with oncologic control. Our aim was to identify whether the neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), and other variables are prognostic factors for survival in patients with metastatic gastric cancer in a Mexican population. MATERIAL AND METHODS: Patients diagnosed with metastatic gastric adenocarcinoma, hospitalized within the time frame of December 2011 to 2021, were analyzed. The NLR, PLR, and albumin and hemoglobin levels obtained from blood samples were calculated. Functional status (ECOG and Karnofsky), sex, histology, and the presence of signet ring cells were also considered possible prognostic factors. Each factor's prognostic value for overall survival was determined through univariate and multivariate analyses. RESULTS: The study included 956 patients diagnosed with metastatic gastric cancer, of whom 494 (51.7%) were men and 462 (48.3%) were women. The main histologic finding was diffuse adenocarcinoma (n = 619, 64.7%), followed by intestinal adenocarcinoma (n = 293, 30.6%), and the presence of signet ring cells was found in 659 (68.9%) patients. Diagnostic laparoscopy was performed on 238 patients (24.9%) to confirm peritoneal carcinomatosis. The multivariate analysis showed that an NLR above 3.2 (HR 1.51, 95% CI 1.27-1.8; p < 0.001), albumin below 3.5 g/dl (HR 1.25, CI 1.06-1.47; p = 0.006), and an ECOG performance status of 2 or higher (HR 1.39, CI 1.10-1.76; p = 0.005) were independent factors that predicted a lower survival rate, whereas a Karnofsky score above 70% (HR 0.69, CI 0.53-0.91; p = 0.008) was associated with a better survival rate. Lastly, the PLR was not statistically significant in the multivariate analysis. CONCLUSIONS: The NLR, nutritional status assessed through albumin measurement, and functional status can act as independent prognostic survival factors in hospitalized Mexican patients diagnosed with metastatic gastric adenocarcinoma and be taken into account during therapeutic decision-making.

Environmental sampling methods' influence on detection of pathogens in cage-free aviary housing.

The environmental sampling of layer housing systems is essential to identifying potential pathogens that are of concern to human health. To identify the natural occurrence of pathogens (Listeria, Campylobacter, and Salmonella) at various locations in a cage-free aviary housing system, swabs were collected when hens were 22 to 39 wks of age. Duplicate environmental swabs were taken and inoculated with a low dose (101 cfu) Salmonella Enteritidis (SE) and examined for the recovery of SE from environmental samples. Detection of Listeria (P < 0.0001) and Campylobacter (P < 0.0001) varied between the environmental sample types taken: concrete dust, drag swabs, egg belt dust, manure belt scraper swabs, and wall dust. Detection of Listeria (P < 0.0001) was the highest (70.0%) at the beginning of the study (22 wk) and decreased over time. Detection of Campylobacter (P < 0.001) was also the highest at 22 wk, however the decrease over time was more gradual. Interestingly, detection of Campylobacter (P < 0.0001) was the greatest in concrete dust samples (96.25%), which can be attributed to the presence of rodent excreta in the samples. Drag swabs and manure belt scraper swabs were the best sampling types for high detection of Listeria and Campylobacter. It should be noted that Listeria recovered was not of human health concern. No naturally occurring Salmonella was identified in this study. The recovery of the SE inoculum increased over time, reaching the greatest recovery in drag (81.25%; P < 0.0001), egg belt dust (100.00%; P < 0.0001) and wall dust swabs (100.00%; P < 0.0001) by 39 wk. This high rate of SE recovery occurred just before US mandatory SE environmental monitoring at 40 to 45 wks of age. Based on this study, the use of drag and manure belt scraper swabs are effective in detecting Listeria and Campylobacter in cage-free aviary housing. Along with good pest management, the occurrence of pathogens could be monitored and reduced in laying hen flocks.

Reproductive alterations of Biomphalaria glabrata (Say, 1818) infected with Angiostrongylus cantonensis (Chen, 1935) and exposed to Euphorbia milii var. hislopii latex.

The natural phytochemical latex of Euphorbia milii var. hislopii is one of the most promising natural molluscicides for the control of Biomphalaria glabrata, and has been widely studied under laboratory conditions for selective control of schistosomiasis transmission. However, the effect of this product on B. glabrata infected by other helminths had not yet been investigated. The present study reports evaluation of the effect of E. milii var. hislopii latex on the survival and reproductive activity of B. glabrata infected by Angiostrongylus cantonensis. For this purpose, the following groups were formed: control (C), exposed (E), infected (I) and infected and exposed for different time intervals (1 day - I+E-1D, 7 days - I+E-7D, 14 days - I+E-14D, 21 days - I+E-21D and 28 days - I+E-28D). The experimental infection was performed with L1 larvae of A. cantonensis and exposure to 0.08 mg/L (LC50) of E. milii latex for a period of 24 hours. We analyzed the effects of E. milii latex on the survival of snails during four weeks, reproductive parameters and possible histophysiopathological changes in the gonad and albumen gland of the snails. Regarding survival, there was reduction of more than 50% in the groups exposed to latex (E and I + E) compared to the control group. As for the number of ovigerous masses, eggs, and average number of hatched snails, significant increases were observed in the I+E-1D group at the 4th week compared to the control group and the other weeks in the same group. Angiostrongylus cantonensis larvae were observed in the gonad and albumen gland from day 21 and 28 of infection in groups I and I+E, respectively, with granuloma-like formation. At these observation periods and in these groups, an increase in galactogen was observed in the albumen gland, which influenced egg laying, suggesting the existence of a fecundity compensation mechanism phenomenon. It was possible to conclude that both stressors - A. cantonensis infection and exposure to E. milii latex - directly influenced the survival and reproductive parameters of B. glabrata.

First record of Bulimulus tenuissimus (Mollusca) as potential experimental intermediate host of Angiostrongylus cantonensis (Nematoda) / Primeiro registro de Bulimulus tenuissimus (Mollusca) como hospedeiro intermediário de Angiostrongylus cantonensis (Nematoda)

Abstract Snails are essential to complete the life cycle of the metastrongylid nematode Angiostrongylus cantonensis, the causative agent of infections in domestic and wild animals, mainly rodents, and also of neural angiostrongyliasis or eosinophilic meningitis in humans. There are many reports of mollusks that can act as intermediate hosts of this parasite, especially freshwater snails and the African giant Achatina fulica. The terrestrial gastropod Bulimulus tenuissimus is widely distributed in Brazil and other species of the same genus occur in Brazil and other countries, overlapping regions in which there are reports of the occurrence of A. cantonensis and angiostrongyliasis. In spite of this, there are no records in the literature of this species performing the role of intermediate host to A. cantonensis. The present study analyzed the experimental infection with first-stage larvae of A. cantonensis, under laboratory conditions, of B. tenuissimus, by using histology and electron microscopy techniques. Three weeks after exposure to L1 larvae, it was possible to recover L3 larvae in small numbers from the infected snails. Developing larvae were observed in the cephalopedal mass (foot), ovotestis, and mantle tissues, being located inside a granulomatous structure composed of hemocyte infiltration, but there was no calcium or collagen deposition in these structures in significant amounts. In the third week post exposure, it was possible observe a sheath around the developing larvae. The infected snails presented reduction in the fibrous muscular tissue in the foot region, loss of the acinar organization in the digestive gland, with increase of amorphous material inside the acini and loss of epithelial pattern of nuclear organization in the acinar cells. However, the ovotestis seemed unaffected by the infection, since there was a large number of developing oocytes and spermatozoa in different stages of formation. The digestion of infected snails allows us the third-stage recovery rate of 17.25%, at 14 days post exposure to the L1. These L3 recovered from B. tenuissimus were used to infect rats experimentally, and 43 days post infection first-stage (L1) larvae of A. cantonensis were recovered from fresh feces. The results presented constituted the first report of the role of B. tenuissimus as an experimental intermediate host to A. cantonensis and shed some light on a possible problem, since the overlapping distribution of B. tenuissimus and A. cantonensis in Brazil and other countries where different species of Bulimulus occur enables the establishment and maintenance of the life cycle of this parasite in nature, with wild rodents as reservoirs, acting as a source of infection to humans, causing neural angiostrongyliasis.

Resumo Os moluscos são um requisito essencial para a conclusão do ciclo de vida pelo nematoide metastrogilídeo Angiostrongylus cantonensis, o agente causador de infecções em animais domésticos e selvagens, principalmente roedores, e também de angiostrongilíase neural ou meningite eosinofílica em humanos. Há muitos relatos de moluscos que podem atuar como hospedeiro para este parasito, sendo o foco dado aos moluscos de água doce e no gigante africano Achatina fulica. O gastrópode terrestre Bulimulus tenuissimus é amplamente distribuído no território brasileiro e há outras espécies do mesmo gênero que ocorrem no Brasil e outros países, sobrepondo-se às regiões em que há relatos à ocorrência de A. cantonensis e angiostornigilíase. Apesar disso, não há registro na literatura, acerca desta espécie como hospedeiro intermediário para A. cantonensis. O presente estudo teve como objetivo verificar a possibilidade de infectar experimentalmente, utilizando larvas L1 de A. cantonensis, em condições laboratoriais, o molusco B. tenuissimus, utilizando técnicas de histologia e microscopia eletrônica. Três semanas após a exposição às larvas L1, foi possível recuperar larvas L3 dos moluscos infectados, em pequena quantidade. As larvas em desenvolvimento foram observadas na massa cefalopediosa (pé), ovotestis e nos tecidos do manto, sendo localizadas dentro de uma estrutura granulomatosa constituída por infiltração hemocitária, mas não houve deposição de cálcio ou colágeno nessas estruturas em quantidade significativa. Na terceira semana pós exposição, foi possível observar uma bainha ao redor das larvas em desenvolvimento. Os caracóis infectados apresentaram redução no tecido muscular fibroso na região do pé, perda da organização acinar na glândula digestiva, com aumento de material amorfo dentro dos ácinos e perda do padrão epitelial da organização nuclear nas células acinares. No entanto, o ovotestis, pareceu não ser afetado pela infecção, uma vez que houve um grande número de oócitos em desenvolvimento e espermatozóides em diferentes estágios de formação. A digestão dos moluscos infectados nos permitiu a recuperação de larvas de terceiro estágio (17,25%), aos 14 dias após a exposição à L1 de A. cantonensis . Estas L3 recuperadas de B. tenuissimus foram utilizados para infectar ratos experimentalmente, e 43 dias após a infecção, as larvas do primeiro estágio (L1) foram recuperadas de fezes frescas. Os resultados apresentados representam o primeiro registro do papel de B. tenuissimus como hospedeiro intermediário experimental de A. cantonensis e trazem alguma luz a um problema, até então silencioso, uma vez que a sobreposição da distribuição de B. tenuissimus e A. cantonensis no Brasil, e outros países, onde as diferentes espécies de Bulimulus ocorrem, torna possível o estabelecimento e manutenção do ciclo de vida deste parasito na natureza, com roedores selvagens como reservatório, agindo como fonte de infecção para humanos e causando a angiostrongilíase neural.

First record of Bulimulus tenuissimus (Mollusca) as potential experimental intermediate host of Angiostrongylus cantonensis (Nematoda).

Snails are essential to complete the life cycle of the metastrongylid nematode Angiostrongylus cantonensis, the causative agent of infections in domestic and wild animals, mainly rodents, and also of neural angiostrongyliasis or eosinophilic meningitis in humans. There are many reports of mollusks that can act as intermediate hosts of this parasite, especially freshwater snails and the African giant Achatina fulica. The terrestrial gastropod Bulimulus tenuissimus is widely distributed in Brazil and other species of the same genus occur in Brazil and other countries, overlapping regions in which there are reports of the occurrence of A. cantonensis and angiostrongyliasis. In spite of this, there are no records in the literature of this species performing the role of intermediate host to A. cantonensis. The present study analyzed the experimental infection with first-stage larvae of A. cantonensis, under laboratory conditions, of B. tenuissimus, by using histology and electron microscopy techniques. Three weeks after exposure to L1 larvae, it was possible to recover L3 larvae in small numbers from the infected snails. Developing larvae were observed in the cephalopedal mass (foot), ovotestis, and mantle tissues, being located inside a granulomatous structure composed of hemocyte infiltration, but there was no calcium or collagen deposition in these structures in significant amounts. In the third week post exposure, it was possible observe a sheath around the developing larvae. The infected snails presented reduction in the fibrous muscular tissue in the foot region, loss of the acinar organization in the digestive gland, with increase of amorphous material inside the acini and loss of epithelial pattern of nuclear organization in the acinar cells. However, the ovotestis seemed unaffected by the infection, since there was a large number of developing oocytes and spermatozoa in different stages of formation. The digestion of infected snails allows us the third-stage recovery rate of 17.25%, at 14 days post exposure to the L1. These L3 recovered from B. tenuissimus were used to infect rats experimentally, and 43 days post infection first-stage (L1) larvae of A. cantonensis were recovered from fresh feces. The results presented constituted the first report of the role of B. tenuissimus as an experimental intermediate host to A. cantonensis and shed some light on a possible problem, since the overlapping distribution of B. tenuissimus and A. cantonensis in Brazil and other countries where different species of Bulimulus occur enables the establishment and maintenance of the life cycle of this parasite in nature, with wild rodents as reservoirs, acting as a source of infection to humans, causing neural angiostrongyliasis.

Expression of nutrient transporters and host defense peptides in Campylobacter challenged broilers.

Campylobacter is a bacterium that colonizes the lower gastrointestinal tract of poultry and may influence the intestinal environment to promote its survival. The objective of this study was to characterize the effects of Campylobacter challenge on the mRNA abundance of nutrient transporters and host defense peptides (HDP), such as the avian ß-defensins (AvBD) and liver expressed antimicrobial peptide 2 (LEAP2). On the day of hatch, broiler chicks were challenged with one of three (106, 107, 108 colony-forming units, cfu) levels of Campylobacter jejuni. Quantitative PCR analysis revealed that there were dose-, tissue-, and age-specific changes in gene expression for both nutrient transporters and HDP. Expression of zinc transporter 1 (ZnT1) mRNA increased on d 7 in the duodenum, ileum, and cecum of birds challenged with 106 cfu of C. jejuni. At d 14, there was upregulation of the amino acid transporter bo,+AT mRNA in the duodenum, jejunum, and ileum of birds challenged with 106 cfu of C. jejuni. Other transporters such as EAAT3, GLUT2, SGLT1, and ZnT1 showed upregulation of mRNA in the ileum of the 106 cfu challenged group. There was a delayed response of the HDP to the C. jejuni challenge, with only a few HDP changed at d 7 but all HDP changed at d 14. At d 7, there was upregulation of AvBD10 mRNA in the duodenum of the 106 cfu challenged group but downregulation of AvBD10 in the ileum and AvBD12 and LEAP2 in the cecum of the 108 cfu challenged group. At d 14, there was upregulation of AvBD1, AvBD6, AvBD8, AvBD10, AvBD11, AvBD12, and AvBD13 mRNA in the ileum and cecum of the 106 cfu challenged group but not the 107 and 108 cfu challenged groups compared to control. These results indicated that at a low dose (106 cfu) of C. jejuni, intestinal cells increased nutrient transporter and AvBD mRNA abundance to try to counter the infection, but that at higher doses the cellular response was suppressed.

Midostaurin for the treatment of adult patients with newly diagnosed acute myeloid leukemia that is FLT3 mutation-positive.

Midostaurin is a multitargeted tyrosine kinase inhibitor (TKI) that potently inhibits activated fms-related tyrosine kinase 3 (FLT3) in the nanomolar range and other kinases including platelet-derived growth factor receptors α (PDGFR- α) and ß (PDGFR- ß), cyclin-dependent kinase, proto-oncogene tyrosine-protein kinase Src, tyrosine-protein kinase Fgr, spleen tyrosine kinase (Syk), KIT proto-oncogene receptor tyrosine kinase and the major vascular endothelial growth factor receptor (VEGFR). Activating mutations in FLT3, which is one of the more common acute myeloid leukemia (AML) mutations, particularly those that result in an FLT3-ITD (internal tandem duplication) mutation, confer poor prognosis and represent a therapeutic target. Small molecule TKIs that vary in potency and selectivity for FLT3 are under investigation. Here, we provide a comprehensive review of the preclinical and clinical activity of midostaurin, a recently approved drug indicated to be used in combination with cytarabine and daunorubicin induction and cytarabine consolidation chemotherapy for the treatment of AML featuring an FLT3 mutation.

Lysolecithin as feed additive enhances collagen expression and villus length in the jejunum of broiler chickens.

Adding lysolecithin to feed has reportedly improved the performance of broiler chickens. Lysolecithin is generated by phospholipase catalyzed hydrolysis of lecithin. The enzymatic reaction converts various phospholipids into the corresponding lysophospholipids, with lysophosphatidylcholine (LPC) one of the primary products. Here we compared supplementation with a commercial lysolecithin (Lysoforte®) with comparable levels of highly purified LPC for effects on broilers. Despite no differences in weight gain during the starter period, we discovered a significant increase in average villus length with lysolecithin and an increase in villus width with purified LPC. High-throughput gene expression microarray analyses revealed many more genes were regulated in the epithelium of the jejunum by lysolecithin compared to purified LPC. The most up-regulated genes and pathways were for collagen, extracellular matrix, and integrins. Staining sections of the jejunum with Picrosirius Red confirmed the increased deposition of collagen fibrils in the villi of broilers fed lysolecithin, but not purified LPC. Thus, lysolecithin elicits gene expression in the intestinal epithelium, leading to enhanced collagen deposition and villus length. Purified LPC alone as a supplement does not mimic these responses. Feed supplementation with lysolecithin triggers changes in the intestinal epithelium with the potential to improve overall gut health and performance.

First record of Bulimulus tenuissimus (Mollusca) as potential experimental intermediate host of Angiostrongylus cantonensis (Nematoda)

Abstract Snails are essential to complete the life cycle of the metastrongylid nematode Angiostrongylus cantonensis, the causative agent of infections in domestic and wild animals, mainly rodents, and also of neural angiostrongyliasis or eosinophilic meningitis in humans. There are many reports of mollusks that can act as intermediate hosts of this parasite, especially freshwater snails and the African giant Achatina fulica. The terrestrial gastropod Bulimulus tenuissimus is widely distributed in Brazil and other species of the same genus occur in Brazil and other countries, overlapping regions in which there are reports of the occurrence of A. cantonensis and angiostrongyliasis. In spite of this, there are no records in the literature of this species performing the role of intermediate host to A. cantonensis. The present study analyzed the experimental infection with first-stage larvae of A. cantonensis, under laboratory conditions, of B. tenuissimus, by using histology and electron microscopy techniques. Three weeks after exposure to L1 larvae, it was possible to recover L3 larvae in small numbers from the infected snails. Developing larvae were observed in the cephalopedal mass (foot), ovotestis, and mantle tissues, being located inside a granulomatous structure composed of hemocyte infiltration, but there was no calcium or collagen deposition in these structures in significant amounts. In the third week post exposure, it was possible observe a sheath around the developing larvae. The infected snails presented reduction in the fibrous muscular tissue in the foot region, loss of the acinar organization in the digestive gland, with increase of amorphous material inside the acini and loss of epithelial pattern of nuclear organization in the acinar cells. However, the ovotestis seemed unaffected by the infection, since there was a large number of developing oocytes and spermatozoa in different stages of formation. The digestion of infected snails allows us the third-stage recovery rate of 17.25%, at 14 days post exposure to the L1. These L3 recovered from B. tenuissimus were used to infect rats experimentally, and 43 days post infection first-stage (L1) larvae of A. cantonensis were recovered from fresh feces. The results presented constituted the first report of the role of B. tenuissimus as an experimental intermediate host to A. cantonensis and shed some light on a possible problem, since the overlapping distribution of B. tenuissimus and A. cantonensis in Brazil and other countries where different species of Bulimulus occur enables the establishment and maintenance of the life cycle of this parasite in nature, with wild rodents as reservoirs, acting as a source of infection to humans, causing neural angiostrongyliasis.

Resumo Os moluscos são um requisito essencial para a conclusão do ciclo de vida pelo nematoide metastrogilídeo Angiostrongylus cantonensis, o agente causador de infecções em animais domésticos e selvagens, principalmente roedores, e também de angiostrongilíase neural ou meningite eosinofílica em humanos. Há muitos relatos de moluscos que podem atuar como hospedeiro para este parasito, sendo o foco dado aos moluscos de água doce e no gigante africano Achatina fulica. O gastrópode terrestre Bulimulus tenuissimus é amplamente distribuído no território brasileiro e há outras espécies do mesmo gênero que ocorrem no Brasil e outros países, sobrepondo-se às regiões em que há relatos à ocorrência de A. cantonensis e angiostornigilíase. Apesar disso, não há registro na literatura, acerca desta espécie como hospedeiro intermediário para A. cantonensis. O presente estudo teve como objetivo verificar a possibilidade de infectar experimentalmente, utilizando larvas L1 de A. cantonensis, em condições laboratoriais, o molusco B. tenuissimus, utilizando técnicas de histologia e microscopia eletrônica. Três semanas após a exposição às larvas L1, foi possível recuperar larvas L3 dos moluscos infectados, em pequena quantidade. As larvas em desenvolvimento foram observadas na massa cefalopediosa (pé), ovotestis e nos tecidos do manto, sendo localizadas dentro de uma estrutura granulomatosa constituída por infiltração hemocitária, mas não houve deposição de cálcio ou colágeno nessas estruturas em quantidade significativa. Na terceira semana pós exposição, foi possível observar uma bainha ao redor das larvas em desenvolvimento. Os caracóis infectados apresentaram redução no tecido muscular fibroso na região do pé, perda da organização acinar na glândula digestiva, com aumento de material amorfo dentro dos ácinos e perda do padrão epitelial da organização nuclear nas células acinares. No entanto, o ovotestis, pareceu não ser afetado pela infecção, uma vez que houve um grande número de oócitos em desenvolvimento e espermatozóides em diferentes estágios de formação. A digestão dos moluscos infectados nos permitiu a recuperação de larvas de terceiro estágio (17,25%), aos 14 dias após a exposição à L1 de A. cantonensis . Estas L3 recuperadas de B. tenuissimus foram utilizados para infectar ratos experimentalmente, e 43 dias após a infecção, as larvas do primeiro estágio (L1) foram recuperadas de fezes frescas. Os resultados apresentados representam o primeiro registro do papel de B. tenuissimus como hospedeiro intermediário experimental de A. cantonensis e trazem alguma luz a um problema, até então silencioso, uma vez que a sobreposição da distribuição de B. tenuissimus e A. cantonensis no Brasil, e outros países, onde as diferentes espécies de Bulimulus ocorrem, torna possível o estabelecimento e manutenção do ciclo de vida deste parasito na natureza, com roedores selvagens como reservatório, agindo como fonte de infecção para humanos e causando a angiostrongilíase neural.

Electrophoresis and spectrometric analyses of adaptation-related proteins in thermally stressed Chromobacterium violaceum.

Chromobacterium violaceum is a Gram-negative proteobacteria found in water and soil; it is widely distributed in tropical and subtropical regions, such as the Amazon rainforest. We examined protein expression changes that occur in C. violaceum at different growth temperatures using electrophoresis and mass spectrometry. The total number of spots detected was 1985; the number ranged from 99 to 380 in each assay. The proteins that were identified spectrometrically were categorized as chaperones, proteins expressed exclusively under heat stress, enzymes involved in the respiratory and fermentation cycles, ribosomal proteins, and proteins related to transport and secretion. Controlling inverted repeat of chaperone expression and inverted repeat DNA binding sequences, as well as regions recognized by sigma factor 32, elements involved in the genetic regulation of the bacterial stress response, were identified in the promoter regions of several of the genes coding proteins, involved in the C. violaceum stress response. We found that 30 °C is the optimal growth temperature for C. violaceum, whereas 25, 35, and 40 °C are stressful temperatures that trigger the expression of chaperones, superoxide dismutase, a probable small heat shock protein, a probable phasing, ferrichrome-iron receptor protein, elongation factor P, and an ornithine carbamoyltransferase catabolite. This information improves our comprehension of the mechanisms involved in stress adaptation by C. violaceum.

Haematological alterations in Rattus norvegicus (Wistar) experimentally infected with Echinostoma paraensei (Trematoda: Echinostomatidae).

Laboratory rats (Rattus norvegicus) were infected with Echinostoma paraensei (Trematoda: Echinostomatidae). The rodents received 150 metacercariae each and blood samples were collected weekly until the fifth week of infection. The blood samples were analyzed for determination of haematocrit, total red blood cells with their dimensions, haemoglobin and haematimetric index (mean corpuscular volume, MCV; mean corpuscular haemoglobin, MCH; and mean corpuscular haemoglobin concentration, MCHC) and platelets. Red blood cells, haematocrit and haemoglobin in the first week had significantly lower levels than those of uninfected (control) rats, suggesting the development of normocytic and normocromic anaemia with anisocytic alteration. The number of eosinophils did not increase significantly among the groups. We concluded that E. paraensei produces haematological alterations in R. norvegicus, causing regenerative anaemia. This system can therefore be a useful model to study the direct and indirect effects of gastrointestinal infections.

An improved protocol for electroporation of Oenococcus oeni ATCC BAA-1163 using ethanol as immediate membrane fluidizing agent.

AIMS: To finalize an effective and reproducible electroporation procedure to transform Oenococcus oeni ATCC BAA-1163 strain. METHODS AND RESULTS: The vector pGID052 was selected to optimize the electroporation procedure. Transformation efficiency was 5.8 x 10(3) per microg of DNA. Transformation was improved when competent cells were prepared with exponential phase cultures; optimum electroporation parameters were an electric pulse of 125 kV cm(-1), under a resistance of 200 omega and the presence of 10% (v/v) ethanol in the electroporation buffer (EPB). CONCLUSIONS: An effective protocol to transform O. oeni ATCC BAA-1163 strain by electroporation has been obtained by addition of ethanol to the EPB. A heterologous expression was obtained in O. oeni ATCC BAA-1163 by introducing a recombinant vector encoding a truncated form of ClpL2 protein. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report of a successful electroporation of O. oeni ATCC BAA-1163. The major improvement was the addition of ethanol to the EPB, which has never been reported before as means of enhancing the incorporation of foreign DNA molecules into prokaryote cells by electroporation. This method constitutes a useful tool for the genetic study of this lactic bacterium.

Autologous serum eyedrops in the treatment of aniridic keratopathy.

OBJECTIVE: To study the effect of autologous serum eyedrop application in aniridic keratopathy. DESIGN: Prospective, consecutive, comparative, interventional case series. PARTICIPANTS: Twenty-six eyes from 13 patients (7 males and 6 females) with aniridic keratopathy treated with autologous serum eyedrops. METHODS: All patients underwent a complete ophthalmic examination. The ocular surface examinations included corneal impression cytologic analysis and tear film evaluation. The eyes were divided into 4 groups according to the Mackman classification. Ocular surface photography was used to evaluate the corneal surface and tear film before treatment and every 2 or 3 days until serum drops were stopped. Tear films were evaluated by tear film break-up time (BUT) (normal, 10 seconds or more), Schirmer's test with anesthesia (normal, 10 mm/5 minutes or more), tear meniscus level (normal, 0.5 mm or more), and rose bengal and fluorescein staining pattern of the cornea. Impression cytologic analysis was carried out both before starting the serum eyedrops treatment and a few days after its finalization. MAIN OUTCOMES MEASURES: Tear film production and stability, corneal epithelialization, and corneal epithelium squamous metaplasia. RESULTS: There were no local side effects from autologous serum treatment. Clinical manifestations and slit-lamp findings were in relation to the severity of keratopathy. All patients showed a subjective improvement of keratopathy symptoms after the autologous serum applications. The corneal epithelialization, corneal epithelial cell squamous metaplasia, and tear stability improved significantly with the treatment, but visual acuity, regression of vascular pannus, and subepithelial scarring showed only slight improvement with treatment. CONCLUSIONS: Autologous serum eyedrops improved the aniridic keratopathy in all patients, particularly in patients with light or moderate severity. In these patients, use of autologous serum eyedrops was superior to conventional therapy with substitute tears for improving the ocular surface and subjective comfort.

Different conjunctival adaptive response in patients with aqueous-deficient and with mucous-deficient dry eyes.

PURPOSE: To describe the different cellular adaptive patterns found in the conjunctival epithelium from patients with aqueous-deficient and mucous-deficient dry eyes. METHODS: The authors studied different conjunctival areas, by impression cytology and by biopsy, 50 eyes with facial nerve paralysis (FNP), 50 eyes with ocular cicatricial pemphigoid (OCP), and 50 eyes from patients with primarily Sjögren syndrome (1SS). RESULTS: Eyes with FNP from the first clinical grade showed a progressive alteration of the nonsecretory cells, with a significant decrease in density goblet cells, generally with a PAS-positive staining. Eyes with OCP, during clinical grades 1 and 2, showed a slow deterioration of the nonsecretory cells; but from clinical grade 3, there was a significant increase of the cellular size and the thickness of the conjunctiva. Goblet cells showed a significant decrease in density from clinical grade 1, generally with a PAS-negative staining. Eyes with 1SS during clinical grades 1 and 2 showed a progressive alteration of the nonsecretory cells, with a significant decrease in density goblet cells, and a PAS-positive staining. From clinical grade 3 appeared a significant increase of nonsecretory cellular size and thickness of conjunctiva, with a significant decrease in goblet cell counts, and a PAS-negative staining. CONCLUSIONS: Patients with FNP (a primarily aqueous-deficient alteration) follow completely the squamous metaplasia process. Patients with OCP (a primarily mucous-deficient syndrome) have a hypertrophy and hyperplasia process along the ocular surface. Patients with 1SS (a primarily aqueous-deficient and mucin-deficient alteration) have a squamous metaplasia process, but from clinical grade 3 also appears a hypertrophy and hyperplasia process.

[Use of autologous serum in ophthalmic practice]. / Aplicaciones del suero autólogo en oftalmología.

OBJECTIVE: To establish a protocol for the use of autologous serum in the ophthalmic practice. METHODS: Personal experience and a literature review. RESULTS: The use of autologous serum, in eye drop form, has been reported as a new treatment for several ocular surfaces diseases. These products have biomechanical and biochemical properties similar to normal tears. They contain components such as fibronectin, vitamin A and growth factors that have an epitheliotrophic effect on the ocular surface epithelial cells. CONCLUSIONS: The clinical studies performed showed a variable efficacy, with the preparative process and use of the autologous serum eye drops varying considerably between different studies.

Analysis of corneal surface evolution after moderate alkaline burns by using impression cytology.

PURPOSE: To compare corneal surface evolution after moderate alkaline burns by impression cytology in patients treated with medical therapy or with amniotic membrane transplantation (AMT). METHODS: A prospective study of 24 eyes from 18 patients (13 men and 5 women) with moderate alkaline burns was performed. All patients were divided according to the clinical ocular severity and the therapy used. Twelve eyes were treated surgically with AMT and the other 12 eyes received only medical therapy. Corneal cytology was obtained immediately after the burns, and 1, 2, 5, and 9 months later. We differentiated between samples obtained from affected areas and areas not affected by the burns. Cellular size, nuclear size, and nuclear-cytoplasmic (N:C) ratio were examined in corneal epithelial cells, as was the presence of goblet cells in corneal epithelium. RESULTS: Nuclear size, cellular size, and N:C ratio in non-burn-affected corneal areas had no significant alterations in comparison with normal eyes. In contrast, in burn-affected corneal areas, these parameters were significantly worse, and the presence of goblet cells in corneal epithelium was frequent 1 month after severe burns. Cellular size, nuclear size, N:C ratio, and corneal conjunctivalization improved during the study in all patients, but corneal reepithelialization occurred earlier in patients treated with AMT than in patients with only medical therapy. CONCLUSION: Morphologic and morphometric analysis of corneal cells by impression cytology after ocular burns permits the establishment of cellular reepithelialization patterns in relation with limbal deficiency level and with clinical ocular severity. AMT improves corneal reepithelialization earlier than medical therapy in moderate alkaline burns.

[Congenital aniridia keratopathy treatment]. / Manejo terapéutico de la queratopatía asociada a aniridia congénita.

OBJECTIVE: To attempt to review the aniridia keratopathy pathogenesis and establish a treatment protocol according to the severity of the symptoms. METHODS: Personal experience in aniridic keratopathy management and a bibliography review. RESULTS: The ocular manifestations of this anomaly include defects of the cornea, glaucoma, lens subluxation, cataracts, hypoplasia of the iris, fovea and optic nerve, amblyopia and nystagmus. The keratopathy occurs in a 20% of patients with aniridia. The correct PAX6 expression is necessary for normal corneal development, limbal stem cell activity and correct corneal epithelial cell migration and adhesion. CONCLUSIONS: The management of ocular surface diseases due to limbal stem cell deficiency in aniridia is complex but has changed in recent years, as an understanding of the limbal stem cells and their microenvironment has modified the therapeutic approach. The use of autologous serum eye drops, amniotic membrane transplantation, limbal transplantation or cultivated limbal cell transplantation have all been reported as a treatment for several ocular surface diseases.

Manejo terapéutico de la queratopatía asociada a aniridia congénita / Congenital aniridia keratopathy treatment

Objetivo: Revisar la patogenia de la queratopatía por aniridia y establecer un protocolo de tratamiento según su gravedad. Métodos: Revisión bibliográfica y experiencia personal en el manejo de pacientes con queratopatía por aniridia. Resultados: Las manifestaciones oculares de esta anomalía incluyen defectos a nivel corneal, glaucoma, subluxación del cristalino y cataratas, hipoplasia de iris, fóvea y nervio óptico, ambliopía y nistagmus. La queratopatía por aniridia ocurre en un 20% de los pacientes. La correcta expresión del gen PAX6 es necesaria para el normal desarrollo corneal y actividad de las células limbares así como la correcta migración y adhesión de las células epiteliales. Conclusiones: El manejo de los trastornos oculares debidos a la deficiencia limbal asociada a la aniridia es complejo pero ha cambiado en los últimos años. El concepto de célula madre limbal y de su microambiente ha modificado la aproximación terapéutica de estos pacientes. El empleo del suero autólogo, trasplante de membrana amniótica, trasplante de limbo o trasplante de células limbares cultivadas han sido utilizados en el tratamiento de distintos trastornos de la superficie ocular

Objective: To attempt to review the aniridia keratopathy pathogenesis and establish a treatment protocol according to the severity of the symptoms. Methods: Personal experience in aniridic keratopathy management and a bibliography review. Results: The ocular manifestations of this anomaly include defects of the cornea, glaucoma, lens subluxation, cataracts, hypoplasia of the iris, fovea and optic nerve, amblyopia and nystagmus. The keratopathy occurs in a 20% of patients with aniridia. The correct PAX6 expression is necessary for normal corneal development, limbal stem cell activity and correct corneal epithelial cell migration and adhesion. Conclusions: The management of ocular surface diseases due to limbal stem cell deficiency in aniridia is complex but has changed in recent years, as an understanding of the limbal stem cells and their microenvironment has modified the therapeutic approach. The use of autologous serum eye drops, amniotic membrane transplantation, limbal transplantation or cultivated limbal cell transplantation have all been reported as a treatment for several ocular surface diseases

[Corneal epithelium squamous metaplasia determination as diagnostic factor in limbal deficiency]. / Determinación del grado de metaplasia escamosa del epitelio corneal como factor diagnóstico de insuficiencia limbal.

PURPOSE: To determine the correlation between the squamous metaplasia grade of the corneal surface and the clinical severity of the limbal deficiency in patients with this disorder. METHODS: We studied 98 eyes of patients with limbal deficiency by impression cytology. These patients were divided into four groups in relation to their clinical severity: 14 eyes had no symptoms, 34 eyes showed a mild grade of limbal deficiency, 28 eyes had a moderate grade and 22 eyes had severe limbal deficiency. Corneal cytology was performed in each patient. Cellular size, nuclear size, nuclear changes and the nuclear-cytoplasmic (N:C) ratio were defined in corneal epithelial cells, in addition to evaluation for the presence of goblet cells in the corneal epithelium. RESULTS: In patients with limbal deficiency without symptoms, we found that the cellular size was 477 (SD140) microm2 and the N:C ratio was 1:5.25 (SD1.5). In patients with mild limbal deficiency, the cellular size was 764 (SD122.6) microm2 and the N:C ratio was 1:8.2 (SD1.4). These patients did not show corneal conjunctivalization. In patients with moderate limbal deficiency the cellular size was 1,162 (SD340.2) microm2 and the N:C ratio was 1:13.2 (SD3.6). Some patients showed corneal conjunctivalization that was defined by the presence of goblet cells in the corneal epithelium. In patients with severe limbal deficiency, the cellular size was 2,036 (SD382) microm2 and the N:C ratio was 1:23.6 (SD4.0). Corneal conjunctivalization was present in all of these patients. CONCLUSIONS: There was a correlation between limbal deficiency severity and squamous metaplasia grade defined by impression cytology. Impression cytology was very useful for the diagnosis of cases of mild or subclinical limbal deficiency.

Determinación del grado de metaplasia escamosa del epitelio corneal como factor diagnóstico de insuficiencia limbal / Corneal epithelium squamous metaplasia determination as diagnostic factor in limbal deficiency

Objetivos: Determinar si existe relación entre elgrado de metaplasia escamosa de la superficie cornealmediante citología de impresión y la gravedadclínica en pacientes con insuficiencia limbal.Métodos: Hemos estudiado mediante citología deimpresión 98 ojos de pacientes con insuficiencialimbal que fueron divididos en cuatro grupos segúnla gravedad clínica: 14 ojos no presentaban sintomatologíaalguna, 34 ojos presentaron un gradoleve de insuficiencia limbal, 28 ojos un gradomoderado y 22 ojos una insuficiencia limbal grave.A todos los pacientes se les realizó una citologíacorneal en la que se estudió el tamaño celular, tamañonuclear, alteraciones nucleares y relaciónnúcleo-citoplasma (N:C), así como la presencia decélulas caliciformes.Resultados: Los pacientes con insuficiencia limbalasintomática presentaron un tamaño celular mediode 477 DE 140 μm2, con una relación N:C de 1:5,25DE 1,5. Los pacientes con insuficiencia limbal levepresentaron un tamaño celular medio de 764 DE122,6 μm2 y una relación N:C de 1:8,2 DE 1,4. Ninguno de estos pacientes presentó conjuntivalizacióndel epitelio corneal. Los pacientes con insuficiencialimbal moderada presentaron un tamaño celularmedio de 1.162 DE 340 μm2 y una relación N:C de1:13,2 DE 3,6. Estos pacientes presentaron concierta frecuencia conjuntivalización del epitelio corneal,definida por la presencia de células caliciformes.Los pacientes con insuficiencia limbal gravepresentaron un tamaño celular medio de 2.036 DE382 μm2 y una relación N:C de 1:23,6 DE 4,0. Enestos pacientes la presencia de conjuntivalizaciónen la citología corneal fue la norma.Conclusiones: Existe una correlación entre la gravedadclínica de la insuficiencia limbal y el gradode metaplasia escamosa determinado mediante citologíade impresión corneal. La citología de impresiónes particularmente útil para el diagnóstico enlos casos de déficit limbal leve o subclínico

Purpose: To determine the correlation between the ;;squamous metaplasia grade of the corneal surface ;;and the clinical severity of the limbal deficiency in ;;patients with this disorder. ;;Methods: We studied 98 eyes of patients with limbal ;;deficiency by impression cytology. These ;;patients were divided into four groups in relation to ;;their clinical severity: 14 eyes had no symptoms, 34 ;;eyes showed a mild grade of limbal deficiency, 28 ;;eyes had a moderate grade and 22 eyes had severe ;;limbal deficiency. Corneal cytology was performed ;;in each patient. Cellular size, nuclear size, nuclear ;;changes and the nuclear-cytoplasmic (N:C) ratio ;;were defined in corneal epithelial cells, in addition ;;to evaluation for the presence of goblet cells in the ;;corneal epithelium. ;;Results: In patients with limbal deficiency without ;;symptoms, we found that the cellular size was 477 ;;(SD140) μm2 and the N:C ratio was 1:5.25 (SD1.5). ;;In patients with mild limbal deficiency, the cellular ;;size was 764 (SD122.6) μm2 and the N:C ratio was ;;1:8.2 (SD1.4). These patients did not show corneal conjunctivalization. In patients with moderate limbal ;;deficiency the cellular size was 1,162 (SD340.2) ;;μm2 and the N:C ratio was 1:13.2 (SD3.6). Some ;;patients showed corneal conjunctivalization that ;;was defined by the presence of goblet cells in the ;;corneal epithelium. In patients with severe limbal ;;deficiency, the cellular size was 2,036 (SD382) μm2 ;;and the N:C ratio was 1:23.6 (SD4.0). Corneal conjunctivalization ;;was present in all of these patients. ;;Conclusions: There was a correlation between limbal ;;deficiency severity and squamous metaplasia ;;grade defined by impression cytology. Impression ;;cytology was very useful for the diagnosis of cases ;;of mild or subclinical limbal deficiency