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AIMS: Identify novel bacterial taxa that could increase the availability of branched-chain amino acids and the amount of distinctive volatiles during skim milk fermentation. METHODS AND RESULTS: We recovered 344 bacterial isolates from stool samples of healthy and breastfed infants. Five were selected based on their ability to produce branched-chain amino acids. Three strains were identified as Escherichia coli, one as Klebsiella pneumoniae and other as Klebsiella variicola by molecular and biochemical methods. HPLC and solid-phase microextraction with GC-MS were used for the determination of free amino acids and volatile compounds respectively. The consortium formed by K. variicola and four Lactobacillus species showed the highest production of Leu and Ile in skim milk fermentation. In addition, the production of volatile compounds, such as acetoin, ethanol, 2-nonanone, and acetic, hexanoic and octanoic acids, increased in comparison to commercial yogurt, Emmental and Gouda cheese. Also, distinctive volatiles, such as 2,3-butanediol, 4-methyl-2- hexanone and octanol, were identified. CONCLUSION: The use of K. variicola in combination with probiotic Lactobacillus species enhances the availability of Leu and Ile and the amount of distinctive volatiles during skim milk fermentation. SIGNIFICANCE AND IMPACT OF THE STUDY: The identified consortium increases the functional potential of fermented dairy products.

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Objetivos. Tanto para cirugía laparoscópica como para cirugía abierta la analgesia multimodal puede ayudar a controlar el dolor postoperatorio. La colocación de un catéter en la herida quirúrgica de manera intraoperatoria tras cirugía de colon podría optimizar el control del dolor con menor consumo de opiáceos y menos efectos secundarios. Método. Realizamos un estudio prospectivo, aleatorizado de pacientes reclutados para cirugía de colon laparoscópica en el Hospital de Galdakao-Usansolo de enero de 2012 a enero de 2013. Los pacientes fueron asignados aleatoriamente al grupo del catéter o al grupo de la analgesia postoperatoria estándar. Un miembro de la Unidad de dolor agudo monitorizó todos los pacientes a lo largo de 48 h tras la cirugía. Las variables principales analizadas fueron la escala numérica verbal y la cantidad de morfina intravenosa utilizada por cada paciente mediante PCA. Resultados. Se incluyeron 92 pacientes en el estudio, 43 en el grupo con catéter y 49 en el estándar. Se observaron diferencias estadísticamente significativas en el consumo de morfina entre ambos grupos a lo largo de todo el periodo. La cantidad total de morfina consumida en el grupo del catéter fue de 5,63 ± 5,02 mg y de 21,86 ± 17,88 mg en el grupo de analgesia estándar (p = 0,0001). Los pacientes con catéter presentaban menores valores en la escala numérica verbal. No se encontraron efectos adversos asociados a la colocación del catéter y la administración de anestésico local. El grupo de catéter presentó menor estancia hospitalaria respecto al otro grupo (p = 0,02). Conclusión. En los pacientes intervenidos de cirugía de colon laparoscópico una infusión continua de anestésico local a través de un catéter interfascial durante 48 h tras la cirugía reduce la percepción del dolor y el consumo de morfina intravenosa, disminuyendo la estancia hospitalaria (AU)

Objectives. For major laparoscopic surgery, as with open surgery, a multimodal analgesia plan can help to control postoperative pain. Placing a wound catheter intraoperatively following colon surgery could optimize the control of acute pain with less consumption of opioids and few adverse effects. Methods. We conducted a prospective, randomized, study of patients scheduled to undergo laparoscopic colon surgery for cancer in Galdakao-Usansolo Hospital from January 2012 to January 2013. Patients were recruited and randomly allocated to wound catheter placement plus standard postoperative analgesia or standard postoperative analgesia alone. A physician from the acute pain management unit monitored all patients for pain at multiple points over the first 48 hours after surgery. The primary outcome variables were verbal numeric pain scale scores and amount of intravenous morphine used via patient controlled infusion. Results. 92 patients were included in the study, 43 had a wound catheter implanted and 49 did not. Statistically significant differences in morphine consumption were observed between groups throughout the course of the treatment period. The mean total morphine consumption at the end of the study was 5.63 ± 5.02 mg among wound catheter patients and 21. 86 ± 17.88 mg among control patients (P = .0001). Wound catheter patients had lower pain scale scores than control patients throughout the observation period. No adverse effects associated with the wound catheter technique were observed. The wound catheter group showed lower hospital stays with statistically significant difference (P = .02). Conclusions. In patients undergoing laparoscopic colon surgery, continuous infusion of local anaesthetics through interfascial wound catheters during the first 48 h aftersurgery reduced the level of perceived pain and also reduced parenteral morphine consumption with no associated adverse effects and lower hospital stays (AU)

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OBJECTIVES: For major laparoscopic surgery, as with open surgery, a multimodal analgesia plan can help to control postoperative pain. Placing a wound catheter intraoperatively following colon surgery could optimize the control of acute pain with less consumption of opioids and few adverse effects. METHODS: We conducted a prospective, randomized, study of patients scheduled to undergo laparoscopic colon surgery for cancer in Galdakao-Usansolo Hospital from January 2012 to January 2013. Patients were recruited and randomly allocated to wound catheter placement plus standard postoperative analgesia or standard postoperative analgesia alone. A physician from the acute pain management unit monitored all patients for pain at multiple points over the first 48 hours after surgery. The primary outcome variables were verbal numeric pain scale scores and amount of intravenous morphine used via patient controlled infusion. RESULTS: 92 patients were included in the study, 43 had a wound catheter implanted and 49 did not. Statistically significant differences in morphine consumption were observed between groups throughout the course of the treatment period. The mean total morphine consumption at the end of the study was 5.63±5.02mg among wound catheter patients and 21. 86±17.88mg among control patients (P=.0001). Wound catheter patients had lower pain scale scores than control patients throughout the observation period. No adverse effects associated with the wound catheter technique were observed. The wound catheter group showed lower hospital stays with statistically significant difference (P=.02). CONCLUSIONS: In patients undergoing laparoscopic colon surgery, continuous infusion of local anaesthetics through interfascial wound catheters during the first 48h aftersurgery reduced the level of perceived pain and also reduced parenteral morphine consumption with no associated adverse effects and lower hospital stays.

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OBJECTIVE: All-trans-retinoic acid (ATRA) promotes cell differentiation. We have studied its effect on the local recurrence and metastatic spreading of an experimental rhabdomyosarcoma in rats. DESIGN: syngenic rhabdomyosarcoma cells (S4MH) were inoculated s.c. in male WAG/RijCrl rats. After 25 days tumors were excised and a 40% hepatectomy was performed for all animals. Ten days later the rats were sacrificed and a thorough necropsy was performed. The animals were randomly allocated to receive daily doses of ATRA (5 mg/kg, i.p.) or its solvent (Clinoleic/ethanol 90/10), starting three days before surgery until the end of the experiment. RESULTS: ATRA reduced the incidence of local recurrence from 70 to 33% (p < 0.05), but the tumor size was not altered (1.8 vs. 2.0 cc). Regarding inguinal metastasis, there was a six-fold decrease (0.2 vs. 1.2 cc; p < 0.05) in mean tumor volume, although the rate of this proliferation increased sharply (86 vs. 29%; p < 0.05) for treated animals. The volume of the retroperitoneal tumor masses also decreased with ATRA (0.7 vs. 5.1 cc; p < 0.05), but the difference in rate was not significant (71 vs. 67%). Lung metastases, which were present in 100% of control animals, were found in only 33% of treated rats, while the mean number of metastatic foci dropped from 26.7 to 5.7 (p < 0.05). CONCLUSION: Protocols including retinoid administration prior to and following primary tumor excision could help in controlling both recurrence and metastatic progression in surgically treated rhabdomyosarcoma.

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Objetivo: el ácido trans-retinoico (ATRA, "all-trans retinoic acid") es un agente inductor de la diferenciación celular. Se estudia su efecto sobre la recidiva local y la diseminación metastásica de los tumores sólidos. Diseño experimental: mediante inoculación subcutánea de células de rabdomiosarcoma (S4MH) se ha inducido un tumor en ratas WAG/RijCrl. Tras 25 días, se practicó una tumorectomía y simultáneamente hepatectomía del 40%. Los animales se sacrificaron el día 35, y fueron sometidos a estudio necrópsico. La mitad de los animales fueron tratados con ATRA (5 mg/kg, i.p.) desde el día 22 al 35, mientras que los controles recibieron el excipiente (Clinoleico®/etanol 90/10). Resultados: el tratamiento redujo la tasa de recidiva local del 70 al 33% (p < 0,05), aunque no afectó a su tamaño (1,8 vs. 2,0 cc). El volumen medio de las metástasis inguinales se redujo a la sexta parte (0,2 vs. 1,2 cc; p < 0,05), si bien su frecuencia aumentó con el ATRA (86 vs. 29%; p < 0,05). La extensión retroperitoneal del rabdomisoarcoma también se redujo (0,7 vs. 5,1 cc; p < 0,05), aunque no hubo variación en la incidencia (71 vs. 67%). La incidencia de afectación pulmonar (100% en controles) se redujo hasta el 33% (p < 0,05), a la vez que el número medio de focos en el pulmón pasó de 26,7 a 5,7 (p < 0,05). Conclusión: la estrategia terapéutica fundamentada en el tratamiento pre y postextirpación quirúrgica con retinoides podría favorecer el control local de rabdomiosarcomas sometidos a cirugía

Objective: all-trans-retinoic acid (ATRA) promotes cell differentiation. We have studied its effect on the local recurrence and metastatic spreading of an experimental rhabdomyosarcoma in rats. Design: syngenic rhabdomyosarcoma cells (S4MH) were inoculated s.c. in male WAG/RijCrl rats. After 25 days tumors were excised and a 40% hepatectomy was performed for all animals. Ten days later the rats were sacrificed and a thorough necropsy was performed. The animals were randomly allocated to receive daily doses of ATRA (5 mg/kg, i.p.) or its solvent (Clinoleic®/ethanol 90/10), starting three days before surgery until the end of the experiment. Results: ATRA reduced the incidence of local recurrence from 70 to 33% (p < 0.05), but the tumor size was not altered (1.8 vs. 2.0 cc). Regarding inguinal metastasis, there was a six-fold decrease (0.2 vs. 1.2 cc; p < 0.05) in mean tumor volume, although the rate of this proliferation increased sharply (86 vs. 29%; p < 0.05) for treated animals. The volume of the retroperitoneal tumor masses also decreased with ATRA (0.7 vs. 5.1 cc; p < 0.05), but the difference in rate was not significant (71 vs. 67%). Lung metastases, which were present in 100% of control animals, were found in only 33% of treated rats, while the mean number of metastatic foci dropped from 26.7 to 5.7 (p < 0.05). Conclusion: protocols including retinoid administration prior to and following primary tumor excision could help in controlling both recurrence and metastatic progression in surgically treated rhabdomyosarcoma

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In the early stages of metastasis, development of the disease is dependent on growth factors produced by the host. There are clinical situations associated with an increase in these factors, such as partial resection of metastasized liver. Given the important role of hepatotrophic factors in liver regeneration, we have studied the effect of partial hepatectomy on the development of residual micrometastases in the liver, and on the neoplastic process as a whole. We used a murine model in which a rabdomiosarcoma was established by subcutaneous inoculation of syngeneic tumor cells in male Wag rats. Subsequently, the primary tumor was resected and/or a 40% hepatectomy was performed. The effect of these two surgical procedures on the tumor process was analyzed on the 25th and 35th days post-inoculation, and the percentage of regenerating hepatocytes was assessed. Both the tumorectomy and liver resection, when not combined, produced an increase in regional adenopathies without modifying the evolution of metastasis in the liver. However, when tumor excision and partial hepatectomy were performed simultaneously, there was a net increase in the metastatic process. In addition to a rapid spread of the disease (lung, mediastinum, retroperitoneum), the number of liver metastases increased by 300%. This development coincided with a steep rise in the percentage of regenerating hepatocytes, which nearly doubled that of the group subjected only to liver resection. We conclude that liver resection, alone or combined with excision of the primary tumor, may enhance tumor progression, both locally and at the metastasic level.

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An in vitro presymbiotic system between mesquite [Prosopis laevigata(Willd.) M.C. Johnst], a semi-arid leguminous plant, and pregerminated spores of Gigaspora rosea Nicol. & Schenck was established. After characteristic hyphal branching, high performance liquid chromatographic analyses of methanol extracts from P. laevigata roots revealed a concentration change in one ultraviolet-detectable product. This product was identified by nuclear magnetic resonance and mass spectrometry as trigonelline, a pyridine alkaloid. The concentration of trigonelline was constant in the aerial parts of the plant with or without G. rosea, but its concentration in the roots increased 1.8-fold when G. roseawas present. Trigonelline may be a regulatory factor during early signal events in the establishment of the arbuscular mycorrhizal symbiosis in P. laevigata.

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For the first time, the production of transgenic plants of the forage grass blue grama, Bouteloua gracilis [H.B.K.] Lag. ex Steud., is reported. Transgenic plants containing a gus Colon, two colons nptll fusion driven by a double CaMV35S promoter were obtained by microprojectile bombardment of the highly chlorophyllous embryogenic cell line 'TIANSJ98'. Transformed B. gracilis cell lines resisted a lethal concentration of 160 mg/l of kanamycin for at least 8 months. Chlorophyll development and growth rate were used as useful criteria for discriminating transformed from non-transformed clones. Stable integration of the transgene in the blue grama genome was demonstrated by PCR and Southern-hybridization analysis. Expression of the NPTll protein in transgenic plants grown under greenhouse conditions was confirmed indirectly by spraying kanamycin (150-250 mg/l) on plant foliage, and directly by ELISA immunological tests. Control plants sprayed with kanamycin showed foliar necrosis and reduced growth (tillering) compared to plants containing the transgene. NPTll was found in transgenic plants in levels ranging between 12.6 and 29.6 ng/mg FW of cells, as determined by ELISA reactions.

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A finely dispersed, homogeneous and highly chlorophyllous cell suspension (TIANSJ98 cell line) was obtained from shoot apices of Bouteloua gracilis (H.B.K.) Lag. ex Steud. cultured on MPC medium containing MS salts supplemented with 2,4-D (1 mg/l), BAP (2 mg/l) and adenine (40 mg/l). When the TIANSJ98 cell line was grown in this medium with shaking at 180 rpm it had doubling times of 7.2 and 3.7 days in terms of fresh and dry weight, respectively. Total chlorophyll content in this cell culture ranged from 121.6 to 18.3 µg/g FW at 12 and 21 days following culture initiation. Plants regenerated from the TIANSJ98 cell line, via somatic embryogenesis, were grown to maturity and produced seeds. Although different cell culture systems have been described for cereals and grasses, to the best of our knowledge this is the first report of a highly chlorophyllous and regenerable cell suspension in Poaceae.

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OBJECTIVE: An experimental model to monitor hemodynamics during intestinal reperfusion syndrome is presented. EXPERIMENTAL DESIGN: A) Three groups of rats were monitored during four hours: 1) CONTROL: sham operated, 2) Ischemic: clamping the superior mesenteric artery for 60' and 3) Saline: adding 2 ml of saline during the last 15' minutes of ischemia. Blood mean pressure, heart rate, breath rate and diuresis are monitored, while mortality is assessed 48 hours later, B) Blood gases, haematimetry and electrolytes were assessed both in control animals and in rats subjected to intestinal ischemia and reperfusion (at the beginning and concluding the ischemic period, and 5', 60' and 180' following reperfusion). RESULTS: Mortality rate was 0% for control animals, while 83% for both ischemic-reperfused groups (p < 0.01). Hemodynamic parameters remained stable during intestinal ischemia. Starting reperfusion, blood pressure and diuresis dropped sharply, while heart and breath rates showed a steady increase. The other parameters assessed also remained constant during ischemia. However, during reperfusion acidosis progressed (pH = 7.15, p < 0.01; HCO3 = 8, p < 0.01), hyperkalaemia appeared (6 mEq/l, p < 0.01), and haematocrit rose (56%, p < 0.01). CONCLUSION: This experimental model is easy to establish, allows an appropriate hemodynamic monitoring of shock during intestinal reperfusion syndrome, and seems appropriate for considering the impact of drug intervention.

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Among isolates of Rhizoctonia solani Kühn collected as sclerotia from potato (Solanum tuberosum L.) tubers harvested from the Toluca Valley in 1995 was one isolate (MexND) of anastomosis group 7 (AG-7). Virulence of this and other isolates of R. solani representing AG-3 (four isolates) or AG-7 (four isolates), was determined on developing potato plants (cv. Russet Burbank) in growth chamber studies at 10 and 21°C by methods used previously (1). A control treatment was inoculated with sterile disks of potato dextrose agar (PDA). Seed tubers were placed on a layer of pasturized sand-soil mix near the bottom of 6- × 25-cm black plastic tubes, then covered with 2 to 3 cm of the sand-soil mix. Seed tubers used in virulence tests carried no R. solani as they had been surface disinfested by immersing for 2 min in a 1.85% solution of formaldehyde. Inoculum, in the form of one 7-mm agar disk cut from the growing edge of appropriate colonies growing on PDA, was placed 2 to 3 cm above the seed and covered with 7 to 10 cm of sand-soil mix (1). Each treatment had five replications. Plants were harvested after control plants had emerged, 29 days at 21°C and 36 days at 10°C, and damage to roots and shoots was assessed according to Carling and Leiner (1). All four isolates of AG-3, one from the U.S. and three from Mexico, caused major damage to roots and shoots at both temperatures. The two isolates of AG-7 from the U.S. caused superficial discoloration to shoots at both temperatures but the two isolates of AG-7 from Japan caused no damage to shoots. None of the four isolates of AG-7 damaged roots at either temperature. MexND caused no damage to roots or shoots at 21°C but produced lesions (< 1 mm in diameter) on roots and shoots at 10°C. Thus, MexND caused less damage to potato than AG-3, more damage than other isolates of AG-7, and appears more aggressive at lower temperatures. This is the first reported occurrence of R. solani AG-7 in Mexico and its first reported recovery from a potato tuber-borne sclerotium. Reference: (1) D. E. Carling and R. H. Leiner. Phytopathology 80:930, 1990.

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PURPOSE OF THE STUDY: To determine whether Langerhans' islets exert a beneficial effect on splenic hepatocellular transplantation. HYPOTHESIS: the addition of pancreatic islets to hepatocytes would improve their normal function through the support of insulin and glucagon in the same proportions as they usually receive in the hepatic environment. EXPERIMENTAL DESIGN: Potential improvement in the hepatocellular function to be evaluated by means of regenerative activity recorded after applying a specific stimulus (partial hepatectomy and/or cyclosporine A). METHODS: An immunohistochemical technique (antinuslin antibodies) was used to confirm the presence of active islets of Langerhans in the spleen. Microcytophotometry was used to quantify hepatocyte regeneration. RESULTS: Although demonstrating the presence of active (insulin-positive) islets together with hepatocytes in the spleen was feasible, the expected positive influence on the regenerative activity of hepatocytes was impossible to prove. CONCLUSIONS: Further study is needed to reach definitive conclusions about the real usefulness of pancreatic islets in hepatocyte transplantation.

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Mycorrhizal enhancement of drought resistance of two woody plant species, loblolly pine (Pinus taeda L.) and rose (Rosa hybrida L. cv. Ferdy), occurred independently of phosphorus nutrition. Mycorrhizae tended to alter root morphology and carbon allocation patterns of shoots and roots. Increased drought resistance of mycorrhizal plants was in part attributed to drought-induced colonization by mycorrhizae and the ability of the mycorrhizal plants to maintain high transpiration rates as a result of greater lateral root formation and lower shoot mass (in ectomycorrhizal loblolly pine), and a higher root/shoot ratio and leaf abscission (in endomycorrhizal roses). Neither the endo- nor ectomycorrhizal symbionts affected osmotic adjustment of droughted plants.

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Hepatic ischemia hinders the proliferative response of hepatocytes, necessary to restore the liver/body ratio after liver resection/transplantation. Folinic acid administered during the ischemic period following 70% hepatectomy plus 15 min of normothermic liver ischemia has restored the regenerative response to the levels of normoperfused livers. This unexpected finding has guided us to design the present study in order to find out whether the folinic acid is an hepatotrophic substance or not. Sprague-Dawley rats submitted to partial (40 or 70%) hepatectomies were used. Saline (2 cc) or folinic acid (2.5 mg/kg) have been administered i.v. Forty-eight hours after hepatectomy the hepatocyte's DNA content has been assessed by means of a cytophotometric technique, and the percentage of regenerating hepatocytes (PRH) has been calculated. Folinic acid administration has significantly increased the PRH in both resting (5.1 vs 1.2) and regenerating livers (70% hepatectomy) (22.2 vs 41) when compared with nontreated groups. Folinic acid administration after liver ischemia plus hepatectomy has shown similar results, corroborating our previous study. Although its mechanisms of augmentation of liver regeneration remain unclear and further studies are required, folinic acid seems to be a promising therapeutic tool in liver surgery.

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OBJECTIVE: To find out the incidence of wound infection in patients with HIV and reduced counts of CD4 lymphocytes. DESIGN: Open study. SETTING: University hospital, Spain. SUBJECTS: 70 patients with HIV infection and enlarged lymph nodes. INTERVENTIONS: Biopsy of lymph nodes and withdrawal of a sample of blood for counts of CD4 lymphocytes and neutrophils. MAIN OUTCOME MEASURE: Development of infection at the biopsy site, and correlation of infecting organism with culture taken at the time of biopsy. RESULTS: Patients were divided into three groups depending on their CD4 count: more than 500 cells/ml (n = 26), 200-500 cells/ml (n = 24), and less than 200 cells/ml (n = 20). Their neutrophil counts were 5.1, 3.8, and 2.5 x 10(9)/1, respectively. There were found four wound infections (6%); 2 were in the group with more than 500 CD4 cells/ml, and these were caused by Staphylococcus aureus (which had been grown from nodes in 6 patients at the time of biopsy). The other 2 were in the group with less than 500 cells/ml and these were caused by Mycobacterium tuberculosis; cultures of the nodes had shown Staphylococcus epidermidis (n = 3) and M tuberculosis (n = 17). There were no infections in the group with 200-500 CD4 cells/ml, in which S epidermidis (n = 5) and M tuberculosis (n = 8) had been cultured from the lymph nodes. CONCLUSIONS: The CD4 count was of no prognostic importance in the development of wound infection, but severe depression of the CD4 count may increase the risk of atypical wound infections.

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Liver regeneration plays a key role in restoring the liver/body ratio after partial liver transplantation. However, hepatic ischemia hinders the proliferative response of the hepatocytes. In this study, different ways of improving the regenerating capacity of ischemic hepatocytes are tested. Following 70% hepatectomy and 15 min of normothermic liver ischemia, the percentage of regenerating hepatocytes and the regenerative gradient are assessed. Cyclosporine A (hepatotrophic agent), superoxide dismutase and folinic acid (antioxidants), administered during the ischemic period, have significantly increased these indices. The later drug has restored the regenerative response to the levels of normoperfused livers.

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The aim of this study was to evaluate the efficacy of certain antioxidant and/or hepatotrophic drugs on the sensitivity to ischemia of hepatocytes implanted into the spleen. Twenty four hours after hepatocellular transplantation, animals were submitted to 15 minutes of normotermic ischemia followed by 70% hepatectomy. Hepatocytic function was assessed 24 h later by measuring the intensity of the regenerative response, both in the liver and in the spleen. All drugs increased the percentage of regenerating hepatocytes, but only allopurinol and cyclosporine achieved significance. However none of the drugs were useful for hepatocytes implanted in the spleen, allopurinol having a deleterious effect.

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An experimental model for the induction of hepatic metastasis by means of the subcutaneous injection of rhabdomyosarcoma cultured cells (S4MH) is described. The growth of the primary tumor and the dissemination process (local, ganglionar and hematogenous) are studied. The particular ability of the tumor to produce liver metastasis is assessed. The microscopic foci may be found in all of the specimens on day 25th post-injection. The maximum metastatic load is achieved by day 35 th. In contrast, the ganglionar and pulmonary metastasis were only found on the last step of the process.

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Ischemia is a common situation, not always desirable, in liver surgery. In any case, it implies cellular damage that would be worth avoiding. In the present study, different antioxidant drugs (superoxide-dismutase, allopurinol and folinic acid) are administered prior to liver reperfusion in order to reduce ischemic damage. Liver regeneration, following a 70% hepatectomy and 15 minutes of normothermic hepatic ischemia, serves as an indirect functional test of the reperfused liver. SOD (6 mg/kg) and allopurinol (50 mg/kg) have accelerated hepatocytic DNA synthesis without increasing the number or percentage of activated hepatocytes. However, the folinic acid has proved to be very effective, counteracting the deleterious effect of liver ischemia on hepatocytic regeneration.

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