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Biomedical subjects

C Lopez

Publications and source records attributed to C Lopez.

At least 19 recordsLinked to original sources

Functional interaction between Rh proteins and the spectrin-based skeleton in erythroid and epithelial cells.

We summarize the different experimental approaches which provide evidence that direct interaction of Rh and RhAG to ankyrin-R constitutes, together with the AE-1 (Band 3)-ankyrin-protein 4.2 and GPC-protein 4.1-p55 complexes, another major anchoring site between the red cell membrane bilayer and the underlying spectrin-based skeleton. The observations that some residues of the ankyrin binding site are mutated in Rh and RhAG proteins from some weak D and Rh(null) variants, respectively, suggest that the Rh-RhAG/ankyrin-R interaction plays a crucial role in the biosynthesis and/or the stability of the Rh complex in the red cell membrane. Similarly, binding to ankyrin G is required for cell surface expression of the non-erythroid member of the Rh protein family, RhBG, at the basolateral membrane domain of polarized epithelial cells. The next challenge will be to determine whether binding to the membrane skeleton may be critical for the emerging ammonium transport function of Rh proteins in erythroid and non-erythroid cells.

Animals↗

Clinical features and outcome of spontaneous bacterial peritonitis in HIV-infected cirrhotic patients: a case-control study.

The aim of this study was to evaluate the clinical characteristics and outcome of spontaneous bacterial peritonitis, a serious complication in patients with cirrhosis and ascites, in an HIV-infected cirrhotic population. Thirty-five HIV-infected cirrhotic patients who developed spontaneous bacterial peritonitis during a 12-year period were compared with 70 non-HIV-infected cirrhotic subjects. Patients were matched according to the date of the first episode of spontaneous bacterial peritonitis. A bacteriological diagnosis was made in 37 of 47 (79%) and in 50 of 97 (52%) episodes in the HIV group and in the non-HIV group, respectively (p=0.003), and Streptococcus pneumoniae was isolated more frequently in the HIV group (22 vs. 8%, p=0.02). Median survival after the initial diagnosis of spontaneous bacterial peritonitis was 2.9 and 14.0 months in the HIV group and non-HIV group, respectively. Age (hazard ratio [HR] 1.04; 95%CI 1.01-1.07), male sex (HR 2.55; 95%CI 1.34-4.83), Child-Pugh score at first spontaneous bacterial peritonitis episode (HR 1.29; 95%CI 1.10-1.54), renal impairment at first spontaneous bacterial peritonitis episode (HR 2.61; 95%CI 1.49-4.62), and HIV infection (HR 9.81; 95%CI 4.03-23.84) were independently associated with higher long-term mortality after the first diagnosis of spontaneous bacterial peritonitis. In conclusion, HIV-infected cirrhotic patients with spontaneous bacterial peritonitis have a higher rate of bacteriological diagnosis and a more frequent pneumococcal etiology than non-HIV-infected subjects. Life expectancy in these patients, once spontaneous bacterial peritonitis has developed, is poor. These data are particularly relevant for determining the optimal time for liver transplantation in this population.

Adult↗

Endogenous processes during long-term starvation in activated sludge performing enhanced biological phosphorus removal.

In many biological wastewater treatment systems, bacterial growth and the amount of active biomass are limited by the availability of substrate. Under these low growth conditions, endogenous processes have a significant influence on the amount of active biomass and therefore, the overall system performance. In enhanced biological phosphorus removal (EBPR) systems endogenous processes can also influence the levels of the internal storage compounds of the polyphosphate accumulating organisms (PAO), directly affecting phosphorus removal performance. The purpose of this study was to evaluate the significance of different endogenous processes that occur during the long-term starvation of EBPR sludge under aerobic and anaerobic conditions. Activated sludge obtained from a laboratory sequencing batch reactor was used to perform a series of batch starvation experiments. Under aerobic starvation conditions we observed a significant decay of PAO (first-order decay rate of 0.15/d) together with a rapid utilization of polyhydroxyalkanoates (PHA) and a slower utilization of glycogen and polyphosphate to generate maintenance energy. On the other hand, anaerobic starvation was best described by maintenance processes that rapidly reduce the levels of polyphosphate and glycogen under starvation conditions while no significant decay of PAO was observed. The endogenous utilization of glycogen for maintenance purposes is currently not included in available EBPR models. Our experimental results suggest that mathematical models for in EBPR should differentiate between aerobic and anaerobic endogenous processes, as they influence active biomass and storage products differently.

Aerobiosis↗

Altering renneting pH changes microstructure, cell distribution, and lysis of Lactococcus lactis AM2 in cheese made from ultrafiltered milk.

The objective of this study was to investigate the lysis of a highly autolytic strain of Lactococcus lactis ssp. cremoris AM2 in a model cheese made from concentrated ultrafiltered milk. From the same initial ultrafiltered retentate inoculated with L. lactis AM2, 5 cheeses were made by the addition of rennet at different pH values (6.6, 6.2, 5.8, 5.4, and 5.2). Lysis was monitored by measurement of the release of lactate dehydrogenase, an intracellular marker enzyme, and by immunodetection of intracellular proteins with species-specific antibodies. Confocal scanning laser microscopy (CSLM) was used to investigate the cheese microstructure by staining for protein and fat. Dual staining with a bacterial viability kit with CSLM was performed to reveal the integrity and localization of the bacterial cells. Levels of soluble calcium significantly increased when the pH at which the rennet was added decreased. In cheese renneted at pH 6.6, CSLM revealed an open porous structure containing a dense protein network with fat globules of different sizes distributed in the aqueous phase. In cheese renneted at pH 5.2, the protein network was homogeneous, with a less dense protein network, and an even distribution of fat globules. On d 1, bacterial cells were organized into colonies in cheese renneted at pH 6.6, whereas in cheeses renneted at pH 5.2, bacteria were evenly dispersed as single cells throughout the protein network. Lysis was detected on d 1 in cheeses renneted at high pH values and continued to increase throughout ripening, whereas induction of lysis was delayed in cheeses renneted at lower pH values until the end of ripening. This study demonstrates that alterations in the microstructure of the cheese and the distribution of cells play a role in lysis induction of L. lactis AM2.

Amino Acids↗

The linear relationship between the proportion of fresh grass in the cow diet, milk fatty acid composition, and butter properties.

Fresh grass in the cow diet improves the rheological and nutritional properties of butter. However, the relationship between the proportion of fresh grass in the diet and these properties is still unknown. The objective of the study was to determine the relationship between the proportion of fresh grass in the diet and the properties of milk and butter. Four groups of 2 cows were fed 4 isoenergetic diets characterized by increasing amounts of fresh grass (0, 30, 60, and 100% dry matter of forage) according to a Youden square design. Energy levels were similar among all diets. Thus, no effect of mobilization was observed and the results were only due to the proportion of fresh grass in the diet. Milk yield linearly increased with the proportion of fresh grass in the diet (+0.21 kg/d per 10% of grass). Fat yield remained unchanged. Thus, by effect of dilution, increasing the proportion of fresh grass in the diet induced a linear decrease in fat content. Milk fat globule size decreased by 0.29 mum when the proportion of grass reached 30% in the diet. Increasing the proportion of fresh grass in the diet induced a linear increase in unsaturated fatty acids percentages at the expense of saturated fatty acids. Relationships were +0.38, +0.12, +0.05 and -0.69 points/10% of fresh grass in the diet for C18:1 trans-11, C18:2 cis-9,trans-11, C18:3n-3, and C16:0, respectively. These modifications in fatty acid composition, and in particular in the spreadability index, C16:0/C18:1, were responsible for linear decreases in final melting temperature and solid fat content in butter fat, perceived in sensory analysis by a linear decrease in firmness in mouth. The nutritional value of butter was also linearly improved by the proportion of fresh grass in the diet by halving the atherogenicity index.

Animal Feed↗

Milk fat thermal properties and solid fat content in emmental cheese: a differential scanning calorimetry study.

The experiments reported in this study give deeper insight into the crystallization of milk fat in Emmental cheese, which is the most widely consumed hard cheese in France. Differential scanning calorimetry (DSC) was used to monitor the thermal properties of milk fat after the main stages involved during manufacture of Emmental cheese. By heating the samples to 60 degrees C to eliminate their thermal history and cooling them at 2 degrees C/min, the liquid --> solid phase transition of fat was investigated. Confocal laser scanning microscopy was used to characterize in situ the supramolecular organization of milk fat dispersed in the casein matrix. The destabilization of fat globules by aggregation or coalescence and the formation of free fat during the manufacture altered the thermal properties of milk fat by increasing the initial temperature of crystallization and by the formation of 2 overlapping exotherms. The melting properties of the crystalline structures formed by fat at the temperatures used for ripening (12, 21, and 4 degrees C) were examined. Differential scanning calorimetry was used to determine the ratio of solid to liquid fat; that is, the amount of fat that is crystallized, by dividing the partial enthalpy of melting of the fat for ripening temperature by the total enthalpy of melting of the same fat extracted from cheese. This study shows, for the first time, that milk fat is partially crystallized in Emmental cheese: about 55.7 +/- 3.5% of fat is solid at 4 degrees C at the end of ripening. Polymorphic phase transitions of milk fat are also suggested during ripening of Emmental cheese.

Animals↗

Using cDNA and genomic sequences as tools to develop SNP strategies in cassava (Manihot esculenta Crantz).

Single nucleotide polymorphisms (SNP) are the most abundant type of DNA polymorphism found in animal and plant genomes. They provide an important new source of molecular markers that are useful in genetic mapping, map-based positional cloning, quantitative trait locus mapping and the assessment of genetic distances between individuals. Very little is known on the frequency of SNPs in cassava. We have exploited the recently-developed collection of cassava expressed sequence tags (ESTs) to detect SNPs in the five cultivars of cassava used to generate the sequences. The frequency of intra-cultivar and inter-cultivar SNPs after analysis of 111 contigs was one polymorphism per 905 and one per 1,032 bp, respectively; totaling 1 each 509 bp. We have obtained further information on the frequency of SNPs in six cassava cultivars by analysis of 33 amplicons obtained from 3' EST and BAC end sequences. Overall, about 11 kb of DNA sequence was obtained for each cultivar. A total of 186 SNPs (136 and 50 from ESTs and BAC ends, respectively) were identified. Among these, 146 were intra-cultivar polymorphisms, while 80 were inter-cultivar polymorphisms. Thus the total frequency of SNPs was one per 62 bp. This information will help to develop new strategies for EST mapping as well as their association with phenotypic characteristics.

Computational Biology↗

Thermal and structural behavior of anhydrous milk fat. 3. Influence of cooling rate.

The crystallization behavior of anhydrous milk fat has been examined with a new instrument coupling time-resolved synchrotron x-ray diffraction as a function of temperature (XRDT) at both small and wide angles and high-sensitivity differential scanning calorimetry. Crystallizations were monitored at cooling rates of 3 and 1 degrees C/ min from 60 to -10 degrees C to determine the triacylglycerol organizations formed. Simultaneous thermal analysis permitted the correlation of the formation/melting of the different crystalline species monitored by XRDT to the thermal events recorded by differential scanning calorimetry. At intermediate cooling rates, milk fat triacylglycerols sequentially crystallize in 3 different lamellar structures with double-chain length of 46 and 38.5 A and a triple-chain length of 72 A stackings of alpha type, which are correlated to 2 exothermic peaks at 17.2 and 13.7 degrees C, respectively. A time-dependent slow sub-alpha <--> alpha reversible transition is observed at -10 degrees C. Subsequent heating at 2 degrees C/min has shown numerous structural rearrangements of the alpha varieties into a single beta' form before final melting. This polymorphic evolution on heating, as well as the final melting point observed (approximately 39 degrees C), confirmed that cooling at 3 degrees C/min leads to the formation of crystalline varieties that are not at equilibrium. An overall comparison of the thermal and structural properties of the crystalline species formed as a function of the cooling rate and stabilization time is presented. The influence on crystal size of the cooling rates applied in situ using temperature-controlled polarized microscopy is also determined for comparison.

Animals↗

Evaluation of microscopic techniques (epifluorescence microscopy, CLSM, TPE-LSM) as a basis for the quantitative image analysis of activated sludge.

Microscopic techniques ranging from epifluorescence microscopy to confocal laser scanning microscopy (CLSM) and two photon excitation laser scanning microscopy (TPE-LSM) combined with fluorescent stains can help to evaluate complex microbial aggregates such as activated sludge flocs. To determine the application limits of these microscopic techniques, activated sludge samples from three different sources were evaluated after staining with a fluorescent viability indicator (Baclight Bacterial Viability Kit, Molecular Probes). Image analysis routines were developed to quantify overall amounts of red and green stained cells, location of stained cells within the flocs, and the spatial organization in clusters and filaments. It was found that the selection of the appropriate microscopic technique depends strongly on the type of microbial aggregates being analyzed. For flocs with high cell density, the use of TPE-LSM is preferred, since it provides a clearer image of the internal structure of the aggregate. Epifluorescence microscopy did not allow to reliably quantify red stained cells in dense aggregates. CLSM did not adequately image the internal filamentous structure and the location of stained cells within dense flocs. However, for typical activated sludge flocs epifluorescence and CLSM proved adequate.

Flocculation↗

[Cryptorchidism: the point of view of a pediatric urologist].

Cryptorchidism is a very common anomaly of testicular migration, different from ectopy, anorchidy or retractile testis. Physiopathology of cryptorchidism is now well known. Hypogonadotrophic hypogonadism during pregnancy is mostly admitted. Recent epidemiologic studies show a significant increase due to toxic environmental factors, in particular pesticide. Cryptorchidism is most often isolated but polymalformatives entities must be eliminated, requiring sometimes hormonal or genetic investigations. Diagnosis is firstly clinical but coelioscopy adds accuracy and fiability. Infertility and cancer are the two majors risks of cryptorchidism, but their physiopathological mechanisms are still debated. It is not yet clear, in particular as far as the risk of infertility is concerned, if the testicular damage is primitive or (and) deteriorates with age. Medical and surgical treatments are now rather well codified. If, so far, there is no consensus about the age of management of cryptorchidism, many data of the literature lead pediatric urologists to manage these boys as soon as they are two or three.

Cryptorchidism↗

Energetic and kinematic consequences of weighting the distal limb.

REASON FOR PERFORMING STUDY: It is well known that adding a load to a horse's back increases its energetic costs of locomotion, but the magnitude of increase obtained by loading the most distal portion of limb has not been measured. OBJECTIVES: To measure oxygen consumption in horses with mass added to the back and hooves. Because such mass distribution alters inertial parameters of the limbs, kinematic measurements were made to quantify the magnitude of change in limb movement. METHODS: Steady-state oxygen consumption was measured in 6 horses with a load of 2.4 kg. The load was either carried on the back or distributed equally between the 4 limbs. Modified bell boots kept the mass at the level of P3. Horses trotted on a treadmill at speeds ranging from 2 to 5 m/sec (in 0.5 m/sec increments). High-speed (250 Hz) digital images were recorded in a sagittal plane and the positions of retroreflective markers located on standard positions on the limbs were digitised for kinematic analysis. RESULTS: Loading of the distal limbs produced a 6.7% increase in metabolic rate, an order of magnitude higher than when the mass was added over the back. Although the stride period was 2% longer in horses with loads on the distal limbs, time of contact and duty factor were not different. Distal limb loading increased the range of motion in hind- but not forelimbs. CONCLUSIONS: The costs of swinging the limbs in the horse are considerable and the addition of weights to the distal limb can have a profound effect on not only the energetics of locomotion but also the kinematics, at least in the hindlimb. POTENTIAL RELEVANCE: The use of weighted shoes, intended to increase animation of the gait, increases the metabolic effort of performance horses a disproportionate amount. The additional mass also increases the joint range of motion and, potentially, the likelihood of injury.

Animals↗

Oxidative stress and activation of proteasome protease during serum deprivation-induced apoptosis in rat hepatoma cells; inhibition of cell death by melatonin.

Growth factor deprivation-induced apoptosis has been shown in various cell systems and is recognized as one of the standard models for the study of programmed cell death. The mechanism of induction of apoptosis by serum deprivation is still not clear. The objective of the present study was to investigate if serum-deprivation causes oxidative stress, which then leads to apoptotic death. We have demonstrated that indeed, there was a significant increase in reactive oxygen species following serum deprivation of 5123tc hepatoma cells. Furthermore, treatment with anti-oxidants; melatonin or vitamin E, prevented cell death caused by serum-deprivation. We also demonstrated that there was activation of proteasome proteases and decrease in glutathione levels following serum deprivation. Interestingly, melatonin treatment blocked these changes and rescued the cells from apoptosis induced by serum-deprivation. These results indicated that oxidative stress may play a causal role in the induction of apoptosis induced by serum deprivation.

Animals↗

Endothoracic nodules in patients who under-went nephrectomy for renal cell carcinoma. Results of surgical resection.

AIM: A review of our experience with surgical resection of endothoracic nodules in patients who underwent nephrectomy for renal cell carcinoma (RCC) is presented, to evaluate the incidence of metastases in our series and the effectiveness and the opportunity of surgical treatment in this sort of patients. METHODS: Between January 1988 and January 2002, 41 consecutive patients (33 men, 8 women) underwent resection for suspected endothoracic metastases from RCC; 1 more male patient for metastases from an occult renal cancer. Mean age was 62 y (range: 43-80 y). Mean time between nephrectomy and 1st pulmonary resection in 41 patients was 29 mo (range: 0-120 mo). Nineteen patients had solitary lesions, 11 multiple unilateral and 12 bilateral. Antero-lateral thoracotomy was performed in 37 patients, median sternotomy in 1, simultaneous bilateral thoracotomy (clam-shell) in 2, sterno-laparotomy in 1, thoracofrenolaparotomy in 1. Wedge excision was performed in 36 patients, lobectomy with lymphadenectomy in 5, mediastinal limphadenectomy in 1. Six patients had repeat resection for recurrent metastases. RESULTS: Only 24 patients (57%) had histologic diagnosis of pulmonary metastases from RCC; 11 (26%) had benign lesions; 7 (17%) primary lung cancer. Mean follow-up was 25 mo (range: 1-91 mo). Overall, 4-y survival was 50%. Patients with solitary metastasis had a lower survival than those with 4 and more lesions. CONCLUSION: The evidence of pulmonary nodules in patients submitted to nephrectomy for RCC is not necessarily indicative of metastatic disease. Pulmonary resection for RCC metastases, even bilateral and recurrent, may help prolong survival in selected patients.

Adult↗

[Rare vesical malformations].

Ultrasonography has radically changed the approach of these rare malformations, still diagnosed with difficulties in spite of modern iconography. Bladder anomalies may easily be discovered when associated with otherwise obvious malformations of genitalia. The rest must be suspected for non-specific signs as micturition troubles, multiple urinary tract infections, chronic renal failure. The abnormal bladder is the only common feature of a polymorphic spectrum of diseases including complete or partial duplication or septum, congenital diverticula, vesicovaginal fistula, agenesis and urachal anomalies. Treatment must be accorded to each case and it needs perfect knowledge of open and endoscopic bladder surgery.

Female↗

[Anesthesia induction with sevoflurane in adult patients with predictive signs of difficult intubation].

OBJECTIVE: This work was carried out to study induction with sevoflurane in adult patients with predictive signs of difficult intubation. STUDY DESIGN: Randomised prospective study. PATIENTS AND METHODS: The study had two parts. Part I: 15 patients without predictive signs of difficult intubation but with a cervical collar. Eight patients were anaesthetised with propofol 3 mg.kg-1 and fentanyl 2 micrograms.kg-1, seven with sevoflurane 8%. Part II: 20 patients with predictive signs of difficult intubation anaesthetised with sevoflurane 8%. RESULTS: In part I, all patients were intubated, the time for intubation was longer with sevoflurane, 6 vs 4 min. They were apneic only in the propofol group. After intubation, 7 cases of coughing (4 severe) occurred in the propofol group and 3 moderate coughing in the sevoflurane group. In part II, one patient experienced considerable agitation after oral airway insertion and was excluded. Other patients were intubated with sevoflurane. Seven patients were intubated with a bougie, three patients through an intubating LMA and one patient with a rigid bronchoscope. The other patients were intubated with a Macintosh blade. The mean time for intubation was 10 +/- 7 min and end tidal sevoflurane concentration after intubation was 4 +/- 0.6%. After intubation, 7 cases of coughing (3 severe) occurred but no desaturation < 95%. No significant haemodynamic variations occurred. CONCLUSION: Induction with sevoflurane 8% allowed tracheal intubation without major incidents. All patients breathed spontaneously. Sevoflurane can be recommended for induction in cases of predictive difficult intubation.

Adult↗

Transcriptional regulation of the KEL gene and Kell protein expression in erythroid and non-erythroid cells.

The Kell blood-group antigen was originally reported to be a protein expressed in erythroid tissue only. Transcriptional analysis of the KEL promoter activity in human erythroleukaemia K562 and epithelial HeLa cells by electrophoretic mobility-shift and supershift assays, chloramphenicol acetyltransferase assays, co-transfection studies and site-directed mutagenesis provided the following results: (i) the KEL promoter exhibits a strong transcriptional activity in K562 cells and, unexpectedly, a basal non-erythroid activity in HeLa cells, (ii) up-regulation of the 5' distal promoter activity occurs only in the erythroid context, and (iii) two motifs localized in the exon 1 region, which bind the Sp1/Sp3 and the human GATA-1/Ku70/80 factors, were required for down-regulation of the promoter activity, but inhibition of the promoter activity by the repressing factors in HeLa cells was incomplete. KEL expression in HeLa cells was performed further by primer-extension analysis, which revealed the presence of a low amount of Kell transcript correlating with basal expression of the Kell protein in these cells, as shown by immunopurification and Western-blot analysis. DNA sequencing of the transcript revealed a sequence identical to that obtained from erythroid tissue. In human tissues, KEL expression was investigated by dot-blot analysis and revealed high levels of Kell mRNAs, particularly in brain tissues, testis and lymphoid tissues. Moreover, most tissues analysed exhibited low levels of Kell transcripts. The Kell protein was also detected by immunohistochemistry in the Sertoli cells of the testis and in lymphoid tissues like spleen and tonsil, specifically localized in the follicular dendritic cells. Altogether, the results indicated that KEL expression is not restricted to erythroid tissue.

Antigens, Nuclear↗

Immunological and hematological response in experimental Toxocara canis-infected pigs.

The relationship between the immunological and hematological response to infection was studied in pigs inoculated experimentally with Toxocara canis. Two groups of four pigs were infected with doses of 1000 and 2000 infective eggs, respectively. Two uninfected animals were used as negative controls. Blood samples were collected from each pig once a week. Serological examination by ELISA to determine antibody levels against T. canis L2/L3 excretory-secretory (ES) antigens showed values higher than the positive cut-off point (1:32) for both the infected groups. These values increased from day 7 p.i. and remained high during the experimental period until day 56. Significant differences were recorded for the two inoculating doses (p</=0.01) from day 14 to 42 p.i. Western blots run on day 7 p.i. with serum from the 2000 eggs group showed two polypeptide bands of 70 and 55kDa. These same bands were observed for both infected groups from day 15 p.i. until the end of the experiment. Eosinophil counts were significantly higher (p</=0.05) for both inoculated groups between weeks 2 and 7 p.i. as compared with the control group. No significant changes were observed for other blood parameters. The pig is a useful animal model since T. canis induces an early immunological response and ELISA can detect the presence of these antibodies against T. canis L(2)/L(3) ES antigens in animals inoculated with low doses of Toxocara eggs.

Animals↗

The Composition of the Milk Fat Globule Surface Alters the Structural Characteristics of the Coagulum.

The effects of the composition of a fat globule surface in reconstituted milks on the properties of rennet-induced coagulums were studied by rheological measurements and by front-face fluorescence spectroscopy in combination with a multivariate statistical method to investigate, at a molecular level, the evolution of the structure during the milk coagulation process. Reconstituted milks used in this study were prepared from different fat-in-water emulsions stabilized by whole skim-milk proteins, beta-casein, or beta-lactoglobulin. Coagulation of milk reconstituted with natural fat globules was also investigated. The study showed that the fat droplet/water interface influences the rheological properties (G' modulus) of the reconstituted milks during the coagulation process. The tryptophan fluorescence emission spectra of proteins were recorded during the kinetics of coagulation. The results of the principal component analysis performed on the spectral data showed a discrimination in the different systems investigated. It was shown that the fluorescence properties of protein tryptophans and, consequently, the structures of the protein networks were different for the investigated systems. The development of fluorescence transfer between protein tryptophans and fat-globule vitamin A during the coagulation kinetics agreed with the interactions between the protein network and fat globules. Copyright 2001 Academic Press.

Journal Article↗