935 resultados para Nacl
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Oxidative metabolism of isolated toad skin epithelium (Bufo viridis) was investigated in vitro under open-circuit conditions using the spectrophotometric oxyhemoglobin micromethod. This highly sensitive technique has been adapted for studying several epithelia in parallel and for detecting possible regional variations of oxygen uptake in individual epithelium. Changes in the proportion of mitochondria-rich cells (MRC) by ionic acclimation affected oxidative metabolism under nontransporting condition. After acclimation of animals to either NaNO3 or NaCl solutions (100 mmol/l, for greater than 2 wk), the number of MRC per square millimeter in epithelia from nonacclimated and NaNO3- and NaCl-acclimated animals was 350 +/- 113, 460 +/- 196, and 107 +/- 52, respectively. O2 uptake of nonacclimated and NaNO3-acclimated epithelia was significantly higher than that of NaCl-acclimated epithelia (i.e., 0.89 and 0.90 vs. 0.57 nmol O2.h-1.mm-2, respectively). The correlation established between O2 uptake and number of MRC allowed evaluation of the respiration rate of one single MRC, i.e., approximately 1 pmol O2/h. The lowest mitochondrial oxidative activity was found in the epithelia from NaCl-acclimated toads where the uncoupler 2,4-dinitrophenol (50 mumols/l) had the highest relative stimulatory effect (+114%). Acetazolamide (50 mumols/l), a potent inhibitor of carbonic anhydrase mainly present in the MRC, reduced selectively by 31% O2 uptake of the MRC-rich epithelia (NaNO3 acclimated). O2 uptake increased significantly by approximately 80% when basolateral pH increased from 5.8 to 7.8, but did not depend on apical pH. These findings indicate that under nontransporting (open-circuit) conditions, aerobic metabolism of the isolated toad skin epithelium is related to the density and/or characteristics of the MRC.(ABSTRACT TRUNCATED AT 250 WORDS)
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Regulation of sodium balance is a critical factor in the maintenance of euvolemia, and dysregulation of renal sodium excretion results in disorders of altered intravascular volume, such as hypertension. The amiloride-sensitive epithelial sodium channel (ENaC) is thought to be the only mechanism for sodium transport in the cortical collecting duct (CCD) of the kidney. However, it has been found that much of the sodium absorption in the CCD is actually amiloride insensitive and sensitive to thiazide diuretics, which also block the Na-Cl cotransporter (NCC) located in the distal convoluted tubule. In this study, we have demonstrated the presence of electroneutral, amiloride-resistant, thiazide-sensitive, transepithelial NaCl absorption in mouse CCDs, which persists even with genetic disruption of ENaC. Furthermore, hydrochlorothiazide (HCTZ) increased excretion of Na+ and Cl- in mice devoid of the thiazide target NCC, suggesting that an additional mechanism might account for this effect. Studies on isolated CCDs suggested that the parallel action of the Na+-driven Cl-/HCO3- exchanger (NDCBE/SLC4A8) and the Na+-independent Cl-/HCO3- exchanger (pendrin/SLC26A4) accounted for the electroneutral thiazide-sensitive sodium transport. Furthermore, genetic ablation of SLC4A8 abolished thiazide-sensitive NaCl transport in the CCD. These studies establish what we believe to be a novel role for NDCBE in mediating substantial Na+ reabsorption in the CCD and suggest a role for this transporter in the regulation of fluid homeostasis in mice.
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The purpose of this study was to design microspheres combining sustained delivery and enhanced intracellular penetration for ocular administration of antisense oligonucleotides. Nanosized complexes of antisense TGF-beta2 phosphorothioate oligonucleotides (PS-ODN) with polyethylenimine (PEI), and naked PS-ODN were encapsulated into poly(lactide-co-glycolide) microspheres prepared by the double-emulsion solvent evaporation method. The PS-ODN was introduced either naked or complexed in the inner aqueous phase of the first emulsion. We observed a marked influence of microsphere composition on porosity, size distribution and PS-ODN encapsulation efficiency. Mainly, the presence of PEI induced the formation of large pores observed onto microsphere surface. Introduction of NaCl in the outer aqueous phase increased the encapsulation efficiency and reduced microsphere porosity. In vitro release kinetic of PS-ODN was also investigated. Clearly, the higher the porosity, the faster was the release and the higher was the burst effect. Using an analytical solution of Fick's second law of diffusion, it was shown that the early phase of PS-ODN and PS-ODN-PEI complex release was primarily controlled by pure diffusion, irrespectively of the type of microsphere. Finally, microspheres containing antisense TGF-beta2 nanosized complexes were shown, after subconjunctival administration to rabbit, to significantly increase intracellular penetration of ODN in conjunctival cells and subsequently to improve bleb survival in a rabbit experimental model of filtering surgery. These results open up interesting prospective for the local controlled delivery of genetic material into the eye.
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PURPOSE: To test the ability of two preparations of FGF2-saporin, either FGF2 chemically conjugated to saporin (FGF2-SAP) or genetically engineered FGF2-saporin (rFGF2-SAP) to inhibit the growth of bovine epithelial lens (BEL) cells in vitro when in solution and when immobilized on heparin surface-modified (HSM) polymethylmethacrylate (PMMA) intraocular lenses (IOLs). METHOD: Bovine epithelial lens cells were incubated with various concentrations FGF2-saporin for as long as 4 days. The number of surviving cells was determined by counting the number of nuclei. Because FGF2 binds to heparin, FGF2-saporin was incubated with HSM PMMA IOLs; excess toxin was washed off, and the BEL cells were grown on the FGF2-saporin-treated IOLs (HSM and non-HSM) for 4 days. Cell density was determined by image analysis. RESULTS: Both FGF2-SAP and rFGF2-SAP were highly cytotoxic (nM range), with rFGF2-SAP 10 times less active than FGF2-SAP. FGF2-saporin bound to the surface of HSM IOLs and eluted by 2M NaCl retained its activity. Toxin bound to HSM IOLs killed more than 90% of the BEL cells placed on the IOL surface within 4 days. The ability of FGF2-saporin to prevent the growth of cells on the IOL surface was strictly dependent on the presence of heparin on the IOL. CONCLUSIONS: FGF2-saporin is bound to HSM PMMA IOLs and prevents the growth of epithelial cells on the surface of the lens.
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O presente estudo foi realizado com o objetivo de identificar a prevalência de colonização pelo Staphylococcus aureus em profissionais de enfermagem de um hospital universitário de Pernambuco, bem como avaliar o perfil de resistência deles isoladamente. Para isso, foi realizado um estudo transversal, no qual foram coletadas amostras biológicas das mãos e da cavidade nasal. A identificação do S. aureus foi realizada por meio do semeio em agar-sangue, agar manitol-salgado e através dos testes de catalase e coagulase. O perfil de sensibilidade foi determinado pela técnica de Kirby Bauer e para determinação da resistência à meticilina foi realizado o screening em placa com oxacilina com adição de 4% de NaCl. Dos 151 profissionais avaliados, 39 se encontravam colonizados, o que demonstrou uma prevalência de 25,8%. Dentre as variáveis estudadas, a faixa etária e a quantidade de EPI apresentaram-se associadas à colonização pelo microrganismo. De todas as linhagens isoladas, apenas cinco apresentaram resistência à meticilina.
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Estudo experimental para aferição do potencial hidrogeniônico (pH) dos antimicrobianos ceftriaxona sódica, cloridrato de vancomicina, metronidazol, penicilina G potássica e sulfato de amicacina, após reconstituição, diluição com NaCl 0,9% (SF) e soro glicosado 5% (SG), em oito momentos distintos e sob condições cotidianas de luminosidade e temperatura ambiente de unidade hospitalar não climatizada. O objetivo deste estudo foi verificar alterações no comportamento ácido-básico das soluções, indicativas de instabilidade química ou relacionadas a complicações da terapia intravenosa. Nos 186 valores de pH analisados, não foram identificadas variações maiores que 1,0 valor nem alterações físicas visíveis a olho nu. Todas as soluções tiveram pH menor que 7 e não houve diferença considerável para a prática clínica segundo o diluente. As médias dos valores de pH após a diluição em SF e SG, do cloridrato de vancomicina, metronidazol e sulfato de amicacina constituem fator de risco para o desenvolvimento de complicações intravenosas devido a sua extrema acidez.
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Zeta potential is a physico-chemical parameter of particular importance to describe sorption of contaminants at the surface of gas bubbles. Nevertheless, the interpretation of electrophoretic mobilities of gas bubbles is complex. This is due to the specific behavior of the gas at interface and to the excess of electrical charge at interface, which is responsible for surface conductivity. We developed a surface complexation model based on the presence of negative surface sites because the balance of accepting and donating hydrogen bonds is broken at interface. By considering protons adsorbed on these sites followed by a diffuse layer, the electrical potential at the head-end of the diffuse layer is computed and considered to be equal to the zeta potential. The predicted zeta potential values are in very good agreement with the experimental data of H-2 bubbles for a broad range of pH and NaCl concentrations. This implies that the shear plane is located at the head-end of the diffuse layer, contradicting the assumption of the presence of a stagnant diffuse layer at the gas/water interface. Our model also successfully predicts the surface tension of air bubbles in a KCl solution. (c) 2012 Elsevier Inc. All rights reserved.
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Introduction : Au CHUV, les contrôles microbiologiques des préparations pharmaceutiques stériles produites par la pharmacie de l'hôpital se basent sur l'essai de stérilité de la Pharmacopée Européenne. Avant 2000, une méthode en circuit ouvert était utilisée, puis, dès l'année 2000, une nouvelle méthode développée par H. Ing du service de pharmacie des HUG a été adoptée (méthode « Ing »). Cette dernière permet d'opérer en circuit fermé et de filtrer le milieu de culture. De plus, elle utilise du matériel bon marché (trousse de perfusion, filtres à usage unique). Objectifs : Le présent travail avait pour but : 1) l'évaluation préliminaire de cette méthode (validation). 2) l'évaluation du bénéfice apporté en terme d'incidence de faux positifs sur les préparations stériles filtrables. Matériel et méthode : La validation a été effectuée en analysant des flexs de NaCl 0.9% préalablement inoculés avec 10-100 CFU de 6 souches microbiennes décrites dans la Pharmacopée pour le test de validation, ainsi qu'un flex « contrôle » non inoculé. Le bénéfice de la méthode a été évalué à partir des résultats des essais de routine effectués au laboratoire. Un taux de faux positifs imputable à chaque méthode a ainsi pu être déterminé (i.e. croissance microbienne due à une contamination lors de l'essai et non à une contamination initiale de la préparation pharmaceutique) et la comparaison a été effectuée à l'aide du test statistique de Fisher. Résultats : Une croissance a été observée dans toutes les préparations préalablement inoculées par des micro-organismes. La méthode a donc pu être implantée dans le laboratoire pour les analyses de routine dès février 2000. L'analyse rétrospective des résultats des essais de stérilité effectués sur une période de plus de 4 ans (2 ans avec l'ancienne méthode (système ouvert) et de presque 3 ans avec la nouvelle méthode) montre que l'ancienne méthode produisait un taux de faux positifs de 1.57 %, alors que ce taux n'est que de 0.21% avec la méthode « Ing ». Cette dernière se caractérise donc par un taux de faux positifs significativement plus bas que celui de l'ancienne méthode (p < 0.0001). Conclusion : La méthode « Ing » constitue une technique bien adaptée à l'essai de stérilité pour l'hôpital, suffisamment sensible et conforme aux recommandations de la Pharmacopée. En maintenant le produit dans un espace clos, elle permet de diminuer les risques de contamination susceptibles de se produire lors de l'essai.
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Salt taste in mammals can trigger two divergent behavioural responses. In general, concentrated saline solutions elicit robust behavioural aversion, whereas low concentrations of NaCl are typically attractive, particularly after sodium depletion. Notably, the attractive salt pathway is selectively responsive to sodium and inhibited by amiloride, whereas the aversive one functions as a non-selective detector for a wide range of salts. Because amiloride is a potent inhibitor of the epithelial sodium channel (ENaC), ENaC has been proposed to function as a component of the salt-taste-receptor system. Previously, we showed that four of the five basic taste qualities-sweet, sour, bitter and umami-are mediated by separate taste-receptor cells (TRCs) each tuned to a single taste modality, and wired to elicit stereotypical behavioural responses. Here we show that sodium sensing is also mediated by a dedicated population of TRCs. These taste cells express the epithelial sodium channel ENaC, and mediate behavioural attraction to NaCl. We genetically engineered mice lacking ENaCalpha in TRCs, and produced animals exhibiting a complete loss of salt attraction and sodium taste responses. Together, these studies substantiate independent cellular substrates for all five basic taste qualities, and validate the essential role of ENaC for sodium taste in mice.
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Neste trabalho estudou-se a lixiviação de um concentrado de sulfureto de zinco e a recuperação de Fe(III) de soluções sulfúricas por extracção líquido-líquido. Nos estudos de lixiviação utilizou-se o ião férrico (sulfato de ferro ou cloreto de ferro) como agente oxidante e avaliaram-se os efeitos da razão sólido/líquido, concentração do ião Fe(III) e o tipo do meio (sulfúrico e clorídrico) na eficiência da lixiviação. Os resultados mostraram que para uma razão sólido/líquido de 5% foi possível lixiviar 48% de zinco e 15% do cobre em 2h de lixiviação com uma solução de 0,5 M H2SO4 e 0,11 M Fe2(SO4)3 a 80 ºC. A lixiviação do cobre é favorecida em meio clorídrico onde cerca de 23% de cobre foi lixiviado, após 2h, com uma razão sólido/líquido de 20% e uma solução de 0,5 M FeCl3, 2 M HCl e 1 M NaCl a 80 ºC, quando em meio sulfúrico tinha sido possível lixiviar apenas 6% deste metal. A realização de dois andares de lixiviação, sendo o primeiro em meio sulfúrico e o segundo em meio clorídrico, permitiu aumentar as taxas de lixiviação dos metais, tendo-se obtido um rendimento global de 45% de Zn e 23% de Cu nas seguintes condições experimentais: s/l=20% (p/v), T=80ºC, v=350 rpm, 1º andar com [Fe2(SO4)3]=0,25 M; [H2SO4]=2 M e 2º andar com [FeCl3]=0,5 M; [HCl]=2 M; [NaCl]=1 M. Nos estudos de extracção líquido-líquido para a recuperação de Fe(III) de meios sulfúricos utilizaram-se misturas de extractantes, i.e. um ácido organofosforado (Ionquest ou DEHPA) com uma amina primária (JMT). Foi, ainda, testada a presença de um modificador (isotridecanol) no solvente. As várias misturas revelaram uma elevada afinidade para o ferro na gama de pH’s 1,04 a 1,74 e elevada selectividade para a separação ferro/zinco. Os factores de separação Fe/Zn atingem valores na gama 1000-8000, sendo favorecidos pelas elevadas concentrações de ferro na fase orgânica. A presença de modificador no solvente dificultou a extracção de ferro da fase aquosa mas favoreceu a respectiva reextracção da fase orgânica carregada que foi efectuada com uma solução 50 g/L de H2SO4.
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BACKGROUND: Sodium wasting during the night has been postulated as a potential pathophysiological mechanism in patients suffering from orthostatic hypotension due to severe autonomic deficiency. METHODS: In this study, the diurnal variations in creatinine clearance, sodium excretion and segmental renal tubular handling of sodium were evaluated in 18 healthy subjects and 20 young patients with orthostatic hypotension (OH). In addition, 24-hour ambulatory blood pressure and the neuro-hormonal response to changes in posture were determined. The patients and their controls were studied on a free sodium intake. In a second protocol, 10 controls and 10 patients were similarly investigated after one week of a high salt diet (regular diet + 6 g NaCl/day). RESULTS: Our results demonstrate that, in contrast to normal subjects in whom no significant changes in glomerular filtration, sodium excretion and segmental sodium reabsorption were observed throughout the day, patients with OH were characterized by a significant increase in glomerular filtration rate during the nighttime (P = 0.03) and significant increases in urinary lithium excretion (P < 0.05) and lithium clearance (P = 0.05) during the night, suggesting a decreased proximal reabsorption of sodium. On a high sodium diet, the symptoms of orthostatic hypotension and the circadian variations in sodium reabsorption were significantly blunted. CONCLUSIONS: These results suggest that, while the patient is in a supine position the effective blood volume of those with OH becomes excessive due to the increased venous return. Hence, the kidney responds with an increase in glomerular filtration and a relative escape of sodium from the proximal tubular segments. These circadian variations in renal sodium handling may contribute to the maintenance of the orthostatic syndrome.
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Le rein joue un rôle essential dans le maintien de l'homéostasie des fluides extracellulaires (FEC) et la pression artérielle. L'objectif de notre groupe est d'identifier de nouveaux mécanismes impliqués dans le contrôle de l'homéostasie des FEC et de la pression artérielle par le rein. Projet 1) Caractérisation du rôle fonctionnel du récepteur à l'a-cétogluatarate Oxgrl dans le rein Oxgrl est le récepteur spécifique de l'a-cétogluatarate, une moléule intermédiaire du cycle de l'acide citrique, filtrée par le rein et réabsorbée ou secrétée au niveau des tubules proximaux. Le rôle fonctionnel de ces deux récepteurs reste inconnu. Nos résultats montrent qu'Oxgrl est localisé au niveau des cellules intercalaires du tube collecteur (CCD). Des souris (Oxgrr/_) montrent une diminution du pH urinaire ,une augmentation de la concentration de l'acide urinaire titrable et une augmentation des niveaux d'a-cétoglutarate. Le traitement au Na-bicarbonate provoque une augmentation plus prononcée de l'alcalose métabolique chez les souris Oxgrl"7"' accompagnée d'une augmentation de la concentration de bicarbonate et une diminution du niveau de chlore plasmatique. En parallèle, des études de microperfusion ont montré que a-cétoglutarate stimule la réabsorption éléctroneutre de NaCl dans le CCD des souris de type sauvage mais pas des souris Oxgrl"7". En résumé, ces résultats montrent que l'a-cétoglutarate joue un rôle de molécule messagère du tubule proximal jusqu'au tube collecteur au niveau du rein et qu'Oxgrl pourrait être impliqué dans la régulation de l'échange Cl/bicarbonate et la réabsorption du NaCl dans les cellules intercalées. Projet 2) Rôle du système circadien dans les cellules productrices de rénine. Le système chronologique circadien est un mécanisme moléculaire ubiquitaire qui permet à l'organisme de coordonner ses fonctions principales en fonction du temps géophysique. Comme l'activité de la rénine plasmatique montre une rythmicité circadienne nette chez l'homme et la souris ; dans ce projet, nous avons abordé la question à savoir dans quelle mesure le système circadien est impliqué dans cette variabilité circadienne. Pour cela, le gène Bmall, élément principal de l'horloge moléculaire, a été perturbé dans les cellules granulaires productrices de rénine par le système Cre-LoxP. Nos résultats montrent que les souris Renld- Cre/Bmalllox/lox (cKO) présentent de faibles taux d'ARNm de Reni, altèrent la dynamique d'expression de la protéine rénine, mais il le niveau de concentration plasmatique de la rénine reste le même. Cependant, les souris cKO montrent une réduction significative de la concentration plasmatique de l'aldostérone. Nos analyses de l'urine récupérée dans des intervalles de temps de 24 et 1 heure montrent une augmentation du volume urinaire, une tendance à une hypercalciurie, ainsi qu'une altération de la dynamique d'excrétion urinaire de sodium chez les souris cKO. Plusieurs gènes impliqués dans la production/sécrétion de la rénine et dans le contrôle de la fonction rénale montrent une altération de l'expression circadienne d'ARNm. Par ailleurs, les souris cKO montrent une baisse significative de la pression artérielle. Nos résultats suggèrent que l'horloge intrinsèque des cellules productrices de la rénine joue un rôle important dans le control des FEC et l'homéostasie de la pression artérielle via régulation de la fonction rénale.
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Os Estafilococos Coagulase Negativa (SCN) são bactérias isoladas a partir de uma ampla variedade de produtos cárneos fermentados. Os SCN possuem diversas propriedades tecnológicas importantes, das quais se destaca a capacidade de multiplicação em diferentes condições de temperatura, cloreto de sódio e pH. Mas também podem apresentar aspetos negativos, como a presença de resistências a antibióticos. Assim, o presente trabalho teve como objetivo selecionar estirpes sem qualquer resistência secundária a antibióticos e avaliar a capacidade desses selecionados para desenvolverem em temperaturas de 7 ºC, 15 ºC e 25 ºC, na presença de 2 %, 4 % e 6 % de NaCl e a valores de pH 4,5, 5,5 e 6,5. A partir dos resultados de antibioresistências, foram escolhidos 5 isolados de Staphylococcus xylosus (C1C3, L1M2, P1B2, S2M7 e S3M3) e 4 isolados de Staphylococcus equorum (Ch3C2, Cv1C2, P1B6 e SG3C5). Face às diferentes condições de estudo, dos resultados obtidos concluiu-se que as estirpes L1M2, S2M7, P1B2, Ch3C2 e Cv1C2, pelas suas capacidades de multiplicação e adaptação, são as mais indicadas para serem utilizadas em culturas de arranque
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Studies assessing skin irritation to chemicals have traditionally used laboratory animals; however, such methods are questionable regarding their relevance for humans. New in vitro methods have been validated, such as the reconstructed human epidermis (RHE) model (Episkin®, Epiderm®). The comparison (accuracy) with in vivo results such as the 4-h human patch test (HPT) is 76% at best (Epiderm®). There is a need to develop an in vitro method that better simulates the anatomo-pathological changes encountered in vivo. To develop an in vitro method to determine skin irritation using human viable skin through histopathology, and compare the results of 4 tested substances to the main in vitro methods and in vivo animal method (Draize test). Human skin removed during surgery was dermatomed and mounted on an in vitro flow-through diffusion cell system. Ten chemicals with known non-irritant (heptylbutyrate, hexylsalicylate, butylmethacrylate, isoproturon, bentazon, DEHP and methylisothiazolinone (MI)) and irritant properties (folpet, 1-bromohexane and methylchloroisothiazolinone (MCI/MI)), a negative control (sodiumchloride) and a positive control (sodiumlaurylsulphate) were applied. The skin was exposed at least for 4h. Histopathology was performed to investigate irritation signs (spongiosis, necrosis, vacuolization). We obtained 100% accuracy with the HPT model; 75% with the RHE models and 50% with the Draize test for 4 tested substances. The coefficients of variation (CV) between our three test batches were <0.1, showing good reproducibility. Furthermore, we reported objectively histopathological irritation signs (irritation scale): strong (folpet), significant (1-bromohexane), slight (MCI/MI at 750/250ppm) and none (isoproturon, bentazon, DEHP and MI). This new in vitro test method presented effective results for the tested chemicals. It should be further validated using a greater number of substances; and tested in different laboratories in order to suitably evaluate reproducibility.
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Para el estudio de la microalga Dunaliella salina Teodoresco, se colectaron muestras de 2 lagunas hipersalinas; las Salinas de Chimus y las Salinas de Chilca. La metodología usada fue la técnica de micropipeta, tratamiento con antibiótico y la técnica de sedimentación algal, obteniendo cultivos unialgales y axénicos. El medio usado fue medio Johnson modificado en agua de mar, utilizado concentraciones de NaCl (1 – 5M), éste permitió evaluar el crecimiento, densidad y velocidad algal a través de conteo en cámara de Newbahuer y lecturas de absorvancia en espectrofotómetro. La relación entre los métodos de conteo, se realizó con el análisis de regresión potencial. Las mejores densidades algales se observaron en cultivo de concentraciones 1M, 1,5M y 3,5M de NaCl, para las cepas de las salinas de Chimus y Chilca. La mayor densidad algal ( 4,603 x 106 cels. ml-1 equivalente al 56.4 % para la cepa de Chimus) se observó en el cultivo de 1,5M de NaCl. La velocidad de crecimiento durante la fase exponencial para ambas cepas, estuvieron entre 0,56-0,83 div. día-1, con un Td de 29-43 horas, en cámara de Newbahuer como en espectrofotómetro y a las mismas concentraciones de cultivo.