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1.
BMC Plant Biol ; 24(1): 733, 2024 Jul 31.
Artículo en Inglés | MEDLINE | ID: mdl-39085764

RESUMEN

BACKGROUND: Improving the quality and shelf life of groundnut oil is one of the foremost objectives of groundnut breeding programmes. This can be achieved by marker-assisted introgression, a technique that efficiently and precisely enables breeders to develop plants with enhanced qualities. This study focused on improving the oleic acid content of an elite groundnut variety, TMV 7, by introgressing a recessive mutation responsible for the increase in oleic acid from ICG 15419. Hybridization was performed between the donor and recurrent parents to develop the F1, BC1F1, BC2F1 and BC2F2 populations. Introgressed lines with increased oleic acid in the genetic background of TMV 7 were identified using allele-specific marker, F435-F, F435SUB-R and a set of SSR markers were employed to recover the genome of the recurrent parent. RESULTS: With two backcrosses, a total of ten homozygous plants in the BC2F2 population were identified with oleic acid content ranging from 54.23 to 57.72% causing an increase of 36% over the recurrent parent. Among the ten lines, the line IL-23 exhibited the highest level of recurrent parent genome recovery of 91.12%. CONCLUSIONS: The phenotypic evaluation of 10 homozygous introgressed lines indicated fewer differences for all other traits under study compared to the recurrent parent, except for oleic acid and linoleic acid content confirming the genetic background of the recurrent parent. The identified lines will be subjected to multilocation trials before their commercial release.


Asunto(s)
Arachis , Ácido Oléico , Fitomejoramiento , Ácido Oléico/metabolismo , Arachis/genética , Arachis/metabolismo , Fitomejoramiento/métodos , Marcadores Genéticos , Introgresión Genética , Aceites de Plantas/metabolismo
2.
J Environ Manage ; 351: 119917, 2024 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-38183950

RESUMEN

Air conditioners alleviate the discomfort of human beings from heat waves that are consequences of climate change caused by anthropogenic activities. With each passing year, the effects of global warming worsen, increasing the growth of air conditioning industry. Air conditioning units produce substantial amounts of non-nutritive and (generally) neglected condensate water and greenhouse gases. Considering this, the study explored the potential of using air conditioner condensate water (ACW) to cultivate Chlorella sorokiniana, producing biomass, and sequestering carbon dioxide (CO2). The maximum biomass production was obtained in the BG11 medium (1.45 g L-1), followed by ACW-50 (1.3 g L-1). Similarly, the highest chlorophyll-a content was observed in the BG11 medium (11 µg mL-1), followed by ACW-50 (9.11 µg mL-1). The ACW-50 cultures proved to be better adapted to physiological stress (Fv/Fm > 0.5) and can be suitable for achieving maximum biomass with adequate lipid, protein, and carbohydrate production. Moreover, C. sorokiniana demonstrated higher lipid and carbohydrate yields in the ACW-50 medium, while biomass production and protein yields were comparable to the BG11 medium. The lipid, protein, and carbohydrate productivity were 23.43, 32.9, and 23.19 mg L-1 d-1, respectively for ACW-50. Estimation of carbon capture potential through this approach equals to 9.5% of the total emissions which is an added advantage The results indicated that ACW could be effectively utilized for microalgae cultivation, reducing the reliance on freshwater for large-scale microalgal biomass production and reduce the carbon footprints of the air conditioning industry.


Asunto(s)
Chlorella , Microalgas , Humanos , Dióxido de Carbono/metabolismo , Microalgas/metabolismo , Lípidos , Agua/metabolismo , Biomasa , Carbohidratos
3.
Mol Vis ; 28: 11-20, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-35400989

RESUMEN

Purpose: Glutathione peroxidase 1 (GPX1) and catalase are expressed in the lens epithelial cells and cortical fiber cells, where they detoxify H2O2 to reduce oxidative stress, which is a major cause for cataractogenesis. We sought to find out, between these two enzymes, which is critical for transparency and homeostasis in the aging lens by investigating alterations in the lens's refractive property, transparency, and gap junction coupling (GJC) resistance. Methods: Wild-type (C57BL/6J), GPX1 knockout (GPX1-/-) and catalase knockout (CAT-/-) mice were used. Lens transparency was quantified using dark-field images and ImageJ software. For optical aberration evaluation, each lens was placed over a copper electron microscopy specimen grid; the grid image was captured through the lens using a digital camera attached to a dark-field binocular microscope. Optical aberrations were assessed by the quality of the magnified gridlines. Microelectrode-based intact lens intracellular impedance was measured to determine GJC resistance. Results: In contrast to wild-type (WT) and CAT-/- lenses, GPX1-/- lenses developed accelerated age-related cataracts. While two-month-old lenses were normal, at nine months of age, GPX1-/- mice started to show the development of abnormal optical distortion aberrations and loss of transparency. At 12 months of age, GPX1-/- lenses developed significant opacity and abnormal optical distortion aberrations compared to CAT-/- and WT (p<0.001); these aberrations gradually increased with age and matured into cataracts by 24 months of age. There was also a significant increase (p<0.001) in GJC resistance in the differentiating and mature fiber cells of GPX1-/- lenses at 12 months of age compared to that in similar areas of age-matched CAT-/- and WT lenses. Conclusions: Changes in the refractive and physiological properties of the lens occurred before cataract formation in GPX1-/- lenses but not in CAT-/- lenses. GPX1 is more critical than catalase for lens transparency, optical quality, and homeostasis in the aging lens under normal physiological conditions. GPX1 could be a promising therapeutic target for developing potential strategies to reduce adverse oxidative stress and delay/treat/prevent age-related cataracts.


Asunto(s)
Catarata , Cristalino , Envejecimiento , Animales , Catalasa/genética , Catarata/genética , Glutatión Peroxidasa , Peróxido de Hidrógeno , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Glutatión Peroxidasa GPX1
4.
Anim Genet ; 53(1): 68-79, 2022 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-34729794

RESUMEN

The live attenuated classical swine fever (CSF) vaccine has been successfully used to prevent and control CSF outbreaks for 6 decades. However, the immune response mechanisms against the vaccine remain poorly understood. Moreover, very few reports exist regarding the breed differences in the response to CSF vaccine. In this study, we generated the peripheral blood mononuclear cell transcriptomes of indigenous Ghurrah and commercial Landrace pig breeds, before and 7 days after CSF vaccination. Subsequently, between and within-breed differential gene expression analyses were carried out. Results revealed large differences in pre-vaccination peripheral blood mononuclear cell transcriptome profiles of the two breeds, which were homogenised 7 days after vaccination. Before vaccination, gene set enrichment analysis showed that pathways related to antigen sensing and innate immune response were enriched in Ghurrah, while pathways related to adaptive immunity were enriched in Landrace. Ghurrah exhibited greater immunomodulation compared to Landrace following the vaccination. In Ghurrah, cell-cycle processes and T-cell response pathways were upregulated after vaccination. However, no pathways were upregulated in Landrace after vaccination. Pathways related to inflammation were downregulated in both the breeds after vaccination. Key regulators of inflammation such as IL1A, IL1B, NFKBIA and TNF genes were strongly downregulated in both the breeds after vaccination. Overall, our results have elucidated the mechanisms of host immune response against CSF vaccination in two distinct breeds and revealed common key genes instrumental in the global immune response to the vaccine.


Asunto(s)
Peste Porcina Clásica/inmunología , Inmunidad Innata , Transcriptoma/inmunología , Vacunas Virales/administración & dosificación , Animales , Femenino , Especificidad de la Especie , Sus scrofa , Porcinos
5.
Exp Parasitol ; 234: 108216, 2022 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-35104468

RESUMEN

Infections caused by protozoan parasites are a major public health concern globally. These infections are commonly diagnosed during water-borne outbreaks, necessitating accurate and highly sensitive detection procedures to assure public health protection. Current molecular techniques are challenged by several factors, such as low parasite concentration, inefficient DNA extraction methods, and inhibitors in environmental samples. This study focused on the development and validation of a molecular protocol for DNA extraction, efficient protozoan (oo)cyst recovery and quantification of protozoan parasites from wastewater using droplet digital polymerase chain reaction (ddPCR). Five DNA extraction methods, including commercial kits, custom phenol-chloroform, and in-house modified methods, were evaluated. The efficiency of each method was assessed via spectrophotometric analysis and ddPCR amplification using specific primers. Lastly, the developed protocol was evaluated for the detection and quantification of Cryptosporidium parvum in wastewater from different regions in South Africa. The conventional phenol-chloroform extraction method yielded the highest DNA concentration of 223 (±0.71) ng/µl and detected the highest number of Cryptosporidium parvum (1807 (±0.30) copies/ddPCR reaction) compared to other methods evaluated in this study. Additionally, the phenol-chloroform method demonstrated high sensitivity in extracting DNA from as few as one cyst/L of Cryptosporidium parvum, corresponding to 5.93 copies/ddPCR reaction. It was also observed that analysis of both the filtered supernatant and pellets after centrifugation improves the recovery efficiency of oocysts from wastewater by 10.5%, resulting in a total recovery of 64.1%. This optimized protocol was successfully applied to measure protozoan concentration in wastewater from different regions in South Africa. The improved DNA extraction and quantification method proposed in this study would be effective in monitoring protozoan concentration in the environment, which will help in instituting mitigation measures to reduce water-borne infections.


Asunto(s)
Cryptosporidium/aislamiento & purificación , ADN Protozoario/aislamiento & purificación , Aguas Residuales/parasitología , Centrifugación , Cryptosporidium/genética , Cryptosporidium/crecimiento & desarrollo , Cartilla de ADN/normas , Filtración , Límite de Detección , Reacción en Cadena de la Polimerasa/métodos , Sensibilidad y Especificidad
6.
Exp Eye Res ; 209: 108645, 2021 08.
Artículo en Inglés | MEDLINE | ID: mdl-34087204

RESUMEN

Lens-specific beaded filament (BF) proteins CP49 and filensin interact with the C-terminus of the water channel protein Aquaporin 0 (AQP0). Previously we have reported that a C-terminally end-deleted AQP0-expressing transgenic mouse model AQP0ΔC/ΔC developed abnormal optical aberrations in the lens. This investigation was undertaken to find out whether the total loss of the BF structural proteins alter the optical properties of the lens and cause optical aberrations similar to those in AQP0ΔC/ΔC lenses; also, to map the changes in the optical quality as a function of age in the single or double BF protein knockouts as well as to assess whether there is any significant change in the water channel function of AQP0 in these knockouts. A double knockout mouse (2xKO) model for CP49 and filensin was developed by crossing CP49-KO and filensin-KO mice. Wild type, CP49-KO, filensin-KO, and 2xKO lenses at different ages, and AQP0ΔC/ΔC lenses at postnatal day-17 were imaged through the optical axis and compared for optical quality and focusing property. All three knockout models showed loss of transparency, and development of abnormal optical distortion aberration similar to that in AQP0ΔC/ΔC. Copper grid focusing by the lenses at 6, 9 and 12 months of age showed an increase in aberrations as age advanced. With progression in age, the grid images produced by the lenses of all KO models showed a transition from a positive barrel distortion aberration to a pincushion distortion aberration with the formation of three distinct aberration zones similar to those produced by AQP0ΔC/ΔC lenses. Water permeability of fiber cell membrane vesicles prepared from CP49-KO, filensin-KO and 2xKO models, measured using the osmotic shrinking method, remained similar to that of the wild type without any statistically significant alteration (P > 0.05). Western blotting and quantification revealed the expression of comparable quantities of AQP0 in all three BF protein KOs. Our study reveals that loss of single or both beaded filament proteins significantly affect lens refractive index gradient, transparency and focusing ability in an age-dependent manner and the interaction of BF proteins with AQP0 is critical for the proper functioning of the lens. The presence of BF proteins is necessary to prevent abnormal optical aberrations and maintain homeostasis in the aging lens.


Asunto(s)
Acuaporinas/genética , Catarata/genética , Proteínas del Ojo/genética , Regulación de la Expresión Génica , Proteínas de Filamentos Intermediarios/genética , Cristalino/metabolismo , ARN/genética , Animales , Acuaporinas/biosíntesis , Western Blotting , Catarata/metabolismo , Catarata/fisiopatología , Modelos Animales de Enfermedad , Proteínas del Ojo/biosíntesis , Proteínas de Filamentos Intermediarios/biosíntesis , Cristalino/patología , Cristalino/fisiopatología , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados
7.
J Appl Microbiol ; 130(4): 1217-1231, 2021 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-33025721

RESUMEN

AIM: This study aims to establish the role of quorum sensing (QS) system on the regulation of naphthalene ring cleaving gene ndo (encoding naphthalene dioxygenase) in biofilm forming marine bacterium Pseudomonas aeruginosa N6P6 for naphthalene degradation. METHODS AND RESULTS: Total cell count of P. aeruginosa N6P6 during biofilm mode of growth was slightly higher (7·3 × 108  CFU per ml) than its planktonic mid-exponential phase culture (4·7 × 108  CFU per ml). Naphthalene degradation in 20h by biofilm (48-h old) and planktonic culture was 99·4 ± 0·002% and 77 ± 3·25%, respectively. Pseudomonas aeruginosa N6P6 was able to degrade 64·3 ± 4·7% naphthalene in sterile soil microcosm in 24 h. The bacterium showed the presence of 136 bp ndo gene which was upregulated in a dose-dependent manner in presence of naphthalene. QS inhibitor (QSI) tannic acid downregulated the expression of ndo gene, naphthalene 1, 2-dioxygenase (N12O) enzyme activity and naphthalene degradation (by biofilm culture). CONCLUSIONS: P. aeruginosa N6P6 shows chemotaxis towards naphthalene and adapts well in terrestrial environment for naphthalene degradation. SIGNIFICANCE AND IMPACT THE OF STUDY: This study provides the information that the QS plays crucial role in biofilm formation in P. aeruginosa N6P6 and QS regulatory genes subsequently control the ndo gene for enzymatic degradation of naphthalene.


Asunto(s)
Biopelículas , Dioxigenasas/genética , Complejos Multienzimáticos/genética , Naftalenos/metabolismo , Pseudomonas aeruginosa/fisiología , Percepción de Quorum/fisiología , Biodegradación Ambiental , Biopelículas/efectos de los fármacos , Biopelículas/crecimiento & desarrollo , Dioxigenasas/metabolismo , Regulación Bacteriana de la Expresión Génica/efectos de los fármacos , Complejos Multienzimáticos/metabolismo , Pseudomonas aeruginosa/genética , Pseudomonas aeruginosa/crecimiento & desarrollo , Pseudomonas aeruginosa/metabolismo , Percepción de Quorum/efectos de los fármacos , Contaminantes del Suelo/metabolismo , Taninos/farmacología
8.
J Postgrad Med ; 67(1): 39-42, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33533746

RESUMEN

An elderly hypertensive lady presented with fever, respiratory symptoms, and mild abdominal discomfort and was diagnosed to have COVID-19 pneumonia. Respiratory symptoms improved with steroids, awake proning, high flow nasal cannula oxygen therapy and antibiotics. After 4 days, she developed non-occlusive superior mesenteric artery thrombosis, which initially responded to anticoagulants but was complicated on tenth day by intestinal obstruction necessitating emergency surgery. Challenges encountered perioperatively were multi systemic involvement, pneumonia, ventilation- perfusion mismatch, sepsis along with technical difficulties like fogging of goggles, stuck expiratory valve on anesthesia machine, inaudibility through stethoscope and discomfort due to personal protective equipment. Perioperative focus should be on infection prevention, maintenance of hemodynamics, and optimization of oxygenation with preoperative high flow nasal cannula oxygen therapy. Ultrasound lung helps in correct placement of endotracheal tube. We recommend daily machine check, taping of N95 mask to face and ambient operation theatre temperatures of 20-22°C to reduce technical problems.


Asunto(s)
Anestesia General/métodos , COVID-19/complicaciones , Enfermedades del Íleon/cirugía , Obstrucción Intestinal/cirugía , Laparotomía , COVID-19/diagnóstico , COVID-19/terapia , Urgencias Médicas , Femenino , Humanos , Enfermedades del Íleon/diagnóstico , Enfermedades del Íleon/virología , Obstrucción Intestinal/diagnóstico , Obstrucción Intestinal/virología , Persona de Mediana Edad
9.
Biochem Biophys Res Commun ; 524(4): 1025-1029, 2020 04 16.
Artículo en Inglés | MEDLINE | ID: mdl-32063362

RESUMEN

High levels of reactive oxygen species such as hydrogen peroxide (H2O2) cause oxidative stress in the lens and lead to cataractogenesis. The present investigation was undertaken to find out whether the mammalian lens aquaporins (AQPs) 0, 1, and 5 perform H2O2 transport across the plasma membrane to reduce oxidative stress. Our in vitro cell culture and ex vivo lens experiments demonstrated that in addition to the established water transport role, mouse AQP0, AQP1 and AQP5 facilitate transmembrane H2O2 transport and function as peroxiporins. Human lens epithelial cells expressing AQP1, AQP5 and AQP8, when treated with 50 µM HgCl2 water channel inhibitor showed a significant reduction in H2O2 transport. Data obtained from the experiments involving H2O2-degrading enzyme glutathione peroxidase 1 (GPX1) knockout lenses showed H2O2 accumulation, suggesting H2O2 transport level by AQPs in the lens is regulated by GPX1. Under hyperglycemic conditions, there was an increased loss of transparency, and enhanced production and retention of H2O2 in AQP5-/- lenses compared to similarly-treated WT lenses. Overall, the results show that lens AQPs function as peroxiporins and cooperate with GPX1 to maintain lens H2O2 homeostasis to prevent oxidative stress, highlighting AQPs and GPX1 as promising therapeutic drug targets to delay/treat/prevent age-related lens cataracts.


Asunto(s)
Acuaporinas/metabolismo , Peróxido de Hidrógeno/metabolismo , Cristalino/metabolismo , Animales , Acuaporina 1/metabolismo , Acuaporina 5/metabolismo , Transporte Biológico , Línea Celular , Membrana Celular/metabolismo , Perros , Proteínas del Ojo/metabolismo , Humanos , Células de Riñón Canino Madin Darby , Ratones Endogámicos C57BL
10.
Chaos ; 30(12): 123115, 2020 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-33380015

RESUMEN

We present a new four-step feedback procedure to study the full dynamics of a nonlinear dynamical system, namely, the logistic map. We show that by using this procedure, the chaotic behavior of the logistic map can be controlled easily and rapidly or the system can be made stable for higher values of the population growth parameter. We utilize various dynamical techniques (orbit evolution, time series analysis, bifurcation diagrams, and Lyapunov exponents) to analyze the dynamics of the logistic map. Additionally, we adopt the switching strategy to control chaos or to increase the stability performance of the logistic map. Finally, we propose a modified traffic control model to enable rapid control of unexpected traffic on the road. The results of this model are supported by a physical interpretation. The model is found to be more efficient than existing models of Lo and Cho [J. Franklin Inst. 342, 839-851 (2005)] and Ashish et al. [Nonlinear Dyn. 94, 959-975 (2018)]. This work provides a novel feedback procedure that facilitates rapid control of chaotic behavior and increases the range of stability of dynamical systems.

11.
Biochem Biophys Res Commun ; 511(3): 626-630, 2019 04 09.
Artículo en Inglés | MEDLINE | ID: mdl-30826060

RESUMEN

The purpose of this investigation was to find out whether C-terminally end-cleaved aquaporin 0 (AQP0), that is present predominantly in the lens mature fiber cells of the WT, functions as a water channel and a cell-to-cell adhesion (CTCA) protein in a knockin (KI) mouse model (AQP0ΔC/ΔC) that does not express intact AQP0. A genetically engineered KI mouse model, AQP0ΔC/ΔC, expressing only end-cleaved AQP0 was developed. This model expresses 1-246 amino acids of AQP0, instead of the full length 1-263 amino acids. Lens transparency of postnatal day 10 (P10) was analyzed qualitatively by dark field imaging. WT, AQP0+/- and AQP0+/ΔC lenses were transparent; AQP0-/- and AQP0ΔC/ΔC mouse lenses displayed loss of transparency. Lens fiber cell membrane vesicles (FCMVs) were prepared from wild type (WT), AQP0 heterozygous (AQP0+/-), AQP0 knockout (AQP0-/-), AQP0+/ΔC and AQP0ΔC/ΔC; water permeability (Pf) was measured using the osmotic shrinking method. CTCA assay was performed using adhesion-deficient L-cells and FCMVs prepared from the abovementioned genotypes. FCMVs of AQP0+/- and AQP0-/- showed a statistically significant reduction (P < 0.001) in Pf and CTCA compared to those of WT. AQP0+/ΔC and AQP0ΔC/ΔC FCMVs exhibited no statistically significant alteration (P > 0.05) in Pf compared to those of WT. However, CTCA of AQP0+/ΔC AQP0ΔC/ΔC FCMVs was significantly higher (P < 0.001) than that of WT FCMVs. Our experiments clearly show that C-terminally end-cleaved AQP0 can function both as a water channel and a CTCA molecule in the lens fiber cell membranes. Also, end-truncation plays an important role in increasing the CTCA between fiber cells.


Asunto(s)
Acuaporinas/metabolismo , Proteínas del Ojo/metabolismo , Cristalino/metabolismo , Animales , Acuaporinas/química , Acuaporinas/genética , Catarata/genética , Catarata/metabolismo , Adhesión Celular , Permeabilidad de la Membrana Celular , Proteínas del Ojo/química , Proteínas del Ojo/genética , Cristalino/citología , Ratones , Ratones Endogámicos C57BL , Mutación , Agua/metabolismo
12.
Opt Express ; 27(10): 14684-14694, 2019 May 13.
Artículo en Inglés | MEDLINE | ID: mdl-31163913

RESUMEN

Applications of wavelet analysis in ultra-thin film transient reflectivity (TR) measurements have been investigated. Advantages of utilizing different localized wavelet bases, in position and time, have been addressed on the residual TR signals. Morse wavelets have been used to obtain information from the abrupt oscillatory modes in the signal, which are not distinguishable with conventional methods such as Fourier transforms. These abrupt oscillatory modes are caused by the surface, interface, or any short-lived oscillatory modes which are suppressed in the TR signal in ultra-thin films. It is demonstrated that by choosing different Morse wavelets, information regarding different oscillatory modes in the TR signal of a heterostructure thin film is achievable. Moreover, by performing wavelet analysis on multiferroic heterostructures, oscillatory modes with very close energy ranges are easily distinguishable. For illustration, residuals of the TR signals have been obtained by a pump-probe setup in reflectivity mode on La0.7Sr0.3MnO3/SrTiO3 and BaTiO3/La0.7Sr0.3MnO3/SrTiO3 samples, where sufficient signal to noise ratios have been achieved by taking multiple scans. The residual signals have been analyzed with Morse wavelets, and multiple oscillatory modes with close energy ranges have been observed and distinguished. This approach can isolate the location of various oscillatory modes at the surface, interface and in the bulk of the heterostructure sample.

13.
Sensors (Basel) ; 19(19)2019 Sep 27.
Artículo en Inglés | MEDLINE | ID: mdl-31569806

RESUMEN

In the present paper, a new kind of concave shaped refractive index sensor (CSRIS) exploiting localized surface plasmon resonance (LSPR) is proposed and numerically optimized. The LSPR effect between polaritons and the core guided mode of designed CSRIS is used to enhance the sensing performance. The sensor is characterized for two types of sensing structures coated with gold (Au) film and Au nanowires (AuNWs), respectively. The influence of structural parameters such as the distance (D) of the concave shaped channel (CSC) from the core, the diameter of the nanowire (dn) and the size (s) of the CSC are investigated here. In comparison to Au film, the AuNWs are shown to significantly enhance the sensitivity and the performance of the designed sensor. An enhanced sensitivity of 4471 nm/RIU (refractive index unit) is obtained with AuNWs, for a wide range of analytes refractive index (na) varying between 1.33 to 1.38. However, for conventional Au film; the sensitivity of 808.57 nm/RIU is obtained for the same range of analytes.

14.
Biochem Biophys Res Commun ; 506(1): 284-289, 2018 11 17.
Artículo en Inglés | MEDLINE | ID: mdl-30348525

RESUMEN

Cell-to-cell adhesion (CTCA), which is key for establishing lens transparency, is a critical function of Aquaporin 0 (AQP0). The aim of this investigation was to find out the possible mechanism by which AQP0 exerts CTCA between fiber cells, since there are two proposals currently, either an AQP0-AQP0 interaction or an AQP0-lipid interaction. We studied the mechanism of AQP0-induced CTCA in intact AQP0 and C-terminally cleaved AQP0 (CTC-AQP0). Assays showed CTCA between L-cells transfected with intact AQP0 or CTC-AQP0 and parental L-cells indicating AQP0-membrane interaction. Both forms of AQP0 significantly (P < 0.001) promoted adhesion to negatively charged l-α-phosphatidylserine lipid vesicles signifying AQP0-lipid interaction. AQP0-expressing L-cells also promoted adhesion of WT and AQP0-KO mouse lens fiber cell membrane vesicles (FCMVs) significantly (P < 0.001). However, when FCMVs of WT or AQP0-KO were plated over parental L-cells, only WT vesicles adhered significantly, corroborating AQP0-membrane interaction. After incubating with extracellular domain-specific AQP0 antibody, L-cells expressing intact AQP0 or CTC-AQP0 showed a significant reduction (P < 0.001) in the adhesion of AQP0-KO FCMVs indicating extracellular loop involvement in CTCA. WT FCMVs from outer cortex and inner cortex promoted adhesion to parental L-cells, without any statistically significant difference in adhesion efficiency (P > 0.05). Ultrastructure studies of WT, AQP0-KO and transgenic lenses showed AQP0 is critical for fiber CTCA and compaction. The data collected clearly demonstrate that the positively charged amino acids in the AQP0 extracellular loop domains interact with the negatively charged lipids in the plasma membrane to promote CTCA for compaction of fiber cells.


Asunto(s)
Acuaporinas/metabolismo , Adhesión Celular/efectos de los fármacos , Proteínas del Ojo/metabolismo , Cristalino/citología , Animales , Liposomas/metabolismo , Membranas Artificiales , Ratones , Fosfatidilserinas/metabolismo , Electricidad Estática
15.
Exp Eye Res ; 172: 152-158, 2018 07.
Artículo en Inglés | MEDLINE | ID: mdl-29660329

RESUMEN

Aquaporins (AQPs), ordinarily regarded as water channels, have recently been shown to participate in other cellular functions such as cell-to-cell adhesion, cell migration, cell proliferation etc. The current investigation was undertaken to find out whether AQP5 water channel plays a role in corneal epithelial wound healing. Expression of AQP5 in mouse cornea and transfected Madin-Darby canine kidney (MDCK) cells was detected using immunofluorescence or EGFP tag. Cell migration and proliferation, the two major events in wound healing, were studied in vitro using cell culture scratch-wound healing model and cell proliferation assay, in vivo by conducting wound healing experiments on corneas of wild-type and AQP5 knockout mouse model and ex vivo on corneal epithelial cells isolated from wild type and AQP5 knockout mice. MDCK cells stably expressing AQP5 showed significantly higher levels of cell migration and proliferation compared to control cells. Likewise, corneal epithelial cells of wild type mouse with innate AQP5 exhibited faster wound healing than those of AQP5 knockout in vivo and under ex vivo culture conditions. In vitro, in vivo and ex vivo studies showed that presence of AQP5 improved cell migration, proliferation and wound healing. The data collected suggest that AQP5 plays a significant role in corneal epithelial wound healing.


Asunto(s)
Acuaporina 5/fisiología , Movimiento Celular/fisiología , Repitelización/fisiología , Cicatrización de Heridas/fisiología , Animales , Western Blotting , Técnicas de Cultivo de Célula , Proliferación Celular/fisiología , Córnea/metabolismo , Perros , Epitelio Corneal/fisiología , Técnica del Anticuerpo Fluorescente Indirecta , Proteínas Fluorescentes Verdes/metabolismo , Células de Riñón Canino Madin Darby/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Transfección
16.
Bioorg Med Chem ; 26(8): 1996-2008, 2018 05 01.
Artículo en Inglés | MEDLINE | ID: mdl-29525336

RESUMEN

A facile one-pot method for the synthesis of new phenanthrene fused-dihydrodibenzo-quinolinone derivatives has been successfully accomplished by employing sulfamic acid as catalyst. These new compounds were evaluated for their in vitro cytotoxic potential against human lung (A549), prostate (PC-3 and DU145), breast (MCF-7) and colon (HT-29 and HCT-116) cancer cell lines. Among all the tested compounds, one of the derivatives 8p showed good anti-proliferative activity against A549 lung cancer cell line with an IC50 of 3.17 ±â€¯0.52 µM. Flow cytometric analyses revealed that compound 8p arrested both Sub G1 and G2/M phases of cell cycle in a dose dependent manner. The compound 8p also displayed significant inhibition of tubulin polymerization and disruption of microtubule network (IC50 of 5.15 ±â€¯0.15 µM). Molecular docking studies revealed that compound 8p efficiently interacted with critical amino acid Cys241 of the α/ß-tubulin by a hydrogen bond (SH…O = 2.4 Å). Further, the effect of 8p on cell viability was also studied by AO/EB, DCFDA and DAPI staining. The apoptotic characteristic features revealed that 8p inhibited cell proliferation effectively through apoptosis by inducing the ROS generation. Analysis of mitochondrial membrane potential through JC-1 staining and annexin V binding assay indicated the extent of apoptosis in A549 cancer cells.


Asunto(s)
Antineoplásicos/síntesis química , Apoptosis/efectos de los fármacos , Quinolonas/química , Ácidos Sulfónicos/química , Moduladores de Tubulina/síntesis química , Tubulina (Proteína)/metabolismo , Células A549 , Antineoplásicos/metabolismo , Antineoplásicos/farmacología , Sitios de Unión , Puntos de Control del Ciclo Celular/efectos de los fármacos , Ensayos de Selección de Medicamentos Antitumorales , Humanos , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Simulación del Acoplamiento Molecular , Fenantrenos/química , Estructura Terciaria de Proteína , Quinolonas/metabolismo , Quinolonas/farmacología , Especies Reactivas de Oxígeno/metabolismo , Relación Estructura-Actividad , Tubulina (Proteína)/química , Moduladores de Tubulina/metabolismo , Moduladores de Tubulina/farmacología
18.
J Med Syst ; 43(1): 1, 2018 Nov 15.
Artículo en Inglés | MEDLINE | ID: mdl-30456688

RESUMEN

Diabetes, a metabolic disorder due to high blood glycemic index in the human body. The glycemic index varies in the human of improper diet and eating pattern such as junk foods, variation in the quantity of food, swallowing of food without chewing and stress. However, the diagnose of increase or decrease in the glycemic index is a challenging task. Similarly, the regulation of glycemic index without regular exercise is a major problem in day to day life. In this paper, we propose a novel SCS method to regulate glycemic index without exercise through changing the eating method. The proposed SCS eating method consists of Size of the food, Chewing style and Swallow time (SCS) of the food to regulate glycemic index. Furthermore, the proposed SCS method evaluate and validate through the acoustic signal acquired and processed with deep learning algorithm to analyze the chewing pattern of food to formulate a standard procedure for eating style and to reduce the glycemic level. The validation of diabetes done by measurement of blood glycemic through AccuChek Instant S Glucometer. Furthermore, the SCS method of eating style from 50 diabetes persons reduces the blood glucose level drastically by 85% after following the proposed method of eating style.


Asunto(s)
Glucemia/metabolismo , Aprendizaje Profundo , Deglución/fisiología , Diabetes Mellitus/fisiopatología , Índice Glucémico/fisiología , Masticación/fisiología , Algoritmos , Humanos , Sistemas Microelectromecánicos , Saliva/metabolismo
19.
Morphologie ; 102(336): 21-24, 2018 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-28927574

RESUMEN

Muscles of flexor compartment of forearm have a common origin from medial epicondyle of humerus. Additional bellies of flexor muscles are commonly reported but presence of supernumerary muscles is an infrequent phenomenon. The present study describes an unusual muscle mass in flexor compartment of forearm simulating pronator teres. During routine dissection the upper limb of a 50 years old male cadaver, a supernumerary muscle was found on left side of the upper limb in the flexor compartment. The origin of the muscle was 2cm wide and aponeurotic in nature and attached to an oblique line extending from the inferior surface of the medial epicondyle and the medial surface of the trochlea. It was inserted on an oblique line 2.5cm wide on the radius in area between supinator superiorly and flexor digitorum profundus inferiorly. Existence of accessory muscles, which connect flexor muscles, could be explained embryologically by incomplete cleavage of flexor mass during development. The flexor muscles of the forearm develop from the flexor mass which subsequently divides into two layers: superficial and deep. The deep layer gives rise to flexor digitorum superficialis, flexor digitorum profundus and flexor pollicis longus. These supernumerary muscles are extremely rare entities and probably represent deranged embryological development or the process of atavism in which the anomalous part persist for a longer time in the tree of evolution.


Asunto(s)
Variación Anatómica , Antebrazo/anatomía & histología , Músculo Esquelético/anomalías , Animales , Cadáver , Humanos , Masculino , Persona de Mediana Edad
20.
Biomarkers ; 22(5): 446-454, 2017 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-27775440

RESUMEN

OBJECTIVE: This study was designed to determine the genotoxic effect of exposure to a mixture of pesticides in 106 female agricultural workers employed in cotton fields from India. METHODS: Comet, micronucleus and chromosomal aberrations tests were carried out in peripheral blood lymphocytes. Micronucleus test was also performed in buccal epithelial cells. Levels of antioxidant enzymes, RBC acetylcholinesterase and hematological parameters were analyzed in the blood samples of the study subjects. RESULTS: The results indicated significant DNA damage, increased frequency of micronuclei and chromosomal aberrations in the exposed subjects (p < 0.05). The levels of antioxidant enzymes were significantly lowered and the rate of lipid peroxidation was elevated in the exposed subjects. CONCLUSION: The outcome of the study revealed an increased risk of genotoxicity and health implications in female agricultural workers.


Asunto(s)
Agricultura , Pruebas de Mutagenicidad/métodos , Exposición Profesional/análisis , Plaguicidas/farmacología , Acetilcolinesterasa/metabolismo , Adolescente , Adulto , Antioxidantes/metabolismo , Aberraciones Cromosómicas/inducido químicamente , Daño del ADN/efectos de los fármacos , Femenino , Humanos , India , Peroxidación de Lípido/efectos de los fármacos , Linfocitos/química , Pruebas de Micronúcleos , Oxidorreductasas/metabolismo , Plaguicidas/toxicidad , Adulto Joven
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