Evaluation of the OPTC gene in primary open angle glaucoma: functional significance of a silent change

BACKGROUND: We investigated the molecular basis of primary open-angle glaucoma (POAG) using Opticin (OPTC) as a candidate gene on the basis of its expression in the trabecular meshwork cells involved in the disease pathogenesis. Two hundred POAG patients and 100 controls were enrolled in this study. The coding sequence of OPTC was amplified by PCR from genomic DNA of POAG patients, followed by SSCP, DHPLC and DNA sequencing. Subsequent bioinformatic analysis, site-directed mutagenesis, quantitative RT-PCR and western blot experiments were performed to address the functional significance of a 'silent' change in the OPTC coding region while screening for mutations in POAG patients. RESULTS: We detected two missense (p.Glu66Gly & p.Ile89Thr) and one silent change (p.Phe162Phe; c.602 C>T) that was present in 3 different patients but in none of the 100 controls screened. The mutant (c.602T) mRNA was predicted to have remarkably different secondary structure compared to the wild-type transcript by in silico approaches. Subsequent wet-lab experiments showed lower expression of the gene both at the mRNA and protein levels. CONCLUSION: Our study suggests OPTC as a candidate gene for POAG. Further, it highlights the importance of investigating the 'silent' variations for functional implication that might not be apparent from only in silico analysis

LncRNA VEAL2 regulates PRKCB2 to modulate endothelial permeability in diabetic retinopathy

Long non‐coding RNAs (lncRNAs) are emerging as key regulators of endothelial cell function. Here, we investigated the role of a novel vascular endothelial‐associated lncRNA (VEAL2) in regulating endothelial permeability. Precise editing of veal2 loci in zebrafish (veal2 (gib005Δ8/+)) induced cranial hemorrhage. In vitro and in vivo studies revealed that veal2 competes with diacylglycerol for interaction with protein kinase C beta‐b (Prkcbb) and regulates its kinase activity. Using PRKCB2 as bait, we identified functional ortholog of veal2 in humans from HUVECs and named it as VEAL2. Overexpression and knockdown of VEAL2 affected tubulogenesis and permeability in HUVECs. VEAL2 was differentially expressed in choroid tissue in eye and blood from patients with diabetic retinopathy, a disease where PRKCB2 is known to be hyperactivated. Further, VEAL2 could rescue the effects of PRKCB2‐mediated turnover of endothelial junctional proteins thus reducing hyperpermeability in hyperglycemic HUVEC model of diabetic retinopathy. Based on evidence from zebrafish and hyperglycemic HUVEC models and diabetic retinopathy patients, we report a hitherto unknown VEAL2 lncRNA‐mediated regulation of PRKCB2, for modulating junctional dynamics and maintenance of endothelial permeability

Laccase mediated delignification of pineapple leaf waste: an ecofriendly sustainable attempt towards valorization

Abstract Background Escalating energy security, burgeoning population and rising costs of fossil fuels have focussed our attention on tapping renewable energy sources. As the utilization of food crops for biofuel production culminates into food vs. fuel dilemma, there is an intensive need for alternatives. Production of biofuels from lignocellulosic biomass owing to its profuse availability and high holocellulose content is a promising area for research. Results In the present study, pineapple leaf, an agro-industrial waste was pretreated with laccase to enhance the enzymatic digestibility of the substrate for improved production of reducing sugar. Variables determining enzymatic delignification of pineapple leaf waste have been optimized by response surface methodology based on central composite design. Maximum delignification of 78.57%(w/w) resulted in reducing sugar of 492.33 ± 3.1 mg/g in 5.30 h. The structural changes in pineapple leaf waste, after laccase treatment, were studied through Fourier transformed infrared spectroscopy, X-ray diffraction and Scanning electron microscopy. Specific surface area, pore volume, and pore diameter of the substrate were studied using the Brunauer–Emmett–Teller and Barrett–Joyner–Halenda methods and found a significant increase in the aforementioned parameters after delignification. Conclusion Laccase mediated delignification of pineapple leaf waste is a cleaner sustainable process for enhanced production of reducing sugar which can accomplish the demand for biofuels

Chronic female pelvic pain

Chronic pelvic pain (CPP) is defined as nonmalignant pain perceived in the structures related to the pelvis that has been present for more than 6 months or a non acute pain mechanism of shorter duration. Pain in the pelvic region can arise from musculoskeletal, gynaecological, urologic, gastrointestinal and or neurologic conditions. Key gynaecological conditions that contribute to CPP include pelvic inflammatory disease (PID), endometriosis, adnexa pathologies (ovarian cysts, ovarian remnant syndrome), uterine pathologies (leiomyoma, adenomyosis) and pelvic girdle pain associated with pregnancy. Several major and minor sexually transmitted diseases (STD) can cause pelvic and vulvar pain. A common painful condition of the urinary system is Interstitial cystitis(IC. A second urologic condition that can lead to development of CPP is urethral syndrome. Irritable bowel syndrome (IBS) is associated with dysmenorrhoea in 60% of cases. Other bowel conditions contributing to pelvic pain include diverticular disease,Crohn′s disease ulcerative colitis and chronic appendicitis. Musculoskeletal pathologies that can cause pelvic pain include sacroiliac joint (SIJ) dysfunction, symphysis pubis and sacro-coccygeal joint dysfunction, coccyx injury or malposition and neuropathic structures in the lower thoracic, lumbar and sacral plexus. Prolonged pelvic girdle pain, lasting more than 6 months postpartum is estimated in 3% to 30% of women. Nerve irritation or entrapment as a cause of pelvic pain can be related to injury of the upper lumbar segments giving rise to irritation of the sensory nerves to the ventral trunk or from direct trauma from abdominal incisions or retractors used during abdominal surgical procedures. Afflictions of the iliohypogastric, ilioinguinal, genitofemoral, pudendal and obturator nerves are of greatest concern in patients with pelvic pain. Patient education about the disease and treatment involved is paramount. A knowledge of the differential diagnosis of the pain generators leads to a diagnosis specific management of the pain condition. Using a multidisciplinary approach can improve outcomes for patients suffering from the condition and minimize the associated disability

Design, evaluation and statistical optimisation of a controlled release multiparticulate Acyclovir delivery system

El presente trabajo tiene por objeto el diseño y la evaluación de una forma de liberación controlada de Acyclovir utilizando etilcelulosa como polímero formador de la matriz y empleando la técnica de evaporación del solvente para la microencapsulación. El producto fue caracterizado por parámetros fisicoquímicos tales como el rendimiento (51%-86%), la eficiencia de retención de la droga (88% - 96%), el tamaño de partícula (principalmente tamiz de malla 30 ASTM), la topografía superficial, la compatibilidad droga-excipiente y la liberación in vitro (11% - 81% después de 8 h). El perfil de liberación controlada fue optimizado utilizando una mezcla de látex sencilla diseñada para lograr la combinación más adecuada de las micropartículas (proporción de partículas de relación droga-polímero 1:1 a 1:2 = 68%:32% de peso), que es el más apropiado para el perfil de liberación. El error entre el blanco perseguido (% de liberación a las 2 h, 6 h y 8 h de 40, 60 y 75, respectivamente) y la combinación óptima de las micropartículas fue menor que el 10%. El estudio demuestra la utilidad y ventajas del diseño experimental en la investigación de la liberación controlada de drogas.This work aims at designing and evaluating a multiparticulate controlled release dosage form of Acyclovir using Ethyl Cellulose as the matrix-forming polymer employing the solvent evaporation technique of microencapsulation. The product was characterized by physicochemical parameters such as yield (51%-86%), drug entrapment efficiency (88% - 96%), particle size (mainly #30 mesh ASTM), surface topography, drug-excipient compatibility and in vitro release (11% - 81% after 8 h). The controlled release profile was optimized using a simplex lattice mixture design for achieving the correct blend of microparticles (proportion of particles of drug-polymer ratio 1:1 to 1:2 = 68%: 32% by weight) that closely matches the target release profile. The error between the target (% release at 2nd , 6th and 8th h of 40, 60 and 75 respectively) and optimum blend of the microparticles was less than 10%. The study illustrates the utility and advantage of designed experimentation in controlled drug delivery research.Colegio de Farmacéuticos de la Provincia de Buenos Aire

Design, evaluation and statistical optimisation of a controlled release multiparticulate Acyclovir delivery system

El presente trabajo tiene por objeto el diseño y la evaluación de una forma de liberación controlada de Acyclovir utilizando etilcelulosa como polímero formador de la matriz y empleando la técnica de evaporación del solvente para la microencapsulación. El producto fue caracterizado por parámetros fisicoquímicos tales como el rendimiento (51%-86%), la eficiencia de retención de la droga (88% - 96%), el tamaño de partícula (principalmente tamiz de malla 30 ASTM), la topografía superficial, la compatibilidad droga-excipiente y la liberación in vitro (11% - 81% después de 8 h). El perfil de liberación controlada fue optimizado utilizando una mezcla de látex sencilla diseñada para lograr la combinación más adecuada de las micropartículas (proporción de partículas de relación droga-polímero 1:1 a 1:2 = 68%:32% de peso), que es el más apropiado para el perfil de liberación. El error entre el blanco perseguido (% de liberación a las 2 h, 6 h y 8 h de 40, 60 y 75, respectivamente) y la combinación óptima de las micropartículas fue menor que el 10%. El estudio demuestra la utilidad y ventajas del diseño experimental en la investigación de la liberación controlada de drogas.This work aims at designing and evaluating a multiparticulate controlled release dosage form of Acyclovir using Ethyl Cellulose as the matrix-forming polymer employing the solvent evaporation technique of microencapsulation. The product was characterized by physicochemical parameters such as yield (51%-86%), drug entrapment efficiency (88% - 96%), particle size (mainly #30 mesh ASTM), surface topography, drug-excipient compatibility and in vitro release (11% - 81% after 8 h). The controlled release profile was optimized using a simplex lattice mixture design for achieving the correct blend of microparticles (proportion of particles of drug-polymer ratio 1:1 to 1:2 = 68%: 32% by weight) that closely matches the target release profile. The error between the target (% release at 2nd , 6th and 8th h of 40, 60 and 75 respectively) and optimum blend of the microparticles was less than 10%. The study illustrates the utility and advantage of designed experimentation in controlled drug delivery research.Colegio de Farmacéuticos de la Provincia de Buenos Aire