Highly Luminescent Europium(III) Complexes in Solution and PMMA-Doped Films for Bright Red Electroluminescent Devices.

This paper reports the synthesis, structure, photophysical, and optoelectronic properties of five eight-coordinate Europium(III) ternary complexes, namely, [Eu(hth)3(L)2], bearing 4,4,5,5,6,6,6-heptafluoro-1-(2-thienyl)-1,3-hexanedione (hth) as a sensitizer and L = H2O (1), dpso (diphenyl sulphoxide, 2), dpsoCH3 (4,4'-dimethyl diphenyl sulfoxide, 3), dpsoCl (bis(4-chlorophenyl)sulphoxide, 4), and tppo (triphenylphosphine oxide, 5) as co-ligands. The NMR and the crystal structure analysis confirmed the eight-coordinate structures of the complexes in solution and in a solid state. Upon UV-excitation on the absorption band of the ß-diketonate ligand hth, all complexes showed the characteristic bright red luminescence of the Europium ion. The tppo derivative (5) displayed the highest quantum yield (up to 66%). As a result, an organic light-emitting device, OLED, was fabricated with a multi-layered structure-ITO/MoO3/mCP/SF3PO:[complex 5] (10%)/TPBi:[complex 5] (10%)/TmPyPB/LiF/Al-using complex 5 as the emitting component.

A2ML1 and otitis media: novel variants, differential expression, and relevant pathways.

A genetic basis for otitis media is established, however, the role of rare variants in disease etiology is largely unknown. Previously a duplication variant within A2ML1 was identified as a significant risk factor for otitis media in an indigenous Filipino population and in US children. In this report exome and Sanger sequencing was performed using DNA samples from the indigenous Filipino population, Filipino cochlear implantees, US probands, Finnish, and Pakistani families with otitis media. Sixteen novel, damaging A2ML1 variants identified in otitis media patients were rare or low-frequency in population-matched controls. In the indigenous population, both gingivitis and A2ML1 variants including the known duplication variant and the novel splice variant c.4061 + 1 G>C were independently associated with otitis media. Sequencing of salivary RNA samples from indigenous Filipinos demonstrated lower A2ML1 expression according to the carriage of A2ML1 variants. Sequencing of additional salivary RNA samples from US patients with otitis media revealed differentially expressed genes that are highly correlated with A2ML1 expression levels. In particular, RND3 is upregulated in both A2ML1 variant carriers and high-A2ML1 expressors. These findings support a role for A2ML1 in keratinocyte differentiation within the middle ear as part of otitis media pathology and the potential application of ROCK inhibition in otitis media.

Molecular genetics of MARVELD2 and clinical phenotype in Pakistani and Slovak families segregating DFNB49 hearing loss.

Pathogenic mutations of MARVELD2, encoding tricellulin, a tricelluar tight junction protein, cause autosomal recessive non-syndromic hearing loss (DFNB49) in families of Pakistan and Czech Roma origin. In fact, they are a significant cause of prelingual hearing loss in the Czech Roma, second only to GJB2 variants. Previously, we reported that mice homozygous for p.Arg497* variant of Marveld2 had a broad phenotypic spectrum, where defects were observed in the inner ear, heart, mandibular salivary gland, thyroid gland and olfactory epithelium. The current study describes the types and frequencies of MARVELD2 alleles and clinically reexamines members of DFNB49 families. We found that MARVELD2 variants are responsible for about 1.5 % (95 % CI 0.8-2.6) of non-syndromic hearing loss in our cohort of 800 Pakistani families. The c.1331+2T>C allele is recurrent. In addition, we identified a novel large deletion in a single family, which appears to have resulted from non-allelic homologous recombination between two similar Alu short interspersed elements. Finally, we observed no other clinical manifestations co-segregating with hearing loss in DFNB49 human families, and hypothesize that the additional abnormalities in the Marveld2 mutant mouse indicates a critical non-redundant function for tricellulin in other organ systems.

Improving corneal permeability of dexamethasone using penetration enhancing agents: First step towards achieving topical drug delivery to the retina.

With an ever-increasing burden of vision loss caused by diseases of the posterior ocular segment, there is an unmet clinical need for non-invasive treatment strategies. Topical drug application using eye drops suffers from low to negligible bioavailability to the posterior segment as a result of static and dynamic defensive ocular barriers to penetration, while invasive delivery systems are expensive to administer and suffer potentially severe complications. As the cornea is the main anatomical barrier to uptake of topically applied drugs from the ocular surface, we present an approach to increase corneal permeability of a corticosteroid, dexamethasone sodium-phosphate (DSP), using a novel penetration enhancing agent (PEA). We synthesised a novel polyacetylene (pAc) polymer and compared its activity to two previously described cell penetrating peptide (CPP) based PEAs, TAT and penetratin, with respect to increasing transcorneal permeability of DSP in a rapid ex-vivo porcine corneal assay over 60 min. The transcorneal apparent permeability coefficients (Papp) for diffusion of pAc, and fluorescein isothiocyanate (FITC) conjugated TAT and penetratin were up to 5 times higher (p < 0.001), when compared to controls. When pAc was used in formulation with DSP, an almost 5-fold significant increase was observed in Papp of DSP across the cornea (p = 0.0130), a significant 6-fold increase with TAT (p = 0.0377), and almost 7-fold mean increase with penetratin (p = 0.9540). Furthermore, we investigated whether the PEAs caused any irreversible damage to the barrier integrity of the corneal epithelium by measuring transepithelial electrical resistance (TEER) and immunostaining of tight junction proteins using zonula occludens-1 (ZO-1) and occludin antibodies. There was no damage or structural toxicity, and the barrier integrity was preserved after PEA application. Finally, an in-vitro cytotoxicity assessment of all PEAs in human retinal pigment epithelium cells (ARPE-19) demonstrated that all PEAs were very well-tolerated, with IC50 values of 64.79 mM for pAc and 1335.45 µM and 87.26 µM for TAT and penetratin, respectively. Our results suggest that this drug delivery technology could potentially be used to achieve a significantly higher intraocular therapeutic bioavailability after topical eye drop administration, than currently afforded.

Design of liposomal nanocarriers with a potential for combined dexamethasone and bevacizumab delivery to the eye.

Clinicians face numerous challenges when delivering medications to the eyes topically because of physiological barriers, that can inhibit the complete dose from getting to the intended location. Due to their small size, the ability to deliver drugs of different polarities simultaneously, and their biocompatibility, liposomes hold great promise for ocular drug delivery. This study aimed to develop and characterise a dual loaded liposome formulation encapsulating Bevacizumab (BEV) and Dexamethasone (DEX) that possessed the physicochemical attributes suitable for topical ocular delivery. Liposomes were prepared by using thin film hydration followed by extrusion, and the formulations were optimised using a design of experiments approach. Physicochemical characterisation along with cytocompatibility and bioactivity of the formulations were assessed. Liposomes were successfully prepared with a particle size of 139 ± 2 nm, PDI 0.03 ± 0.01 and zeta potential -2 ± 0.7 mV for the optimised formulation. BEV and DEX were successfully encapsulated into the liposomes with an encapsulation efficiency of 97 ± 0.5 % and 26 ± 0.5 %, respectively. A sustained release of BEV was observed from the liposomes and the bioactivity of the formulation was confirmed using a wound healing assay. In summary, a potential topical eye drop drug delivery system, which can co-load DEX and BEV was developed and characterised for its potential to be used in ocular drug delivery.

Ingestion and impacts of water-borne polypropylene microplastics on Daphnia similis.

Polypropylene microplastics are the leading contaminant in aquatic environments, although research on their toxicity remains scarce. The proposed research focuses on the harmful consequences of acute exposure to polypropylene microplastics in Daphnia similis. This work converts widely available polypropylene bags into microplastics using xylene. FTIR findings demonstrated the lack of xylene residue in the produced polypropylene microplastic particles, which were spherical and ranged in size from 11.86 to 44.62 µm (FE-SEM). The results indicate that acute exposure to polypropylene microplastics causes immobility in D. similis. Ingestion of microplastics enhances the generation of reactive oxygen species (ROS), as shown by biochemical studies. Due to the production of free radicals in D. similis, the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione-S-transferase (GST) and a non-antioxidant enzyme of reduced glutathione (GSH) and also oxidative stress effects in lipid (lipid peroxidation - LPO), protein (carbonyl protein - CP) were increased. Additionally, the amount of the neurotransmitter enzyme acetylcholinesterase (AChE) activity was decreased. These findings indicate that the accumulation of polypropylene microplastics in the bodies of filter-feeding organisms should aggravate toxicity in the freshwater environment.

Variable expressivity of FGF3 mutations associated with deafness and LAMM syndrome.

BACKGROUND: Recessive mutations of fibroblast growth factor 3 (FGF3) can cause LAMM syndrome (OMIM 610706), characterized by fully penetrant complete labyrinthine aplasia, microtia and microdontia. METHODS: We performed a prospective molecular genetic and clinical study of families segregating hearing loss linked to FGF3 mutations. Ten affected individuals from three large Pakistani families segregating FGF3 mutations were imaged with CT, MRI, or both to detect inner ear abnormalities. We also modeled the three dimensional structure of FGF3 to better understand the structural consequences of the three missense mutations. RESULTS: Two families segregated reported mutations (p.R104X and p.R95W) and one family segregated a novel mutation (p.R132GfsX26) of FGF3. All individuals homozygous for p.R104X or p.R132GfsX26 had fully penetrant features of LAMM syndrome. However, recessive p.R95W mutations were associated with nearly normal looking auricles and variable inner ear structural phenotypes, similar to that reported for a Somali family also segregating p.R95W. This suggests that the mild phenotype is not entirely due to genetic background. Molecular modeling result suggests a less drastic effect of p.R95W on FGF3 function compared with known missense mutations detected in fully penetrant LAMM syndrome. Since we detected significant intrafamilial variability of the inner ear structural phenotype in the family segregating p.R95W, we also sequenced FGF10 as a likely candidate for a modifier. However, we did not find any sequence variation, pointing out that a larger sample size will be needed to map and identify a modifier. We also observed a mild to moderate bilateral conductive hearing loss in three carriers of p.R95W, suggesting either a semi-dominant effect of this mutant allele of FGF3, otitis media, or a consequence of genetic background in these three family members. CONCLUSIONS: We noted a less prominent dental and external ear phenotype in association with the homozygous p.R95W. Therefore, we conclude that the manifestations of recessive FGF3 mutations range from fully penetrant LAMM syndrome to deafness with residual inner ear structures and, by extension, with minimal syndromic features, an observation with implications for cochlear implantation candidacy.

Carcinosarcoma of the esophagus.

Carcinosarcoma of the esophagus is a rare neoplasm characterized histologically by presence of carcinomatous and sarcomatous elements. Case report of carcinosarcoma of the esophagogastric junction whose morphological and immunohistochemical features makes it quite distinctive from other tumours is presented. It was an ulcerated lesion diagnosed in an elderly Afghan lady located 34 cms from the incisor teeth. The patient was a smoker.

Multi-omic studies on missense PLG variants in families with otitis media.

Otitis media (OM), a very common disease in young children, can result in hearing loss. In order to potentially replicate previously reported associations between OM and PLG, exome and Sanger sequencing, RNA-sequencing of saliva and middle ear samples, 16S rRNA sequencing, molecular modeling, and statistical analyses including transmission disequilibrium tests (TDT) were performed in a multi-ethnic cohort of 718 families and simplex cases with OM. We identified four rare PLG variants c.112A > G (p.Lys38Glu), c.782G > A (p.Arg261His), c.1481C > T (p.Ala494Val) and c.2045 T > A (p.Ile682Asn), and one common variant c.1414G > A (p.Asp472Asn). However TDT analyses for these PLG variants did not demonstrate association with OM in 314 families. Additionally PLG expression is very low or absent in normal or diseased middle ear in mouse and human, and salivary expression and microbial α-diversity were non-significant in c.1414G > A (p.Asp472Asn) carriers. Based on molecular modeling, the novel rare variants particularly c.782G > A (p.Arg261His) and c.2045 T > A (p.Ile682Asn) were predicted to affect protein structure. Exploration of other potential disease mechanisms will help elucidate how PLG contributes to OM susceptibility in humans. Our results underline the importance of following up findings from genome-wide association through replication studies, preferably using multi-omic datasets.

Investigation of transmigrated mandibular canines.

INTRODUCTION: Canine transmigration is a rare orthodontic condition and it is relevant to perform the proper diagnose at an early age. OBJECTIVE: The aim of the current study was to find out the frequency of transmigrated mandibular canines (TRC) in orthodontic patients obtained from South Asian population of Pakistan origin. METHODS: Panoramic radiographs of 2,550 untreated orthodontic patients (1,248 males; 1,302 females) were included, to investigate the presence, site, and type of TRC. Any permanent mandibular canine that was found to be crossing the midline in panoramic radiographs was considered as TRC. RESULTS: The frequency of TRC was found to be 0.98%. TRC were only found unilaterally. No significance regarding gender and side was found. Nineteen TRC displayed a type 1 transmigratory pattern, while type 2 and type 5 transmigratory patterns were encountered in three patients. CONCLUSION: Frequency of transmigrated mandibular canines in the studied sample was 0.98%.

A versatile reducible polycation-based system for efficient delivery of a broad range of nucleic acids.

Synthetic vectors based on reducible polycations consisting of histidine and polylysine residues (HIS RPCs) were evaluated for their ability to deliver nucleic acids. Initial experiments showed that RPC-based vectors with at least 70% histidine content mediated efficient levels of gene transfer without requirement for the endosomolytic agent chloroquine. Significant gene transfer was observed in a range of cell types achieving up to a 5-fold increase in the percentage of transfected cells compared to 25 kDa PEI, a gold standard synthetic vector. In contrast to 25 kDa PEI, HIS RPCs also mediated efficient transfer of other nucleic acids, including mRNA encoding green fluorescent protein in PC-3 cells and siRNA directed against the neurotrophin receptor p75(NTR) in post-mitotic cultures of rat dorsal root ganglion cell neurons. Experiments to elevate intracellular glutathione and linear profiling of cell images captured by multiphoton fluorescent microscopy highlighted that parameters such as the molecular weight and rate of cleavage of HIS RPCs were important factors in determining transfection activity. Altogether, these results demonstrate that HIS RPCs represent a novel and versatile type of vector that can be used for efficient cytoplasmic delivery of a broad range of nucleic acids. This should enable different or a combination of therapeutic strategies to be evaluated using a single type of polycation-based vector.

Mutations in SLC25A46, encoding a UGO1-like protein, cause an optic atrophy spectrum disorder.

Dominant optic atrophy (DOA) and axonal peripheral neuropathy (Charcot-Marie-Tooth type 2, or CMT2) are hereditary neurodegenerative disorders most commonly caused by mutations in the canonical mitochondrial fusion genes OPA1 and MFN2, respectively. In yeast, homologs of OPA1 (Mgm1) and MFN2 (Fzo1) work in concert with Ugo1, for which no human equivalent has been identified thus far. By whole-exome sequencing of patients with optic atrophy and CMT2, we identified four families with recessive mutations in SLC25A46. We demonstrate that SLC25A46, like Ugo1, is a modified carrier protein that has been recruited to the outer mitochondrial membrane and interacts with the inner membrane remodeling protein mitofilin (Fcj1). Loss of function in cultured cells and in zebrafish unexpectedly leads to increased mitochondrial connectivity, while severely affecting the development and maintenance of neurons in the fish. The discovery of SLC25A46 strengthens the genetic overlap between optic atrophy and CMT2 while exemplifying a new class of modified solute transporters linked to mitochondrial dynamics.

Stabilisation of cables of fibronectin with micromolar concentrations of copper: in vitro cell substrate properties.

We have previously described the production of large cables of fibronectin, a large extracellular matrix cell adhesion glycoprotein, which has a potential application in tissue engineering. Here we have stabilised these cables for longer survival and looked at their ultrastructural cell-substrate behaviour in vitro. Dissolution experiments showed that low concentrations of copper not only caused significant material stabilisation but left pores which could promote cell ingrowth, as we have previously reported with Fn-mats. Indeed, the greatest amount of cell ingrowth was observed for copper treated cables. Immunostaining showed S-100(+) multi-layers of cells around the edge of cables while ultrastructural analysis confirmed the presence of a mixture of fibroblasts and bipolar cells associated with fragments of basal lamina, which is a Schwann cell phenotype. Interestingly, the outermost layers of cells consisted of S-100(-) cells, presumed fibroblasts, apparently 'capping' the Schwann cells. Toxicity tests revealed that Schwann cells were only able to grow at the lowest concentration of copper used (1microM) while fibroblasts grew at all concentrations tested. These results could be used to design biomaterials with optimum properties for promoting cellular ingrowth and survival in tissue engineered grafts which may be used to improve peripheral nerve repair.

Investigation of transmigrated mandibular canines

ABSTRACT Introduction: Canine transmigration is a rare orthodontic condition and it is relevant to perform the proper diagnose at an early age. Objective: The aim of the current study was to find out the frequency of transmigrated mandibular canines (TRC) in orthodontic patients obtained from South Asian population of Pakistan origin. Methods: Panoramic radiographs of 2,550 untreated orthodontic patients (1,248 males; 1,302 females) were included, to investigate the presence, site, and type of TRC. Any permanent mandibular canine that was found to be crossing the midline in panoramic radiographs was considered as TRC. Results: The frequency of TRC was found to be 0.98%. TRC were only found unilaterally. No significance regarding gender and side was found. Nineteen TRC displayed a type 1 transmigratory pattern, while type 2 and type 5 transmigratory patterns were encountered in three patients. Conclusion: Frequency of transmigrated mandibular canines in the studied sample was 0.98%.

RESUMO Introdução: a transmigração de caninos é uma condição ortodôntica rara, e é importante realizar seu adequado diagnóstico ainda em idade precoce. Objetivo: o objetivo do presente estudo foi avaliar a frequência da transmigração de caninos inferiores (TCI) em pacientes ortodônticos de origem paquistanesa. Métodos: com o objetivo de analisar a presença, o local e o tipo de TCI, foram avaliadas 2.550 radiografias panorâmicas pré-tratamento (1.248 homens, 1.302 mulheres) de pacientes ortodônticos. Foram considerados como TCI os caninos inferiores permanentes que, na radiografia, estivessem cruzando a linha média. Resultados: a frequência de TCI foi de 0,98%. Os achados demonstraram a presença de apenas TCI unilaterais, sem diferenças significativas em relação ao sexo e ao lado da arcada. O tipo 1 de transmigração foi encontrado em 19 pacientes e os tipos 2 e 5, em três pacientes. Conclusão: na presente amostra de origem paquistanesa, a frequência da transmigração de caninos inferiores foi de 0,98%.

Biodegradation of toluene using Candida tropicalis immobilized on polymer matrices in fluidized bed bioreactors.

A yeast strain, Candida tropicalis, was whole-cell-immobilized on polymer matrices of polyethylene glycol (PEG) and polyethylene glycol/activated carbon/alginate (PACA). The polymer matrices were used as fluidized materials in bubble-column bioreactors for the biodegradation of toluene. Simultaneously, another bubble-column bioreactor using granular activated carbon (GAC) and a conventional compost biofilter were operated for comparison. In the compost biofilter, the toluene removal efficiency gradually deteriorated due to the limitation of microbial activity. The toluene removal in the GAC bioreactor was relatively high because of an increase of toluene mass transfer. However, low toluene removal efficiencies were observed in the PEG bioreactor, presumably because the synthetic polymer alone was not suitable for yeast cell immobilization. In the PACA bioreactor, toluene removal was found to be greater than 95% overall. The CO(2) yield coefficient calculated at the highest toluene loading condition for the PACA bioreactor was found to be higher than those observed in the other bioreactors. Furthermore, almost complete elimination capacities were observed in the PACA bioreactor at short-term toluene loading up to 180 g/m(3)/h. In conclusion, the immobilization of C. tropicalis in the PACA matrix resulted in enhanced toluene biodegradation because of the increases of both mass transfer and microbial activity.

Modified palatal stiffening procedure for snoring.

OBJECTIVE: To evaluate the efficacy of a modified procedure of palatal stiffening for the treatment of snoring. STUDY DESIGN: A quasi-experimental study. PLACE AND DURATION OF STUDY: ENT, and Head and Neck Surgery Departments of Pakistan Air Force (PAF) Hospital Masroor, Combined Military Hospital (CMH) Rawalpindi and PAF Hospital Sargodha, from January 2000 to January 2005. METHODOLOGY: Patients of either gender and above 20 years of age having snoring as their primary complaint were registered. Patients with an obvious cause of upper airway obstruction like oropharyngeal swelling, growths and obstructive sleep apnea were excluded. A grading system was designed to classify the patients according to severity and criteria for successful outcome defined; grades were equated with VAS. Palatal stiffening was achieved by using an insulated 22 gauge LP needle for infiltration cautery. Results were analyzed at 6 weeks post operatively on SPSS-16. Paired t-test was applied to compare the improvement in snoring and significance was tested at p-value less than 0.05. RESULTS: Forty four (44) patients underwent the palatal stiffening procedure. Male to female ratio was 42:2. Twenty eight (28) patients (63%) had complete resolution of symptoms after first surgery. Out of the remaining, ten more patients (22.72%) became symptom free after 1st revision surgery while six patients (12.5%) needed 2nd revision. Pain in throat was the main post operative complaint which settled in 10 to 14 days. CONCLUSION: Palatal stiffening procedure causing relatively less morbidity, simple and minimally invasive, but effective at the same time.

Rapid sarcomatous transformation of an ameloblastic fibroma of the mandible: case report and literature review.

Ameloblastic fibrosarcoma (AFS) is a rare malignant odontogenic tumour regarded as the malignant counterpart of ameloblastic fibroma. It is characterized by a benign epithelial component within a malignant fibrous stroma. AFS is a locally aggressive neoplasm with extremely low potential for metastasis. We report an extremely rare, rapidly progressive, and fatal case originating in the posterior mandible of a 20-year old female patient. Initially histopathologically diagnosed as a benign lesion, it rapidly recurred with apparent transformation into a high-grade sarcoma over a period of 6 months. Subsequent intracranial and pulmonary metastases were noted, and the patient died within 15 months of initial consultation. This case emphasizes the need for a high element of suspicion about clinically ambiguous lesions. We recommend more extensive or radical, primary excisions in lesions that have a known potential for recurrence or malignancy.

Effects of the internal recycling rate on biological nutrient removal and microbial community structure in a sequential anoxic/anaerobic membrane bioreactor.

This study investigated the effects of the internal recycling rate on nutrients removal in a sequential anoxic/anaerobic membrane bioreactor (SAM). Microbial community structure in sludge from the SAM was studied using quinone profile method. Above 98% COD, 68% nitrogen, and 55% phosphorus removal efficiencies were achieved when the internal recycling rate was 2.5 times influent flow. At that rate, the optimum specific nitrate loading rate and COD/NO(3)-N ratio were found to be 2.24 mgNO(3)-N g(-1) MLSS h(-1) and 9.13, respectively. Batch tests demonstrated that anoxic condition suppressed phosphorus release, and that denitrification was also influenced by initial substrate concentration. Denitrification appeared to have some priority over phosphorus release for substrate uptake. Microbial community analysis revealed a predominance of the subclass beta-Proteobacteria. Furthermore, it was found that Rhodocyclus-related bacteria were efficient at phosphorus removal than Actinobacteria.