The Design of Novel Nano-Sized Polyanion-Polycation Complexes for Oral Protein Delivery

Introduction Oral delivery of proteins faces numerous challenges due to their enzymatic susceptibility and instability in the gastrointestinal tract. In recent years, the polyelectrolyte complexes have been explored for their ability to complex protein and protect them against chemical and enzymatic degradation. However, most of the conventional binary polyelectrolyte complexes (PECs) are formed by polycations which are associated with toxicity and non-specific bio-interactions. The aim of this thesis was to prepare a series of ternary polyelectrolyte complexes (APECs) by introduction of a polyanion in the binary complexes to alleviate the aforementioned limitations. Method Eight non-insulin loaded ternary complexes (NIL APECs) were spontaneously formed upon mixing a polycation [polyallylamine (PAH), palmitoyl grafted-PAH (Pa2.5), dimethylamino-1-naphthalenesulfonyl grafted-PAH (Da10) or quaternised palmitoyl-PAH (QPa2.5)] with a polyanion [dextran sulphate (DS) or polyacrylic acid (PAA)] at 2:1 ratio, in the presence of ZnSO4 (4μM). A model protein i.e., insulin was added to a polycation, prior to addition of a polyanion and ZnSO4 to form eight insulin loaded (IL) APECs. PECs were used as a control to compare APECs. The complexes were characterised by dynamic light scattering (DLS) and transmission electron microscope (TEM). In vitro stability of the complexes was investigated at pH (1.2-7.4), temperature (25˚C, 37˚C and 45˚C) and ionic strength (NaCl-68mM, 103mM and 145mM). Insulin complexation efficiency was assessed by using bovine insulin ELISA assay kit. The in vitro cytotoxicity was investigated on CaCo2 and J774 cells by MTT (3-4,5 dimethyl thialzol2,5 diphenyl tetrazolium bromide) assay. All complexes were evaluated for their haemocompatibility by using haemolysis assay, oxidative stress by reactive oxygen species (ROS) assay and immunotoxicity by in vitro and in vivo cytokine generation assay. The potential of the uptake of complexes across CaCo2 cells was determined by flow cytometry and fluorescent microscopy. The underlying mechanism of transport of complexes was determined by TEER measurement, assessment of FITC-Dextran and insulin transport across CaCo2 cells. 15 Results NIL QPa2.5 APECs (except IL QPa2.5-DS) exhibited larger hydrodynamic sizes (228-468nm) than all other APECs, due to the presence of bulky quaternary ammonium moieties. QPa2.5 APECs exhibited lower insulin association efficiency (≤40%) than other APECs (≥55%) due to a competition between the polyanion and insulin for QPa2.5 leading to reduced association of insulin in the complexes. DS based APECs generally offered higher insulin association efficiency (≥75%) than PAA based APECs (≤55%) due to higher molecular weight (6-10kDa) of DS. In comparison to other complexes, Pa2.5 PECs and APECs were more stable at varying temperature, ionic strength and pH due to the presence of long palmitoyl alkyl chain (C16) which reduced the chain flexibility and provided stronger hydrophobic association. The cytotoxicity of polycations on CaCo2 and J774 cells is rated as PAH>Da10=Pa2.5>QPa2.5. The introduction of PAA in Pa2.5 and Da10 brought most significant improvement in IC50 i.e., 14 fold and 16 fold respectively on CaCo2 cells; 9.3 fold and 3.73 fold respectively on J774 cells. In comparison to other complexes, Da10 (8mgml-1) induced higher haemolytic activity (~37%) due to a higher hydrophobic load of 10 percent mole grafting of dansyl pendants. The entire range of APECs displayed ≤12% ROS generation by the CaCo2 cells. The degree of in vitro TNFα production (QPa2.5≥Da10≥Pa2.5=PAH) and in vitro IL-6 generation (QPa2.5≥Pa2.5=PAH≥Da10) by J774 cells established an inverse relationship of cytotoxicity with the cytokine generation. Similar to MTT data, the introduction of PAA in APECs brought more significant reduction in in vitro cytokine secretion than DS based APECs. Pa2.5-PAA brought the most significant reduction in both in vitro and in vivo cytokine generation. All the formulations were able to significantly reduce original TEER, however did not demonstrate appreciable paracellular permeation of a hydrophilic macromolecular tracer of paracellular transport i.e., FITC Dextran. The uptake study revealed internalisation of APECs predominantly by a transcellular route. Transcellular uptake of IL QPa2.5 (≤73%), IL QPa2.5-DS (67%) was higher than their NIL counterparts, whereas the uptake of NIL Pa2.5 (≤89%), NIL Pa2.5-PAA (42%) was higher than their IL counterparts. Conclusion In essence, amphiphilic APECs have shown polyanion dependent ability to reduce polycation associated toxicity and they are able to facilitate transcellular uptake of insulin across CaCo2 cells

Rational use of antibiotics--a quality improvement initiative in hospital setting

Objectives: To minimise irrational use of antibiotics by implementing guidelines for antibiotic usage in obstetrics and Gynaeocology.Methods: The observational study was conducted from January to December 2010 at the maternity unit of Aga Khan Hospital for Women and children, Kharadar, a secondary care facility in Karachi, Pakistan. Data was collected from medical records related to the study period. Prophylactic antibiotics were given according to the American College of Obstetricians and Gynaecologists recommendation 2009. Surveillance was done by surgical site infection rates and infectious morbidity. Data was analysed on SPSS 13.Results: Therapeutic antibiotic use was rationalized, reducing the use of therapeutic antibiotics from 97% (n= 160/165) in January 2010 to 8% (n=10/125) in December 2010. Surgical site infection rates were less than 5%. Cost of antibiotics per patient decreased by 90%. Decrease in the length of stay and workload on nursing staff was also observed.Conclusion: Implementing guidelines for antibiotic use in obstetrics and gynaecology and translating it into our protocols was effective in decreasing the irrational antibiotic consumption and increasing the rational use of antibiotics in the hospital

Impact of global financial crisis on socially innovative microfinance institutions in Pakistan

The wave of global financial crises (2007 – 2008) caused a surge in the capital flows of developed countries particularly, between developed and developing countries. The crisis has hit all financial sectors with unexpected severity and speed. This paper determines the impact of global financial crisis (2007 – 2008) on socially innovative microfinance institutions operating in Pakistan by using descriptive ratio analysis and the Wilcoxon Signed Ranks Test. This paper analyzes performance of MFIs for 15 years i.e., from 2000 – 2014 in three waves: before, during and after the financial crisis. The results show that financial crisis affected performance of all selected MFIs but Thardeep Rural Development Programme (TRDP) showed major changes in three waves of crises. The output of the Wilcoxon Signed Ranks Test confirms that the financial crisis worsened the operations of MFIs in Pakistan. This study will assist microfinance practitioners, policy makers, rural financial institutions, and microfinance institutions in maintaining and developing more effective strategies to survive in such crisis in the future.peer-reviewe

Women’s Social Empowerment and Microfinance: A Brief Review of Literature

Women’s empowerment is essential for socio-economic development as it is considered as one of the major factors for increasing worldwide development efforts. The influence of microfinance on women\u27s empowerment is a debatable topic, as past studies have documented. Some researchers claim that microfinance facilitates women to increase their earning capacity, leading to greater control to tackle cultural irregularities. Others contend that their spouses mostly control micro-loans provided to women, causing severe demotion of women. In this paper, we examined the findings of past studies through word cloud analysis to determine the influence of microfinance on the decision-making power of women and their social empowerment. Among the 54 studies reviewed in this paper, most research studies were found to conclude positive associations between microfinance and women\u27s empowerment. In several studies, results varied due to the measure of empowerment applied gender or socio-political differences. This study is one of its kind in that it evaluated past literature for assessing the relationship between microfinance and women\u27s empowerment

The Effects of Variable Viscosity on the Peristaltic Flow of Non-Newtonian Fluid through a Porous Medium in an Inclined Channel with Slip Boundary Conditions

The present paper investigates the peristaltic motion of an incompressible non-Newtonian fluid with variable viscosity through a porous medium in an inclined symmetric channel under the effect of the slip condition. A long wavelength approximation is used in mathematical modeling. The system of the governing nonlinear partial differential equation has been solved by using the regular perturbation method and the analytical solutions for velocity and pressure rise have been obtained in the form of stream function. In the obtained solution expressions, the long wavelength and low Reynolds number assumptions are utilized. The salient features of pumping and trapping phenomena are discussed explicitly. The flow is investigated in a wave frame of reference moving with velocity of the wave. The features of the flow characteristics are analyzed by plotting the graphs of various values of the physical parameters in detail

Spectrum of neuromuscular injuries in victims of bomb blasts

Introduction: Bomb blast (BB) injuries outside war zones were a rare phenomenon until recently. With the spreading wave of terrorism across the globe, BB related trauma is resurfacing. Explosions can produce unique patterns of neuromuscular injury. No recent data exists categorizing such injuries. Methods: Medical records and electrodiagnostic findings of 20 patients with BB related neuromuscular injuries (NMI) were reviewed retrospectively. Results: Most common site of injury was the lower extremities (55%) with majority presenting clinically with foot drop (40%) or weakness (30%). 9/17 patients (45%) had associated shrapnel or penetrating trauma. 6/20 patients had associated fractures. The most common finding was of mononeuropathies. Discussion: As we re-enter an era of war, we need to recreate awareness of the possible spectrum of NMI. Awareness of such injuries will lead to early identification of nerve trauma and the possibility of reduction in overall disability if treated appropriately soon after the injury

Spectrum of Spinal Dysraphism in Pediatric Patients in a Tertiary Care Hospital

Objectives:  To report the spectrum of spinal dysraphism presenting in pediatric patients admitted to the Department of Neurosurgery Lady reading hospital Peshawar. Material & Methods:  A descriptive case series was conducted and total of 89 patients (age between 2 months to 12 years) were included who underwent the surgical treatment. All patients were examined for clinical and radiological diagnosis of spinal dysraphism. Results:  The average age at treatment was 23 ± 39.77 months. The most recurring (52.8%) presenting symptom was the swelling on the back followed by lower limb weakness. Meningocele was reported in 11.2%, myelomeningocele in 47.2%, myelomeningocele & hydrocephalus in 12.4%, tethered cord syndrome in 25.8%, and diastematomyelia 3.4% of patients. Excision combined with the repair was done in 58.4%, release & repair done in 29.2% and endoscopic third ventriculostomy/ventriculoperitoneal shunts with the repair were done in 12.4% patients. Cerebrospinal fluid leak was reported in 3.7%, wound infection in 4.5% and mortality was reported in 3.4% patients. Conclusion:  Overall, a good outcome was reported in the majority of our patients. Surgical procedures like myelomeningocele’s excision & repair, tethered cord’s release & repair, and ETV/VP shunt in patients with hydrocephalus can lead to satisfactory clinical outcomes

Hip fragility fractures: Anaemia, calcium and vitamin D supplementation

Objective: To assess the burden of anaemia and osteoporosis in hip fracture patients, to determine the rate of blood transfusion per-operatively, to assess the use of calcium and vitamin D supplements and the use of anti-osteoporotic medications postoperatively.Methods: A retrospective study was conducted at Aga Khan University and Hospital (AKUH) between June 2009 and May 2011, and comprised record of patients treated for intertrochanteric and femur neck fractures. Patients with associated pathological, open or long bone fractures were excluded. The main study outcome measures were the use of pre-operative and post-operative calcium, vitamin D, bisphosphonates and management of anaemia pre- and post-operatively. Demographic data was also collected including age, gender, and co-morbids. Statistical analysis was performed using SPSS 19.Results: Of the 129 patients, 65(50.4%) were women. The overall mean age was 67.2±15.5 years. Mean pre-operative haemoglobin level was 12.3±1.5 gm/dl and 39(30%) patients were anaemic, while post-operative haemoglobin was 10.2±1.71 with 90(70%) anaemic patients, but only 14(10.9%) patients received per-operative blood transfusion. Pre-operative and post-operative vitamin D supplementation was advised in 3(2.3%) and 18(14%) patients respectively, whereas pre-operative and post-operative bisphosphonate supplementation was advised in 3(2.3%) and 1(0.8%) patients.Conclusions: There is a strong need to pay attention to the management of peri-operative anaemia and calcium, vitamin D and bisphosphonate supplementation in the discharge medications of patients with hip fragility fractures

Impact of Authentic Material on ESL Learners’ Receptive Vocabulary Size

In Pakistani pedagogical process, English is taught as foreign or second language and as a compulsory subject from class 1 to bachelor level. But the results are unsatisfactory in the whole process. Various studies (Heflin, & Alaimo, 2007) have been conducted to find out the reasons and remedies but vocabulary being the most important part of a language is not given due concern in pedagogical process. This research study is undertaken to observe the impacts of authentic material i.e. newspaper on ESL learners’ receptive vocabulary size at intermediate level. This is basically a quantitative research which borrowed the framework of an experimental research. The population was the ESL learners at bachelor level in Lahore. Through convenience sampling technique, the researchers have selected 50 students and formed them into control and experimental groups. The data were collected with pre-test and post-test tools. Later, data were analyzed statistically and shown in charts. In this research study, the researchers have found that good results can be obtained by teaching ESL learners with the help of authentic material i.e. newspaper as compared to the traditional methods of text book teaching. The EFL process can be enhanced with living and practical settings

Impact of Authentic Material on ESL Learners’ Receptive Vocabulary Size

In Pakistani pedagogical process, English is taught as foreign or second language and as a compulsory subject from class 1 to bachelor level. But the results are unsatisfactory in the whole process. Various studies (Heflin, & Alaimo, 2007) have been conducted to find out the reasons and remedies but vocabulary being the most important part of a language is not given due concern in pedagogical process. This research study is undertaken to observe the impacts of authentic material i.e. newspaper on ESL learners’ receptive vocabulary size at intermediate level. This is basically a quantitative research which borrowed the framework of an experimental research. The population was the ESL learners at bachelor level in Lahore. Through convenience sampling technique, the researchers have selected 50 students and formed them into control and experimental groups. The data were collected with pre-test and post-test tools. Later, data were analyzed statistically and shown in charts. In this research study, the researchers have found that good results can be obtained by teaching ESL learners with the help of authentic material i.e. newspaper as compared to the traditional methods of text book teaching. The EFL process can be enhanced with living and practical settings