Human plasma derived exosomes: Impact of active and passive drug loading approaches on drug delivery.

The aim of the current study was to explore the potential of human plasma-derived exosomes as versatile carriers for drug delivery by employing various active and passive loading methods. Exosomes were isolated from human plasma using differential centrifugation and ultrafiltration method. Drug loading was achieved by employing sonication and freeze thaw methods, facilitating effective drug encapsulation within exosomes for delivery. Each approach was examined for its effectiveness, loading efficiency and ability to preserve membrane stability. Methotrexate (MTX), a weak acid model drug was loaded at a concentration of 2.2 µM to exosomes underwent characterization using various techniques such as particle size analysis, transmission electron microscopy and drug loading capacity. Human plasma derived exosomes showed a mean size of 162.15 ± 28.21 nm and zeta potential of -30.6 ± 0.71 mV. These exosomes were successfully loaded with MTX demonstrated a better drug encapsulation of 64.538 ± 1.54 % by freeze thaw method in comparison 55.515 ± 1.907 % by sonication. In-vitro drug release displayed 60 % loaded drug released within 72 h by freeze thaw method that was significantly different from that by sonication method i.e., 99 % within 72 h (p value 0.0045). Moreover, cell viability of exosomes loaded by freeze thaw method was significantly higher than that by sonication method (p value 0.0091) suggested that there was membrane disruption by sonication method. In conclusion, this study offers valuable insights into the potential of human plasma-derived exosomes loaded by freeze thaw method suggest as a promising carrier for improved drug loading and maintenance of exosomal membrane integrity.

Pharmacological exploration of traditional plants for the treatment of neurodegenerative disorders.

Alzheimer's disease (AD) is clinically characterized as memory deficits, altered behavior and impaired cognitive functions. The most important risk factor for AD is aging and mounting. Evidences suggested in different studies that traditionally used plants in Asia, China, and Europe significantly affect aging and AD involved neurodegeneration pathways. Research into ethnobotanicals for impaired memory and cognition has been burgeoned in last decades. The inclusion and exclusion criteria for the plant selection were based on reputed herbs recommended for treatment of neurological disorders and their scientific validation to cure neurodegenerative disorders. A range of traditional plants imparts effects via acetylcholinesterase activity, ß-amyloid peptide formation in plaques, neurotrophic factors and through antioxidant activity. On one side preclinical investigations identified promising drug candidates for AD, on the other side, clinical evidences are still pending. Presently, according to WHO, around more than 80% world population relay on natural remedies to cure their health related issues. Plants contain rich source of primary and secondary metabolites for improving health problems. Pharmaceutical industry is facing intriguing challenges like elevated cost and unendurable risk management due to the high burden of neurodegenerative disorders. A significant shift of drug discovery is being witnessed from synthetic moieties to herbal formulation.

The improvement on the skin surface by a new type of dermocosmetic loaded plant extract: A split face skin topographic study.

Various extrinsic and intrinsic origins slant skins and pledge evident vicissitudes of the skin surface. We explored the effects of dermocosmetic loaded medical Cannabis seed extract and evaluate the improvement on the skin surface in Asian and male volunteers in a split face topographic study. Dermocosmetic and base (without extract) fabricated were directed to apply by volunteers (Asian male) on their right and left cheek, respectively, in the split face skin topographic study up to three months. Efficacy of dermocosmetic versus base was assessed by non- invasive diagnostic technique focusing on skin texture parameters (energy, contrast and variance) and surface evaluation of the living skin (SELS), SEr (skin roughness), SEsc (skin scaliness), SEsm (skin smoothness), SEw (skin wrinkles). Unlike base, dermocosmetic showed significant effects on skin texture parameters (energy, variance and contrast) and SELS (SEr, SEsc, SEsm and SEw) parameters in Asian male volunteers when ANOVA applied. The level of significance was 5%. Dermocosmetic ultimately improved on skin surface and advocacies for anti-aging effects on skin appearance.

Vicissitudes in polyphenolic extract-based high internal phase creams (HIPCs)- effect of storage temperature dependent characteristics.

The purpose of this investigation was to evaluate the vicissitudes in polyphenolic extract- based high internal phase creams (HIPCs) and effect of storage temperature dependent characteristics. Rheological parameters, that is, power law and IPC analysis with its physical characteristics were exploredat different storage temperatures (8°, 25°, 40° and 40° with 75% relative humidity- RH) with different time intervals up to 2 months of newly formulated poly-phenolic extract- based high internal phase cream and its comparison with base. Polyphenolic- based HIPCs showed non-Newtonian-pseudo plastic tendencies in vicissitudes with time and storage temperatures. Data analysis with Power Law and IPC paste was found to fit to all the rheograms. Flow index, shear sensitivity factor, consistency Index and 10 RPM of freshly prepared HIPCs with and without encapsulated polyphenolic extract were found to be 0.5,0.53, 386.4 cP, and 432.9 cP, respectively.The viscosities were fallen with rise in shear stress.There was no change in color, electrical conductivity, liquefaction and phase separation after centrifugation in any sample of polyphenolic extract-based HIPCs and its base. Polyphenolic- based extract HIPCs behaved non-Newtonian- pseudo plastic tendencies and showed stability up to 2 months and can be directed absolutely to shield skin against ultraviolet radiation (UV) intervened oxidative mutilation.

Effect of varying quantities of polymer on preparation and stability evaluation of carbamazepine cocrystals with dicarboxylic acid coformers.

The current study is an attempt to explore the effect of varying quantities of hydroxypropyl cellulose (HPC) polymer on carbamazepine (CBZ) cocrystal formation with dicarboxylic acid coformers i.e., malonic acid (MA), succinic acid (SA), glutaric acid (GA), and adipic acid (AA). The cocrystals were first prepared without polymer by slurry crystallization method and then tried with different quantities of the polymer. The prepared samples were characterized by Powder X-ray Diffraction (XRPD). The characterization results indicate that in methanol pure carbamazepine-malonic (CBZ-MA) and carbamazepine-adipic acid (CBZ-AA) cocrystal can be prepared, while in ethanol and acetone pure carbamazepine-succinic (CBZ-SA) and carbamazepine-glutaric acid (CBZ-GA) cocrystals can be obtained respectively. The same cocrystals were tried using HPC polymer in three different quantities. The characterization results showed that a higher quantity of HPC polymer transforms CBZ-MA cocrystal polymorph-I to polymorph-II. The CBZ-SA and CBZ-GA cocrystal formation somehow inhibited as the concentration of HPC polymer increases. But on the other side, the formation of CBZ-AA cocrystal utterly not inhibited in the presence of varying quantities of HPC polymer. Furthermore, 11 different quantities of HPC were tried to know about the inhibitory concentration of HPC on CBZ-AA cocrystal formation. The CBZ-AA cocrystal preparation was not inhibited even at higher quantities of HPC compared to the coformer. Additionally, the effect of three different quantities of HPC on the thermal stability of the CBZ-AA cocrystal was investigated. Moreover, the stability of pure CBZ at 92% relative humidity (RH) condition was compared to CBZ-AA cocrystal with and without HPC polymer. The CBZ-AA cocrystal with and without HPC polymer was more stable than pure CBZ.

Evaluation of different Pakistani medicinal plants for inhibitory potential against Echis carinatus induced Phospholipase A2 toxicity.

Medicinal plants of Pakistan are known for their curative properties against snake bite as rural people have been using natural herbs for such injuries for hundreds to thousands of years. People of rural areas of Pakistan are prone to snakebite, and on the whole death due to snakebite has been increasing worldwide. The objective of this study was to test the neutralizing potential of 17 Pakistani medicinal plant extracts against phospholipase A2 activity in Echis carinatus venom. Plant material was extracted by simple maceration and fractionation of active plant extracts. Venom was collected by manual massage of the venom glands. The PLA2 enzymatic assay was performed to map out the venomous activity of Echis carinatus envenomation. Snake venom released fatty acids at different concentrations (0.1-5 mg/ml) of venom in a dose-dependent manner. Reduction of pH by 01 correlated with 133 µmol of fatty acids released at 5mg/ml of venom. All plants extract inhibited PLA2 activity, however, Curcuma longa, Citrullus colocynthis and Rubia cordifolia inhibited maximum of PLA2 activity (⁓78%) comparable to the standard antidote (p>0.5). Medicinal plants possess secondary metabolites and many active compounds that may have neutralizing or inhibiting properties against the PLA2 activity of Echis venom. Further studies such as compound analysis could provide an alternative against snakebites injuries resulting from Echis carinatus venom.

Improved in vitro and in vivo performance of carbamazepine enabled by using a succinic acid cocrystal in a stable suspension formulation.

Currently cocrystals are considered as an established approach for making crystalline solids with overall improved physico-chemical properties. However, some otherwise well behaving cocrystals undergo rapid dissociation during dissolution, with ultimate conversion to parent drug and thus apparent loss of improved solubility. The polymeric carriers are long known to manipulate this conversion during dissolution to parent crystalline drug, which may hinder or accelerate the dissolution process if used in a dosage form. The goal of this study was to deliver in vivo a more soluble carbamazepine-succinic acid (CBZ-SUC) cocrystal in suspension formulation utilizing Hydroxypropyl methyl cellulose (HPMC-AS) as a crystallization inhibitor and Polyvinyl carpolactam-polyvinyl acetate-polyethylene glycol graft copolymer ® as solubilizer. The concentration of these polymers were systemically varied during in vitro dissolution studies, while selected formulations from dissolution studies were tested in vivo. Pharmacokinetic studies (PK) in rabbits demonstrated that formulation F7-X (1% cocrystal, 1% HPMC-AS and 2% Polyvinyl carpolactam-polyvinyl acetatepolyethylene glycol graft co-polymer®) caused almost 6fold improvement in AUC0-72 (***P k 0.05) as well as much higher Cmax of 4.73µ.mL-1 to that of 1.07µ.mL-1 of unformulated 'neat' cocrystal given orally. When reference formulation of CBZ (F5-X) with similar composition to F7-X were given to rabbits, cocrystal formulation gave 1.37fold (***P k 0.05) bioavailability than CBZ reference formulation. Cmax of reference formulation observed was 3.9µmL-1.

PERCEPTION OF ACADEMIC PHARMACISTS TOWARDS THEIR ROLE IN HEALTHCARE SYSTEM OF A DEVELOPING COUNTRY, PAKISTAN: A QUANTITATIVE SIGHT.

To investigate perception of academic pharmacists towards their role in healthcare system of a developing country, Pakistan, was the aim of this study. The study participants consisted of academic pharmacists from the government and private universities of Pakistan. Study was conducted for a period of three months, from January to March 2011, in Pakistan. Academic pharmacists were informed regarding the aim, objective and nature of the study. Verbal consent was given and execution of the study took place. Main sections of the questionnaire were comprised of series of statements pertaining to pharmacist perception and experience with the pharmacists. During the period, of January to April, the total number of questionnaires received from different government and private universities was 113. The response rate based on the 205 academic pharmacists working in universities during the study period was 63.9%. A majority (93.9%) of the respondents were satisfied working as academic pharmacist, 68.7% expressed that their interest in research is the reason to embark carrier as an academic pharmacist. Only half (55.7%) of the respondents agreed about pharmacy curriculum standard in Pakistan, and their satisfaction towards curriculum is less, as after the implementation of Pharm. D. (Doctor of Pharmacy). In the country, the curriculum is still focusing more towards theoretical knowledge rather than practical. In this case significant difference was noted with respect to university (p = 0.012). Academic pharmacists do have concern about the pharmacy profession in Pakistan. They ascertain the key issues facing the profession and its educators. Recommendations have been made to improve the current pharmacy curricula in order that future pharmacists in Pakistan practice effectively with the collaboration of other healthcare professionals.

Anti-Aging Potential of Phytoextract Loaded-Pharmaceutical Creams for Human Skin Cell Longetivity.

The exposure to ultraviolet radiations (UVR) is the key source of skin sunburn; it may produce harmful entities, reactive oxygen species (ROS), leading to aging. The skin can be treated and protected from the injurious effects of ROS by using various pharmaceutical formulations, such as cream. Cream can be loaded with antioxidants to quench ROS leading to photo-protective effects. Moreover, modern medicines depend on ethnobotanicals for protection or treatment of human diseases. This review article summarizes various in vivo antioxidant studies on herbal creams loaded with phyto-extracts. These formulations may serve as cosmeceuticals to protect skin against injurious effects of UVR. The botanicals studied for dermatologic use in cream form include Acacia nilotica, Benincasa hispida, Calendula officinalis, Camellia sinensis, Camellia sinensis, Nelumbo nucifera, Capparis decidua, Castanea sativa, Coffea arabica, Crocus sativus, Emblica officinalis Gaertn, Foeniculum vulgare, Hippophae rhamnoides, Lithospermum erythrorhizon, Malus domestica, Matricaria chamomilla L., Moringa oleifera, Morus alba, Ocimum basilicum, Oryza sativa, Polygonum minus, Punica granatum, Silybum marianum, Tagetes erecta Linn., Terminalia chebula, Trigonella foenum-graecum, and Vitis vinifera. The observed anti-aging effects of cream formulations could be an outcome of a coordinating action of multiple constituents. Of numerous botanicals, the phenolic acids and flavonoids appear effective against UVR-induced damage; however the evidence-based studies for their anti-aging effects are still needed.

Biodistribution and kinetic studies of technetium-99m labeled Naja naja karachiensis venom via gamma scintigraphic and SPECT images.

Naja naja karachiensis have been responsible for plentiful deaths in Pakistan. To investigate bio distribution and blood kinetics, venom was labeled with the radiotracer (technetium-99m) by following the method of direct labeling technique. Its maximum labeling percentage was 97.7% (pH 6, 100 µg stannous chloride dihydrate) which was higher than some other reported venom. Radio labeled venom was stable for more than 4 hours both in vivo (96%) and in vitro (serum 94.1%, saline 94.3%) experimentations. Intravenous doses of venom (250 µg, 0.5 mCi) were found to be evenly distributed (having R/L ratio=1.0) in all parts of sacrificed rabbits. Kidneys (53.75% activity/g) and urinary bladder (23.70% activity/g) were found with the copious quantity of injected dose of venom. Rest of all other organs was found with subsequent remaining dose of venom. Among them, lungs (14.2% activity/g), liver (4.32% activity/g), bones (1.38% activity/g), heart (0.8% activity/g), blood (0.56% activity/g), skin (0.45% activity/g), intestines (0.35% activity/g), skeleton muscles (0.3% activity/g), brain (0.14% activity/g) and stomach (0.05% activity/g) are included. After 24 hours of injection, poisoned blood of rabbits was almost cleared from venom. Gamma scintigraphic images (up to 2 hours) along with bio distribution suggest that kidneys are main organs of excretion in rabbits. Elimination started immediately after administration of venom however, possible sites for metabolism of venom are liver and lungs. More accumulation of venom in heart compared to brain suggests its higher affinity (thus possible higher toxicity) to cardiac muscles as compared to brain tissues.

Preparation and radiochemical control of 99mTc labeled blood pool agent for in vivo labelling of the red blood cells.

Our aim was to prepare cheap blood pool imaging kits by simplified method to overcome the burden on purchase department of MINAR, Nishtar Hospital, Multan, Pakistan. Secondarily, prompt supply of kits should save the time of patient during transportation. A total of 24 subjects selected for this study were equally divided into two groups. Mixture of stannous chloride and sodium pyrophosphate solution at pH 7 was injected to these subjects. Various concentrations (ranging from 200 to 800 microg) of stannous chloride dihydrate were injected to group one, followed by intravenous administration of technetium-99m (99mTc) pertechnetate at 30 min interval in 12 subjects. Labeling percentage of each sample was calculated afterwards followed by imaging under gamma camera. Each parameter was tested on three different patients and average of these three was calculated. In second set of experiments done on group two the same procedure was repeated in another 12 subjects, while keeping the concentration of Sn PYP constant at 400 microg. In this case, 99mTc was administered at different time intervals in different subjects ranging from 15 to 120 min (15, 30, 60 and 120 min) followed by calculation of labeling percentage and imaging under gamma camera. In group one, average percentage values of binding of red blood cells with 99mTc were 23.24, 84.88, 83.78 and 60.33% for concentrations of 200, 400, 600 and 800 microg, respectively. In group two, average percentage binging values of 22.26, 84.36. 55.54 and 28.67% were calculated at time intervals of 15, 30, 60 and 120 min, respectively. It is concluded from the results that the best blood pool imaging under gamma camera was observed for the concentration of 400 microg and the time interval of 30 min. The maximum percentage binding of red blood cells with 99mTc was calculated at concentration of 400 microg after 30 min interval that also correlated with imaging results.

Caffeic acid phenethyl ester and therapeutic potentials.

Caffeic acid phenethyl ester (CAPE) is a bioactive compound of propolis extract. The literature search elaborates that CAPE possesses antimicrobial, antioxidant, anti-inflammatory, and cytotoxic properties. The principal objective of this review article is to sum up and critically assess the existing data about therapeutic effects of CAPE in different disorders. The findings elaborate that CAPE is a versatile therapeutically active polyphenol and an effective adjuvant of chemotherapy for enhancing therapeutic efficacy and diminishing chemotherapy-induced toxicities.

Gist of medicinal plants of Pakistan having ethnobotanical evidences to crush renal calculi (kidney stones).

Human civilization is facing the problem of kidney stones since ancient ages. Although mortality rate is not so high, yet it affects the victim's quality of life. The patient suffers from intense pain and many other symptoms modifying his life style and affecting his socioeconomic status. Many drugs and invasive methods have also been developed for the treatment, but these are highly costly and unaffordable for poor people and the rate of reoccurrence is also high. The use of medicinal plants is both affordable and effective in this respect. In this article, 35 medicinal plants of Pakistan origin and their crucial information have been enumerated in alphabetical order of plant's scientific name, family, place (distribution), part used, local name, habit, major constituents and references. It can also be seen that all parts are used for the treatment of kidney stones. Leaves represent 28% contribution, whole plants and seeds 12%, fruits and roots 11% contribution in this respect. Flowers contribute 8% in the treatment of kidney stone while branches, bark, bushes, buds, milk and shoots contribute only 3% in the removal of kidney stones. Habits of plants were also taken under consideration. It was noticed that herbs are the most useful life form in this regard which contributed 63% for the removal of kidney stone. Shrubs contributed 20%, trees 11% while bushes and weeds contributed 3% for the removal of kidney stones.

Interaction analysis of aspirin with selective amino acids.

This study was conducted to assess the compatibility of aspirin with selective amino acids by studying the effect of amino acids on the solubility of aspirin, so that the attention could be paid towards the use of proteinous foods along with aspirin. Two different types of dissolution media, i.e., 0.5% solution of each amino acid and 100 mL of distilled water (100 mL each), were prepared. Then, 1 g of aspirin was added in both media and shaked gently. Ten milliliters of sample was withdrawn at different time intervals, i.e., 10, 20, 30, 40, 50 and 60 min and analyzed spectrophotometrically at 265 nm. It is evident from results that the absorbance of aspirin increased with the addition of amino acids and this increase was significant (p < 0.05). Absorbance after adding amino acid like glycine, tyrosine, glutamic acid, tartaric acid and aspartic acid was observed to be 2.98, 2.96, 2.92, 3.23 and 3.28, respectively, as compared to that of aspirin alone. The increase in absorbance of aspirin in the presence of tartaric acid and aspartic acid was non-significantly (p > 0.05) greater than that in the presence of other amino acids like glycine, tyrosine and glutamic acid. The absorbance of aspirin in the presence of tartaric acid and aspartic acid was 3.23 and 3.28, respectively, while the absorbance of aspirin in the presence of glycine, tyrosine and glutamic acid was 2.98, 2.96 and 2.92, respectively. This study elaborates that the solubility of aspirin increases with concomitant administration of amino acids, thus the use of amino acids (proteinous foods) with aspirin should be prohibited or low dose of aspirin should be recommended in such situation.