Sohail Ejaz

Effect of aged garlic extract on wound healing: a new frontier in wound management.

Successful wound healing depends upon angiogenesis, and impaired angiogenesis is a hallmark of the chronic wounds encountered with diabetes and venous or arterial insufficiency. To intervene and improve wound closure, it is essential to investigate the effects of different natural remedies in wound healing. The chicken dorsum skin excisional wound assay was used to investigate the influence of different concentrations of aged garlic solution (AGS) on wound healing. Gross, histopathology, scanning electron microscopy (SEM) and computer-based three-dimensional (3D) image-probing techniques were utilized to determine the effects of AGS on wound closure, re-epithelialization, dermal matrix regeneration, and angiogenesis. Ninety chicks, aged 1 week and divided in 6 groups, were topically exposed to different concentrations of AGS for 6 days: control (group A), 1% (group B), 5% (group C), 10% (group D), 15% (group E), and skin lotion (group F). Different patterns, ranging from incomplete to almost complete wound closure, were observed among different groups with highly significant results (P < 0.001) in group E. Histological investigations revealed a positive augment in the re-epithelialization of all AGS exposed wounds. An increase in the number of new loosely packed collagen and maturation of collagen bundles was observed in all treated wounds at days 4 and 6 post-wounding, respectively. Similar results were achieved through SEM of treated wounds. Histological investigations revealed the profuse dose-dependent neovascularization among AGS-treated wounds. Abbott curve, angular spectrum, and different parameters of 3D surface roughness of wounds were also measured for the precise quantification of angiogenesis. A very highly significant (P < 0.001) increase in angiogenesis was observed among all treated groups. No significant change was observed among control and skin lotion–treated groups. These observations substantiate the beneficial use of AGS in the treatment of wounds. Additional studies are needed to study the specific wound-healing mechanisms of chemical, or group of chemicals, present in AGS.

Ecotoxicological risks associated with tannery effluent wastewater.

The problem of water pollution acquires greater relevance in the context of a developing agrarian economy like Pakistan. Even though, the leather industry is a leading economic sector in Pakistan, there is an increasing environmental concern regarding tanneries because they produce large amounts of potentially toxic wastewater containing both trivalent and hexavalent chromium, which are equally hazardous for human population, aquaculture and agricultural activities in the area. Therefore, we defined the scope of the present study as to employ different bioassays to determine the eco-toxic potential of tannery effluent wastewater (TW) and its chromium based components, i.e., potassium dichromate (K(2)Cr(2)O(7)) and chromium sulfate Cr(2)(SO(4))(3). Particle-induced X-ray emission (PIXE) analysis of TW was carried out to determine the concentration of chromium in TW and then equal concentrations of hexavalent (K(2)Cr(2)O(7)) and trivalent chromium Cr(2)(SO(4))(3) were obtained for this study. Cytotoxicity assay, artemia bioassay and phytotoxicity assay was utilized to investigate the eco-toxicological potential of different concentrations of TW, K(2)Cr(2)O(7) and Cr(2)(SO(4))(3). All the dilutions of TW, K(2)Cr(2)O(7) and Cr(2)(SO(4))(3) presented concentration dependent cytotoxic effects in these assays. The data clearly represents that among all three tested materials, different dilutions of K(2)Cr(2)O(7) caused significantly more damage (P<0.001) to vero cell, brine shrimp and germination of maize seeds. Interestingly, the overall toxicity effects of TW treated groups were subsequent to K(2)Cr(2)O(7) treated group. Based on biological evidences presented in this article, it is concluded that hexavalent chromium (K(2)Cr(2)O(7)) and TW has got significant eco-damaging potential clearly elaborating that environmental burden in district Kasur is numerous and high levels of chromium is posing a considerable risk to the human population, aquaculture and agricultural industry that can obliterate ecosystem surrounding the tanneries.

A novel image probing system for precise quantification of angiogenesis.

The rapid development of clinical diagnostic imaging technology, in tandem with medical and angiogenesis research, has led to some major advances in healthcare. The chorioallantoic membrane assay is commonly used for studying normal angiogenesis as well as putative angiogenic and antiangiogenic substances. Despite the progress, it is generally recognized that a major problem is the lack of a suitable quantitative bioassay for angiogenesis. Image probing is a novel solution to this problem, which, together with its associated discipline of evaluating angiogenesis, is showing great potential not only for accurate measurement of even very small blood vessels but also for detailed three-dimensional quantification of blood vessels and surface characterization. This technique could be a helpful tool for quantification in angiogenesis research.

Toxicological overview of cigarette smoking on angiogenesis.

Angiogenesis is the process of generating new capillary blood vessels. It occurs under tight regulation in the female reproductive system, during wound healing and during embryogenesis. Angiogenesis also plays an important role in the pregnancy-associated changes in the reproductive tract. Cigarette smoke inhibits processes that may hinder normal process of angiogenesis resulting in abnormal blood supply to tissues, decreased repair and remodeling. This report summarizes the evidences of the causal association between tobacco smoking and disruption of angiogenesis. Application of small amount of nicotine on day 5 old chorioallantoic membranes (CAMs) did not disrupt the process of angiogenesis, while application of mainstream smokes (MSS) solutions to CAMs caused varying levels of disruption on normal process of angiogenesis and adversely affect capillary plexus formation, diameters of secondary and tertiary vessels. We have also observed that at equivalent doses, sidestream smoke (SSS) can significantly be more potent than MSS and can alter the normal process of angiogenesis more drastically than MSS. It suggests that SSS either contains a toxicant(s) not present in MSS or that the toxicant(s) that produces these effects is present in higher concentration in SSS than in MSS. Therefore, it is undisputed that smoking can interfere the normal process of angiogenesis, which is a vital process to maintain pregnancy and development of fetus. Smoking during pregnancy is harmful to fetal development and is associated with an increased risk of miscarriage, perinatal death and sudden infant death syndrome. Smoking-cessation programs remain a crucial strategy for preventing poor birth outcomes and decreasing the social and financial costs of smoking during pregnancy.

Toxicological Screening for the Effects of Short-Term Exposure of Sidestream Cigarette Smoke on Angiogenesis

Angiogenesis is a fundamental process that ensures adequate metabolic supply to tissues in numerous biological and pathological states, including tumorigenesis. Cigarette smoking causes numerous adverse effects, some of which are associated with disruption of the normal process of angiogenesis. It is believed that sidestream cigarette smoke severely affects different components that help in the normal process of angiogenesis. Sidestream cigarette smoke is composed primarily of smoke that emanates from the burning end of a cigarette, and contaminants that diffuse through the cigarette paper. The aim of the current study was to ascertain the toxicological effects of different sidestream cigarette smoke solutions (SSCSS) on angiogenesis by using chicken chorioallantoic membrane (CAM) assay. Decrease in total vascular area, diameter of secondary and tertiary blood vessels, cell proliferation, migration of blood vessels toward ectoderm, and number of capillary plexuses formation was observed by application of SSCSS. Scanning microscopy also revealed deviation in pattern formed by the major capillary plexuses and the fibrillar elements of the mesoderm in SSCSS treated CAMs. It is concluded that SSCSS may hinder the normal process of angiogenesis resulting in abnormal blood supply to tissues and decreased repair and remodeling, which are common problems among smoke-exposed individuals. Further study is required to delineate the effects of different chemicals in SSCSS on angiogenesis.

Cortical Selective Neuronal Loss, Impaired Behavior, and Normal Magnetic Resonance Imaging in a New Rat Model of True Transient Ischemic Attacks

Background and Purpose— New-definition transient ischemic attacks (TIAs) are frequent but difficult to diagnose because magnetic resonance imaging (MRI)-negative by definition. However, hidden underlying cell damage might be present and account for the reported long-lasting cognitive impairment after TIAs. Most prior rodent models of true TIA targeted the striatum or have not been fully characterized. Here we present the MRI, behavioral, and quantitative cell changes characterizing a new rodent model of true TIA targeting the more behaviorally relevant cerebral cortex. Methods— Fifteen-minute distal middle cerebral artery occlusion was performed in 29 spontaneously hypertensive rats allowed to survive for 7 to 60 days. Behavior was assessed serially using both global neurological and fine sensorimotor tests. Diffusion- and T2-weighted MRI was obtained 20 min postreperfusion and again 7 to 60 days later, and then changes in neurons and microglia were quantified across the middle cerebral artery territory using immunohistochemistry. Results— No MRI changes or pan-necrosis were observed at any time point, but patchy cortical selective neuronal loss affected 28/29 rats, regardless of survival interval, together with topographically congruent microglial activation that gradually declined over time. The Neuroscore was unchanged, but there was marked contralateral sensorimotor impairment, still recovering by day 28. Conclusions— Our new rodent model mimicking true cortical TIA is characterized by normal MRI, but consistent cortical selective neuronal loss and microglial activation and long-lasting sensorimotor deficits. By causing selective neuronal loss, TIAs and silent microemboli might affect neuronal reserve, thereby increasing long-term cognitive impairment risk. Selective neuronal loss and microglial activation might represent novel therapeutic targets that could be detectable in vivo after TIAs using appropriate imaging tracers.

Characterization of tannery effluent wastewater by proton-induced X-ray emission (PIXE) analysis to investigate their role in water pollution.

IntroductionOver the last few decades, the chromium-based tanning industry has shown rapid growth in Pakistan. However, the rules and regulations promulgated by the government are not strictly followed for processing the effluent discharge from the tanneries. Consequently, tannery effluents have become a great source of water pollution in surrounding areas.Materials and methodsIn this case study, characterization of tannery effluent wastewater (TW), shallow groundwater (SW), and deep groundwater (DW) samples was carried out to determine the source of water pollution in the district of Kasur, Pakistan.ResultsThe concentrations of calcium (Ca), chlorine (Cl), chromium (Cr), iron (Fe), potassium (K), Mg, sulfur (S), silicon (Si), and Sr in TW were significantly higher than SW and DW, which also exceeded the international limits. In addition, increased concentrations of major toxic elements (Cl, Cr, Fe, K, Ni, and Si) were also observed in SW, which were higher in comparison to DW. Strikingly, the concentrations of Cr and Si in various DW samples were also beyond World Health Organization (WHO) safe limit, which reinforced the trend that water pollution in the area is directly linked to the distance from the source (TW). The particle-induced X-ray emission (PIXE) indices also suggested that TW is a main contributory source of water-based pollution in the area, which is imposing great threat to local inhabitants due to known hazardous and carcinogenic potential of these elements.ConclusionProtecting the water resources will be a formidable challenge in the study area, which requires modernization of tannery industry, thereby improving the recovery and recycling of TW. Moreover, PIXE analysis presented here as a successful tool, could serve as landmark for the contemporary research in environmental toxicology.

Regional distribution of selective neuronal loss and microglial activation across the MCA territory after transient focal ischemia: quantitative versus semiquantitative systematic immunohistochemical assessment.

Histopathologic assessment in transient middle cerebral artery occlusion (MCAo) rodent models generally lacks comprehensiveness and exposes to interobserver bias. Here we compared a novel quantitative assessment of regional infarction, selective neuronal loss (SNL) and microglial activation (MA) across the MCA territory to a previously published semiquantitative visual protocol. NeuN and OX42 immunohistochemistry was applied after either 15 or 45 minutes distal MCAo to maximize SNL and infarction, respectively. Survival times varied from 28 to 60 days to cover potential biases such as delayed tissue shrinkage. Damage was assessed using a template of 44 cytoarchitectonic regions of interest (ROIs) mapped onto a subset of digitized coronal sections spanning the MCA territory. For each ROI were obtained a semiquantitative visually determined index of histopathologic changes (method 1), and lpsilateral/contralesional ratios of remaining neurons and activated microglia cell counts (method 2). There was excellent agreement between the two methods for 28-day survival for both MCAo durations, whereas method 2 more sensitively detected subtle SNL and MA at 45 days and 60 days after 15-minute MCAo. Thus the visual method is accurate for usual degrees of ischemic damage, but absolute cell quantification is superior to detect subtle changes and should therefore be preferred in brief MCAo models, although requires optimal staining quality.

Cigarette smoke condensate and total particulate matter severely disrupts physiological angiogenesis.

The cigarette smoke condensate (CSC) and total particulate matter (TPM) in cigarette smoke is extremely toxic and may produce several pathologies. In our study, we used a chicken chorioallantoic membrane assay (CAM) to study the toxicological effects of CSC and TPM on different aspects of angiogenesis. CSC and TPM from four different commercial filtered cigarettes were applied to the CAMs on day 6 of incubation. Macroscopic vascular transformations were observed among all treated CAMs. The application of CSC disks caused obliteration of main blood vessels, while the entire architecture of the secondary and tertiary vasculature was completely destroyed. Likewise, the application of TPM from all brands of cigarette caused the disproportionate thinning of all primary and secondary blood vessels. A reduction in the total area and diameter of the primary, secondary, and tertiary blood vessels was observed after treatment with CSC and TPM. Histological evaluations revealed the loss of ectodermal and mesodermal integrity with both types of treatments. We also noted a profound inflammatory reaction restricted to the disk area with a novel filopodial deformity of the endoderm in the CSC treated groups. Scanty capillary plexus formation, deterioration of the extracellular matrix, and delayed migration of blood vessels were prominent findings among all treated groups. Results obtained from the CSC treated groups were more localized, while more generalized results were recorded in the TPM treated groups. Special caution should be taken for the presence of CSC and TPM while smoking during pregnancy or after surgery because it may severely affect the process of angiogenesis, which is vital to the maintenance of pregnancy and wound healing.

Assessment of the cytotoxic and anti-viral potential of aqueous extracts from different parts of Acacia nilotica (Linn) Delile against Peste des petits ruminants virus

Peste des petits ruminants virus (PPRV); a negative sense single stranded RNA enveloped virus that causes Peste des petits ruminants (PPR), is dramatically affecting the health status of ruminants all around the world resulting in extensive economical losses in livestock sector. Acacia nilotica (Linn) Delile; a tannin-rich medicinal plant distributed throughout subcontinent, is traditionally used as food for ruminants and possesses anti-viral potential against different RNA viruses. In the current study, aqueous extracts from the bark, leaves and pods of A. nilotica (Linn) Delile indica were evaluated for their cytotoxicity and anti-viral activities against PPRV by adopting MTT colorimetric assay and anti-viral assay using Vero cell line. Aqueous extract from the leaves presented significantly better (P<0.05) anti-PPRV activities in comparison to pods extract. On the contrary, bark extract did not show any anti-viral activity. The data presented in the study could pave a way toward the discovery of novel anti-viral chemicals in the plants against PPRV and other viral diseases.