Phloroglucinol ameliorated methylglyoxal induced harmful effects in rats.

Glycation is among the underlying mechanisms attributed to ageing and associated morbidities. There is no drug available to combat this deleterious phenomenon. The present study aimed to explore phloroglucinol (PHL) for its anti-glycation potential at preclinical level. The rats were treated with methylglyoxal (MGO, 17.25 mg/kg, i.p. for 14 days) to induce glvcative stress. The treatment groups received additional administration of test drug (PHL; 0.25mg/kg, 0.5mg/kg, and 1mg/kg) or standard aminoguanidine (AG, 50 mg/kg) or saline (control, 5ml/kg). During 14 days, the weight and food intake was noted. Afterwards, the cognitive function was evaluated using Morris Water Maze (MWM) while hepatic and renal functions were assessed through liver function test (bilirubin, alkaline phosphatase, SGPT, and SGOT) and creatinine respectively, using chemical analyzer. The carboxymethyllysine (CML) levels were quantified in the blood using ELISA technique. Histopathological study was performed on the brain, liver, and kidney using H&E staining. Additionally, the qPCR was used to quantify the expression of TNF-α, RAGE and BACE-1 (brain), RAGE, TNF-α, and glyoxalase-I (liver) and RAGE, TNF-α, and VEGF (kidney), while glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as a reference housekeeping gene. The data regarding weight and food intake did not reveal significant alterations. In MWM, the MGO treatment caused significant increase in the time to reach target quadrant, while decrease in the time spent in target quadrant and number of crossings through platform position. All these effects were inhibited by both AG and PHL. The navigation maps also exhibit that the retention of spatial memory. Additionally, the MGO-induced alteration in hepatic and renal function indicators was ameliorated by both AG and PHL treatments. The plasma CML levels were found to be elevated following MGO treatment, while the concomitant administration of AG and PHL has resisted this raise. Histopathological assessment revealed no specific pathology in liver kidney and brain tissues. The qPCR data revealed enhanced expression of all genes, especially TNF-α and BACE, which were found to be reduced following both AG and PHL treatments. PHL prevented the brain, hepatic, and renal impairments caused by MGO induced glycative stress. Hence, the PHL, a clinically used anti-spasmodic drug, presents itself as a potential candidate to be repurposed as anti-glycation drug.

Investigation of wound healing potential of photo-active curcumin-ZnO-nanoconjugates in excisional wound model.

Wound healing, being a dynamic process consisting of hemostasis, inflammation, proliferation, and remodeling, involves the complicated interplay of various growth mediators and the cells associated repair system. Current wound healing therapies usually fail to completely regain skin integrity and functionality. Traditionally, curcumin is considered a potent natural wound healing agent as it possesses antibacterial, antioxidant, and anti-inflammatory properties. It is also known that zinc oxide (ZnO) nanoparticles (NPs) have photocatalytic properties, including the generation of reactive oxygen species. ZnO nanoaprticles are also Food and Drug Administration (FDA) approved as safe substances. While ZnO oxide requires illumination with ultraviolet light to become photocatalytically active, dye-sensitized ZnO can be activated by illumination with visible light. In the present study, we explored the wound healing potential of ZnO nanoparticles sensitized with curcumin (Cu+ZnO Nps) and illuminated with visible (blue) light generated by an array of high power LEDs. We studied the antibacterial effect of our conjugates by percentage reduction in bacterial growth and biofilm formation. The wound healing potential was analyzed by percentage wound contraction, biochemical parameters, and histopathological analysis of the wounded site. Additionally, angiogenesis and wound associated cytokines was evaluated by immunohistochemistry of CD31 and gene expression analysis of IL-1ß, TNF-α, and MMP-9 after 16 days of post-wound treatment, respectively. Our study suggests that the therapeutic effect of Cu+ZnO NPs with LED illumination increases its wound healing potential by producing an antibacterial and anti-inflammatory effect. Moreover, the treatment strategy of using a nano formulation in combination with LED illumination further increases its efficacy. It was concluded that the anti-inflammatory and bactericidal effects of the LED illuminated Cu+ZnO Np showed accelerated wound healing with increased wound contraction, collagen deposition, angiogenesis, and re-epithelialization.

Neuroprotective effect of diclofenac on chlorpromazine induced catalepsy in rats.

Neuroinflammation plays a key role in progressive degeneration of dopaminergic cells. Upregulation of prostaglandins and free radicals formation are involved in the mechanisms of cell death in Parkinson's disease (PD). The present study aimed to investigate the neuroprotective effect of diclofenac against chlorpromazine (CPZ) induced catalepsy and motor impairment in mice. Adult Wistar rats treated with CPZ (3 mg/kg/day, IP) were orally dosed with diclofenac and L-dopa/carbidopa for 21 days. Catalepsy was measured after 21 days of dosing by using standard bar test at 30, 60, 90, 120 and 180 min then motor performances were assessed via open field test and wire hanging test. Histopathological investigation and determination of dopamine (DA) and 3,4-Dihydroxyphenylacetic acid (DOPAC) levels of rat's brain was also carried out. We found that CPZ treated group exhibited reduced motor impairment after 21 days of treatment in open field and wire hanging test (P < 0.01) as compared to control group. The cataleptic scores of CPZ treated rats were also significantly increased (P < 0.01) after 21 days of chronic dosing, however diclofenac treated groups showed significant reduction in cataleptic scores with improved motor performances. Histopathology of CPZ treated rats showed marked degeneration with architecture distortion in the mid brain region. Dopaminergic degeneration is confirmed by neurochemical results that showed reduced amount of dopamine and DOPAC levels in mid brain. Moreover, histopathological slides of diclofenac treated rats showed improved architecture with reduced gliosis of mid brain region as well as improved dopamine and DOPAC levels were achieved after 21 days dosing of diclofenac. Taken together, the present work provide an evidence that diclofenac ameliorated behavioral performances by mediating neuroprotection against CPZ induced PD via preventing dopaminergic neuronal cell death.

Opuntioside, opuntiol and its metallic nanoparticles attenuate adjuvant-induced arthritis: Novel suppressors of Toll-like receptors -2 and -4.

Rheumatoid arthritis (RA) is a chronic autoimmune disease of synovial inflammation and joint destruction. This study reports anti-arthritic potential of opuntioside-I opuntiol, and its gold and silver nanoparticles (NPs) against Complete Freund's Adjuvant (CFA)-induced arthritic rats. The mechanistic studies were performed targeting TLRs (TLR-2 and TLR-4) and cytokines (IL-1ß and TNF-α) expressions to validate their anti-inflammatory and immuno-modulatory response. The nano-formulations were successfully characterized employing Atomic Force Microscopy (AFM) and Dynamic Light Scattering (DLS) analysis. Opuntiol and opuntioside (OP and OPG: 10, 50 and 100 mg/kg) and opuntiol-coated silver and gold NPs (OP-AgNPs and OP-AuNPs: 0.5, 1 and 3 mg/kg) treatments in arthritic rat have shown minimal arthritic score exhibiting mild to moderate articular changes and tissue swelling in ankle joints. Radiographic examination reveals significant reduction in synovitis with improvement in joints degenarative changes in the presence of aforementioned treatments. Likewise, histology of rat ankle joints depicted comparatively lesser influx of inflammatory cells and diminished granulamatous inflammation. Moreover, treatment groups suppressed protein and mRNA expressions of TLRs (TLR-2 and TLR-4) and cytokines (IL-1ß and TNF-α) levels were also significantly declined in the presence of OPG, OP and its NPs comparing to arthritic control. This investigation concludes, the tested compounds and nano-formulations successfully restored the disease progression in CFA-induced arthritic rat owing to their immunomodulatory and anti-inflammatory potentials and can be considered for RA targeted therapy to address the utmost challenges of the disease.

Anticataleptic activity of Zamzam water in chlorpromazine induced animal model of Parkinson disease.

Parkinsonism is characterized by rest tremor, inflexibility, balance debilitation, slow motion and dementia. It is known to be caused by the deficiency of dopaminergic neurons in nigrostriatal pathway. Different studies propose that oxidative burden may be included in the apoptotic process in parkisnons disease. Zamzam water being alkaline in composition may diminish the oxidative stress and hence relieve the symptoms. Therefore, the purpose of this study was to explore the neuroprotective effect of zamzam water in chlorpromazine induced animal model of Parkinsonism. Results revealed that zamzam water did not show significant anticataleptic effect after 21 days as compared to chlorpromazine treated group. However, after 30 days of giving zamzam water showed highly significant decrease (p<0.001) in cataleptic score as compared to chlorpromazine treated group that is negative control. After 30 days of dosing, cataleptic scores by zamzam water were closer to standard drug but standard drug (levodopa/carbidopa) still showed better results than zamzam water. Results from histopathological study of rat's brain also revealed regenerative changes by zamzam treated water when compared with negative control. This regenerative change after zamzam water treatment might play a positive role in future if administered continuously. These results also suggest that zamzam water can be used in combination with standard drug to produce synergistic effect in the management of parkinsons disease.

Gum acacia stabilized silver nanoparticles based nano-cargo for enhanced anti-arthritic potentials of hesperidin in adjuvant induced arthritic rats.

Nanomedicines anticipate drug delivery to inflamed tissues in rheumatoid arthritis (RA) with greater efficacy and lesser side effects. This study investigates the anti-arthritic potentials of Hesperidin (HP) loaded in gum acacia (GA) stabilized green silver nanoparticles (AgNPs). Synthesized GA-AgNPs were characterized through UV-vis spectrophotometer, zetasizer and atomic force microscope (AFM). The HP and its loaded NPs were tested for RA in Complete Freund's adjuvant (CFA) induced arthritis model. GA-AgNPs were found in nano-range size with negative charge, spherical shape and loaded increased HP amount. HP loaded GA-AgNPs showed minimal arthritic score exhibiting mild to moderate tissue swelling, reduced degenerative changes along with mild articular changes. Histopathological analysis revealed comparatively lesser influx of inflammatory cells and diminished granulamatous inflammation in ankle joints tissues in the presence of HP loaded GA-AgNPs. RT-PCR revealed that HP loaded GA-AgNPs significantly reduced the TLRs mRNA expression. Results validate GA stabilized green AgNPs as stable nano-cargos for targeted delivery of HP for restoring the progression of RA.

Bergenin loaded gum xanthan stabilized silver nanoparticles suppress synovial inflammation through modulation of the immune response and oxidative stress in adjuvant induced arthritic rats.

Bergenin (BG) is a naturally occurring C-glycoside with demonstrated anti-arthritic potential. Its therapeutic efficacy is compromised due to its lower absorption and instability at neutral-basic pH. The present study reports fabrication of gum xanthan (GX) stabilized silver nanoparticles (AgNPs) with BG for anti-arthritic activity in a CFA-induced arthritis model targeting ROS, cytokines and TLR expression. NPs were characterized through UV-vis, zetasizer, FT-IR and AFM. Oral administration of BG loaded NPs (1 mg kg-1) exhibited potent anti-arthritic activity with a minimal arthritic score, mild to moderate paw tissue swelling, reduced degenerative changes along with mild articular changes and less influx of inflammatory cells in macroscopic X-ray and histological examination. Administration of BG and its NPs suppressed the levels of reactive oxygen species (ROS) significantly as compared to the arthritic control group. Moreover, increased production of O2˙- in human neutrophils, stimulated by opsonized zymosan (OZ) and phorbol-12-myristate-13-acetate (PMA) was also suppressed. BG and its loaded NPs were revealed to antagonize the oxidative stress via interference with the NADPH oxidase metabolic pathway. Their anti-oxidant activity was further assessed by their inhibitory effect against TLR (TRL-2 & -4) and cytokine (IL-1ß, IL-6 and TNF-α) production. The current investigation validates GX stabilized AgNPs as stable and promising multi-targeted therapeutic nano-cargo for BG delivery with efficient treatment of RA.

Increased Tumour Infiltration of CD4+ and CD8+ T-Lymphocytes in Patients with Triple Negative Breast Cancer Suggests Susceptibility to Immune Therapy

Background: Patients with triple negative breast cancer (TNBC) have limited therapeutic options, largely because the complex tumour environment is not well-characterized. These patients are potential, but largely un-fathomed, candidates for immunotherapy. It is therefore highly relevant to characterize leukocyte complexity in TNBCs. Objective: To investigate leukocyte complexity in tumour environment of patients with TNBCs. Materials and methods: A total of 104 consecutive breast cancer patients undergoing mastectomy were recruited in the study after ethical approval. Clinico-pathological parameters were recorded and H and E staining was performed to investigate tumour morphology. Receptor status was investigated using antibodies against ER, PgR and Her-2, and patients were classified as having TNBC or non-TNBC tumours (including Luminal A, Luminal B and Her2 overexpressing tumours). Immune-cell infiltration was investigated using special stains and antibodies: α-CD3 (T-lymphocytes), α-CD20 (B-lymphocytes), α-CD4 (helper T-lymphocytes) and α-CD8 (cytotoxic T-lymphocytes). Immune cell densities were quantified as cell/ mm2 using the CAP guidelines. Results: Of the 104 breast cancer patients investigated, a total of 27 (26%) had TNBC and 77(74%) non-TNBC. Patients with TNBC showed significantly increased tumour infiltration of lymphocytes (T and B-lymphocytes) compared to the patients with non-TNBC, while myelocytic infiltration was not significantly different in the two groups. Within the TNBC group, infiltration of T-lymphocytes (equal densities of CD4+ and CD8+ T-lymphocytes) was significantly higher compared to B-lymphocytes. Conclusion: Patients with TNBC show increased lymphocytic infiltration (more T-lymphocytes compared to B-lymphocytes). This suggests higher immunogenicity of TNBCs and may indicate a higher responsiveness of these cancers to immunotherapy.

Beneficial anti-Parkinson effects of camel milk in Chlorpromazineinduced animal model: Behavioural and histopathological study.

Potential roles of natural products have been identified for preventing or treating various diseases. Our aim was to investigate the effectiveness of camel milk in an animal model of Parkinson's disease and compare it with standard treatment (levodopa + carbidopa combination). 40 Wistar albino rats weighing 200-250 gram were divided into four groups of 10 animals each. Group I was kept on water and served as normal control, group II served as negative control, treated with chlorpromazine (5mg/kg i.p.), group III was given camel milk (33ml/kg p.o) and group IV the standard combination of levodopa + carbidopa (100+10mg/kg) respectively, 30 minutes after chlorpromazine treatment. All animals were subjected to the drugs treatment for 30 days. Catalepsy was assessed by Bar test on day 21 and day 30 at 30, 60, 90 and 120 minutes interval. On 30th day animals were sacrificed and whole brains were examined for histopathological changes. The results revealed highly significant (p<0.001) anti-cataleptic effect of camel milk on day 21 and 30 in comparison to chlorpromazine. When compared with standard therapy, the results showed that anti-Parkinson's activity of camel milk was significant (p<0.01) on day 21. However, the difference in activity was non-significant on day 30. Histopathology of the brain showed that administration of camel milk reveals intact architecture with mild degenerative changes than chlorpromazine and levodopa + carbidopa treated animals. In conclusion, camel milk possesses anti-Parkinson's activity. However, its long term efficacy and safety needs to be evaluated clinically.

Cancer patterns in Karachi (all districts), Pakistan: First results (2010-2015) from a Pathology based cancer registry of the largest government-run diagnostic and reference center of Karachi.

National level population-based cancer data have never been published from Pakistan in seven decades since independence (1947). Therefore, generation of high-quality regional data becomes highly relevant. Cancer data for the period of 2010-2015 representing the population from all districts of Karachi (14.6 million) are presented herein. After institutional approval (Ref no. IRB-459/DUHS/-14), a Pathology based cancer registry was established at the largest government-run diagnostic and reference center of Karachi. During 2010-2015, a total of 13,508 cancers (including 686 non-melanoma-skin-cancers (NMSC)) were diagnosed. Of these, 5665 (41.9%) were in males while 7843 (58.1%) were in females. Incidence rates for all cancers (excluding NMSC) were 66.7 per 100,000 (crude) and 105.1 per 100,000 (ASR) for males and 112.0 per 100,000 (crude) and 175.8 per 100,000 (ASR) for females. In males, cancer of lip and oral cavity was the most frequently diagnosed cancer (30.8%, ASR 33.1), followed by NMSC (7.7%, ASR 9.5) and colorectum (7%, ASR 7.3). In females, breast cancer was the most frequently recorded malignancy (49.5%, ASR 87.9), followed by lip and oral cavity (11.2%, ASR 22.0) and oesophagus (5.6%, ASR 10.7). We report that Karachi has the highest incidence of cancers of breast, lip and oral cavity, oesophagus and larynx in females and cancer of lip and oral cavity and larynx (2nd only to Turkey) in males compared to any of the Asian populations. Notably, incidence of tobacco associated cancers is very high in Karachi, demanding urgent attention by relevant authorities to address the un-controlled and drastically high consumption of various forms of tobacco in the city.

Resilience Through Squamous Cell Carcinoma and Rhabdomyosarcoma of Oral Cavity: A Unique Case Report.

Rhabdomyosarcoma (RMS) is a soft tissue neoplasm of mesenchymal origin. It is a commonly encountered malignant tumor amongst pediatric patients, yet relatively rare in adults. It usually involves the head and neck region, genitourinary organs and retroperitoneal structures. In adults, the most commonly affected area is the head and neck region. We present here a case of a 30-year-old male patient with a primary squamous cell carcinoma of the tongue (T1, N0, M1), successfully cured with surgery and chemoradiotherapy and later on development of metachronous ipsilateral lesion on the left lower alveolus. Biopsy was consistent with spindle cell RMS. Immunohistochemistry demonstrated positivity for desmin, vimentin and myogenin, thus confirming the mesenchymal origin. With the best of our literature search, this is an exceptional case presenting two malignant lesions with diverse genetic origins, diagnosed at stage 1 and giving a favorable outcome.

Differential immune cell densities in ductal carcinoma In-Situ and invasive breast cancer: Possible role of leukocytes in early stages of carcinogenesis.

OBJECTIVES: To investigate immune cell densities in pre-neoplastic (DCIS), cancer (IDC) and control breast tissues. METHODS: A total of four preneoplastic, 104 cancer and 104 control samples were analyzed. Morphological classification and prognostic scoring along with quantification of immune cells/mm(2) was performed. Data were entered and analyzed using SPSS version 16. Correlation of immune cell densities with various tumour sub-types was investigated using paired t-test and ANOVA. A p-value of <0.05 was considered as significant. RESULTS: Our data show increased infiltration of lymphocytes (mean lymphocytes = 287.6cells/mm(2)) as well as myelocytes (mean lymphocytes = 117.1cells/mm(2)) in pre-neoplastic tissues. This infiltration was significantly high compared to cancer (p-value<0.001) as well as control tissues (p-value <0.001). Moreover, we report increased infiltration of lymphocytes in cancer tissues compared to controls (p-value<0.001). There was no difference in lymphocyte densities within various tumour sub-types (all p-values >0.05). CONCLUSION: Leukocytes may play a role in early stages of breast carcinogenesis.