Chia seed-mediated fabrication of ZnO/Ag/Ag2O nanocomposites: structural, antioxidant, anticancer, and wound healing studies.

Plant extract-mediated fabrication of metal nanocomposites is used in cell proliferation inhibition and topical wound treatment, demonstrating significant effectiveness. Salvia hispanica L. (chia) seed extract (CE) is used as the reaction medium for the green fabrication of ecofriendly ZnO(CE) nanoparticles (NPs) and Ag/Ag2O(CE) and ZnO/Ag/Ag2O(CE) nanocomposites. The resultant nanoparticles and nanocomposite materials were characterized using UV-visible, Fourier-transform infrared (FTIR) spectroscopy, scanning electron microscopy (SEM), X-ray diffraction (XRD), and energy-dispersive X-ray (EDX) techniques. In the context of antioxidant studies, ZnO/Ag/Ag2O(CE) exhibited 57% reducing power and 86% 2,2, diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging. All three materials showed strong antibacterial activity against Staphylococcus aureus (S. aureus), Escherichia coli (E.coli), and Bacillus subtilis (B. subtilis) bacterial strains. Additionally, ZnO(CE), Ag/Ag2O(CE), and ZnO/Ag/Ag2O(CE) also revealed 64.47%, 42.56%, and 75.27% in vitro Michigan Cancer Foundation-7 (MCF7) cancer cell line inhibition, respectively, at a concentration of 100 µg/mL. Selectively, the most effective composite material, ZnO/Ag/Ag2O(CE), was used to evaluate in vivo wound healing potential in rat models. The study revealed 96% wound closure in 10 days, which was quite rapid healing compared to wound healing using clinically available ointment. Therefore, in conclusion, the ZnO/Ag/Ag2O(CE) nanocomposite material could be considered for further testing and formulation as a good anticancer and wound healing agent.

Frequency of incidental ocular findings during pre-employment screening at a tertiary care Eye hospital.

OBJECTIVE: To highlight the prevalence of incidentally discovered ocular findings harvested amongst candidates of different age groups presented for pre-employment screening in a tertiary care eye hospital. METHODS: This Cross sectional prospective study was conducted in Armed Forces Institute of Ophthalmology, Rawalpindi, from Jun 2018 to Dec 2019. Data was collected using non-probability consecutive sampling technique. All candidates who appeared for medical fitness examination were included. Candidates belonged to various regions of Pakistan. Complete ophthalmic checkup including visual acuity, best corrected visual acuity, anterior and posterior segment examination was performed. The data analysis was done by IBM SPSS 2.0 software. RESULTS: One thousand and five hundred (1500) candidates underwent ophthalmic medical fitness examination during Jun 2018 to Dec 2019, out of these 86% (1290) were males and 14% (210) were females. Mean age of the candidates was 23.14 ± 5.66 years. The most common incidental ocular findings were amblyopia 24.6% (369), strabismus 10% (150), cataract 7.3% (110), macular scar 6.5% (100) and colour vision deficiencies 5.5% (82). CONCLUSION: The study demonstrates that out of total patients, 77% (1095) were found to be asymptomatic and 23% (405) were symptomatic. The study provides frequency for prevailing diseases and can help in improvement of eye care screening.

Real-Time Polymerase chain reaction trends in COVID-19 patients.

OBJECTIVE: To assess trends of real-time Polymerase Chain Reaction test in Coronavirus infected Patients. METHODOLOGY: This cross-sectional analytical study was conducted at Tertiary Care Institute, Rawalpindi from March 2020 to June 2020. All patients confirmed COVID positive by real-time Polymerase Chain Reaction (PCR) with recent travel history, close contact with known diagnosed patients and had symptoms of fever or upper respiratory tract with body aches. Nasopharyngeal swabs were taken and results generated within 48 hours. Positive PCR was admission criteria follow up was carried out at 7th and 8th day, with negative PCR were discharged. However, those who had persistent positive PCR on the 8th day were tested again on 11th and 12th day. Those with persistent positive results beyond 12th day were shifted to specialized quarantine centres. RESULTS: A total of three hundred and ninety-two patients with mild to moderate illness, PCR positive for COVID 19 were included study with age range 9 - 45 and mean 33.22±7.98 years. A total of 8 (2%) patients were females and 384(98%) males. The duration of the negative test result was Mean ± Std. Deviation 9.05±2.00 with 7 - 8 days 152(38.8%)in and 11 - 12 days in 160(40.8%). PCR results on Day 7 and 8 were negative in 144(36.7%) patients whereas positive in 248(63.3%). PCR results on Day 11 and 12 were negative in 312(79.6%) patients whereas positive in 80 (20.4%). CONCLUSION: To conclude Real-Time Polymerase Chain Reaction (rT-PCR) inclines to give false negative results additionally can stay positive in asymptomatic patients for moderately longer-term. Hence decision to discharge ought to be intricately adjusted between RT-PCR, clinical judgement, radiological examinations, and biochemical assays.

Establishment of Diagnostic Protocols for COVID-19 Patients.

OBJECTIVE: To evaluate the oscillation of laboratory parameters among indoor patients infected with COVID-19. METHODS: This cross-sectional analytical study was conducted at Tertiary Care Institute, Rawalpindi from 01 March 2020 to 20 May 2020. Three hundred and ninety-two patients with mild to moderate illness, PCR positive for COVID 19 were included. Prevalence of typical symptoms of coronavirus disease cough, fever, sore throat and shortness of breath was recorded.PCR was repeated after seven days of admission, if declared negative, another executed on consecutive day. Discharge Criteria was two consecutive negative PCR. RESULTS: A total of 392 patients enrolled in the study with age range 9-45 and mean 33.22±7.98 years. A total of 8 (2%) patients were females and 384(98%) males. 296(75.5%) did not have a cough whereas 96(24.5%) presented with the trait. 296 (75.5%) patients did not have associated fever whereas 96(24.5%) manifested with fever. Chest x-ray had a bilateral patch in 96 (24.5%) patients only. Ferritin was raised in 96 (24.5%) patients however were recorded within normal limits in 296(75.5%) patients. Coagulation Profile was deranged in 64(16.3%) patients whereas was within range in 328(83.7%) patients. Serum Bilirubin, Serum Alkaline phosphatases, Serum Albumin, Serum Urea, Serum Potassium were essentially in typical tolls in 392(100%) patients. However Serum Alanine Aminotransferase was raised in 32(8.2%), Serum C reactive Protein was elevated in 48(12.2%). An exaggerated values of serum creatinine and serum sodium were noticed in 24(6.1%) and 16(4.1%) respectively. CONCLUSION: Routine haematological tests, biochemical tests, serological tests, and radiographic tests are crucial to conclusion, foundation and progression of ailment in COVID-19 contaminated patients.

Targeted lipid nanoparticle delivery of calcitriol to human monocyte-derived macrophages in vitro and in vivo: investigation of the anti-inflammatory effects of calcitriol.

BACKGROUND: Vitamin D3 possesses anti-inflammatory and modulatory properties in addition to its role in calcium and phosphate homeostasis. Upon activation, macrophages (M) can initiate and sustain pro-inflammatory cytokine production in inflammatory disorders and play a pathogenic role in certain cancers. PURPOSE: The main purpose of this study was to encapsulate and specifically target calcitriol to macrophages and investigate the anti-inflammatory properties of calcitriol in vitro and in vivo. METHODS: In this study we have designed and developed near-infrared calcitriol PEGylated nanoparticles (PEG-LNP(Cal)) using a microfluidic mixing technique and modified lipid nanoparticles (LNPs) to target the M specific endocytic receptor CD163. We have investigated LNP cellular uptake and anti-inflammatory effect in LPS-induced M in vitro by flow cytometry, confocal microscopy and gene expression analyses. LNP pharmacodynamics, bio-distribution and organ specific LNP accumulation was also investigated in mice in vivo. RESULTS: In vitro, we observed the specific uptake of PEG-LNP(Cal)-hCD163 in human M, which was significantly higher than the non-specific uptake of control PEG-LNP(Cal)-IgG(h) in M. Pretreatment with encapsulated calcitriol was able to attenuate intracellular TNF-expression, and M surface marker HLA-DR expression more efficiently than free calcitriol in LPS-induced M in vitro. Encapsulated calcitriol diminished mRNA gene levels of TNF-, NF-B, MCP-1 and IL-6, while upregulating IL-10. TNF- and IL-6 protein secretion also decreased. In mice, an in vivo pharmacodynamic study of PEG-LNP(Cal) showed a rapid clearance of IgG and CD163 modified LNPs compared to PEG-LNP(Cal). Antibody modified PEG-LNP(Cal) accumulated in the liver, spleen and kidney, whereas unmodified PEG-LNP(Cal) accumulation was only observed in the liver. CONCLUSION: Our results show that calcitriol can be effectively targeted to M. Our data confirms the anti-inflammatory properties of calcitriol and this may be a potential way to deliver high dose bioactive calcitriol to M during inflammation in vivo.

25(OH)D3 and 1.25(OH)2D3 inhibits TNF-α expression in human monocyte derived macrophages.

PURPOSE: We wanted to investigate effects of vitamin D3 (25(OH)D3 and 1.25(OH)2D3) on inflammatory cytokine expression in both activated and non-activated Mφ. MATERIALS AND METHODS: Mononuclear cells, isolated from healthy donor buffy coats were cultured for a 6-day differentiation-period. Fully differentiated Mφ were pre-treated with either 25(OH)D3 or 1.25(OH)2D3 for (4, 12 or 24 hours) +/-LPS challenge for 4 hours. Gene expression analyses of VDR, Cyp27b1 and pro-inflammatory markers TNF-α, IL-6, NF-κB, MCP-1, was performed using RT-quantitative PCR. TNF-α protein levels from Mφ culture media were analysed by ELISA. RESULTS: Both 25(OH)D3 and 1.25(OH)2D3 significantly inhibited TNF-α expression in both LPS-stimulated and unstimulated Mφ. Also, NF-κB, and to a lesser extend IL-6 and MCP-1 were inhibited. LPS up-regulated Cyp27b1 gene expression which was partly reverted by 1.25(OH)2D3. CONCLUSION: These data show anti-inflammatory effects of vitamin D3 (25(OH)D3 and 1.25(OH)2D3) in human macrophages, and support, that means for targeting high dose vitamin D3 to the immune system may have beneficial clinical effect in inflammatory conditions.

Increased concentrations of the soluble mannose receptor in serum from patients with pneumococcal bacteraemia, and prediction of survival.

BACKGROUND: The soluble mannose receptor (sMR) is a new serum marker of macrophage activation. The aim of the present study was to investigate sMR as a prognostic marker in patients with invasive pneumococcal disease (IPD), and compare it to other inflammatory biomarkers. METHODS: Samples from 128 patients with IPD were collected at the time of first positive blood culture and analysed using an in-house sMR assay. Clinical data were retrieved from patient files. The main outcome investigated was in-hospital mortality. RESULTS: The median sMR concentration in the entire group of patients was 0.77 mg/L. There was a significant difference in sMR concentration between patients below (n = 92, sMR = 0.82 mg/L) or above (n = 36, sMR = 0.73 mg/L) the age of 75 (p < 0.001). In the entire group there was a significant difference in sMR concentrations between survivors (n = 107, sMR = 0.72 mg/L) and non-survivors (n = 21, sMR = 1.38 mg/L), but for patients 75 years or older this difference was not statistically significant. For prediction of survival sMR seemed most promising (area under the receiver operating characteristic curve (AUC) = 0.79) compared with sCD163 (AUC = 0.70) and CRP (AUC = 0.73). In patients 75 years or older the AUC was lower for all three markers (sMR = 0.56, sCD163 = 0.38, CRP = 0.66). CONCLUSIONS: The results of this study designate sMR as a potential new biomarker in infectious disease. Additionally, it emphasizes the importance of research into macrophage malfunction in elderly patients.

A soluble form of the macrophage-related mannose receptor (MR/CD206) is present in human serum and elevated in critical illness.

BACKGROUND: This study tests the hypothesis that the mannose receptor (MR/CD206), which is expressed primarily by macrophages and dendritic cells, can be found in a soluble form (sMR, sMR) in human serum. Furthermore, we wished to establish and validate an enzyme-linked immunosorbent assay (ELISA) for sMR and to perform initial studies exploring the potential of sMR as a biomarker. METHODS: Western blotting identified a single band of approximately 170 kDa in human serum, and MALDI MS/MS of the purified protein confirmed it to be sMR. An ELISA was established and validated with a measurement range of 1-256 µg/L. RESULTS: The 95% reference interval was 0.10-0.43 mg/L based on measurements of serum samples from healthy individuals (n=217). Samples from hospitalised patients (n=219) revealed that more than 50% of patients had concentrations above 0.43 mg/L. Very high concentrations (up to 6.2 mg/L) were observed in critically ill patients with sepsis and/or severe liver disease. CONCLUSIONS: This study documents, for the first time, the presence of sMR in human serum and describes an optimised ELISA suitable for quantitative measurements. Levels of sMR are strongly elevated in several disease states, including sepsis and liver disease, and the protein therefore shows promise as a new biomarker.