Investigation on effect of cerium oxide additive in waste plastic oil fueled CI engine.

In this study, the effects of incorporating cerium oxide into diesel and WPO blends were investigated to determine the potential of the blend as a fuel additive. The study aimed to assess engine-performance, emission, and combustion properties of the blend. The experiments utilized a single-cylinder diesel engine, and researchers prepared two different blends of WPO with 25% WPO in diesel and 50% WPO in diesel. Cerium oxide was added to these blends at concentrations of 25 ppm and 50 ppm using an ultrasonicator. The results demonstrated that increasing cerium oxide content in the blend (50 ppm) led to reduced CO, HC, and NOx emissions at higher loads. For instance, B50 + 50 ppm exhibited lower CO and NOx emissions, while B25 + 50 ppm demonstrated lower HC and smoke emissions. Furthermore, raising the CeO2 content from 25 ppm to 50 ppm resulted in a 3% increase in brake thermal efficiency. Moreover, cerium oxide positively impacted combustion and performance properties of the blends. Among the tested blends, the B50 + 50 ppm combination showcased the highest brake thermal efficiency, optimal air-fuel ratio, and the lowest specific fuel consumption. In conclusion, employing cerium oxide as a fuel additive in diesel-WPO blends offers a promising approach for realizing a sustainable and environmentally friendly future.

Exploring the interplay between particulate matter capture, wash-off, and leaf traits in green wall species.

The study investigated inter-species variation in particulate matter (PM) accumulation, wash-off, and retention on green wall plants, with a focus on leaf characteristics. Ten broadleaf plant species were studied in an experimental green wall. Ambient PM concentrations remained relatively stable throughout the measurement period: PM1: 16.60 ± 9.97 µgm-3, PM2.5: 23.27 ± 11.88 µgm-3, and PM10: 39.59 ± 25.72 µgm-3. Leaf samples were taken before and after three rainfall events, and PM deposition was measured using Scanning Electron Microscopy (SEM). Leaf micromorphological traits, including surface roughness, hair density, and stomatal density, exhibited variability among species and leaf surfaces. Notably, I.sempervirens and H.helix had relatively high PM densities across all size fractions. The study underscored the substantial potential of green wall plants for atmospheric PM removal, with higher Wall Leaf Area Index (WLAI) species like A.maritima and T.serpyllum exhibiting increased PM accumulation at plant level. Rainfall led to significant wash-off for smaller particles, whereas larger particles exhibited lower wash-off rates. Leaf micromorphology impacted PM accumulation, although effects varied among species, and parameters such as surface roughness, stomatal density, and leaf size did not consistently affect PM deposition. The composition of deposited particles encompassed natural, vehicular, salt, and unclassified agglomerates, with minimal changes after rainfall. Air Pollution Tolerance Index (APTI) assessments revealed that I.sempervirens displayed the highest air pollution tolerance, while O.vulgare had the lowest. APTI showed a moderate positive correlation with PM deposition across all fractions. The study concluded that the interplay of macro and micromorphology in green wall plant species determines their PM removal potential. Further research is needed to identify the key leaf characteristics for optimal green wall species selection for effective PM removal.

A trait-based investigation into evergreen woody plants for traffic-related air pollution mitigation over time.

This study investigated influences of leaf traits on particulate matter (PM) wash-off and (re)capture (i.e., net removal) over time. Leaf samples were taken before and after three rainfall events from a range of 10 evergreen woody plants (including five different leaf types), which were positioned with an optical particle counter alongside a busy road. Scanning electron microscopy was used to quantify the density (no./mm2), mass (µg/cm2), and elemental composition of deposited particles. To enable leaf area comparison between scale-like leaves and other leaf types, a novel metric (FSA: foliage surface area per unit branch length) was developed, which may be utilised by future research. Vehicle-related particles constituted 15 % of total deposition, and there was a notable 50 % decrease in the proportion of tyre wear particles after rainfall. T. baccata presented the lowest proportion (11.1 %) of vehicle-related particle deposition but the most consistent performance in terms of net PM removal. Only four of the 10 plant specimens (C. japonica, C. lawsoniana, J. chinensis, and T. baccata) presented effective PM wash-off across all particle size fractions and rainfall intensities, with a generally positive relationship observed between rainfall intensity and wash-off. Mass deposition was more significantly determined by particle size than number density. Interestingly, larger particles were also less easily washed off than smaller particles. Some traits typically considered to be advantageous (e.g., greater hairiness) may in fact hinder net removal over time due to retention under rainfall. Small leaf area is one trait that may promote both accumulation and wash-off. However, FSA was found to be the most influential trait, with an inverse relationship between FSA and wash-off efficacy. This finding poses trade-offs and opportunities for green infrastructure design, which are discussed. Finally, numerous areas for future research are recommended, underlining the importance of systems approaches in developing vegetation management frameworks.

A novel methodology to demonstrate vestibulo-ocular reflex using caloric stimulation in undergraduate physiology laboratory.

The study aims to develop a novel methodology to demonstrate the vestibulo-ocular reflex (VOR) and nystagmus by caloric stimulation in an undergraduate medical physiology laboratory. The experimental setup involved two sets of electrodes: one set positioned laterally to both eyes, and another set positioned vertically over either the right or left eye. The caloric method is used to stimulate ears, which involves irrigation of warm (44°C) and cold (30°C) water into the ears while maintaining a temperature difference of approximately ±7°C from the body temperature. The changes in chorioretinal potential were calibrated to angular displacement by a two-point calibration method, and angular velocity was derived after taking the first-time derivative. The results obtained from the digital data acquisition system were compared to the traditional instrument used in our Otorhinolaryngology Department [Interacoustics Videonystagmography (VNG) System for hospitals, medical grade] for the normal subject's data. No significant differences in angular velocity were noted (P > 0.05). The cold stimuli elicit a more pronounced VOR compared to the warm stimuli. It has been consistently observed that the onset of nystagmus occurs approximately 20 s after irrigation, reaching its peak intensity between 45 and 90 s, and gradually diminishing until it ceases after approximately 200 s. Our developed methodology enables the recording and quantification of nystagmus using easily accessible equipment. This study serves the goal of visualizing the physiological process of VOR and thereby fulfills the goal of an effective teaching tool for demonstrating to undergraduate medical students.NEW & NOTEWORTHY We developed a novel methodology to demonstrate and visualize the most common and important physiological phenomenon like the vestibulo-ocular reflex as a teaching module for undergraduate students.

Randomized controlled trial of sustained release tacrolimus vs twice daily tacrolimus in adult living donor liver transplantation.

BACKGROUND: To compare the safety and efficacy of once-daily tacrolimus (ODT) versus twice-daily tacrolimus (BDT) in adult live donor liver transplantation (LDLT). METHODS: In this open-labelled randomized trial, 174 adult patients undergoing LDLT were randomized into ODT or BDT, combined with basiliximab induction and mycophenolate mofetil (steroid-free regimen). Tacrolimus was started at a total dose of 1 mg and the trough level was aimed at 3-7 ng/ml. The primary endpoint was eGFR at 1,3- and 6 months post-transplant, using CKD- EPI equation. Secondary endpoints included biopsy-proven acute rejection (BPAR), metabolic complications, post-operative bilio-vascular complications and patient survival. RESULTS: There was no statistically significant difference in eGFR between the two groups at 6 months (ODT -96 ± 19, BDT -91 ± 21, p value-0.164). BPAR was comparable (18/84 in ODT, 19/88 in BDT, p value-0.981). For a similar dosage of tacrolimus, the median trough tacrolimus levels attained were significantly lower for ODT than BDT during the first-month post-transplant (p value-0.001). Metabolic complications due to immunosuppression, post-operative bilio-vascular complications and patient survival was similar between the two groups at 6 months. CONCLUSION: Once-daily tacrolimus has similar renal safety and efficacy as twice-daily tacrolimus when used in combination with basiliximab induction and mycophenolate in adult LDLT.

Long-term outcomes of laparoscopic Heller's myotomy with angle of His accentuation in patients of achalasia cardia.

BACKGROUND: Laparoscopic Heller's myotomy (LHM) is an established treatment for achalasia cardia. Anti-reflux procedures (ARP) are recommended with LHM to reduce the post-operative reflux though the optimal anti-reflux procedure is still debatable. This study reports on the long-term outcomes of LHM with Angle-of-His accentuation (AOH) in patients of achalasia cardia. METHODS: One hundred thirty-six patients of achalasia cardia undergoing LHM with AOH between January 2010 to October 2021 with a minimum follow-up of one year were evaluated for symptomatic outcomes using Eckardt score (ES), DeMeester heartburn (DMH) score and achalasia disease specific quality of life (A-DsQoL) questionnaire. Upper gastrointestinal endoscopy, high resolution manometry (HRM) and timed barium esophagogram (TBE) were performed when feasible and rates of esophagitis and improvement in HRM and TBE parameters evaluated. Time dependent rates of success were calculated with respect to improvement in ES and dysphagia-, regurgitation- and heartburn-free survival using Kaplan-Meier analysis. RESULTS: At a median follow-up of 65.5 months, the overall success (ES ≤ 3) was 94.1%. There was statistically significant improvement in ES, heartburn score and A-DsQoL score (p < 0.00001, p = 0.002 and p < 0.00001). Significant heartburn (score ≥ 2) was seen in 12.5% subjects with 9.5% patients reporting frequent PPI use (> 3 days per week). LA-B and above esophagitis was seen in 12.7%. HRM and TBE parameters also showed a significant improvement as compared to pre-operative values (IRP: p < 0.0001, column height: p < 0.0001, column width: p = 0.0002). Kaplan-Meier analysis showed dysphagia, regurgitation, and heartburn free survival of 75%, 96.2% and 72.3% respectively at 10 years. CONCLUSIONS: LHM with AOH gives a lasting relief of symptoms in patients of achalasia cardia with heartburn rates similar to that reported in studies using Dor's or Toupet's fundoplication with LHM. Hence, LHM with AOH may be a preferred choice in patients of achalasia cardia given the simplicity of the procedure.

Comparative study of the effect of N95 facemask and Powered Air-purifying Respirator (2 fans, N95 filter) on cardiovascular parameters of healthy individuals during exercise.

N95 masks filter 95% of the small particles and respiratory droplets (>0.3 µm diameter). Therefore, they are widely used both by general public and health workers during pandemic. When physical activity or exercise is performed wearing N95 mask, it induces hypercapnic environment. The heat burden is also increased leading to discomfort and reduced compliance. This study was done to compare physiological effects and subjective perceptions while wearing N95 mask and powered air-purifying respirator (PAPR) (2 fans, N95 filter) during incremental exercise. ECG, respiratory movement, SpO2, temperature inside the mask were recorded and perception of discomfort was also assessed. Heart rate variability (HRV) values during baseline were within normal limits in both the mask conditions signifying that cardiac autonomic tone is comparable. During incremental exercise, fall in SpO2 was significantly lesser in PAPR as compared to N95 mask at 60-70% and 70-80% of maximum achievable heart rate. The temperatures inside both the mask conditions were significantly higher than ambient temperature. The scores of humid, hot, breath resistance and fatigue were significantly lower in PAPR than N95 mask. In conditions where prolonged use of mask is required with strenuous physical exertion or exercise, PAPR could be preferred over N95 mask.

Fluoxetine or Venlafaxine for Early Post Stroke Depression.

Context: Post stroke depression (PSD) is an under diagnosed morbidity of stroke and can negatively affect the prognosis of the patient. Aims: We intended to study the prevalence of PSD and the commonly used anti-depressants and their outcome in patients with PSD. Settings and Design: A prospective observational study was conducted in the patients admitted to the stroke unit of a tertiary care centre. Methods and Materials: Diagnosis of post stroke depression was made by the Hamilton Depression Rating Scale (HDRS) during the two-week period after stroke or in the clinic follow up. A comparison of clinical outcome and adverse events of the two anti-depressants used, i.e. venlafaxine and fluoxetine were done by a follow up of up to 6 months. Statistical Analysis Used: Independent sample test was used for statistical purposes in the study. Results: Out of the 326 stroke patients admitted in the department, 73 had PSD and 60 patients out of this were assigned into the study. Forty patients were males, and the mean age of the sample population was found to be 62.13 ± 11.14. Major risk factors identified were hypertension, diabetes mellitus, and dyslipidemia. Venlafaxine showed better outcome and less adverse events compared to fluoxetine. Major adverse events observed were hyponatremia, headache, insomnia, and anxiety. Conclusions: PSD in the early phase affects a substantial number of the stroke patients. Venlafaxine has got a better outcome and adverse event profile compared to fluoxetine in this group of patients. However, larger multicenter studies will provide more helpful data in this area.

Genome-Wide Association Mapping Reveals Novel Putative Gene Candidates Governing Reproductive Stage Heat Stress Tolerance in Rice.

Temperature rise predicted for the future will severely affect rice productivity because the crop is highly sensitive to heat stress at the reproductive stage. Breeding tolerant varieties is an economically viable option to combat heat stress, for which the knowledge of target genomic regions associated with the reproductive stage heat stress tolerance (RSHT) is essential. A set of 192 rice genotypes of diverse origins were evaluated under natural field conditions through staggered sowings for RSHT using two surrogate traits, spikelet fertility and grain yield, which showed significant reduction under heat stress. These genotypes were genotyped using a 50 k SNP array, and the association analysis identified 10 quantitative trait nucleotides (QTNs) for grain yield, of which one QTN (qHTGY8.1) was consistent across the different models used. Only two out of 10 MTAs coincided with the previously reported QTLs, making the remaing eight novel. A total of 22 QTNs were observed for spikelet fertility, among which qHTSF5.1 was consistently found across three models. Of the QTNs identified, seven coincided with previous reports, while the remaining QTNs were new. The genes near the QTNs were found associated with the protein-protein interaction, protein ubiquitination, stress signal transduction, and so forth, qualifying them to be putative for RSHT. An in silico expression analysis revealed the predominant expression of genes identified for spikelet fertility in reproductive organs. Further validation of the biological relevance of QTNs in conferring heat stress tolerance will enable their utilization in improving the reproductive stage heat stress tolerance in rice.

Potential health risks due to in-car aerosol exposure across ten global cities.

Car microenvironments significantly contribute to the daily pollution exposure of commuters, yet health and socioeconomic studies focused on in-car exposure are rare. This study aims to assess the relationship between air pollution levels and socioeconomic indicators (fuel prices, city-specific GDP, road density, the value of statistical life (VSL), health burden and economic losses resulting from exposure to fine particulate matter ≤2.5 µm; PM2.5) during car journeys in ten cities: Dhaka (Bangladesh); Chennai (India); Guangzhou (China); Medellín (Colombia); São Paulo (Brazil); Cairo (Egypt); Sulaymaniyah (Iraq); Addis Ababa (Ethiopia); Blantyre (Malawi); and Dar-es-Salaam (Tanzania). Data collected by portable laser particle counters were used to develop a proxy of car-user exposure profiles. Hotspots on all city routes displayed higher PM2.5 concentrations and disproportionately high inhaled doses. For instance, the time spent at the hotspots in Guangzhou and Addis Ababa was 26% and 28% of total trip time, but corresponded to 54% and 56%, respectively, of the total PM2.5 inhaled dose. With the exception of Guangzhou, all the cities showed a decrease in per cent length of hotspots with an increase in GDP and VSL. Exposure levels were independent of fuel prices in most cities. The largest health burden related to in-car PM2.5 exposure was estimated for Dar-es-Salam (81.6 ± 39.3 µg m-3), Blantyre (82.9 ± 44.0) and Dhaka (62.3 ± 32.0) with deaths per 100,000 of the car commuting population per year of 2.46 (2.28-2.63), 1.11 (0.97-1.26) and 1.10 (1.05-1.15), respectively. However, the modest health burden of 0.07 (0.06-0.08), 0.10 (0.09-0.12) and 0.02 (0.02-0.03) deaths per 100,000 of the car commuting population per year were estimated for Medellin (23 ± 13.7 µg m-3), São Paulo (25.6 ± 11.7) and Sulaymaniyah (22.4 ± 15.0), respectively. Lower GDP was found to be associated with higher economic losses due to health burdens caused by air pollution in most cities, indicating a socioeconomic discrepancy. This assessment of health and socioeconomic parameters associated with in-car PM2.5 exposure highlights the importance of implementing plausible solutions to make a positive impact on peoples' lives in these cities.

Prolonged Exposure to Platelet Activating Factor Transforms Breast Epithelial Cells.

Lipid species are known to have various biological functions owing to their structural differences, and each of them possesses a specific role to play depending upon their location and distribution in the cell. Some of these lipids interact with proteins on the cell membrane and acts as second messengers. The level of lipid mediators is generally maintained in the cell by feedback mechanisms; however, their improper degradation or enhanced production leads to their accumulation in the tumor microenvironment and disturbs the homeostasis of the cell. Platelet activating factor (PAF) is a known phospholipid mediator secreted upon immunological challenges by platelets, neutrophils, basophils, and macrophages. PAF, as a potent inflammatory molecule, is well studied, and its role in various cancers and cardiovascular diseases has also been investigated. Interestingly, increased levels of PAF have been found in the blood plasma of smokers, and breast cancer cells have shown the accumulation of PAF in presence of cigarette smoke extract. This accumulation was found to increase tumor cell motility that in turn could promote metastasis. Beyond this, however, the effect of PAF on tumorigenesis has not yet been well explored. Here, we show that the continuous exposure of 3D breast acinar cultures to PAF resulted in the activation of various oncogenic signaling pathways leading to transformation. We also found that the presence of PAF in the micro-environment increased the expression of PAF receptor (PAF-R), which corroborated with the higher expression of PAF-R detected in some epithelial cancers, as per literature. Thus, this study impresses on the fact that the presence of PAF alters the cellular microenvironment and eventually triggers irreversible effects that can cumulatively lead to transformation.

Temporary reduction in fine particulate matter due to 'anthropogenic emissions switch-off' during COVID-19 lockdown in Indian cities.

The COVID-19 pandemic elicited a global response to limit associated mortality, with social distancing and lockdowns being imposed. In India, human activities were restricted from late March 2020. This 'anthropogenic emissions switch-off' presented an opportunity to investigate impacts of COVID-19 mitigation measures on ambient air quality in five Indian cities (Chennai, Delhi, Hyderabad, Kolkata, and Mumbai), using in-situ measurements from 2015 to 2020. For each year, we isolated, analysed and compared fine particulate matter (PM2.5) concentration data from 25 March to 11 May, to elucidate the effects of the lockdown. Like other global cities, we observed substantial reductions in PM2.5 concentrations, from 19 to 43% (Chennai), 41-53% (Delhi), 26-54% (Hyderabad), 24-36% (Kolkata), and 10-39% (Mumbai). Generally, cities with larger traffic volumes showed greater reductions. Aerosol loading decreased by 29% (Chennai), 11% (Delhi), 4% (Kolkata), and 1% (Mumbai) against 2019 data. Health and related economic impact assessments indicated 630 prevented premature deaths during lockdown across all five cities, valued at 0.69 billion USD. Improvements in air quality may be considered a temporary lockdown benefit as revitalising the economy could reverse this trend. Regulatory bodies must closely monitor air quality levels, which currently offer a baseline for future mitigation plans.

Quantifying particulate matter reduction and their deposition on the leaves of green infrastructure.

The green infrastructure (GI) is identified as a passive exposure control measure of air pollution. This work examines particulate matter (PM) reduction by a roadside hedge and its deposition on leaves. The objectives of this study are to (i) quantify the relative difference in PM concentration in the presence of GI and at an adjacent clear area; (ii) estimate the total mass and number density of PM deposited on leaves of a hedge; (iii) ascertain variations in PM deposition at adult (1.5m) and child (0.6 m) breathing levels on either side of a hedge; (iv) illustrate the relationship between PM deposition to leaves and ambient PM concentration reductions; and (v) quantify the elemental composition of collected particles of the leaves on different heights and sides of hedge. PM reduction of 2-9% was observed behind hedge compared to a clear area and followed a trend of ΔPM1 >ΔPM10 >ΔPM2.5. Counting of particles was found to be an effective method to quantify deposition than weighting methods. Sub-micron particles (PM1) dominated particle deposition on leaves at all sampling points on both sides of the hedge. PM mass deposition and number concentration to the leaves on traffic-facing side was up to 36% and 58% higher at 0.6m compared with 1.5m height, respectively. Such a difference was absent on the backside of the hedge. The SEM-EDS analysis showed up to 12% higher traffic-originated particles deposited to leaves on the traffic-facing side compared to the backside. The naturally occurring particles dominated in identified particles on leaf samples from all collection points on the hedge. These new evidence expand our understanding of PM reduction of GI in the near-road environment and its variations in particle deposition, depending on height and sides of GI, which could allow a better parameterisation of dispersion-deposition models for GI assessment at micro-scale.

The nexus between air pollution, green infrastructure and human health.

Cities are constantly evolving and so are the living conditions within and between them. Rapid urbanization and the ever-growing need for housing have turned large areas of many cities into concrete landscapes that lack greenery. Green infrastructure can support human health, provide socio-economic and environmental benefits, and bring color to an otherwise grey urban landscape. Sometimes, benefits come with downsides in relation to its impact on air quality and human health, requiring suitable data and guidelines to implement effective greening strategies. Air pollution and human health, as well as green infrastructure and human health, are often studied together. Linking green infrastructure with air quality and human health together is a unique aspect of this article. A holistic understanding of these links is key to enabling policymakers and urban planners to make informed decisions. By critically evaluating the link between green infrastructure and human health via air pollution mitigation, we also discuss if our existing understanding of such interventions is sufficient to inform their uptake in practice. Natural science and epidemiology approach the topic of green infrastructure and human health very differently. The pathways linking health benefits to pollution reduction by urban vegetation remain unclear and the mode of green infrastructure deployment is critical to avoid unintended consequences. Strategic deployment of green infrastructure may reduce downwind pollution exposure. However, the development of bespoke design guidelines is vital to promote and optimize greening benefits, and measuring green infrastructure's socio-economic and health benefits are key for their uptake. Greening cities to mitigate pollution effects is on the rise and these need to be matched by scientific evidence and appropriate guidelines. We conclude that urban vegetation can facilitate broad health benefits, but there is little empirical evidence linking these benefits to air pollution reduction by urban vegetation, and appreciable efforts are needed to establish the underlying policies, design and engineering guidelines governing its deployment.

Gelatin microspheres releasing transforming growth factor drive in vitro chondrogenesis of human periosteum derived cells in micromass culture.

For cartilage tissue engineering, several in vitro culture methodologies have displayed potential for the chondrogenic differentiation of mesenchymal stem cells (MSCs). Micromasses, cell aggregates or pellets, and cell sheets are all structures with high cell density that provides for abundant cell-cell interactions, which have been demonstrated to be important for chondrogenesis. Recently, these culture systems have been improved via the incorporation of growth factor releasing components such as degradable microspheres within the structures, further enhancing chondrogenesis. Herein, we incorporated different amounts of gelatin microspheres releasing transforming growth factor ß1 (TGF-ß1) into micromasses composed of human periosteum derived cells (hPDCs), an MSC-like cell population. The aim of this research was to investigate chondrogenic stimulation by TGF-ß1 delivery from these degradable microspheres in comparison to exogenous supplementation with TGF-ß1 in the culture medium. Microscopy showed that the gelatin microspheres could be successfully incorporated within hPDC micromasses without interfering with the formation of the structure, while biochemical analysis and histology demonstrated increasing DNA content at week 2 and accumulation of glycosaminoglycan and collagen at weeks 2 and 4. Importantly, similar chondrogenesis was achieved when TGF-ß1 was delivered from the microspheres compared to controls with TGF-ß1 in the medium. Increasing the amount of growth factor within the micromasses by increasing the amount of microspheres added did not further improve chondrogenesis of the hPDCs. These findings demonstrate the potential of using cytokine releasing, gelatin microspheres to enhance the chondrogenic capabilities of hPDC micromasses as an alternative to supplementation of the culture medium with growth factors. STATEMENT OF SIGNIFICANCE: Gelatin microspheres are utilized for growth factor delivery to enhance chondrogenesis of mesenchymal stem cells (MSCs) in high cell density culture systems. Herein, we employ a new combination of these microspheres with micromasses of human periosteum-derived cells, which possess ease of isolation, excellent expansion potential, and MSC-like differentiation capabilities. The resulting localized delivery of transforming growth factor ß1 increases glycosaminoglycan and collagen production within the micromasses without exogenous stimulation in the medium. This unique combination is able to drive chondrogenesis up to similar levels as seen in micromasses that do receive exogenous stimulation. The addition of growth factor releasing microspheres to high cell density micromasses has the potential to reduce costs associated with this strategy for cartilage tissue engineering.

In Vitro Screening of Molecularly Engineered Polyethylene Glycol Hydrogels for Cartilage Tissue Engineering using Periosteum-Derived and ATDC5 Cells.

The rapidly growing field of tissue engineering and regenerative medicine has brought about an increase in demand for biomaterials that mimic closely the form and function of biological tissues. Therefore, understanding the cellular response to the changes in material composition moves research one step closer to a successful tissue-engineered product. With this in mind, polyethylene glycol (PEG) hydrogels comprised of different concentrations of polymer (2.5%, 4%, 6.5%, or 8% (w/v)); different protease sensitive, peptide cross-linkers (VPMSMRGG or GPQGIWGQ); and the incorporation or lack of a peptide cell adhesion ligand (RGD) were screened for their ability to support in vitro chondrogenesis. Human periosteum-derived cells (hPDCs), a mesenchymal stem cell (MSC)-like primary cell source, and ATDC5 cells, a murine carcinoma-derived chondrogenic cell line, were encapsulated within the various hydrogels to assess the effects of the different formulations on cellular viability, proliferation, and chondrogenic differentiation while receiving exogenous growth factor stimulation via the medium. Through the results of this screening process, the 6.5% (w/v) PEG constructs, cross-linked with the GPQGIWGQ peptide and containing the RGD cell binding molecule, demonstrated an environment that consistently supported cellular viability and proliferation as well as chondrogenic differentiation.

RGD-functionalized polyethylene glycol hydrogels support proliferation and in vitro chondrogenesis of human periosteum-derived cells.

The combination of progenitor cells with appropriate scaffolds and in vitro culture regimes is a promising area of research in bone and cartilage tissue engineering. Mesenchymal stem cells (MSCs), when encapsulated within hydrogels composed of the necessary cues and/or preconditioned using suitable culture conditions, have been shown to differentiate into bone or cartilage. Here, we utilized human periosteum-derived cells (hPDCs), a progenitor cell population with MSC characteristics, paired with protease-degradable, functionalized polyethylene glycol (PEG) hydrogels to create tissue-engineered constructs. The objective of this study was to investigate the effects of scaffold composition, exploring the addition of the cell-binding motif Arginine-Glycine-Aspartic Acid (RGD), in combination with various in vitro culture conditions on the proliferation, chondrogenic gene expression, and matrix production of encapsulated hPDCs. In growth medium, the hPDCs in the RGD-functionalized hydrogels maintained high levels of viability and demonstrated an enhanced proliferation when compared with hPDCs in non-functionalized hydrogels. Additionally, the RGD-containing hydrogels promoted higher glycosaminoglycan (GAG) synthesis and chondrogenic gene expression of the encapsulated hPDCs, as opposed to the non-functionalized constructs, when cultured in two different chondrogenic media. These results demonstrate the potential of hPDCs in combination with enzymatically degradable PEG hydrogels functionalized with adhesion ligands for cartilage regenerative applications. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 33-42, 2018.

Initiating human articular chondrocyte re-differentiation in a 3D system after 2D expansion.

Cartilage damage affects a large population via acute and chronic injury and disease. Since native cartilage does not self-renew, cartilage tissue engineering has gained traction as a potential treatment. However, a limiting factor is that the primary cell type in cartilage, the articular chondrocyte, tends to de-differentiate when grown on 2D surfaces for in vitro expansion. Thus, 3D systems are being developed and used to counter this loss of chondrogenic capabilities. We hypothesize that a 3D matrix that can be remodeled may be more supportive of the chondrogenic phenotype of encapsulated articular chondrocytes than a 2D surface and may allow for the re-differentiation of chondrocytes after 2D expansion. Hence, in this study, enzymatically degradable polyethylene glycol (PEG) hydrogels containing two different protease degradable peptide segments, with different degradation rates, were tested in combination with chondrogenic medium as a 3D in vitro culture system to better recapitulate the native environment of human articular chondrocytes (hACs). In addition, the effect of incorporation of the integrin binding ligand Arg-Gly-Asp (RGD) in the hydrogels was explored. Hydrogels crosslinked with a slower degrading crosslinker and not functionalized with RGD maintained hAC viability and led to increased GAG production and chondrogenic gene expression over time, suggesting that this system can initiate hAC re-differentiation after 2D expansion.

Characterization of host response to Cryptococcus neoformans through quantitative proteomic analysis of cryptococcal meningitis co-infected with HIV.

Cryptococcal meningitis is the most common opportunistic fungal infection causing morbidity and mortality (>60%) in HIV-associated immunocompromised individuals caused by Cryptococcus neoformans. Molecular mechanisms of cryptococcal infection in brain have been studied using experimental animal models and cell lines. There are limited studies for the molecular understanding of cryptococcal meningitis in human brain. The proteins involved in the process of invasion and infection in human brain still remains obscure. To this end we carried out mass spectrometry-based quantitative proteomics of frontal lobe brain tissues from cryptococcal meningitis patients and controls to identify host proteins that are associated with the pathogenesis of cryptococcal meningitis. We identified 317 proteins to be differentially expressed (≥2-fold) from a total of 3423 human proteins. We found proteins involved in immune response and signal transduction to be differentially expressed in response to cryptococcal infection in human brain. Immune response proteins including complement factors, major histocompatibility proteins, proteins previously known to be involved in fungal invasion to brain such as caveolin 1 and actin were identified to be differentially expressed in cryptococcal meningitis brain tissues co-infected with HIV. We also validated the expression status of 5 proteins using immunohistochemistry. Overexpression of major histocompatibility complexes, class I, B (HLA-B), actin alpha 2 smooth muscle aorta (ACTA2) and caveolin 1 (CAV1) and downregulation of peripheral myelin protein 2 (PMP2) and alpha crystallin B chain (CRYAB) in cryptococcal meningitis were confirmed by IHC-based validation experiments. This study provides the brain proteome profile of cryptococcal meningitis co-infected with HIV for a better understanding of the host response associated with the disease.

Proteomics of human aqueous humor.

The aqueous humor is a colorless, transparent fluid that fills the anterior chamber of the eye. It plays an important role in maintaining the intraocular pressure and providing nourishment to the lens and cornea. The constitution of the aqueous humor is controlled by the blood-aqueous barrier. Though this ocular fluid has been extensively studied, its role in ocular physiology is still not completely understood. In this study, aqueous humor samples were collected from 250 patients undergoing cataract surgery, subjected to multiple fractionation strategies and analyzed on a Fourier transform LTQ-Orbitrap Velos mass spectrometer. In all, we identified 763 proteins, of which 386 have been identified for the first time in this study. Sorbitol dehydrogenase (SORD), filensin (BFSP1), and phakinin (BFSP2) are some of the proteins that have not been previously reported in the aqueous humor. Gene Ontology analysis revealed 35% of the identified proteins to be extracellular, with a majority of them involved in cell communication and signal transduction. This study comprehensively reports 386 novel proteins that have important potential as biomarker candidates for future research into personalized medicine and diagnostics aimed towards improving visual health.