Recent advances in microfluidic-based spectroscopic approaches for pathogen detection.

Rapid identification of pathogens with higher sensitivity and specificity plays a significant role in maintaining public health, environmental monitoring, controlling food quality, and clinical diagnostics. Different methods have been widely used in food testing laboratories, quality control departments in food companies, hospitals, and clinical settings to identify pathogens. Some limitations in current pathogens detection methods are time-consuming, expensive, and laborious sample preparation, making it unsuitable for rapid detection. Microfluidics has emerged as a promising technology for biosensing applications due to its ability to precisely manipulate small volumes of fluids. Microfluidics platforms combined with spectroscopic techniques are capable of developing miniaturized devices that can detect and quantify pathogenic samples. The review focuses on the advancements in microfluidic devices integrated with spectroscopic methods for detecting bacterial microbes over the past five years. The review is based on several spectroscopic techniques, including fluorescence detection, surface-enhanced Raman scattering, and dynamic light scattering methods coupled with microfluidic platforms. The key detection principles of different approaches were discussed and summarized. Finally, the future possible directions and challenges in microfluidic-based spectroscopy for isolating and detecting pathogens using the latest innovations were also discussed.

The Potential of Fecal and Urinary Biomarkers for Early Detection of Pancreatic Ductal Adenocarcinoma: A Systematic Review.

Pancreatic ductal adenocarcinoma (PDAC) is a highly lethal cancer often diagnosed at advanced stages, highlighting the urgent need for early detection strategies. This systematic review explores the potential of fecal and urinary biomarkers for early PDAC detection. A comprehensive search identified eight relevant studies investigating various biomarkers, including proteins, metabolites, microbial profiles, DNA mutations, and non-coding RNAs. Promising findings suggest that urinary biomarkers related to metabolic alterations, inflammatory processes, fecal microbiome profiles, and fecal miRNAs hold diagnostic potential even at early stages of PDAC. Combining biomarkers into panels may enhance diagnostic accuracy. Challenges such as validation in larger cohorts, standardization of protocols, and regulatory approval must be addressed for clinical translation. Despite these hurdles, non-invasive urinary and fecal biomarkers represent a promising avenue for improving PDAC outcomes through early detection.

Insight into carbohydrate metabolism, protein quantification and mineral regulation in wheat (Triticum aestivum L.) by the action of green synthesized silver nanoparticles (AgNPs) against heat stress.

In the present investigation, the role of GS-AgNPs treatment in wheat plants was carried out in reducing heat stress with the aim of facilitating scientists on this topic. The effect of GS-AgNPs against heat stress has rarely been deliberated in wheat plants, and only a few studies have been established earlier in this scenario. This work illustrated the effect of GS-AgNPs on the regulation of carbohydrates metabolism, SOD, proteins, crude fibers, and minerals changes in wheat plants. Data were analysed using PCA analysis, correlation parameters, and normal probability distribution in PAST 3 software. The results indicated that heat stress alone caused severe changes in carbohydrates metabolism, SOD, proteins, crude fibers, and minerals immediately so that plants could not recover without foreign stabilizers such as GS-AgNPs. The application of GS-AgNPs increases the flux of carbohydrates metabolism, SOD, and proteins, including HSPs, crude fibers, and minerals, in wheat plants to reduce the effect of heat stress. The 50 mg/l concentration of GS-AgNPs has shown an increase in carbohydrates metabolism and SOD activity, while crude fibres have shown a significant enhancement at 100 mg/l of GS-AgNPs. The crude and true proteins were also shown pronounced increase in treatment to a concentration of 50 mg/l of GS-AgNPs. GS-AgNPs stimulated HSP production; most importantly, smHSP production was observed in the present results with other HSPs in wheat plants treated with a 50 mg/l concentration of GS-AgNPs. The mineral distribution was also regulated by the respective treatment of GS-AgNPs, and the highest amounts of Ca, P and Fe were found to be highest in wheat under heat stress. In general, we computed the expected model based on GS-AgNPs on the genes/factors that respond to heat stress and their potential role in mitigating heat stress in wheat. In addition, we discussed the prospective signalling pathway triggered by GS-AgNPs in wheat against heat stress. In the future, this work might be helpful in distinguishing the genetic variation due to GS-AgNPs in promoting tolerance in wheat against heat stress.Communicated by Ramaswamy H. Sarma.

Fish collagen peptides' modulating effect on human skin microbiota against pathogenic Staphylococcus aureus.

Aim: The current research aims to design effective strategies to enhance the body's immune system against pathogenic bacteria. Methods: Skin commensals were isolated, identified and cultured in fish collagen peptides (FCPs). Results: After culturing in FCP, the skin commensals were used in a dose-dependent manner for Staphylococcus aureus in a dual-culture test, which showed significant growth inhibition of the pathogenic bacteria, which concluded that FCP induced the immune defense system of skin microbiota against pathogenic strains. Conclusion: Results have validated that fish collagen peptide plays a vital role in the growth of selected human skin flora and induces more defensive immunity against pathogenic S. aureus bacteria in dual-culture experimentation.

Pseudomonas aeruginosa detection based on droplets incubation using an integrated microfluidic chip, laser spectroscopy, and machine learning.

Pseudomonas aeruginosa is an opportunist pathogen responsible for causing several infections in the human body, especially in patients with weak immune systems. The proposed approach reports a novel pathogens detection system based on cultivating microdroplets and acquiring the scattered light signals from the incubated droplets using a microfluidic device. Initially, the microdroplets were generated and incubated to cultivate bacteria inside the microdroplets. The second part of the microfluidic chip is the detection module, embedded with three optical fibers to connect laser light and photosensors. The incubated droplets were reinjected in the detection module and passed through the laser light. The surrounding photosensors were arranged symmetrically at 45° to the flowing channel for acquiring the scattered light signal. The noise was removed from the acquired data, and time-domain waveform features were evaluated. The acquired features were trained using machine learning classifiers to classify P. aeruginosa. The k-nearest neighbors (KNN) showed superior classification performance with 95.6 % accuracy among other classifiers, including logistic regression (LR), support vector machines (SVM), and naïve Bayes (NB). The proposed research was performed to validate the method for pathogens detection with a concentration of 105 CFU/mL. The total duration of 6 h is required to test the sample, including five hours for droplets incubation and one hour for sample preparation and detection using light scattering module. The results indicate that acquiring the light scattering patterns from incubated droplets can detect P. aeruginosa using machine learning classification. The proposed system is anticipated to be helpful as a rapid device for diagnosing pathogenic infections.

Analysis of associated risk factors among recurrent cutaneous leishmaniasis patients: A cross-sectional study in Khyber Pakhtunkhwa, Pakistan.

BACKGROUND: Leishmaniasis is the second and fourth highest cause of mortality and morbidity respectively among all tropical diseases. Recurrence in the onset of leishmaniasis is a major problem that needs to be addressed to reduce the case fatality rate and ensure timely clinical intervention. Here we are investigating the association of risk factors with recurrent cutaneous leishmaniasis to address this issue. MATERIAL AND METHODS: Patients received by Nasser Ullah Khan Babar Hospital in Peshawar, Pakistan from March 2019 to July 2020 were enrolled in this study. Those patients who developed symptoms after completion of treatment were included in Group-A while those who had atypical scars like leishmaniasis but were negative for cutaneous leishmaniasis were included in the comparison group tagged as Group B. All those individuals who had completed six weeks of treatment for CL but had normal complete blood counts (CBC) were included to avoid other underlying immunological pathologies, while we excluded those participants who had co-morbidities like diabetes, liver disease, cardiac disease, and pregnant and lactating women through their history Association was tested between Group-A and Group-B with other explanatory variables through chi-square test. The regression model was proposed to determine the predictors. RESULT: A total of 48 participants of both sexes were included in the study with a mean age of 32.2 ± 15.10. The data suggest that females are overrepresented among the patients with recurrent leishmaniasis [21(53.8 %,); p = 0.07]. Compared to patients; healthy participants had a higher proportion of adults (19-59 years) versus adolescents (13-18 years) [26(66.7 %) vs 07(17.9), p = 0.004]. Multivariate logistic regression analysis shows that females are 2.1 times more prone to infections among cases as compared to healthy individuals [unadjusted OR 2.20, 95 % confidence interval (CI) 1.5-10.6, p = 0.02; adjusted OR 2.1, 95 % CI 1.50-10.69, p = 0.02]. We propose that patients receiving intradermal were less likely to be infected as compared to those receiving intralesional injections [unadjusted OR 0.07.0, 95 % confidence interval (CI) 1.18-3.37, p = 0.03; adjusted OR 0.06, 95 % CI 1.18-3.38, p = 0.03]. CONCLUSION: Old age (adults) and sex (females) were the strongest predictors to be associated with recurrent leishmaniasis. Similarly, the choice of intradermal as compared to intralesional injection and the prolonged treatment duration were strongly associated with greater chances of recurrence.

Recent Progress in Spectroscopic Methods for the Detection of Foodborne Pathogenic Bacteria.

Detection of foodborne pathogens at an early stage is very important to control food quality and improve medical response. Rapid detection of foodborne pathogens with high sensitivity and specificity is becoming an urgent requirement in health safety, medical diagnostics, environmental safety, and controlling food quality. Despite the existing bacterial detection methods being reliable and widely used, these methods are time-consuming, expensive, and cumbersome. Therefore, researchers are trying to find new methods by integrating spectroscopy techniques with artificial intelligence and advanced materials. Within this progress report, advances in the detection of foodborne pathogens using spectroscopy techniques are discussed. This paper presents an overview of the progress and application of spectroscopy techniques for the detection of foodborne pathogens, particularly new trends in the past few years, including surface-enhanced Raman spectroscopy, surface plasmon resonance, fluorescence spectroscopy, multiangle laser light scattering, and imaging analysis. In addition, the applications of artificial intelligence, microfluidics, smartphone-based techniques, and advanced materials related to spectroscopy for the detection of bacterial pathogens are discussed. Finally, we conclude and discuss possible research prospects in aspects of spectroscopy techniques for the identification and classification of pathogens.

Narrative on Hydrogen Therapy and its Clinical Applications: Safety and Efficacy.

Molecular hydrogen proved itself as a novel therapeutic candidate and has been thriving from the beginning with its potential clinical significance, higher affinity, and cellular integrity and permeability. Hydrogen Therapy (HT) has gained scientists' attention with the proven clinical ability to attenuate chronic inflammation, diminish oxidative stress, restrict apoptosis, minimize cellular injury, and refine tissue functioning. Therapeutic Implementation of H2 for disease prevention and treatment is a newly emerging field with limited knowledge available on formulations, tissue-specific effects, efficacy, and safety. This article will discuss HT's therapeutic potential for its efficacy and safety in cardiovascular, respiratory, hematological, metabolic, infectious, and neurodegenerative disorders. In addition to this, the molecular mechanisms and nanotechnological implications of hydrogen therapy will be discussed in detail. Finally, the article will provide insight into advancements and automation, future perspectives, and recommendations. There is a need to study and conduct higher-scale trials targeting personalized treatments under molecular and genetic vitals.

Epidemiologic profile of hard ticks and molecular characterization of Rhipicephalus microplus infesting cattle in central part of Khyber Pakhtunkhwa, Pakistan.

Tick infestation is a major public and animal health concern causing significant financial losses, especially in tropical and subtropical regions of the world. This study aimed at investigating the epidemiologic profile of ticks infesting cattle and molecular identification of R. microplus in the centrally ignored part of Khyber Pakhtunkhwa, Pakistan. A total of 600 cattle from 20 farms were examined for the tick infestation, among them 358 (59.7%) cattle were infested with ticks. A total of 2118 nymph, larvae and adult tick stages were collected and morphologically identified followed by molecular confirmation of Rhipicephalus microplus. Host-based demographic and ecological parameter analysis revealed significantly higher tick infestation in adult, female, exotic, freely grazing, and with irregular/no acaricides treated cattle. The univariate logistic analysis showed that host age, gender, breed, acaricides use, and feeding method were significantly (P < 0.05) associated, whereas multivariate analysis revealed only host breed and feeding method were potential risk factors (P < 0.05) for tick infestation. Microscopy-based examination identified four different species of ticks including R. microplus (44.5%), Hyalomma anatolicum (38.5%), and Hyalomma marginatum (10.5%) and Hyalomma excavatum (6.5%). Tick infestation pattern showed that 55.9% of cattle was found co-infested with R. microplus and H. anatolicum followed by R. microplus and H. anatolicum and H. marginatum (29.3%) then R. microplus, H. anatolicum, H. marginatum, and H. excavatum (11.2%). Sequencing of the second internal transcribed spacer (ITS2-) and 16S rRNA gene fragments also confirmed the molecular identification of Rhipicephalus microplus. Phylogenetic analysis of ITS-2 revealed all sequences clustered in single clade of the R. microplus while the 16S rRNA nucleotide sequences showed that R. microplus in this study was clustered together in clade A along with other isolates from Pakistan, China, and India. The high tick infestation suggests the need for designing strategic and integrated control measures for ticks in order to ensure good health of domestic animals in this region of Pakistan.

Coating Silica Layer on Fe3O4 Magnetic Nanoparticles and Application in Extracting High Quality Nucleic Acids from Blood Sample.

The given research revealed that the size of Fe3O4 magnetic nanoparticles (MNPs) could be controlled by varying the pre-mixing conditions in the solvothermal method. Scanning electron microscopy (SEM) showed that the size of the MNPs gradually increased with increasing the initial temperature at which reaction components were mixed while the reaction component's mixing time was kept constant. The smallest sized MNPs were achieved among the five treatments (25, 50, 75, 100, and 125 °C) when reaction components were mixed at 25 °C, while the larger sized MNPs were synthesized among the five treatments when reaction components were mixed at 125 °C. Then, Stöber method was followed for coating silica layer onto the MNPs. However, ammonium hydroxide was replaced with potassium hydroxide as a catalyst, which significantly increased the speed of silica coating onto MNPs. The Fourier transform infrared (FTIR) spectrometer revealed that the MNPs were successfully covered with silica in five minutes. FTIR spectra exhibited a peak about 1088.8 cm-1, which belonged to the asymmetry stretching vibration of Si-O-Si. Transmission electronic microscopy (TEM) analysis was conducted to confirm the presence of silica layer onto MNPs. Thus, potassium hydroxide was successfully employed as a catalyst for quick silica layer coating onto MNPs. Furthermore, these silica coated MNPs were used to extract high quality nucleic acids from blood sample.

Additive Manufacturing in Orthopedics: A Review.

Additive manufacturing is an advanced manufacturing manner that seems like the industrial revolution. It has the inborn benefit of producing complex formations, which are distinct from traditional machining technology. Its manufacturing strategy is flexible, including a wide range of materials, and its manufacturing cycle is short. Additive manufacturing techniques are progressively used in bone research and orthopedic operation as more innovative materials are developed. This Review lists the recent research results, analyzes the strengths and weaknesses of diverse three-dimensional printing strategies in orthopedics, and sums up the use of varying 3D printing strategies in surgical guides, surgical implants, surgical predictive models, and bone tissue engineering. Moreover, various postprocessing methods for additive manufacturing for orthopedics are described.

Antimicrobial Resistance of Shigella flexneri in Pakistani Pediatric Population Reveals an Increased Trend of Third-Generation Cephalosporin Resistance.

The rapid emergence of resistance to third-generation cephalosporins in Shigella flexneri is crucial in pediatric shigellosis management. Limited studies have been conducted on molecular pattern of antibiotic resistance of S. flexneri in diarrhea endemic areas of Pakistan. The aim of the study was to analyze the antimicrobial resistance of S. flexneri isolated from pediatric diarrheal patients in Peshawar, Pakistan. A total of 199 S. flexneri isolates (clinical, n = 1 55 and non-clinical, n = 44) were investigated for drug resistance and mutational analysis of selected drug resistance genes. All isolates were found to be highly resistant to amoxicillin/clavulanic acid (88%), followed by trimethoprim-sulfamethoxazole (77%), chloramphenicol (43%), and quinolones (41.6%). About 34.5% S. flexneri isolates were found to be resistant to third-generation cephalosporin. None of the isolates was resistant to imipenem, piperacillin-tazobactam, and amikacin. Interestingly high frequency of third-generation cephalosporin resistance was observed in S. flexneri isolated from non-clinical samples (49%) when compared to clinical samples (30.5%). Furthermore, the most prevalent phenotypic-resistant patterns among third-generation cephalosporin-resistant isolates were AMC,CAZ,CPD,CFM,CRO,SXT (13%) followed by OFX,AMC,CAZ,CPD,CFM,CRO,SXT,NA,CIP (10%). The most frequently detected resistance genes were trimethoprim-sulfamethoxazole (sul2 = 84%), beta-lactamase genes (blaOXA = 87%), quinolones (qnrS = 77%), and chloramphenicol (cat = 64%). No mutation was detected in any drug-resistant genes. We are reporting for the first time the sequence of the blaTEM gene in S. flexneri. Furthermore, high third-generation cephalosporin resistance was observed in the patients who practiced self-medication as compared to those who took medication according to physician prescription. This study shows the high emergence of third-generation cephalosporin-resistant S. flexneri isolates, which is a potential threat to the community in the country. This finding will be helpful to develop a suitable antibiotic prescription regime to treat shigellosis.

Photosensitizer-loaded gold nanocages for immunogenic phototherapy of aggressive melanoma.

Malignant melanoma remains the life-threatening form of skin cancer with high mortality and poor prognosis. Thus, an ideal melanoma therapeutic strategy is of immediate importance which can remove the primary tumor, as well as inhibit the metastasis and recurrence. Here, we report the fabrication of adjuvant monophosphoryl lipid A (MPLA) lipid bilayer-enveloped and photosensitizer indocyanine green (ICG)-loaded gold nanocages (MLI-AuNCs) for immunogenic phototherapy of aggressive melanoma. Hollow porous AuNCs are used as carriers to deliver MPLA and ICG, and protect ICG from photodegradation. Both AuNCs and ICG absorb near infrared (NIR) light and can be applied in controllable NIR-triggered photothermal and photodynamic combination therapy (PTT/PDT) of melanoma. MLI-AuNCs coated by thermosensitive lipid bilayer exhibit uniform size, good biocompatibility and bioavailability with prominent tumor accumulation, which further improve the PTT/PDT efficacy. MLI-AuNCs under NIR irradiation not only destroy the primary tumor by PTT/PDT, but also elicit robust antitumor immune response with melanoma associated antigens and MPLA released in situ. The released antigens and MPLA subsequently enhance the recruitment and maturation of dendritic cells, which further activate the effector T cells to inhibit metastases and recurrence of melanoma. This immunomodulatory-boosted PTT/PDT nanoplatform provides a new opportunity for highly aggressive melanoma treatment. STATEMENT OF SIGNIFICANCE: An ideal tumor therapeutic strategy not only can remove the primary tumor, but also inhibit metastasis and recurrence. Here, we introduced a versatile nanoplatform MLI-AuNCs for immunogenic phototherapy of aggressive melanoma. Adjuvant MPLA and photosensitizer ICG can be protected and co-delivered to the tumors by thermosensitive lipid-enveloped AuNCs. MLI-AuNCs exhibited prominent tumor accumulation ability and produced the potent PTT/PDT effect to destroy the primary tumors with a single dose of NIR irradiation, as well as elicited the strong antitumor immunity to inhibit the metastasis and relapse. This study may provide a potential therapeutic vaccination strategy against advanced melanoma and other difficult-to-treat cancers.

Programmable Biosensors Based on RNA-Guided CRISPR/Cas Endonuclease.

Highly infectious illnesses caused by pathogens constitute severe threats to public health and lead to global economic loss. The use of robust and programmable clustered regularly interspaced short palindromic repeat and CRISPR-associated protein (CRISPR-Cas) systems, repurposed from genome-engineering applications has markedly improved traditional nucleic acid detection for precise identification, independently enabling rapid diagnostics of multiplex biomarker with genetic and mutation related to tumors, and microbial pathogens. In this review, we delineate the utility of the current CRISPR-Cas enzyme as biosensors by which these effector toolkits achieve recognition, signaling amplification, and finally, accurate detection. Additionally, we discuss the details of the dominance and hurdles related to expanding this revolutionary technology into an effective and convenient contraption crucial for improving the rational redesign to CRISPR/Cas biosensing. Overall, this review provides an insight into the current status of rapid and POC diagnostic systems by CRISPR/Cas tools.

Rapid detection of Pseudomonas aeruginosa based on lab-on-a-chip platform using immunomagnetic separation, light scattering, and machine learning.

The rapid detection of the pathogenic bacteria in patient samples is crucial to expedient patient care. The proposed approach reports the development of a novel lab-on-a-chip device for the rapid detection of P. aeruginosa based on immunomagnetic separation, optical scattering, and machine learning. The immunomagnetic particles with a diameter of 5 µm were synthesized for isolating P. aeruginosa from the test sample. A microfluidic chip was fabricated, and three optical fibers were embedded for connecting a laser light and two photodetectors. The laser light was pointed towards the channel to pass light through the sample. A pair of photodetectors via optical fibers were arranged symmetrically at 45° to the channel. The photodetectors acquired scattered light from the flowing sample and converted the light to an electrical signal. The sample containing immunomagnetic beads linked with bacteria was injected into the microfluidic chip. The optimized conditions for performing the experiments were characterized for real-time detection of P. aeruginosa. The data acquisition system recorded the real-time light scattering from the test sample. After removing noise from the output waveform, five different time-domain statistical features were extracted from each waveform: standard mean, standard variance, skewness, kurtosis, and coefficient of variation. The pathogens classification was performed by training the discrimination model using extracted features based on machine learning algorithms. The support vector machines (SVM) with a sigmoid function kernel showed superior classification performance with 97.9% accuracy among other classifiers, including k-nearest neighbors (KNN), logistic regression (LR), and naïve Bayes (NB). The method can detect P. aeruginosa specifically and quantitatively with a limit of detection of 102 CFU/mL. The device can classify P. aeruginosa within 10 min with a total assay time of 25 min. The device was used to test its ability to detect pathogen from the serum and sputum specimens spiked with 105 CFU/mL concentration of P. aeruginosa. The results indicate that light scattering combined with machine learning can be used to detect P. aeruginosa. The proposed technique is anticipated to be helpful as a rapid device for diagnosing P. aeruginosa related infections.

Rapid classification of micro-particles using multi-angle dynamic light scatting and machine learning approach.

The rapid classification of micro-particles has a vast range of applications in biomedical sciences and technology. In the given study, a prototype has been developed for the rapid detection of particle size using multi-angle dynamic light scattering and a machine learning approach by applying a support vector machine. The device consisted of three major parts: a laser light, an assembly of twelve sensors, and a data acquisition system. The laser light with a wavelength of 660 nm was directed towards the prepared sample. The twelve different photosensors were arranged symmetrically surrounding the testing sample to acquire the scattered light. The position of the photosensor was based on the Mie scattering theory to detect the maximum light scattering. In this study, three different spherical microparticles with sizes of 1, 2, and 4 µm were analyzed for the classification. The real-time light scattering signals were collected from each sample for 30 min. The power spectrum feature was evaluated from the acquired waveforms, and then recursive feature elimination was utilized to filter the features with the highest correlation. The machine learning classifiers were trained using the features with optimum conditions and the classification accuracies were evaluated. The results showed higher classification accuracies of 94.41%, 94.20%, and 96.12% for the particle sizes of 1, 2, and 4 µm, respectively. The given method depicted an overall classification accuracy of 95.38%. The acquired results showed that the developed system can detect microparticles within the range of 1-4 µm, with detection limit of 0.025 mg/ml. Therefore, the current study validated the performance of the device, and the given technique can be further applied in clinical applications for the detection of microbial particles.

Responses of bimetallic Ag/ZnO alloy nanoparticles and urea on morphological and physiological attributes of wheat.

Wheat (Triticum aestivum L.) is the most important staple food crop globally. According to economic survey 2018-19, agriculture sector of Pakistan grew by 0.85%, with wheat accounting for 8.9% of agriculture and 1.6% of GDP, and its production fell short of the target by 4.9%. Wheat requires beneficial ties to improve its efficiency with the help of modern technology. Nanotechnology modifies conventional agricultural practices as these are stimulating agents for plant growth. Green bimetallic Ag/ZnO alloy nanoparticles (NPs) synthesised from salts reduced by Moringa oleifera and characterised by UV-visible spectroscopy, scanning electron microscopy, and energy-dispersive X-ray spectroscopy are studied herein. Different concentrations of urea and Ag/ZnO alloy NPs were applied exogenously to wheat plants (Pakistan-13 and Galaxy13). A significant effect of 100 mg/L urea and 75 ppm Ag/ZnO alloy NPs was observed on the morphology of wheat, with a maximum increase of 58% plant length, 85% leaf area, 89% plant fresh weight and 76% plant dried weight. In physiological parameters, relative water content and membrane stability index have shown maximum increases of 39% and 77%, while chlorophyll a, b, and total chlorophyll content (TCC) showed maximum increases of 92%, 71%, and 84% respectively. Evidence of the morpho-physiological responses of urea and green synthesised alloy NPs on wheat varieties are reported on.

Novel IL-12Rß1 deficiency-mediates recurrent cutaneous leishmaniasis.

BACKGROUND: The IL-12/IFN-γ axis plays a vital role in the control of intramacrophagic pathogens including Leishmania infections. OBJECTIVE: The aim of this study was to investigate genetic defects in the IL-12/IFN-γ axis in cutaneous leishmaniasis patients, using immunological and genetic evaluation. METHODS: Enzyme-linked immunosorbent assay was used to quantify IFN-γ , while flow cytometry was performed to analyze surface IL-12Rß1/IL-12Rß2 expression and phosphorylation of signal transducers as well as the activator of transcription 4 (pSTAT4). Sequencing was carried out for genetic analysis. RESULTS: The peripheral blood mononuclear cells from the two patients (P1 and P2) demonstrated impaired production of IFN-γ. Furthermore, abolishment of the surface expression of Il-12Rß1 was observed in lymphocytes, with consequent impairment of STAT4 phosphorylation in the lymphocytes of P1 and P2. IL-12Rß1 deficiency was identified, which was caused by a novel homozygous missense mutation (c.485>T/p.P162L) and a novel homozygous nonsense mutation (c.805G>T/P.E269*) in the IL-12Rß2 gene of P1 and P2, respectively. In silico analyses predicted these novel mutations as being pathogenic, causing truncated proteins, with consequent inactivation. CONCLUSION: Our data have expanded the phenotype and mutation spectra associated with IL-12Rß1 deficiency, and suggest that patients with CL should be screened for mutations in genes of the IL-12/IFN-γ axis.

Injectable and pH-Sensitive Hyaluronic Acid-Based Hydrogels with On-Demand Release of Antimicrobial Peptides for Infected Wound Healing.

Antibiotics' abuse in bacteria-infected wounds has threatened patients' lives and burdened medical systems. Hence, antibiotic-free hydrogel-based biomaterials, which exhibit biostability, on-demand release of antibacterial agents, and long-lasting antimicrobial activity, are highly desired for the treatment of chronic bacteria-infected wounds. Herein, we developed a hyaluronic acid (HA)-based composite hydrogel, with an antimicrobial peptide [AMP, KK(SLKL)3KK] as a cross-linking agent through Schiff's base formation, which exhibited an acidity-triggered release of AMP (pathological environment in bacteria-infected wounds, pH ∼ 5.5-5.6). During the self-assembly process, AMP adopted an antiparallel ß-sheet secondary structure due to the alternate arrangement of hydrophobic and hydrophilic residues of amino acids. Owing to Schiff's base formation between the primary amines derived from lysine residues and the aldehydes in oxidized HA, the AMP-HA composite hydrogel exhibited injectability, high biostability, and enhanced mechanical strength. Importantly, both AMP and the AMP-HA composite showed excellent broad-spectrum antibacterial activity in vitro and in vivo. Specifically, the AMP-HA composite hydrogel exhibited on-demand full thickness wound healing in an infected mice model. Therefore, this work provides an efficient strategy to fabricate antibiotic-free hydrogel-based biomaterials for the management of chronic bacteria-infected wounds.