Combined cross-linking of Rhizomucor miehei lipase and Candida antarctica lipase B for the effective enrichment of omega-3 fatty acids in fish oil.

The incorporation of a non-specific lipase and a sn-1,3 specific one in a single immobilized system can be a promising approach for the exploitation of both lipases. A one-step immobilization platform mediated by an isocyanide-based multi-component reaction was applied to create co-cross-linked enzymes (co-CLEs) of lipases from Rhizomucor miehei (sn-1,3 specific) and Candida antarctica (non-specific). Glutaraldehyde was found to be effective cross-linker by producing specific activity of 16.9 U/mg and immobilization yield of 99 %. High activity recovery of up to 404 % was obtained for immobilized derivatives. Leaking experiment showed covalent nature of the cross-linking processes. BSA had considerable effect on the immobilization process, providing 87-100 % immobilization yields and up to 10 times improvement in the specific activity of the immobilized derivatives. Scanning electron microscopy images showed flower-like and rod-like structures for the CLEs prepared by glutaraldehyde and undecanedicarboxylic acid, respectively. The prepared co-CLEs were examined in non-selective enrichment of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) from fish oil, showing capability of releasing up to 100 % of both omega-3 fatty acids within 8 h of the reaction. The reusability of co-CLEs in five successive cycles presented retaining 63-72 % of their initial activities after the fifth reuse cycle in the hydrolysis reaction.

Hyperactivation of lipases by immobilization on superhydrophobic graphene quantum dots inorganic hybrid nanoflower.

Various nanoflowers are synthesized for enzyme immobilization. In order to increase the activity of nanoflowers, in this study, 3D flower-like structured organic-inorganic hybrid nanoflowers (hNFs) with various lipases Rhizomucor miehei lipase (RML), Candida antarctica lipase B (CALB), Humicola insolens lipase (HIL), Thermomyces lanuginosus lipase (TLL), Eversa® Transform 2.0 (ET) a genetically modified enzyme derived of TLL and graphene quantum dots (GQDs) were prepared and characterized.Lipase hNFs [lipase-(Cu/Co)3(PO4)2] and lipase@GQDs hNFs [lipase@GQDs-(Cu/Co)3(PO4)2] were straightforwardly prepared through mixing with metal ion (Cu2+or Co2+) aqueous solutions with or without GQDs. The ET@GQDs-(Cu)3(PO4)2 hNFs demonstrated 687 % higher activity than ET-(Cu)3(PO4)2 hNFs and 650 % higher activity than the free ET. Similar results were also observed with other lipase hybrid nanoflowers. For example, TLL@GQDs-(Cu)3(PO4)2 hNFs exhibited a 557 % higher activity than TLL-(Cu)3(PO4)2 hNFs and a 463 % higher activity than free TLL. Additionally, TLL@GQDs-(Co)3(PO4)2 hNFs showed a 141 % higher activity than TLL-(Co)3(PO4)2 hNFs and a 304 % higher activity than free TLL. Upon examining pH and thermal stability, it was revealed that lipase@GQDs hNFs exhibited higher activity compared to free lipase and other hNFs without GQDs. The effect of metal ions, enzyme concentrations and amount of GQDs on the morphology and enzyme activity of the lipase-hNFs was examined.

Improving performance, reproduction, and immunity in laying Japanese quail with algal derivatives.

We investigated the effect of the Persian Gulf algae derivatives, namely phycocyanin (PC) and fucoidan (FUC), on the performance, reproductive traits, and immune responses of laying Japanese quails. A completely randomized design was used to distribute 250 six-wk-old Japanese quails with an average body weight of 215 ± 10 g into 5 treatments, 5 replicates, and 10 birds in each replicate over a 5-wk period. Unlike the control groups, the treatment groups received drinking water supplemented with PC and FUC at concentrations of 20 or 40 mg/L, denoted as PC20, PC40, FUC20, and FUC40, respectively, while all birds were provided with identical feed. Supplemental algal derivatives notably improved hen day egg production (HDEP), egg mass, and feed conversion ratio (FCR) compared to the control group (P < 0.01). Incorporating PC and FUC had no significant effect on the weight of males' testes or the weight and length of hens' oviducts. Additionally, the experimental treatments had no impact on the chicks' hatching weight. The supplementation of PC and FUC resulted in increased fertility (P = 0.038) and hatchability (P < 0.001) rates, with the exception of fertility in the PC40 group. The effect of the experimental treatments on immune responses was largely not statistically significant, except in the case of ND. Specifically, the experimental treatments resulted in increased (P = 0.033) antibody titers against ND when compared to the control group, with the exception of FUC20. Supplemental algal derivatives significantly (P < 0.01) reduced total cholesterol, creatinine, and triglycerides (except in the case of PC20). Overall, these findings underscore the potential of algal derivatives to enhance quail performance, reproductive traits, and immune responses.

Unraveling the link between insulin resistance and Non-alcoholic fatty liver disease (or metabolic dysfunction-associated steatotic liver disease): A Narrative Review.

Objective: Non-alcoholic fatty liver disease (NAFLD) is rapidly becoming a significant global health concern, representing the leading cause of chronic liver disease and posing a substantial public health challenge. NAFLD is associated with higher insulin resistance (IR) levels, a key pathophysiological mechanism contributing to its development and progression. To counter this growing trend, it is crucial to raise awareness about NAFLD and promote healthy lifestyles to mitigate the impact of this disease. Methods: Relevant studies regarding IR and NAFLD published until May 30, 2023, were extracted from Google PubMed, Scopus, and Web Of Science web databases. The following keywords were used: IR, diabetes mellitus, Non-alcoholic fatty liver disease, and metabolic syndrome. Results: IR leads to an accumulation of fatty acids within liver cells, resulting from increased glycolysis and decreased apolipoprotein B-100. Furthermore, the manifestations of NAFLD extend beyond liver-related morbidity and mortality, affecting multiple organs and giving rise to various non-communicable disorders such as diabetes mellitus, metabolic syndrome, polycystic ovary syndrome, obstructive sleep apnea, and cardiovascular disease. Although lifestyle modification remains the primary treatment approach for NAFLD, alternative therapies, including pharmacological, herbal, and surgical interventions, may be considered. By implementing early and simple measures, cirrhosis, end-stage liver disease, and hepatocellular carcinoma can be prevented. Conclusions: There is a clear association between NAFLD and elevated levels of IR. Several metabolic conditions, such as obesity, type 2 diabetes mellitus, dyslipidemia, and metabolic syndrome, are closely interrelated with NAFLD and IR. Raising awareness about NAFLD and promoting a healthy lifestyle are crucial steps to reverse the impact of this disease.

A Novel Electrokinetic-Based Technique for the Isolation of Circulating Tumor Cells.

The separation of rare cells from complex biofluids has attracted attention in biological research and clinical applications, especially for cancer detection and treatment. In particular, various technologies and methods have been developed for the isolation of circulating tumor cells (CTCs) in the blood. Among them, the induced-charge electrokinetic (ICEK) flow method has shown its high efficacy for cell manipulation where micro-vortices (MVs), generated as a result of induced charges on a polarizable surface, can effectively manipulate particles and cells in complex fluids. While the majority of MVs have been induced by AC electric fields, these vortices have also been observed under a DC electric field generated around a polarizable hurdle. In the present numerical work, the capability of MVs for the manipulation of CTCs and their entrapment in the DC electric field is investigated. First, the numerical results are verified against the available data in the literature. Then, various hurdle geometries are employed to find the most effective geometry for MV-based particle entrapment. The effects of electric field strength (EFS), wall zeta potential magnitude, and the particles' diameter on the trapping efficacy are further investigated. The results demonstrated that the MVs generated around only the rectangular hurdle are capable of trapping particles as large as the size of CTCs. An EFS of about 75 V/cm was shown to be effective for the entrapment of above 90% of CTCs in the MVs. In addition, an EFS of 85 V/cm demonstrated a capability for isolating particles larger than 8 µm from a suspension of particles/cells 1-25 µm in diameter, useful for the enrichment of cancer cells and potentially for the real-time and non-invasive monitoring of drug effectiveness on circulating cancer cells in blood circulation.

Compact all-optical decoder design for optical integrated circuits.

This paper constructs and discusses a 1×2 decoder based on two-dimensional photonic crystals. The designed decoder is a priority decoder with one main input and one enabled input. This logic circuit's structure is very simple and compact, and it employs a photonic crystal structure with dimensions of 11×11 rods built of GaAs. The calculation results reveal that the output power values for logical mode 1 are extremely close to the power of the light source, whereas the power is very low and close to zero in logical mode 0. As a result, the difference between logical values 0 and 1 in the output will be adequate, and the circuit's accuracy will be good. The finite difference-time domain (FDTD) approach is used for time computations and light propagation in waveguides.

Peptiligase, an enzyme for efficient chemo-enzymatic synthesis of aviptadil.

Increasing attention to peptides as prospective therapeutics has created a renaissance in searching for new alternatives to the current peptide synthetic approaches as well as their modification. In this context, it is necessary to develop different approaches for peptide ligation. Using enzymes as a novel strategy and powerful tool for the peptide and protein ligation has recently received a lot of attention. We here designed a fully convergent chemo-enzymatic peptide synthesis (CEPS) process for the synthesis of aviptadil a 28-mer therapeutic peptide with potential therapeutic effects in various medical contexts specially in the treatment of acute respiratory distress syndrome (ARDS) by coupling two peptide segments with four different peptiligase variants in aqueous environments. Our study reveals that peptiligase variants are capable of ligation reaction in 15 min. The overall time of ligation is shorter than those peptides with similar lengths and hinderance to aviptadil which reported for conventional synthesis by full solid-phase peptide synthesis. Yields ranged from 54 % to 76 %.

Comparison of different transient gene expression systems for the production of a new humanized anti-HER2 monoclonal antibody (Hersintuzumab).

BACKGROUND: Producing therapeutic proteins can be done quickly and on a large scale through Transient Gene Expression (TGE). Chinese hamster ovary (CHO) cell lines are commonly used to achieve this. Although there are few comparative studies, TGE has been observed in suspension-adapted CHO cells. OBJECTIVES: We tested TGE's effectiveness in DG-44, CHO-S, and ExpiCHO-S cell lines with four transfection reagents. METHODS: A design of experiments (DoE) was followed to optimize transfection using a recombinant monoclonal antibody (mAb) construct. To evaluate the efficacy, flow cytometry and ELISA were used. Feeding strategies and temperature shifts were implemented to enhance transfection effectiveness. The quality of the mAb was assessed through ELISA, SDS-PAGE, and proliferation inhibition assays. RESULTS: We adapted all cell lines to grow in suspension using a serum-free medium. Our findings from flow cytometry and ELISA tests indicate that PEI and Pmax reagents had a higher rate of transfection and mAb production than the ExpiCHO commercial transfection reagent. While DG-44 cells had better transfection efficiency than CHO-S and ExpiCHO-S, there was no significant difference between CHO-S and ExpiCHO-S. Our TGE system was more productive at 32 °C than at 37 °C. In the optimized TGE of Pmax-based transfection in DG-44 at 37 and 32 °C, the production level of mAb was more than half of the amount of the commercial ExpiCHO-S expression system. Still, the number of transfected cells was three times higher, making it more efficient. The purified mAb from all transfected cell lines had similar structural and functional properties under different conditions. CONCLUSION: Our research shows that using Pmax and DG-44 cells in the TGE system is a cost-effective and efficient way to produce humanized monoclonal antibodies. We discovered that this method outperforms the ExpiCHO-S kit.

Antimicrobial properties of green synthesized novel TiO2 nanoparticles using Iranian propolis extracts.

The oral antimicrobial and cytotoxic properties of green synthesized novel titanium dioxide nanoparticles (TiO2 NPs) using Iranian propolis extracts were investigated on oral bacteria and fibroblast cells. In this study, propolis was sampled, and alcoholic extracts were prepared. The TiO2 NPs were biosynthesized using propolis extracts. The synthesized TiO2 NPs were characterized by scanning electron microscope (SEM), X-ray diffraction analysis, energy-dispersive X-ray (EDX), Fourier transform infrared spectroscopy (FTIR), dynamic light scattering, ultraviolet-visible (UV-Vis), transmission electron microscope, Brunauer-Emmett-Teller, and zeta potential. MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide), minimal inhibitory concentration, minimum bactericidal concentration, minimum fungicidal concentration, biofilm formation, and degradation tests were studied to clarify the oral antimicrobial properties of green synthesized TiO2  NPs. According to the FTIR analysis, the propolis extract contained flavonoids and phenolic compounds in addition to TiO2 NPs. Additionally, UV-Vis revealed that intense bands had formed NPs. EDX spectra and SEM images revealed that the stabilizing agent was in perfect quasi-spherical shapes around 21 nm. An EDX spectrum was used to verify the presence of titanium and oxygen. There were no significant cytotoxicity effects. The antibacterial results showed that Pro1TiO2 (Khalkhal sample) had better effects than Pro2TiO2 (Gilan sample) and TiO2 NPs. The present study presents a new process for synthesizing TiO2 NPs from propolis extracts with less toxic effects and user-friendly, eco-friendly, and economical materials. Pro1TiO2 NPs may be considered the best candidate for clinical application.

Insights into the Synergistic Antibacterial Activity of Silver Nitrate with Potassium Tellurite against Pseudomonas aeruginosa.

The constant, ever-increasing antibiotic resistance crisis leads to the announcement of "urgent, novel antibiotics needed" by the World Health Organization. Our previous works showed a promising synergistic antibacterial activity of silver nitrate with potassium tellurite out of thousands of other metal/metalloid-based antibacterial combinations. The silver-tellurite combined treatment not only is more effective than common antibiotics but also prevents bacterial recovery, decreases the risk of future resistance chance, and decreases the effective concentrations. We demonstrate that the silver-tellurite combination is effective against clinical isolates. Further, this study was conducted to address knowledge gaps in the available data on the antibacterial mechanism of both silver and tellurite, as well as to give insight into how the mixture provides synergism as a combination. Here, we defined the differentially expressed gene profile of Pseudomonas aeruginosa under silver, tellurite, and silver-tellurite combination stress using an RNA sequencing approach to examine the global transcriptional changes in the challenged cultures grown in simulated wound fluid. The study was complemented with metabolomics and biochemistry assays. Both metal ions mainly affected four cellular processes, including sulfur homeostasis, reactive oxygen species response, energy pathways, and the bacterial cell membrane (for silver). Using a Caenorhabditis elegans animal model we showed silver-tellurite has reduced toxicity over individual metal/metalloid salts and provides increased antioxidant properties to the host. This work demonstrates that the addition of tellurite would improve the efficacy of silver in biomedical applications. IMPORTANCE Metals and/or metalloids could represent antimicrobial alternatives for industrial and clinical applications (e.g., surface coatings, livestock, and topical infection control) because of their great properties, such as good stability and long half-life. Silver is the most common antimicrobial metal, but resistance prevalence is high, and it can be toxic to the host above a certain concentration. We found that a silver-tellurite composition has antibacterial synergistic effect and that the combination is beneficial to the host. So, the efficacy and application of silver could increase by adding tellurite in the recommended concentration(s). We used different methods to evaluate the mechanism for how this combination can be so incredibly synergistic, leading to efficacy against antibiotic- and silver-resistant isolates. Our two main findings are that (i) both silver and tellurite mostly target the same pathways and (ii) the coapplication of silver with tellurite tends not to target new pathways but targets the same pathways with an amplified change.

Studying the shelf life of butter containing fucoidan, by evaluating sensory and chemical properties.

Fucoidan powder was added in amounts of 0.05, 0.1,0.3, and 0.5% to sour cream butter and sensory and chemical properties were tested on their shelf life for 60 days during storage. Peroxide levels initially increased until day 40 of storage and then decreased. Butter samples from the control group had the highest amount of peroxide on day 40 (15.25 ± 1.41 meq/kg butter), while samples treated with fucoidan 0.5% had the lowest amount of peroxide (6.35 ± 0.53 meq/kg butter). The acidity of butter treatments increased during storage (p < .05). Butter samples from the control group had the highest acidity at 60 days of storage (0.40 ± 0.033 mg KOH / g butter), while samples treated with 0.5% fucoidan had the lowest acidity (0.17 ± 0.013 mg KOH / g butter). The treated butter samples showed the highest stability. Fucoidan, as an antioxidant, reduces the taste, odor, and discoloration of butter added with fucoidan during storage because it completely removes odorless tasteless powder, and the free radical chain is involved in oxidation and improves product properties. The results showed that there are no significant changes in the acceptance rate of butter treated with fucoidan during 60 days of storage in the refrigerator (p > .05). The sensory scores of the treated butter showed that the sensory properties during the storage period were similar to the control samples, but on day 40 of storage, they decreased. In general, a concentration of 0.5% fucoidan delays the oxidative process and increases shelf life and is selected as a superior treatment in terms of sensory evaluation, and is introduced as a functional food.

Biocatalytic synthesis of oxadiazole thioethers and evaluation of their antitumor activity.

A multicomponent enzyme-catalyzed process is suggested for the synthesis of a novel series of 1,3,4-oxadiazole thioether derivatives with yields ranging from 65 to 94%. Novozym 435, the immobilized form of Candida antarctica lipase B (CALB), was found to efficiently catalyze the reaction. The products were evaluated for antitumor activities against two cancer cell lines, HT-29 (human colorectal cancer cell) and HepG2 (human liver cancer cell), by MTT assays. Among them, two compounds exhibited higher antitumor activities, for both cell lines, compared to doxorubicin. In silico molecular docking and computational ADME analysis were performed to propose a mode of action for the anti-cancer activities and to predict drug-likeness, respectively.

A Novel Fc-Engineered Anti-HER2 Bispecific Antibody With Enhanced Antitumor Activity.

Human epidermal growth factor receptor 2 (HER2) overexpression has been demonstrated in a variety of cancers. Targeted therapy with anti-HER2 monoclonal antibodies (mAbs) has been approved as a therapeutic modality. Despite the efficacy of mAbs in tumor treatment, many patients do not benefit from this therapeutic platform. Fragment crystallizable (Fc) engineering is a common approach to improve the efficacy of therapeutic mAbs. Five Fc-engineered mAbs have so far been approved by FDA. We have recently developed an anti-HER2 bispecific mAb, BiHT, constructed from variable domains of trastuzumab, and our novel humanized anti-HER2 mAb, hersintuzumab. BiHT displayed promising antitumor activity as potently as the combination of the parental mAbs. Here, we aimed to modify the Fc of BiHT to improve its therapeutic efficacy. The Fc-engineered BiHT (MBiHT) bound to recombinant HER2 and its subdomains with an affinity similar to BiHT. It also recognized native HER2 on different cell lines, inhibited their proliferation, downregulated HER2 expression, and suppressed downstream signaling pathways similar to BiHT. Compared with BiHT, MBiHT displayed enhanced antibody-dependent cellular cytotoxicity activity against various tumor cell lines. It also inhibited the growth of ovarian xenograft tumors in nude mice more potently than BiHT. Our findings suggest that MBiHT could be a potent therapeutic candidate for the treatment of HER2-overexpressing cancer types.

The current regenerative medicine approaches of craniofacial diseases: A narrative review.

Craniofacial deformities (CFDs) develop following oncological resection, trauma, or congenital disorders. Trauma is one of the top five causes of death globally, with rates varying from country to country. They result in a non-healing composite tissue wound as they degenerate in soft or hard tissues. Approximately one-third of oral diseases are caused by gum disease. Due to the complexity of anatomical structures in the region and the variety of tissue-specific requirements, CFD treatments present many challenges. Many treatment methods for CFDs are available today, such as drugs, regenerative medicine (RM), surgery, and tissue engineering. Functional restoration of a tissue or an organ after trauma or other chronic diseases is the focus of this emerging field of science. The materials and methodologies used in craniofacial reconstruction have significantly improved in the last few years. A facial fracture requires bone preservation as much as possible, so tiny fragments are removed initially. It is possible to replace bone marrow stem cells with oral stem cells for CFDs due to their excellent potential for bone formation. This review article discusses regenerative approaches for different types of craniofacial diseases.

Biocatalytic stereoselective synthesis of pyrrolidine-2,3-diones containing all-carbon quaternary stereocenters.

Highly functionalized pyrrolidine-2,3-diones can be synthesized efficiently and stereoselectively under mild conditions using a biocatalytic approach. The reaction led to the formation of new all-carbon quaternary stereocenters from Myceliophthora thermophila laccase (Novozym 51003) catalyzed oxidation of catechols to ortho-quinones and subsequent 1,4-addition with 3-hydroxy-1,5-dihydro-2H-pyrrol-2-ones. The reaction was conducted with various substituents on both reactants, resulting in 13 products in moderate to good yields (42-91%). The same 15 reactions were also tested with K3Fe(CN)6 as a catalyst, but here only one reaction resulted in a product (60% yield).

Isocyanide-based multi-component reactions for carrier-free and carrier-bound covalent immobilization of enzymes.

Strategies for the covalent immobilization of enzymes depend on the type of functional group selected to perform the coupling reaction, and on the relative importance of selectivity, loading capacity, immobilization time and activity/stability of the resulting immobilized preparation. However, no single strategy is applicable for all covalent immobilization methods or can meet all these criteria, exemplifying the challenge of introducing a versatile process broadly compatible with the residues on the surface of proteins and the functional groups of common linkers. Here, we describe the use of isocyanide-based multi-component reactions for the carrier-bound and carrier-free covalent immobilization of enzymes. Isocyanide-based multi-component reactions can accept a wide variety of functional groups such as epoxy, acid, amine and aldehyde, as well as many commercially available bi-functional linkers, and are therefore suitable for either covalent coupling of enzymes on a solid support or intermolecular cross-linking of enzymes. In this strategy, an isocyanide is directly added to the reaction medium, the enzyme supplies either the exposed amine or carboxylic acid groups, and the support (in carrier-bound immobilization) or the bi-functional cross-linking agent (in carrier-free immobilization) provides another reactive functional group. The protocol offers operational simplicity, high efficiency and a notable reduction in time over alternative strategies, and can be performed by users with expertise in chemistry or biology. The immobilization reactions typically require 1-24 h.

Diagnostic performance of a novel antigen-capture ELISA for the detection of SARS-CoV-2.

BACKGROUND AND AIMS: The coronavirus disease 2019 (COVID-19) pandemic is a serious health problem worldwide. Early virus detection is essential for disease control and management. Viral antigen detection by ELISA is a cost-effective, rapid, and accurate antigen diagnostic assay which could facilitate early viral detection. METHOD: An antigen-capture sandwich ELISA was developed using novel nucleocapsid (NP)-specific mouse monoclonal antibodies (MAbs). The clinical performance of the assay was assessed using 403 positive and 150 negative respiratory samples collected during different SARS-CoV-2 variants outbreaks in Iran. RESULTS: The limit of detection of our ELISA assay was found to be 43.3 pg/ml for recombinant NP. The overall sensitivity and specificity of this assay were 70.72% (95% CI: 66.01-75.12) and 100% (95% CI: 97.57-100), respectively, regardless of Ct values and SARS-CoV-2 variants. There was no significant difference in our assay sensitivity for the detection of Omicron subvariants compared to Delta variant. Assay sensitivity for the BA.5 Omicron subvariant was calculated as 91.89% (95% CI: 85.17-96.23) for samples with Ct values < 25 and 82.70% (95% CI: 75.19-88.71) for samples with Ct values < 30. CONCLUSION: Our newly developed ELISA method is reasonably sensitive and highly specific for detection of SARS-CoV-2 regardless of the variants and subvariants of the virus.

Separation of bacteria smaller than 4 µm from other blood components using insulator-based dielectrophoresis: numerical simulation approach.

Bloodstream infection (BSI) is a life-threatening infection that causes more than 80,000 deaths and more than 500,000 infections annually in North America. The rapid diagnosis of infection reduces BSI mortality. We proposed bacterial enrichment and separation approach in the current work that may reduce culturing time and accelerate the diagnosis of infection. Over the last two decades, multiple separation methods have been developed, and among these methods, insulator-based dielectrophoresis (iDEP) is considered a powerful technique for separating biological particles. Bacterial separation in the blood is challenging due to the presence of other blood cells, such as white blood cells, red blood cells, and platelets. In the present study, a model is presented which is capable of blood cells separation and directing each cell to a specific outlet using continuous flows of particles with sizes larger than 8 µm, 8-4 µm, and smaller than 4 µm. Compared to other methods, such as filtration, the main advantage of this model is that particles larger than 8 µm are separated from the flow before other particles, which prevents the accumulation of particles in the channel. The outcomes of simulations demonstrated that the factors such as applied voltage and channel dimensions significantly affect the separation efficiency. If these values are properly selected (for example voltage of 70 V that was causing an electric field of 200 V/cm), the proposed model can completely (100%) separate particles larger than 8 µm and smaller than 4 µm (8-4 µm particles separation efficiency is 95%).

Immobilization of Bacillus amyloliquefaciens protease "Neutrase" as hybrid enzyme inorganic nanoflower particles: A new biocatalyst for aldol-type and multicomponent reactions.

Organic-inorganic hybrid nanoflowers (hNFs) with commercial protease "Neutrase" is proposed and characterized as efficient and green biocatalysts for promiscuous catalysis in aldol-type and multicomponent reactions. Neutrase hNFs [Neutrase-(Cu/Ca/Co/Mn)3(PO4)2] are straightforwardly prepared through mixing metal ion (Cu2+, Ca2+, Co2+ or Mn2+) aqueous solutions with Neutrase in phosphate buffer (pH 7.4, 10 mM) resulting in precipitation (3 days). The hNFs were characterized by various techniques including scanning electron microscopy (SEM), energy dispersive X-ray (EDX), element mapping, X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FTIR). In SEM images, the metal-Neutrase complexes revealed flower-like or granular structures after hybridization. The effect of metal ions and enzyme concentrations on the morphology and enzyme activity of the Neutrase-hNFs was examined. The synthesized Neutrase-Mn hNFs showed superior activity and stability compared to free Neutrase. Traditional organic CC coupling reactions such as aldol condensation, decarboxylative aldol, Knoevenagel, Hantzsch-type reactions and synthesis of 4H-pyran derivatives were used to test the generality and scope of Neutrase promiscuity, while optimizing conditions for the Neutrase-Mn hNF biocatalyst. Briefly, Neutrase-Mn3(PO4)2 hNFs showed excellent enzyme activity, stability and reusability, qualifying as effective reusable catalysts for coupling reactions under mild conditions.

Adherence to practice guidelines for the management of febrile neutropenia in patients undergoing hematopoietic stem cell transplantation: An observational study in a referral center in Iran.

INTRODUCTION: Patients undergoing hematopoietic stem cell transplantation (HSCT) are suspected to develop febrile neutropenia (FN) and severe infections. Therefore, appropriate prescription of antibiotics in these patients is crucial to reduce the rates of morbidity, mortality, and antimicrobial resistance. The present study aimed to evaluate the physicians' prescription and adherence to the FN clinical guidelines among patients undergoing HSCT. METHODS: This prospective observational single-center study was conducted during a 15-month period in a tertiary referral hospital in Iran. The patients with at least one episode of FN following HSCT were included in the current study. The physicians' adherence to the Infectious Diseases Society of America (IDSA) and National Comprehensive Cancer Network (NCCN) clinical guidelines for the management of FN was evaluated using prescription data and medical record reviews. RESULTS: Two hundred and fifteen patients with 297 FN episodes were evaluated. The timing of antibiotics and the selection of the initial regimen were considered guideline-based therapy. However, antibiotic dosing and initial regimen modification were not followed in terms of the guideline recommendations in 58.1% of the patients. In particular, vancomycin was inappropriately given in 83.1% of patients. The overall adherence of physicians to the guidelines was 35.8%. CONCLUSION: Non-adherence to clinical guidelines is high particularly in initial regimen modification and administration of vancomycin, which affects hospital stay and patient's outcome. Implementation of guideline-review sessions to raise the awareness of the physicians and to improve the rational use of antimicrobial agents may be crucial.