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Purpose: This study aimed to assess the effectiveness of erythropoietin (EPO) as a novel treatment for peripheral nerve injury after surgical repair of an induced tibial nerve injury in dogs., Methods: Mongrel dogs (n = 27) were randomly divided into three equal groups. A complete tibial nerve injury was induced and repaired directly by stay sutures and the local application of 1 mL fibrin glue (control group). In the "systemic" group, 20,000 IU of EPO were given subcutaneously immediately after surgery and on the first and second days after surgery. In the "local" group, EPO was mixed with fibrin glue at 1,000 IU/mL. Lameness score, compound muscle action potential of the tibial nerve, and serum biochemical and histopathological examinations were performed to evaluate the treated dogs over the study period (12 weeks)., Results: EPO significantly improved the lameness score and compound muscle action potential in both the systemic and local groups. After 12 weeks, systemic and local groups showed earlier improvement in lameness, reaching scores of -1 and 0, respectively, in comparison with the control group, which did not reach a score of -1. The histological study revealed a normal architecture of the nerve bundles within connective tissue. The axons were aligned in a regular pattern, whereas the control group had disrupted and degenerated nerve axons with large gaps in between., Conclusions: EPO has an accelerating healing effect after tibial nerve surgical repair. Local EPO mimics systemic EPO treatment without systemic adverse effects. These findings indicated that EPO has a potential role in tibial nerve recovery and nerve regeneration., Clinical Relevance: The findings of the present experimental study supported the beneficial effects of systemic and local EPO when combined with peripheral nerve surgical repair, potentially improving functional outcomes and enhancing faster recovery., (Copyright © 2023 American Society for Surgery of the Hand. Published by Elsevier Inc. All rights reserved.)

The overexpression of EGFR has been recognized as the driver mechanism in the development of several human malignancies and the clinical use of EGFR inhibitors currently constitutes the standard of care for a wide range of malignancies, including colorectal cancer. However, the clinical efficacy of EGFR targeted inhibitors is limited by the development of intrinsic or acquired resistance, requiring the discovery of new compounds with different structural characteristics from those already developed. In this context, we explored the replacement of the aminoquinazoline pharmacophore of several FDA-approved EGFR inhibitors by its bioisosteric hydrazinothiazole moiety. A series of 14 new compounds were designed, synthesized, and evaluated as potential EGFR inhibitors. Compound 5i was active against 12 different cell lines in the NCI-60 cell line panel and showed an IC 50 of 6.9 ± 0.013 µM against HCT-116 cells, with no significant toxicity against normal human fibroblasts (WI-38). Further studies showed that this compound showed submicromolar activity against EGFR and was able to induce tumor cell cycle arrest and cell apoptosis. Additionally, docking experiments, molecular dynamics and binding free energy calculations were performed and confirmed the potential of 2-hydrazino-2,3-dihydrothiazole derivatives as new EGFR inhibitors., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Biological cellular materials are an important area of research in Additive manufacturing due to their intricate lightweight designs and forms with high energy absorption characteristics under compressive loading. In this study, we utilize the capability of Additive Manufacturing (AM) technology, experimental testing, and Finite Element Analysis (FEA) to design and investigate the mechanical behavior and energy absorption capabilities of novel Biomorphic Cellular Structures (BCS) inspired by the microstructure of cedar, oak, and palm wood. A comparative study of the elastic properties of the biomorphic cellular structures is carried out. The deformation and failure modes of the different cells were studied, and their performance was also discussed. Nonlinear finite element numerical simulation conducted has shown high accuracy in the prediction of deformation of the samples manufactured using additive manufacturing. The results show that cedar-bcs provides the best mechanical performance compared to the other two biomorphic cellular structures which could be as a result of its more vertical cell wall orientation, nevertheless, the palm-bcs showed a step-wise deformation and improved collapse stress. The obtained results suggest that the unique opportunities offered by the proposed experimental method, in combination with computational models, could serve to provide novel important information for the rational design of additively manufactured porous biomorphic materials., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

New sulfonamide derivatives have been synthesized and tested as antitumor agents. All newly synthesized compounds were tested in vitro against 60 lines of human cancer cells. Compound VIIb shows broad-spectrum activity with a mean inhibition value of 91.67% against all cell lines. It exhibited potent anticancer activity with GI50 values of 1.06-8.92 μM relative to most of the tested cancer cell lines. Compound VIIb has been tested for enzyme inhibition activity toward vascular endothelial growth factor receptor 2, where VEGFR-2 was potently inhibited at a lower IC50 value of 3.6 μM, compared with sorafenib (IC50 = 4.8 μM). Hybrid VIIb was also able to induce cell cycle disturbance and apoptosis in Renal UO-31 cells, as shown by DNA flow cytometry and Annexin V-FITC/PI assays. It has also revealed lower Bcl-2 protein expression anti-apoptotic levels and higher BAX, p53, and caspases 3 expression levels., (Copyright © 2021 Elsevier Inc. All rights reserved.)

In the current medical era, human health is experiencing numerous challenges, particularly the human malignancies. Therefore, the therapeutic arsenal for these malignancies is to be inexorably enhanced with new treatments that target tumor cells in a selective manner. In this regard, the present work aims at developing a new set of small molecules featuring the privileged isatin scaffold conjugated with a thiazolo[3,2-a]benzimidazole (TBI) motif through a cleavable hydrazide linker (7a-e and 10a-i) as potential anticancer CDK2 inhibitors. The large tricyclic TBI motif is anticipated to achieve a plethora of hydrophobic interactions within the CDK2 binding site. The growth of the two examined cell lines was significantly inhibited by most the prepared hybrids with IC 50 ranges; (2.60 ± 1.47-20.90 ± 1.17 µM, against MDA-MB-231) and (1.27 ± 0.06-16.83 ± 0.95 µM, against MCF-7). In particular, hybrids 7a, 7d and 10a displayed potent dual activity against the examined cell lines, and thus selected for further investigations. They exerted a significance alteration in the cell cycle progression, in addition to an apoptosis induction within both MDA-MB-231 and MCF-7 cells. Furthermore, 7a, 7d and 10a displayed potent CDK2 inhibitory action (IC 50  = 96.46 ± 5.3, 26.24 ± 1.4 and 42.95 ± 2.3 nM, respectively). The docking simulations unveiled, as expected, the ability of the TBI ring to well-accommodate and establish several hydrophobic interactions within a hydrophobic pocket in the CDK2 binding site. Also, the docking simulations highlighted the significance of incorporation of the hydrazide linker and isatin unsubstituted (NH) functionality in the H-bonding interactions. Interestingly, the most potent CDK2 inhibitor 7d achieved the best binding score (-11.2 Kcal/mole) and formed the most stable complex with CDK2 enzyme (RMSD = 1.24 Å) in a 100 ns MD simulation. In addition, the MM-PBSA calculations ascribed the lowest binding free energy to the 7d-CDK2 complex (-323.69 ± 15.17 kJ/mol). This could be attributed to an incorporation of the 5-OCH 3 group that was engaged in an extra hydrogen bonding with key THR14 amino acid residue. Finally, these results suggested hybrid 7d as a good candidate for further optimization as promising breast cancer antitumor agent and CDK2 inhibitor., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Novel hybrids of pyridazine-pyrazoline were synthesized aiming to develop new antiproliferative candidates. All compounds were submitted to the National Cancer Institute (NCI), USA, and many were proved to have significant antiproliferative activity. In addition, in vitro studies of the epidermal growth factor receptor (EGFR) inhibition showed that compounds IXn, IXg, IXb and IXl exhibited excellent inhibitory effect (IC 50  = 0.65, 0.75, 0.82 and 0.84 μM, respectively) compared to Erlotinib (IC 50  = 0.95 μM). The mechanistic effectiveness in cell cycle progression, apoptotic induction and gene regulation were assessed for the promising compounds IXg and IXn due to their significant EGFR inhibition. Flow cytometeric analysis indicated that compounds IXg and IXn result in increased cell numbers in phase G2/M, suggesting cell cycle arrest in phase G2/M in UO-31cells. Furthermore, real time PCR assay illustrated that compounds IXg and IXn elevated Bax/Bcl2 ratio which confirmed the mechanistic pathway of them. Moreover, the apoptotic induction of UO-31 renal cancer cells was enhanced effectively through activation of caspase-3 by compounds IXg and IXn. On the other hand, molecular docking study was performed to investigate binding mode of interaction of compounds with EGFR-PK in the active site with the aim of rationalizing its promising inhibitory activity. Finally, based on the aforementioned findings, compounds IXg and IXn could be considered as effective apoptosis modulators and promising leads for future development of new anti-renal cancer agents., (Copyright © 2020 Elsevier Inc. All rights reserved.)

The inter-particle interference of lignocellulosic materials describes the order of the macromolecules at a larger size scale, which can give information about the pore structure, and interface of cellulose and lignin. The pore structure and interface influence the rate of enzymatic hydrolysis and thermal decomposition in cellulosic ethanol manufacturing. In this study, the inter-particle interference of cellulose and lignin of three major categories of lignocellulosic materials: wood-based (cedar and oak), energy crop (bamboo), and agricultural or forestry waste (palm) were evaluated. Scanning electron microscopy (SEM) reveals morphological irregularities in the case of bamboo and palm, which may form nucleation sites for faster accessibility to enzyme molecules. Small-angle X-ray scattering (SAXS) shows increased power-law exponent for palm, suggesting a less clustered structure, which was consistent with the rough surface morphology as detected by the SEM. Differential Scanning Calorimetry (DSC) showed a higher temperature maximum for cedar and oak, which is indicative of higher intermolecular forces within their organic compounds, and could result in slower disintegration of the macromolecules during biochemical processing. This study will help to estimate the activity of the macromolecules and absorption capacity of lignocellulosic materials during biochemical processing., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The study of lignocellulosic materials calls for understanding the structure, and function of different cellulosic materials from diverse sources to scale-up cellulosic ethanol production. For the first time, a systematic assessment of the molecular and supramolecular structure highlighting the similarities and dissimilarities of three major categories of lignocellulosic materials: wood-based (cedar and oak), energy crop (bamboo), and agricultural or forestry waste (palm) are reported. The cellulose, hemicellulose, and lignin constituents were compared for their suitability in cellulosic ethanol production. FTIR showed structural variations within the functional groups with notable OH group in the palm and CC group in cedar. From the X-ray scattering, bamboo exhibited the highest crystallinity (49.5%), and palm showed the lowest crystallinity (22.6%) and crystallite size (2.6 nm). TGA revealed high cellulose amount and stable structure for cedar and oak, and the most unstable structure in the palm, which indicates a better cellulose/hemicellulose accessibility and biodegradability for enzymatic or chemical action in the palm. This comparative assessment can greatly facilitate material selection and component mixture, for developing an efficient and cost-effective biochemical process in ethanol manufacturing., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Large bone defects and bone loss after fractures remain significant challenges for orthopedic surgeons. Our study aims to find an available, applicable and biological treatment for bone regeneration overcoming the limitations in ESC/iPSC technology. We directly reprogrammed the mouse embryonic fibroblast (MEF) into osteoblast cells using different combinations of Yamanaka factors with human lim mineralization protein-3 (hLMP-3). LMP is an intracellular LIM-domain protein acting as an effective positive regulator of the osteoblast differentiation. After transduction, cells were cultured in osteogenic medium, and then examined for osteoblast formation. The expression of osteogenic markers (BMP2, Runx2 and Osterix) during reprogramming and in vitro mineralization assay revealed that the best reprogramming cocktail was (c-Myc - Oct4) with hLMP-3. In addition, both immunofluorescent staining and western blot analysis confirmed that osteocalcin (OCN) expression increased in the cells treated with the c-Myc/Oct4/hLMP3 cocktail than using hLMP-3 alone. Furthermore, this reprogramming cocktail showed efficient healing in an induced femoral bone defect in rat animal model one month after transplantation. In the present study, we reported for the first time the effect of combining Yamanaka factors with hLMP-3 to induce osteoblast cells from MEF both in vitro and in vivo., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The popularity of food produced from animals kept under an organic regimen has increased in recent years. In Germany, turkey meat consumption has increased. Despite several studies assessing the susceptibility of campylobacters to various antibiotics in poultry, no sufficient data exists regarding the antimicrobial resistance of campylobacters in organic-reared turkeys. This study provides information about antibiotic resistance in Campylobacter isolated from turkeys reared on organic farms in Germany. Ninety-six Campylobacter strains (41 C. jejuni and 55 C. coli) were isolated from different free-range turkey flocks. In vitro antimicrobial sensitivity testing was done using a broth microdilution test, and the presence of resistance genes to antibiotics (ciprofloxacin, tetracycline) was investigated. All Campylobacter isolates from organic turkeys (n = 96) were phenotypically sensitive to gentamicin, erythromycin, streptomycin, and chloramphenicol. In this study, the antibiotic susceptibilities of C. jejuni to ciprofloxacin, tetracycline, and naladixic acid were 56.0%, 51.3%, and 56.0%, respectively. In contrast, 44.0%, 73.0%, and 74.6% of C. coli isolates were resistant to tetracycline, ciprofloxacin, and nalidixic acid, respectively. Replacement of the Thr-86→Ile in the gyrA gene, and the presence of the tet(O) gene were the mainly identified resistance mechanisms against fluoroquinolones and tetracycline, respectively.These results also reinforce the need to develop strategies and implement specific control procedures to reduce the development of antimicrobial resistance., (© 2015 Poultry Science Association Inc.)

Infected laying hens regularly excrete large amounts of Campylobacter jejuni with their feces, which represent a reservoir of infection within the flock and for animals in the region. However, the knowledge about survival times of C. jejuni in these feces is still scarce. Therefore, orienting laboratory experiments were carried out under controlled conditions to estimate the survival times of C. jejuni both in artificially and naturally contaminated laying hen feces. In 6 different laying hen flocks (3 Campylobacter-free and 3 Campylobacter-positive flocks), fresh excreta were randomly collected and pooled in 20-g samples per flock. In the laboratory, each of the 3 pooled samples from the Campylobacter-free barns were homogenized and mixed with 10 mL of a freshly prepared C. jejuni suspension (3 × 10(8) cfu/mL). The other 3 samples were homogenized only. The 6 samples were stored at 20 ± 1°C and 40 to 60% RH in 2 different incubators. Specimens of 2 g were taken from all 6 samples 1 h after storage and daily at the same time during the next 10 consecutive days and investigated on culturable C. jejuni. The survival times of culturable C. jejuni ranged from 72 to 96 h in artificially inoculated feces and varied from 120 to 144 h in naturally colonized flocks. The flaA typing by RFLP confirmed that the isolates from the artificially contaminated feces were identical with the added strain. A total of 5 different flaA types were identified from the naturally contaminated feces, and survival of these isolates was dependent on flaA type. The demonstrated survival times indicate that contaminated fresh feces are an important reservoir of C. jejuni, representing a permanent source of infection over at least 6 d after excretion. It shows the considerable potential of fresh feces in transmitting the agent within and between flocks during that period. This 6-d span should be considered when poultry manure is applied to land as organic fertilizer.

Two candidate DNA vaccines based on the proteins CFP10 and CFP21 encoded by regions of difference (RDs) of Mycobacterium tuberculosis were evaluated individually and in multivalent combination with the immunodominant protein Ag85B for induction of protective immune responses against experimental tuberculosis. Experimental DNA vaccines induced substantial levels of cell-mediated immune responses as indicated by marked lymphocyte proliferation, significant release of the Th1 cytokines IFN-gamma and IL-12 (p40), and predominant cytotoxic T cell activity. High levels of antigen-specific IgG1 and IgG2a antibodies observed in the sera of immunized mice depicted strong humoral responses generated by DNA vaccine constructs. The multivalent combination of three DNA vaccine constructs induced maximal T cell and humoral immune responses. All the experimental vaccines imparted significant protection against challenge with M. tuberculosis H(37)Rv (in terms of colony-forming unit reduction in lungs and spleen) as compared to vector controls. The level of protection exhibited by multivalent DNA vaccine formulation was found to be equivalent to that of Mycobacterium bovis BCG observed both at 4 and 8 weeks post-challenge. These results show the protective potential of the multivalent DNA vaccine formulation used in this study.