[Effects of different types of intraocular lens implantation on patient's visual quality and function after phacoemulsification].

Objective: To explore the effects of different types of intraocular lens (IOL) implantation on patient's visual quality and function after phacoemulsification. Methods: The clinical data of patients with monocular cataract who underwent phacoemulsification in the Department of Ophthalmology, People's Hospital Affiliated to Shandong First Medical University between December 2021 and May 2023 were retrospectively analyzed. According to the types of IOL, the patients were divided into monofocal group, bifocal group and depth of focus extension group. Three months later, uncorrected distance visual acuity (UCDVA), best corrected distance visual acuity (BCDVA), uncorrected intermediate visual acuity (UCIVA), best corrected intermediate visual acuity (BCIVA), uncorrected near visual acuity (UCNVA) and best corrected near visual acuity (BCNVA) were detected. Contrast sensitivity and total wavefront aberration were measured by visual function analyzer. Satisfaction with visual quality was evaluated by hospital-made satisfaction questionnaire. Results: A total of 92 patients were included, with 31 males and 61 females, and their age was (61.8±5.2) years. There were 43, 28 and 21 cases in monofocal group, bifocal group and depth of focus extension group, respectively. No statistically significant difference was found in clinical baseline data among the three groups. UCIVA, UCDVA, BCIVA and BCDVA in depth of focus extension group were 1.01±0.13, 0.92±0.18, 1.21±0.19 and 1.20±0.23, respectively, which were higher than those in monofocal group (0.62±0.12, 0.74±0.13, 1.02±0.17, 1.07±0.19, respectively) and bifocal group (0.67±0.15, 0.78±0.14, 1.01±0.16, 1.01±0.18, respectively), while absolute value of spherical equivalent [(-0.42±0.07) D] was lower than that in the other two groups [ (-0.49±0.05) D and (-0.45±0.08) D] (both P<0.05). UCNVA and BCNVA in bifocal group were 0.91±0.18 and 1.25±0.18, which were higher than those in depth of focus extension group (0.63±0.24 and 1.19±0.17) (both P<0.05). There were no significant differences in contrast sensitivity among the three groups under day vision or between monofocal group and bifocal group under night vision (all P>0.05), but the contrast sensitivity was higher in depth of focus extension group under night vision (3.0, 6.0, 12.0 c/d) than other two groups (all P<0.05). The score of ocular discomfort was the highest in bifocal group, followed by depth of focus extension group and monofocal group (both P<0.05). The score of visual interference in bifocal group was lower than that in monofocal group and depth of focus extension group (both P<0.05). The scores of subjective feeling in bifocal group and depth of focus extension group were higher than that in monofocal group (both P<0.05). The reading score was the highest in bifocal group, followed by depth of focus extension group and monofocal group (both P<0.05). There was no significant difference in total low-order aberration among the three groups (P=0.472). The total aberration and higher-order aberration [(0.74±0.35) µm and (0.41±0.12) µm] were the highest in monofocal group, followed by bifocal group [(0.61±0.21) µm and (0.22±0.09) µm] and depth of focus extension group [(0.46±0.13) µm and (0.06±0.09) µm] (all P<0.05). Conclusions: IOL implantation with depth of focus extension can enhance visual range, night vision and contrast sensitivity, and thus effectively improve postoperative visual quality and function in cataract patients. The bifocal IOL can better improve the patient's UCNVA and BCNVA, resulting in high satisfaction with visual quality.

Naturally occurring phenylethanoids and phenylpropanoids: antimalarial potential.

Malaria as an infectious disease is one of the world's most dangerous parasitic diseases. There is an urgent need for the development of new antimalarial drugs. Natural products are a very rich source of new bioactive compounds. Our research aims to shed light on the recent studies which demonstrated the antimalarial potential of phenylpropanoids as a major natural-products class. This study involves an in silico analysis of naturally-occurring phenylpropanoids and phenylethanoids which showed 25 compounds with moderate to strong binding affinity to various amino acid residues lining the active site; P. falciparum kinase (PfPK5), P. falciparum cytochrome bc1 complex (cyt bc1), and P. falciparum lysyl-tRNA synthetase (PfKRS1); of Plasmodium falciparum parasite, a unicellular protozoan which causes the most severe and life-threatening malaria. Furthermore, the study was augmented by the assessment of antiplasmodial activity of glandularin, a naturally occurring dibenzylbutyrolactolic lignan, against chloroquine-sensitive 3D7 strain of P. falciparum using SYBR green I-based fluorescence assay, which showed high antimalarial activity with IC50 value of 11.2 µM after 24 hours of incubation. Our results highlight phenylpropanoids and glandularin in particular as a promising chemical lead for development of antimalarial drugs.

[Clinical value of the MeltPro MTB assays in detection of drug-resistant tuberculosis in paraffin-embedded tissues].

Objective: To evaluate the clinical value of the MeltPro MTB assays in the diagnosis of drug-resistant tuberculosis. Methods: A cross-sectional study design was used to retrospectively collect all 4 551 patients with confirmed tuberculosis between January 2018 and December 2019 at Beijing Chest Hospital, Capital Medical University. Phenotypic drug sensitivity test and GeneXpert MTB/RIF (hereafter referred to as "Xpert") assay were used as gold standards to analyze the accuracy of the probe melting curve method. The clinical value of this technique was also evaluated as a complementary method to conventional assays of drug resistance to increase the detective rate of drug-resistant tuberculosis. Results: By taking the phenotypic drug susceptibility test as the gold standard, the sensitivity of the MeltPro MTB assays to detect resistance to rifampicin, isoniazid, ethambutol and fluoroquinolone was 14/15, 95.7%(22/23), 2/4 and 8/9,respectively; and the specificity was 92.0%(115/125), 93.2%(109/117), 90.4%(123/136) and 93.9%(123/131),respectively; the overall concordance rate was 92.1%(95%CI:89.6%-94.1%),and the Kappa value of the consistency test was 0.63(95%CI:0.55-0.72).By taking the Xpert test results as the reference, the sensitivity of this technology to the detection of rifampicin resistance was 93.6%(44/47), the specificity was100%(310/310), the concordance rate was 99.2%(95%CI:97.6%-99.7%), and the Kappa value of the consistency test was 0.96(95%CI:0.93-0.99). The MeltPro MTB assays had been used in 4 551 confirmed patients; the proportion of patients who obtained effective drug resistance results increased from 83.3% to 87.8%(P<0.01); and detection rate of rifampicin, isoniazid, ethambutol, fluoroquinolone resistance, multidrug and pre-extensive drug resistance cases were increased by 3.2%, 14.7%, 22.2%, 13.7%, 11.2% and 12.5%, respectively. Conclusion: The MeltPro MTB assays show satisfactory accuracy in the diagnosis of drug-resistant tuberculosis. This molecular pathological test is an effective complementary method in improving test positivity of drug-resistant tuberculosis.

Prognosis and molecular characteristics of IBD-associated colorectal cancer: Experience from a French tertiary-care center.

BACKGROUND: Little is known about the prognosis of colorectal cancer associated with inflammatory bowel disease (CRC-IBD) in a real-world cohort in France. METHODS: We conducted a retrospective observational study including all patients presenting CRC-IBD in a French tertiary center. RESULTS: Among 6510 patients, the rate of CRC was 0.8% with a median delay of 19.5 years after IBD diagnosis (median age 46 years, ulcerative colitis 59%, initially localized tumor 69%). There was a previous exposure to immunosuppressants (IS) in 57% and anti-TNF in 29% of the cases. A RAS mutation was observed in only 13% of metastatic patients. OS of the whole cohort was 45 months. OS and PFS of synchronous metastatic patients was 20.4 months and 8.5 months respectively. Among the patients with localized tumor those previously exposed to IS had a better PFS (39 months vs 23 months; p = 0.05) and OS (74 vs 44 months; p = 0.03). The IBD relapse rate was 4%. No unexpected chemotherapy side-effect was observed CONCLUSIONS: OS of CRC-IBD is poor in metastatic patients although IBD is not associated with under-exposure or increased toxicity to chemotherapy. Previous IS exposure may be associated with a better prognosis.

[Relationship between expression of NLRP3 inflammasome and improvement of macular structure in patients with wet age-related macular degeneration after anti-vascular endothelial growth factor therapy].

Objective: To explore the relationship between expression of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) inflammasome and improvement of macular structure in patients with wet age-related macular degeneration (wAMD) after anti-vascular endothelial growth factor (VEGF) therapy. Methods: A before-after study was carried out. A total of 110 patients (110 eyes) with wAMD who were admitted to Department of Ophthalmology, People's Hospital Affiliated to Shandong First Medical University between August 2019 and December 2021 were enrolled, and all patients were given vitreous injection of anti-VEGF drug (ranibizumab or bevacizumab). The aqueous humor was collected to detect mRNA levels of NLRP3, cysteinyl aspartate specific protease-1 (Caspase-1), apoptosis-associated speck-like protein (ASC) and interleukin (IL) 1ß by fluorescence quantitative PCR. The levels of IL-1ß, IL-18, tumor necrosis factor α (TNF-α) and VEGF in aqueous humor were detected by enzyme-linked immunosorbent assay (ELISA). The correlation between the above indexes and central macular thickness (CMT) in wAMD patients was analyzed by multivariate linear regression analysis. Results: In the 110 wAMD patients, there were 68 males and 42 females, with a mean age of (68.7±7.6) years. Compared with those before treatment, mRNA levels of NLRP3 (1.65±0.27, 1.34±0.19 vs 1.97±0.23, both P<0.017), Caspase-1 (1.47±0.15, 1.29±0.17 vs 1.53±0.18, both P<0.017), ASC (1.33±0.14, 1.21±0.18 vs 1.47±0.12, both P<0.017) and IL-1ß (1.78±0.21, 1.46±0.17 vs 2.21±0.24, both P<0.017), and levels of IL-1ß [(26.9±5.7), (20.3±4.6) vs (33.6±8.3) ng/L, both P<0.017], IL-18 [(32.7±7.6), (23.3±6.9) vs (46.4±9.4) ng/L, both P<0.017], TNF-α [(39.4±6.6), (21.7±6.3) vs (52.9±9.1) ng/L, both P<0.017] and VEGF [(35.7±10.2), (23.4±6.7) vs (65.4±19.3) ng/L, both P<0.017] were decreased after the first and second injection. Moreover, the above-mentioned indexes after second injection were lower than those after the first injection (all P<0.017). The results of multivariate linear regression analysis showed that NLRP3 mRNA (the first injection: ß=53.750, P<0.001; the second injection: ß=94.648, P<0.001), IL-1ß (the first injection: ß=1.356, P=0.021; the second injection: ß=2.008, P=0.003), IL-18 (the first injection: ß=1.984, P<0.001; the second injection: ß=1.251, P=0.003) and VEGF (the first injection: ß=1.875, P<0.001; the second injection: ß=2.119, P<0.001) had linear relationships with CMT. Conclusion: The decrease of NLRP3 inflammasome and its products in aqueous humor may be related to the improvement of macular structure in wAMD patients after anti-VEGF therapy.

Metabolomic Profiling, In Vitro Antimalarial Investigation and In Silico Modeling of the Marine Actinobacterium Strain Rhodococcus sp. UR111 Associated with the Soft Coral Nephthea sp.

Malaria is a persistent illness with a great public health concern. To combat this fatal disease, developing effective antimalarial medications has become a necessity. In the present study, we described the actinomycetes associated with the Red Sea soft coral Nephthea sp. and isolated a strain that was sub-cultured in three different media (M1, ISP2, and OLIGO). Actinomycete isolate's phylogenetic analysis of the 16S rRNA gene revealed that it belongs to the genus Rhodococcus. In vitro screening of the antimalarial activity for three extracts against Plasmodium falciparum was carried out. Non-targeted metabolomics for the chemical characterization of the isolated actinomycete species UA111 derived extracts were employed using high-resolution liquid chromatography-mass spectrometry (LC-HR-MS) for dereplication purposes. Additionally, statistical analysis of the vast LC-MS data was performed using MetaboAnalyst 5.0. Finally, an in silico analysis was conducted to investigate the potential chemical compounds that could be the source of the antimalarial potential. The results revealed that ISP2 media extract is the most effective against Plasmodium falciparum, according to antimalarial screening (IC50 8.5 µg/mL), in contrast, OLIGO media extract was inactive. LC-HRMS-based metabolomics identified a range of metabolites, mainly alkaloids, from the genus Rhodococcus. On the other hand, multivariate analysis showed chemical diversity between the analyzed samples, with ISP2 extract being optimal. The docking analysis was able to anticipate the various patterns of interaction of the annotated compounds with three malarial protein targets (P. falciparum kinase, P. falciparum cytochrome bc1 complex, and P. falciparum lysyl-tRNA synthetase). Among all of the test compounds, perlolyrine (11) and 3097-B2 (12) displayed the best docking profiles. In conclusion, this work demonstrated the value of the established method for the metabolic profiling of marine actinomycetes using the data from liquid chromatography-mass spectrometry (LC-MS), which helps to streamline the difficult isolation stages required for their chemical characterization. In addition, the antimalarial efficacy of this strain has intriguing implications for future pharmaceutical development.

[Serotyping methods of Streptococcus pneumonia].

More than 100 serotypes of Streptococcus pneumonia have been identified, which has been one bottleneck problem for pneumococcal disease diagnosis, surveillance, development of pneumococcal vaccine and effectiveness evaluation of pneumococcal vaccines. Three categories of approaches for pneumococcal serotyping will be discussed including phenotyping based on anti-serum, biochemical typing based on pneumococcal capsular characteristics and genotyping based on pneumococcal capsular locus sequences. We reviewed the development and applications of different serotyping of pneumococcus to provide guidance for pneumococcal disease prevention and control.

Cytotoxic potential of Nephthea sp.-derived actinomycetes supported by metabolomics analysis.

Soft corals and associated microorganisms are known to produce leads for anticancer drugs. Keeping this in mind, Nephthea sp.; a Red Sea soft coral was investigated for the first time using the OSMAC approach. Two isolates, Streptomyces sp. UR63 and Micrococcus sp. UR67 were identified. Their extracts revealed the presence of alkaloids, macrolides, quinones, fatty acids and terpenoids. Further comparison through a set of multivariate data analyses revealed their unique chemical profiles. The extracts displayed inhibitory potencies against HepG-2, Caco-2 and MCF-7 tumor cell lines with IC50 values ranging from 11.4 to 38.7 µg/mL when compared with the positive control, doxorubicin. The study not only highlights the cytotoxic potential of soft coral-associated actinomycetes but also shows the advantage of using the OSMAC approach in this regard.

Alginate in corneal tissue engineering.

Corneal blindness is the major cause of vision impairment and the fourth-largest leading cause of blindness worldwide. An allograft corneal transplant is the most routine treatment for visual loss. Further complications can occur, such as transplant rejection, astigmatism, glaucoma, uveitis, retinal detachment, corneal ulceration due to reopening of the surgical wounds, and infection. For patients with autoimmune disorders, allografting for chemical burns and infections is contraindicated because of the risk of disease transmission and further complications. Moreover, corrective eye surgery renders the corneas unsuitable for allografting, further increasing the gap between donor tissue demand and supply. Due to these challenges, other therapeutic strategies such as artificial alternatives to donor corneal tissue are being considered. This review focuses on the use of alginate as a building block of therapeutic drugs or cell delivery systems to enhance drug retention and encourage corneal regeneration. The similarity of alginate hydrogel water content to native corneal tissue makes it a promising support structure. Alginate possess desired drug carrier characteristics, such as mucoadhesiveness and penetration enhancing properties. Whilst alginates have been extensively studied for their application in tissue engineering (TE), with many reviews being published, no reviews exist to our knowledge directly looking at alginates for corneal applications. The role of alginate in drug delivery to the surface of the eye and as a support structure (bioinspired tissue scaffold) for corneal TE is discussed. Biofabrication techniques such as gel casting, electrospinning, and bioprinting to develop tissue precursors and substitutes are compared. Finally, cell and tissue encapsulation in alginate for storage and transport to expand the scope of cell-based therapy for corneal blindness is also discussed in the light of recent applications of alginate in maintaining the function of biofabricated constructs for storage and transport.

Storable Cell-Laden Alginate Based Bioinks for 3D Biofabrication.

Over the last decade, progress in three dimensional (3D) bioprinting has advanced considerably. The ability to fabricate complex 3D structures containing live cells for drug discovery and tissue engineering has huge potential. To realise successful clinical translation, biologistics need to be considered. Refinements in the storage and transportation process from sites of manufacture to the clinic will enhance the success of future clinical translation. One of the most important components for successful 3D printing is the 'bioink', the cell-laden biomaterial used to create the printed structure. Hydrogels are favoured bioinks used in extrusion-based bioprinting. Alginate, a natural biopolymer, has been widely used due to its biocompatibility, tunable properties, rapid gelation, low cost, and easy modification to direct cell behaviour. Alginate has previously demonstrated the ability to preserve cell viability and function during controlled room temperature (CRT) storage and shipment. The novelty of this research lies in the development of a simple and cost-effective hermetic system whereby alginate-encapsulated cells can be stored at CRT before being reformulated into an extrudable bioink for on-demand 3D bioprinting of cell-laden constructs. To our knowledge the use of the same biomaterial (alginate) for storage and on-demand 3D bio-printing of cells has not been previously investigated. A straightforward four-step process was used where crosslinked alginate containing human adipose-derived stem cells was stored at CRT before degelation and subsequent mixing with a second alginate. The printability of the resulting bioink, using an extrusion-based bioprinter, was found to be dependent upon the concentration of the second alginate, with 4 and 5% (w/v) being optimal. Following storage at 15 °C for one week, alginate-encapsulated human adipose-derived stem cells exhibited a high viable cell recovery of 88 ± 18%. Stored cells subsequently printed within 3D lattice constructs, exhibited excellent post-print viability and even distribution. This represents a simple, adaptable method by which room temperature storage and biofabrication can be integrated for on-demand bioprinting.

A clinically relevant combination treatment with doxorubicin and cyclophosphamide does not induce hepatotoxicity in C57BL/6J mice.

BACKGROUND AND AIM: Chronic exposure to chemotherapeutics can lead to severe adverse events including hepatotoxicity. A combination chemotherapy regimen of doxorubicin (DOX) and cyclophosphamide (CPS) is employed in treatment of several cancers such as leukemia, lymphoma, and breast cancer. It is not well understood whether a combination therapy of DOX and CPS can induce hepatotoxicity. We therefore sought to determine whether co-administration of DOX and CPS at their clinically relevant doses and frequency results in hepatotoxicity. METHODS: Male C57BL/6J mice received one intraperitoneal injection of saline or DOX-2mg /kg and CPS-50mg/kg once a week for 4 weeks. After the treatment period, liver histology and various serum biomarkers of hepatotoxicity were assessed. RESULTS: Co-treatment of DOX and CPS did not alter the serum levels of alanine aminotransferase (ALT), alkaline phosphatase (ALP), bilirubin, albumin, globulin, or total protein. Similarly, co-administration of DOX and CPS did not result in a noticeable change in liver histology. However, it was notable that the concomitant treatment with DOX and CPS resulted in a significant increase in serum levels of aspartate aminotransferase (AST). Elevated serum AST levels were also associated with increased serum creatinine kinase (CK) levels, suggesting that the elevated serum AST levels are likely due to muscle injury following the co-administration of DOX and CPS. CONCLUSION: Taken together, our results, for the first time, suggest that co-administration of DOX and CPS, at their clinically relevant doses and frequency does not induce a significant hepatotoxicity in the mice.

[Exploration on the construction of analysis indicators system for antibiotic resistance monitoring].

Antibiotic resistance (AR) is a severe and fast-growing public health challenge with rapid globalization, especially in China. Although some monitoring systems were established in different fields, fragmentation of information failed to show the overall trend and spread of AR. It is necessary to establish a national monitoring system to reveal the occurrence, development, and spread of AR. The new AR monitoring system needs an updated analysis indicators system. We intend to recommend a new analysis indicators system for AR was constructed and applied to AR data monitoring and analysis for humans, animals, the environment, and foods. After investigating and analyzing the 5 Chinese major AR monitoring systems and literature, we have formulated 15 AR monitoring analysis indicators and initially established an evaluation system for the country's new AR monitoring system.

Expression of Rasd1 in mouse endocrine pituitary cells and its response to dexamethasone.

Dexamethasone-induced Ras-related protein 1 (Rasd1) is a member of the Ras superfamily of monomeric G proteins that have a regulatory function in signal transduction. Rasd1, also known as Dexras1 or AGS1, is rapidly induced by dexamethasone (Dex). While prior data indicates that Rasd1 is highly expressed in the pituitary and that the gene may function in regulation of corticotroph activity, its exact cellular localization in this tissue has not been delineated. Nor has it been determined which endocrine pituitary cell type(s) are responsive to Dex-induced expression of Rasd1. We hypothesized that Rasd1 is primarily localized in corticotrophs and furthermore, that its expression in these cells would be upregulated in response to exogenous Dex administration. Rasd1 expression in each pituitary cell type both under basal conditions and 1-hour post Dex treatment were examined in adult male mice. While a proportion of all endocrine pituitary cell types expressed Rasd1, a majority of corticotrophs and thyrotrophs expressed Rasd1 under basal condition. In vehicle treated animals, approximately 50-60% of corticotrophs and thyrotrophs cells expressed Rasd1 while the gene was detected in only 15-30% of lactotrophs, somatotrophs, and gonadotrophs. In Dex treated animals, Rasd1 expression was significantly increased in corticotrophs, somatotrophs, lactotrophs, and gonadotrophs but not thyrotrophs. In Dex treated animals, Rasd1 was detected in 80-95% of gonadotrophs and corticotrophs. In contrast, Dex treatment increased Rasd1 expression to a lesser extent (55-60%) in somatotrophs and lactotrophs. Corticotrophs of the pars intermedia, which lack glucocorticoid receptors, failed to display increased Rasd1 expression in Dex treated animals. Rasd1 is highly expressed in corticotrophs under basal conditions and is further increased after Dex treatment, further supporting its role in glucocorticoid negative feedback. In addition, the presence and Dex-induced expression of Rasd1 in endocrine pituitary cell types, other than corticotrophs, may implicate Rasd1 in novel pituitary functions.

Milliscale Substrate Curvature Promotes Myoblast Self-Organization and Differentiation.

Biological tissues comprise complex structural environments known to influence cell behavior via multiple interdependent sensing and transduction mechanisms. Yet, and despite the predominantly nonplanar geometry of these environments, the impact of tissue-size (milliscale) curvature on cell behavior is largely overlooked or underestimated. This study explores how concave, hemicylinder-shaped surfaces 3-50 mm in diameter affect the migration, proliferation, orientation, and differentiation of C2C12 myoblasts. Notably, these milliscale cues significantly affect cell responses compared with planar substrates, with myoblasts grown on surfaces 7.5-15 mm in diameter showing prevalent migration and alignment parallel to the curvature axis. Moreover, surfaces within this curvature range promote myoblast differentiation and the formation of denser, more compact tissues comprising highly oriented multinucleated myotubes. Based on the similarity of effects, it is further proposed that myoblast susceptibility to substrate curvature depends on mechanotransduction signaling. This model thus supports the notion that cellular responses to substrate curvature and compliance share the same molecular pathways and that control of cell behavior can be achieved via modulation of either individual parameter or in combination. This correlation is relevant for elucidating how muscle tissue forms and heals, as well as for designing better biomaterials and more appropriate cell-surface interfaces.

A metabolomic approach to target antimalarial metabolites in the Artemisia annua fungal endophytes.

Fungal endophytes are a major source of anti-infective agents and other medically relevant compounds. However, their classical blinded-chemical investigation is a challenging process due to their highly complex chemical makeup. Thus, utilizing cheminformatics tools such as metabolomics and computer-aided modelling is of great help deal with such complexity and select the most probable bioactive candidates. In the present study, we have explored the fungal endophytes associated with the well-known antimalarial medicinal plant Artemisia annua for their production of further antimalarial agents. Based on the preliminary antimalarial screening of these endophytes and using LC-HRMS-based metabolomics and multivariate analyses, we suggested different potentially active metabolites (compounds 1-8). Further in silico investigation using the neural-network-based prediction software PASS led to the selection of a group of quinone derivatives (compounds 1-5) as the most possible active hits. Subsequent in vitro validation revealed emodin (1) and physcion (2) to be potent antimalarial candidates with IC50 values of 0.9 and 1.9 µM, respectively. Our approach in the present investigation therefore can be applied as a preliminary evaluation step in the natural products drug discovery, which in turn can facilitate the isolation of selected metabolites notably the biologically active ones.

The Role of the Histone Methyltransferase PfSET10 in Antigenic Variation by Malaria Parasites: a Cautionary Tale.

The virulence of the malaria parasite Plasmodium falciparum is due in large part to its ability to avoid immune destruction through antigenic variation. This results from changes in expression within the multicopy var gene family that encodes the surface antigen P. falciparum erythrocyte protein one (PfEMP1). Understanding the mechanisms underlying this process has been a high-profile research focus for many years. The histone methyltransferase PfSET10 was previously identified as a key enzyme required both for parasite viability and for regulating var gene expression, thus making it a prominent target for developing antimalarial intervention strategies and the subject of considerable research focus. Here, however, we show that disruption of the gene encoding PfSET10 is not lethal and has no effect on var gene expression, in sharp contrast with previously published reports. The contradictory findings highlight the importance of reevaluating previous conclusions when new technologies become available and suggest the possibility of a previously unappreciated plasticity in epigenetic gene regulation in P. falciparumIMPORTANCE The identification of specific epigenetic regulatory proteins in infectious organisms has become a high-profile research topic and a focus for several drug development initiatives. However, studies that define specific roles for different epigenetic modifiers occasionally report differing results, and we similarly provide evidence regarding the histone methyltransferase PfSET10 that is in stark contrast with previously published results. We believe that the conflicting results, rather than suggesting erroneous conclusions, instead reflect the importance of revisiting previous conclusions using newly developed methodologies, as well as caution in interpreting seemingly contrary results in fields that are known to display considerable plasticity, for example metabolism and epigenetics.

Effect of isolation method on human corneal stromal cell behaviour.

Current research on healthy corneal stromal cells will typically use primary cells as they are the most representative of in vivo behaviour. Primary cells are normally isolated from the limbus of discarded donor peripheral corneal tissue left over from transplantation (due to its relative abundance). Therefore, the central part of the cornea is less used in research as this tissue is usually used for transplantation. In some cases, although rare, the whole cornea, can become available for research. It is important to keep in mind that these corneas often have longer storage time, but the use of the central tissue for research is even more interesting, as knowing what cells are being transplanted into recipients would be highly relevant. To this end, stromal cells were extracted from both the limbus and central button of healthy corneas donated for research. This allowed for important comparison between central and limbal cells in culture. Of interest here was the extraction method of stromal cells from the donor tissue. The two most common methods of extraction are enzyme digestion and explant migration. However, no work has been done to understand how each method relatively affects the extracted cells. The extraction method and location from which stromal cells are harvested seems to have a significant effect on the cell adherence, survival, and gene expression of the stromal cells in culture. Enzyme digested cells showed that limbal and central cells had different gene expressions prior to culture, with gene such as ALDH3A1 being much more expressed in limbal cells. Enzyme digesting the limbal ring seems to yield the hardiest populations of stromal cells, a desirable trait in the culture of primary cells.

Use of biomaterials in corneal endothelial repair.

Human corneal endothelium (HCE) is a single layer of hexagonal cells that lines the posterior surface of the cornea. It forms the barrier that separates the aqueous humor from the rest of the corneal layers (stroma and epithelium layer). This layer plays a fundamental role in maintaining the hydration and transparency of the cornea, which in turn ensures a clear vision. In vivo, human corneal endothelial cells (HCECs) are generally believed to be nonproliferating. In many cases, due to their nonproliferative nature, any damage to these cells can lead to further issues with Descemet's membrane (DM), stroma and epithelium which may ultimately lead to hazy vision and blindness. Endothelial keratoplasties such as Descemet's stripping automated endothelial keratoplasty (DSAEK) and Descemet's membrane endothelial keratoplasty (DEK) are the standard surgeries routinely used to restore vision following endothelial failure. Basically, these two similar surgical techniques involve the replacement of the diseased endothelial layer in the center of the cornea by a healthy layer taken from a donor cornea. Globally, eye banks are facing an increased demand to provide corneas that have suitable features for transplantation. Consequently, it can be stated that there is a significant shortage of corneal grafting tissue; for every 70 corneas required, only 1 is available. Nowadays, eye banks face long waiting lists due to shortage of donors, seriously aggravated when compared with previous years, due to the global COVID-19 pandemic. Thus, there is an urgent need to find alternative and more sustainable sources for treating endothelial diseases, such as utilizing bioengineering to use of biomaterials as a remedy. The current review focuses on the use of biomaterials to repair the corneal endothelium. A range of biomaterials have been considered based on their promising results and outstanding features, including previous studies and their key findings in the context of each biomaterial.

Response of human oral mucosal epithelial cells to different storage temperatures: A structural and transcriptional study.

PURPOSE: Seeking to improve the access to regenerative medicine, this study investigated the structural and transcriptional effects of storage temperature on human oral mucosal epithelial cells (OMECs). METHODS: Cells were stored at four different temperatures (4°C, 12°C, 24°C and 37°C) for two weeks. Then, the morphology, cell viability and differential gene expression were examined using light and scanning electron microscopy, trypan blue exclusion test and TaqMan gene expression array cards, respectively. RESULTS: Cells stored at 4°C had the most similar morphology to non-stored controls with the highest viability rate (58%), whereas the 37°C group was most dissimilar with no living cells. The genes involved in stress-induced growth arrest (GADD45B) and cell proliferation inhibition (TGFB2) were upregulated at 12°C and 24°C. Upregulation was also observed in multifunctional genes responsible for morphology, growth, adhesion and motility such as EFEMP1 (12°C) and EPHA4 (4°C-24°C). Among genes used as differentiation markers, PPARA and TP53 (along with its associated gene CDKN1A) were downregulated in all temperature conditions, whereas KRT1 and KRT10 were either unchanged (4°C) or downregulated (24°C and 12°C; and 24°C, respectively), except for upregulation at 12°C for KRT1. CONCLUSIONS: Cells stored at 12°C and 24°C were stressed, although the expression levels of some adhesion-, growth- and apoptosis-related genes were favourable. Collectively, this study suggests that 4°C is the optimal storage temperature for maintenance of structure, viability and function of OMECs after two weeks.

Scale-Up Technologies for the Manufacture of Adherent Cells.

Great importance is being given to the impact our food supply chain and consumers' food habits are having on the environment, human health, and animal welfare. One of the latest developments aiming at positively changing the food ecosystem is represented by cultured meat. This form of cellular agriculture has the objective to generate slaughter-free meat products starting from the cultivation of few cells harvested from the animal tissue of interest. As a consequence, a large number of cells has to be generated at a reasonable cost. Just to give an idea of the scale, there were billions of cells just in a bite of the first cultured-meat burger. Thus, one of the major challenges faced by the scientists involved in this new ambitious and fascinating field, is how to efficiently scale-up cell manufacture. Considering the great potential presented by cultured meat, audiences from different backgrounds are very interested in this topic and eager to be informed of the challenges and possible solutions in this area. In light of this, we will provide an overview of the main existing bioprocessing technologies used to scale-up adherent cells at a small and large scale. Thus, giving a brief technical description of these bioprocesses, with the main associated advantages and disadvantages. Moreover, we will introduce an alternative solution we believe has the potential to revolutionize the way adherent cells are grown, helping cultured meat become a reality.