bluuebunny/crossref_metadata_2025 huggingface dataset:
Obesity and periodontal disease in diabetic pregnant women
Authors: Chapper Ana , Munch Artur , Schermann Camila , Piacentini Carolina Carraro , Fasolo Maria Thereza Martins
Published: January 2006

This cross-sectional study investigated the impact of pregestational overweight and obesity on periodontal status of patients with gestational diabetes mellitus (GDM). Sixty pregnant women with gestational diabetes mellitus (GDM) were recruited for the study. According to the pregestational body mass index (BMI), patients were classified into 3 groups: normal, overweight or obese. The periodontal assessment parameters were the presence of gingival bleeding (GB) and bleeding on probing (BOP) per tooth. Clinical attachment loss (CAL) was assessed per tooth and classified according to following values: 1) absence of attachment loss; 2) between 1 and 2 mm, 3) between 3 and 5 mm; and 4) CAL > 6 mm. The means of individual percentage of teeth with GB and BOP and the means of the individual classified values of CAL were compared through ANOVA. Differences between the groups were established through post hoc Bonferroni test for multiple comparisons (p < 0.05). The analysis revealed significant differences between the normal group and the obese group considering GB (52.76% ± 27.99% and 78.85% ± 27.44%, respectively) and CAL (2.21 ± 0.41 and 2.61 ± 0.54, respectively). Although an increase was found in BOP as the BMI increased (ranging from 55.65% to 75.31%), no statistically significant differences were found among the groups. Patients with GDM and pregestational obesity had significantly more gingivitis and periodontal attachment loss that those with normal pregestational BMI. Periodontal treatment should be considered in the establishment of future recommendations for metabolic control for this special group of patients.
Blood cell attachment to root surfaces treated with EDTA gel
Authors: Leite Fábio Renato Manzolli , Moreira Camila Sala , Theodoro Letícia Helena , Sampaio José Eduardo Cezar
Published: January 2006

Root debridement generates a smear layer which contains microorganisms and toxins that could interfere in periodontal healing. For this reason, different substances have been used to remove it and to expose collagen fibers at the tooth surface. Blood element adhesion to demineralized roots and clot stabilization by collagen fibers are extremely important for the success of periodontal surgery. The aim of this study was to evaluate the different patterns of blood element adsorption and adhesion to root surfaces only irrigated with distilled water and after application of a manipulated or an industrialized EDTA gel. Thirty samples were planed, equally divided into three groups and treated with distilled water (control), a manipulated EDTA gel or an industrialized one. Immediately after, samples were exposed to fresh blood and prepared for scanning electron microscopy. Untreated planed dentin presented the best results with blood cells entrapped in a thick web of fibrin. In the manipulated EDTA group, the web of fibrin was thick with sparse blood elements. The worst result was seen with the industrialized EDTA group, in which no blood elements could be seen. Statistical difference was obtained between control and industrialized EDTA groups. Surfaces only irrigated presented the most organized fibrin network and cell entrapment.
Study of the inflammatory process induced by injection of carrageenan or formalin in the rat temporomandibular joint
Authors: Goulart Alan Cruvinel , Correia Francisco Antônio dos Santos , Sousa Suzana Cantanhede Orsini Machado de , Luz João Gualberto de Cerqueira
Published: January 2006

The aim of this study was to evaluate the effects of the injection of two phlogistic agents, carrageenan and formalin, in the rat TMJ, and the inflammatory process induced by these substances. In this study, a total of 45 adult rats were distributed in two experimental groups and a control group. The animals were sacrificed after three hours, 24 hours, three days, seven days, and 15 days after a single injection of each substance. Histological data initially demonstrated an inflammatory process represented by acute infiltration, which later became mixed, and finally chronic in both experimental groups. Hyperplasia of the synovial membrane was observed after three days, being intense at seven days, and present after 15 days only in the formalin group. Local saline injection in the control group caused no inflammatory reaction. It was concluded that a single local injection of carrageenan or formalin was enough to induce inflammatory reaction in the TMJ and periarticular soft tissues, and that the resulting processes were similar, but more persistent in the formalin group.
Characterization of titanium surfaces for dental implants with inorganic contaminant
Authors: Diniz Marilia Garcia , Pinheiro Marco Antonio Santos , Andrade Junior Antonio Carlos Canabarro , Fischer Ricardo Guimarães
Published: January 2006

The aim of this research was to characterize titanium surfaces blasted with aluminum oxide (Al2O3) particles using the KS 400 digital image processing program. Samples of grade II titanium plates were submitted to blasting processes using particles of Al2O3, and treated with a hydrofluoric acid-based solution. Three digital images from each sample surface were obtained using Scanning Electron Microscopy, and half-quantitative chemical analyses were subsequently performed using Electron Dispersive Spectroscopy (EDS). In addition, parameters related to the alumina phase, such as the concentration level, the area and perimeter of the particles and their circular form factor were measured using KS 400. The mechanical/chemical treatment caused depressions up to 10 µm on homogeneous surfaces. Although the chemical attack significantly removed the alumina phase, residual particles could still be identified by the EDS. The average area occupied by the alumina phase on the samples surfaces was 469.32 (± 284.98) µm², the particle average perimeter was 81.61 (± 27.68) µm, and the mean circular form factor was 0.60 (± 0.05). Characterizing the titanium surface is essential in the evaluation of the material manufacturing process because the presence of residual aluminum particles may have deleterious effects on the formation of the osseous/implant tissue.
Effects of long-term cyclosporin therapy on gingiva of rats: analysis by stereological and biochemical estimation
Authors: Spolidorio Luis Carlos , Spolidorio Denise Madalena Palomari , Holzhausen Marinella , Nassar Patricia Oehlmeyer , Nassar Carlos Augusto
Published: January 2006

Cyclosporin A (CsA) is used as an immunosuppressive agent and its prominent side effect is the induction of gingival overgrowth, which remains a significant problem. The risk factors appraised include the duration of treatment. However, there are no stereological and biochemical studies exploring the effects of long-term CsA therapy on gingival tissue. The purpose of the present study was to investigate the level of TGF-beta1 in saliva and describe the densities of fibroblasts and collagen fibers in the gingival tissue of rats treated with CsA for long periods. Rats were treated for 60, 120, 180 and 240 days with a daily subcutaneous injection of 10 mg/kg of body weight of CsA. At the end of the experimental periods, saliva was collected for the determination of TGF-beta1 levels. After histological processing, the oral epithelium and the connective tissue area were measured as well as the volume densities of fibroblasts (Vf) and collagen fibers (Vcf). After 60 and 120 days of CsA treatment, there was a significant increase in Vf and Vcf as well as a significant increase in TGF-beta1 levels. After 180 and 240 days, reduction in the gingival overgrowth associated with significant decreases in the level of TGF-beta1, and also decreased Vf and Vcf, were observed. The data presented here suggest that after long-term therapy, a decrease in TGF-beta1 levels occurs, which might contribute to an increase in the proteolytic activity of fibroblasts in the gingiva, favoring the normality of extracellular matrix synthesis.
Comparison among dental, skeletal and chronological development in HIV-positive children: a radiographic study
Authors: Holderbaum Rejane Maria , Veeck Elaine Bauer , Oliveira Helena Willhelm , Silva Carmem Lúcia , Fernandes Ângela
Published: January 2006

The goal of this study was to evaluate skeletal, dental and chronological development in an HIV-positive group of children, as compared with a control group, during a four-year period. Panoramic radiographs and hand and wrist radiographs of 60 children were taken. The children, of both sexes, aged 5 years and 2 months to 15 years and 5 months, were selected as follows: 30 HIV-positive volunteers who had acquired the disease vertically, and 30 volunteers who did not present the HIV infection or any other systemic disease. All radiographs were technically standardized and analyzed according to criteria established by Nolla (dental age), Greulich and Pyle (bone age), and Eklöf and Ringertz (bone age). The results were submitted to Student's t-test at a 5% level of significance. Based on the comparison between the chronological age and the dental or the skeletal age, significant differences were observed between HIV-positive and HIV-negative children, both in 1999 and in 2003 (p < 0.05). Considering the results obtained with the methodology used, it was concluded that HIV-positive children of both sexes presented delayed bone development despite the administration of antiretroviral drugs, and that HIV-positive female children presented younger dental ages compared with their chronological ages in 1999 and in 2003; and HIV-positive males, in 1999.
Assessment of methyl methacrylate vapor toxicity on the rat tracheal epithelium
Authors: Parizi José Luiz Santos , Nai Gisele Alborghetti , Batalha Camila Fedatto , Lopes Carla Cristina Barbosa , Rizzo Mariana Fernandez , Falcone Ciro Eduardo , Bertão José Maria
Published: January 2006

Methyl methacrylate (MMA) is a monomer that is polymerized into resin by light and heat, producing a clear, resistant, durable and relatively inert plastic material. Because of these characteristics, MMA is largely used in Medicine as bone cement and in Dentistry, in dental braces and prostheses, thus generating continuous interest in its toxicity. Experimental and clinical studies have documented that monomers may cause a wide range of adverse health effects. The most important occupational exposure route of MMA is by inhalation. This study aims to evaluate the toxicity of MMA to the tracheal epithelium, according to the time of exposure. For this purpose, two experimental groups of rats were exposed to MMA by inhalation under poor ventilation: one group (n = 36) was exposed permanently, and the other (n = 36) was exposed during 8 hours per day, without water and food supply during the exposure period. A control group (n = 8) received normal air supply. Twelve animals of each study group were sacrificed after 5, 8 and 10 days of exposure together with two or four control animals. Twenty-nine (80.5%) of the rats continuously exposed to MMA developed inflammation on the tracheal epithelium, as well as 58.33% (n = 21) of those exposed 8 h/day and 87.5% (n = 7) of the control rats. No association was observed between the inflammatory process and MMA exposure; no significant alterations in the tracheal epithelium thickness were observed. Further studies on longer exposure times and analysis of other parameters will have to be conducted to exclude the possibility of tracheal damage by vapors of MMA.
A Practical Update on the Use of Bortezomib in the Management of Multiple Myeloma
Authors: San Miguel Jesús , Bladé Joan , Boccadoro Mario , Cavenagh Jamie , Glasmacher Axel , Jagannath Sundar , Lonial Sagar , Orlowski Robert Z. , Sonneveld Pieter , Ludwig Heinz
Published: January 2006

Learning Objectives

After completing this course, the reader will be able to: Discuss clinical trial data for treatment with bortezomib monotherapy and bortezomib-based combination therapy in patients with multiple myeloma who have received at least one prior therapy.Discuss the management of the most common adverse events associated with bortezomib, including peripheral neuropathy and thrombocytopenia.Describe prognostic factors in patients with multiple myeloma who have received at least one prior therapy.

Access and take the CME test online and receive 1 AMA PRA category 1 credit at CME.TheOncologist.com
The Molecular Perspective: Tumor Necrosis Factor
Authors: Goodsell David S.
Published: January 2006

Learning Objective

After completing this course, the reader will be able to: Discuss tumor necrosis factor and its role in cancer chemotherapy.

Access and take the CME test online and receive 1 AMA PRA category 1 credit at CME.TheOncologist.com
Management of Gastrointestinal Stromal Tumors in the Imatinib Era: Selected Case Studies
Authors: Benjamin Robert S. , Blanke Charles D. , Blay Jean-Yves , Bonvalot Sylvie , Eisenberg Burton
Published: January 2006

The introduction of imatinib, an orally administered inhibitor of the KIT receptor tyrosine kinase, is prompting revision of the management algorithms that have traditionally guided the treatment of gastrointestinal stromal tumor (GIST). Historically, patients with GISTs have had substantial rates of relapse as well as limited long-term survival even after complete surgical resection of a primary tumor. Imatinib has been shown to induce durable tumor responses in more than half of the patients with malignant metastatic or unresectable GISTs and to halt disease progression in an additional third. These encouraging results have led to the initiation of clinical trials of imatinib as an adjuvant or neoadjuvant therapy with surgery. Until relevant data are reported to provide definitive direction for the management of operable or potentially operable GISTs, treatment decisions must be made on the basis of the available evidence and clinical experience with imatinib. This paper presents selected case studies describing approaches to the combined use of surgery and systemic therapy that have been applied in the treatment of individual GIST patients. The management of GIST in these cases required a coordinated, multidisciplinary approach involving medical oncologists, diagnostic radiologists, gastroenterologists, surgeons, and pathologists.
Why Is the Pediatrician Asking Me About Social Drivers of Health?
Authors: Palakshappa Deepak , Thompson Lindsay A.
Published: February 2024

This JAMA Pediatrics Patient Page describes why pediatric clinics ask families about social drivers of health.
E2F1 promotes cell migration in hepatocellular carcinoma via FNDC3B
Authors: Hua Kate , Wu Chen‐Tang , Lin Chin‐Hui , Chen Chian‐Feng
Published: February 2024

FNDC3B (fibronectin type III domain containing 3B) is highly expressed in hepatocellular carcinoma (HCC) and other cancer types, and fusion genes involving FNDC3B have been identified in HCC and leukemia. Growing evidence suggests the significance of FNDC3B in tumorigenesis, particularly in cell migration and tumor metastasis. However, its regulatory mechanisms remain elusive. In this study, we employed bioinformatic, gene regulation, and protein‐DNA interaction screening to investigate the transcription factors (TFs) involved in regulating FNDC3B. Initially, 338 candidate TFs were selected based on previous chromatin immunoprecipitation (ChIP)‐seq experiments available in public domain databases. Through TF knockdown screening and ChIP coupled with Droplet Digital PCR assays, we identified that E2F1 (E2F transcription factor 1) is crucial for the activation of FNDC3B. Overexpression or knockdown of E2F1 significantly impacts the expression of FNDC3B. In conclusion, our study elucidated the mechanistic link between FNDC3B and E2F1. These findings contribute to a better understanding of FNDC3B in tumorigenesis and provide insights into potential therapeutic targets for cancer treatment.
Unravelling the Factors Influencing Halide Perovskite Based Switchable Photovoltaics
Authors: Xing Xuechao , Ng Si En , Tay Yeow Boon , Yantara Natalia , Lew Wen Siang , Mathews Nripan
Published: February 2024

Lead halide perovskites have revolutionized the field of optoelectronics (such as photovoltaics and light emitting diodes) demonstrating extraordinary material properties despite being formed at low temperatures. However, ion migration in the bulk or at the interfaces results in stability issues especially in devices where metal electrodes directly interface with the perovskite film. Utilizing the switchable photovoltaic phenomenon (SPV) in halide perovskites as a measure of ion migration and electrochemical reactions within them, Cs0.05MA0.15FA0.70PbI2.5Br0.5 triple cation perovskite, widely used in photovoltaics is evaluated. The various factors determining the SPV, including electric field magnitudes, type of metal contacts, Illumination conditions, and temperature is systematically measured. This study reveals the roles of electrode work functions and reactivities on ion migration and local electronic structure modulation. ITO electrodes demonstrated the highest open‐circuit voltage (Voc) about 0.85 V while Ag electrodes developed conductive filaments. However, the Voc distribution for Ti and Cr electrodes shows a more pronounced linear correlation with the poling electric field strength. Insights from this lateral design are directly relevant to transistor and memristor architectures and offer inputs into the design of perovskite‐based photovoltaic/optoelectronic devices.
Single‐Shot Laser‐Induced Switching of an Exchange Biased Antiferromagnet
Authors: Guo Zongxia , Wang Junlin , Malinowski Gregory , Zhang Boyu , Zhang Wei , Wang Hangtian , Lyu Chen , Peng Yi , Vallobra Pierre , Xu Yong , Xu Yongbing , Jenkins Sarah , Chantrell Roy W. , Evans Richard F. L. , Mangin Stéphane , Zhao Weisheng , Hehn Michel
Published: February 2024

Ultrafast manipulation of magnetic order has challenged the understanding of the fundamental and dynamic properties of magnetic materials. So far single‐shot magnetic switching has been limited to ferrimagnetic alloys, multilayers, and designed ferromagnetic (FM) heterostructures. In FM/antiferromagnetic (AFM) bilayers, exchange bias (He) arises from the interfacial exchange coupling between the two layers and reflects the microscopic orientation of the antiferromagnet. Here the possibility of single‐shot switching of the antiferromagnet (change of the sign and amplitude of He) with a single femtosecond laser pulse in IrMn/CoGd bilayers is demonstrated. The manipulation is demonstrated in a wide range of fluences for different layer thicknesses and compositions. Atomistic simulations predict ultrafast switching and recovery of the AFM magnetization on a timescale of 2 ps. The results provide the fastest and the most energy‐efficient method to set the exchange bias and pave the way to potential applications for ultrafast spintronic devices.
Photo‐Induced Electric Field Effects on Water Droplets Generated in a LiNbO3 Opto‐Microfluidic Platform
Authors: Bragato Giovanni , Zaltron Annamaria , Zanardi Michele , Zamboni Riccardo , De Ros Maddalena , Sada Cinzia
Published: February 2024

In this work, droplets elongation as a result of the photo‐induced photovoltaic electric field is presented and modeled. Moving water droplets are generated in microfluidics channels by means of a droplet‐based opto‐microfluidic platform integrated in lithium niobate (LN). A cross‐junction is engraved on the substrate as droplets generator, on top of which a z‐cut iron‐doped lithium niobate (Fe:LN) crystal is placed in order to build up photo‐induced electric field as a result of photovoltaic effect promoted upon suitable illumination. Droplets detection is realized by optical waveguides, designed in a Mach‐Zehnder Interferometer (MZI) configuration on the same substrate. The dynamics of droplet elongation is investigated, highlighting the impact of the photovoltaic electric field on the droplet dimension and surface tension.
Compositionally Sequenced Interfacial Layers for High‐Energy Li‐Metal Batteries
Authors: Lee Jeong‐A , Kim Saehun , Cho Yoonhan , Kweon Seong Hyeon , Kang Haneul , Byun Jeong Hwan , Kwon Eunji , Seo Samuel , Kim Wonkeun , Ryu Kyoung Han , Kwak Sang Kyu , Hong Seungbum , Choi Nam‐Soon
Published: February 2024

Electrolyte additives with multiple functions enable the interfacial engineering of Li‐metal batteries (LMBs). Owing to their unique reduction behavior, additives exhibit a high potential for electrode surface modification that increases the reversibility of Li‐metal anodes by enabling the development of a hierarchical solid electrolyte interphase (SEI). This study confirms that an adequately designed SEI facilitates the homogeneous supply of Li+, nonlocalized Li deposition, and low electrolyte degradation in LMBs while enduring the volume fluctuation of Li‐metal anodes on cycling. An in‐depth analysis of interfacial engineering mechanisms reveals that multilayered SEI structures comprising mechanically robust LiF‐rich species, electron‐rich P–O species, and elastic polymeric species enabled the stable charge and discharge of LMBs. The polymeric outer SEI layer in the as‐fabricated multilayered SEI could accommodate the volume fluctuation of Li‐metal anodes, significantly enhancing the cycling stability Li||LiNi0.8Co0.1Mn0.1O2 full cells with an electrolyte amount of 3.6 g Ah−1 and an areal capacity of 3.2 mAh cm−2. Therefore, this study confirms the ability of interfacial layers formed by electrolyte additives and fluorinated solvents to advance the performance of LMBs and can open new frontiers in the fabrication of high‐performance LMBs through electrolyte‐formulation engineering.
Sharing Perioperative Nursing Expertise Through Publication: A Guide for the Novice Author
Authors: Vortman Rebecca , Darbyshire Pamayla E. , Talatala Ray‐an
Published: February 2024

Perioperative nurses can share their expertise by writing for publication in a peer‐reviewed journal. Writing can help perioperative nurses grow their professional careers and advance the science of the perioperative nursing specialty. Despite the value and importance of publishing, perioperative nurses may lack confidence and fear rejection and negative feedback; increasing their knowledge and understanding of the authoring and publishing processes can assuage these fears. This education article describes concepts associated with scholarly publishing for authors and offers strategies to encourage perioperative nurses to share their practice experiences or research via peer‐reviewed journals. Key steps associated with the writing and publication process are described. The article also explains the editorial and peer‐review processes and provides supportive strategies for authors when a manuscript is not accepted initially.
Self‐assembly of thiourea intercalated graphene oxide based dual IIP for the SPE coupled FAAS method development of Cu(II) and Pb(II) determination
Authors: Islam Aminul , Rais Saman
Published: February 2024

A new dual‐template surface imprinted polymer for Cu(II) and Pb(II) was synthesized in one pot. Magnetic graphene oxide was self‐assembled with low cost and environmentally benign thiourea. Presence of sulfur and nitrogen donor atoms provide hooks for coordination and partial reduction of graphene oxide matrix. It was used as an solid‐phase extraction adsorbent for extraction, preconcentration, and coupled with flame atomic absorption spectrometry to manifest performance comparable with inductively coupled plasma atomic emission spectrometry (ICPAES) both in terms of quantification limit as well as interference. The critical experimental parameters such as pH; 4.6, contact time of 15 min and initial concentration of 777 (Qe; 227 mg g−1) and 800 μg L−1 (Qe; 273 mg g−1) for Cu(II) and Pb(II), respectively, were optimized using RSM‐CCD and artificial neural network. The adsorption process was kinetically faster (50% adsorption in 5 min), following fractal‐like‐pseudo‐second‐order (FLPSO) kinetics and Brouers–Sotolongo isotherm model owing to the heterogenous energy landscape. The imprinting factors were in the range of 4–7 in the presence of all coexisting ions. The proposed method was robust in the determination and removal of Cu(II) and Pb(II) from food, ground water, and industry effluents with low limit of detection (Cu(II); 1.03 μg L−1 & Pb(II); 1.79 μgL−1). Spiking and recovery tests were used to assess the method's accuracy. Cu(II)/Pb(II) loaded dual template IIP (DIIP) was utilized to remove anionic dyes with >95% efficiency. Thorough examination of the method and material selectivity (in binary, ternary, and multielement system), multi fold applications of determination, removal of Cu(II), Pb(II), and removal of anionic dyes makes DIIP a promising candidate for environmental remediation.
Marine protection enhances the resilience of biological communities on temperate rocky reefs
Authors: Sanabria‐Fernández José A. , Alday Josu G.
Published: February 2024

Conservation science faces the urgent challenge of halting the biodiversity loss caused by the biological crisis of the present era. To achieve this, conservation science requires cutting‐edge tools to focus on vital properties of ecosystems, such as the resilience. Resilience informs about the cost of recovering biological communities.

Here, we developed a metric to quantify the ecological assemblage recovery cost based on the dissimilarity between unprotected and partially protected communities compared with totally protected communities in Cabo de Gata Marine Reserve.

Our results show that the biological assemblage composed of fish, macroinvertebrates and cryptic fish, and macroalgae species in unprotected zones requires a higher ecological recovery cost than in partially protected zones when moving towards a fully protected community.

This research contributes to monitoring marine the effectiveness of marine protection from a resilience perspective, with the goal of promoting the use of the recovery cost metric for building resilient coastal ecosystems.
CRISPR/Cas system: A revolutionary tool for crop improvement
Authors: Mishra Ayushi , Pandey Veda P.
Published: February 2024

World's population is elevating at an alarming rate thus, the rising demands of producing crops with better adaptability to biotic and abiotic stresses, superior nutritional as well as morphological qualities, and generation of high‐yielding varieties have led to encourage the development of new plant breeding technologies. The availability and easy accessibility of genome sequences for a number of crop plants as well as the development of various genome editing technologies such as zinc finger nucleases (ZFNs), transcription activator‐like effector nucleases (TALENs) has opened up possibilities to develop new varieties of crop plants with superior desirable traits. However, these approaches has limitation of being more expensive as well as having complex steps and time‐consuming. The CRISPR/Cas genome editing system has been intensively studied for allowing versatile target‐specific modifications of crop genome that fruitfully aid in the generation of novel varieties. It is an advanced and promising technology with the potential to meet hunger needs and contribute to food production for the ever‐growing human population. This review summarizes the usage of novel CRISPR/Cas genome editing tool for targeted crop improvement in stress resistance, yield, quality and nutritional traits in the desired crop plants.
Identification of critical process parameters for expansion of clinical grade human Wharton's jelly‐derived mesenchymal stromal cells in stirred‐tank bioreactors
Authors: López‐Fernández Alba , Garcia‐Gragera Víctor , Lecina Martí , Vives Joaquim
Published: February 2024

Cell therapies based on multipotent mesenchymal stromal cells (MSCs) are traditionally produced using 2D culture systems and platelet lysate‐ or serum‐containing media (SCM). Although cost‐effective for single‐dose autologous treatments, this approach is not suitable for larger scale manufacturing (e.g., multiple‐dose autologous or allogeneic therapies with banked MSCs); automated, scalable and Good Manufacturing Practices (GMP)‐compliant platforms are urgently needed. The feasibility of transitioning was evaluated from an established Wharton's jelly MSCs (WJ‐MSCs) 2D production strategy to a new one with stirred‐tank bioreactors (STRs). Experimental conditions included four GMP‐compliant xeno‐ and serum‐free media (XSFM) screened in 2D conditions and two GMP‐grade microcarriers assessed in 0.25 L‐STRs using SCM. From the screening, a XSFM was selected and compared against SCM using the best‐performing microcarrier. It was observed that SCM outperformed the 2D‐selected medium in STRs, reinforcing the importance of 2D‐to‐3D transition studies before translation into clinical production settings. It was also found that attachment efficiency and microcarrier colonization were essential to attain higher fold expansions, and were therefore defined as critical process parameters. Nevertheless, WJ‐MSCs were readily expanded in STRs with both media, preserving critical quality attributes in terms of identity, viability and differentiation potency, and yielding up to 1.47 × 109 cells in a real‐scale 2.4‐L batch.
Engineering and transcriptome study of Saccharomyces cerevisiae to produce ginsenoside compound K by glycerol
Authors: Zhang Chuanbo , Tian Jinping , Zhang Jiale , Liu Ruixia , Zhao Xiaomeng , Lu Wenyu
Published: February 2024

Synthetic biology‐based engineering of Saccharomyces cerevisiae to produce terpenoid natural products is an effective strategy for their industrial application. Previously, we observed that glycerol addition was beneficial for ginsenoside compound K (CK) production in a S. cerevisiae when it was fermented using the YPD medium. Here, we reconstructed the CK synthesis and glycerol catabolic pathway in a high‐yield protopanaxadiol (PPD) S. cerevisiae strain. Remarkably, our engineered strain exhibited the ability to utilize glycerol as the sole carbon source, resulting in a significantly enhanced production of 433.1 ± 8.3 mg L−1 of CK, which was 2.4 times higher compared to that obtained in glucose medium. Transcriptomic analysis revealed that the transcript levels of several key genes involved in the mevalonate (MVA) pathway and the uridine diphosphate glucose (UDPG) synthesis pathway were up‐regulated in response to glycerol. The addition of glycerol enhanced CK titers by augmenting the flux of the terpene synthesis pathway and facilitating the production of glycosyl donors. These results suggest that glycerol is a promising carbon source in S. cerevisiae, especially for the production of triterpenoid saponins.
Secretory production and characterization of a highly effective chitosanase from Streptomyces coelicolor A3(2) M145 in Pichia pastoris
Authors: Liang Jiayu , He Shengbin , Sun Jian , Bao Haodong , Cui Lanyu
Published: February 2024

In this study, a glycoside hydrolase family 46 chitosanase from Streptomyces coelicolor A3(2) M145 was firstly cloned and expressed in Pichia pastoris GS115 (P. pastoris GS115). The recombinant enzyme (CsnA) showed maximal activity at pH 6.0 and 65°C. Both thermal stability and pH stability of CsnA expressed in P. pastoris GS115 were significantly increased compared with homologous expression in Streptomyces coelicolor A3(2). A stable chitosanase activity of 725.7 ± 9.58 U mL−1 was obtained in fed‐batch fermentation. It's the highest level of CsnA from Streptomyces coelicolor expressed in P. pastoris so far. The hydrolytic process of CsnA showed a time‐dependent manner. Chitosan oligosaccharides (COSs) generated by CsnA showed antifungal activity against Fusarium oxysporum sp. cucumerinum (F. oxysporum sp. cucumerinum). The secreted expression and hydrolytic performance make the enzyme a desirable biocatalyst for industrial controllable production of chitooligosaccharides with specific degree of polymerization, which have potential to control fungi that cause important crop diseases.
A highly sensitive and specific Homo1‐based real‐time qPCR method for quantification of human umbilical cord mesenchymal stem cells in rats
Authors: He Jing , Wang Zhangfan , Ao Chunchun , Tu Chengshu , Zhang Yaqi , Chang Cheng , Xiao Cuihong , Xiang E , Rao Wei , Li Changyong , Wu Dongcheng
Published: February 2024

Background

Owing to the characteristics of easier access in vitro, low immunogenicity, and high plasticity, human umbilical cord‐derived mesenchymal stem cells (UC‐MSCs) are considered as a promising cell‐based drugs for clinical application. No internationally recognized technology exists to evaluate the pharmacokinetics and distribution of cell‐based drugs in vivo.

Methods

We determined the human‐specific gene sequence, Homo1, from differential fragments Homo sapiens mitochondrion and Rattus norvegicus mitochondrion. The expression of Homo1 was utilized to determine the distribution of UC‐MSCs in the normal and diabetic nephropathy (DN) rats.

Results

We observed a significant correlation between the number of UC‐MSCs and the expression level of Homo1. Following intravenous transplantation, the blood levels of UC‐MSCs peaked at 30 min. A large amount of intravenously injected MSCs were trapped in the lungs, but the number of them decreased rapidly after 24 h. Additionally, the distribution of UC‐MSCs in the kidneys of DN rats was significantly higher than that of normal rats.

Conclusions

In this study, we establish a highly sensitive and specific Homo1‐based real‐time quantitative PCR method to quantify the distribution of human UC‐MSCs in rats. The method provides guidelines for the safety research of cells in preclinical stages.
In vitro evolution of diagnostic antibodies targeting native antigens in plasma by sandwich flow cytometry
Authors: Liang Mingxia , Ran Fanlei , Li Li , Hang Haiying , An Lili
Published: February 2024

Monoclonal antibodies (mAbs) that recognize and bind to specific antigens (Ags) have a wide range of applications in research, therapy, and diagnostics. However, many of these antibodies cannot bind well to the native Ags. In this study, based on the Chinese hamster ovary (CHO) cell display platform developed previously in our lab, we reported a novel artificial evolution procedure to improve the affinity of mAb against the native Ag directly using the plasma samples without purification of the native Ag. In this procedure, a pair of antibodies able to bind the Ag in sandwich manner are first confirmed (Ab1/Ab2) and the antibody (Ab) to be affinity‐improved (Ab1) is displayed on CHO cells for Ab mutation. Then the cells were detected and sorted with flow cytometry in the form of Ab1‐Ag‐fluorescence labeled Ab2, which we named sandwich flow cytometry. Here, we used soluble isoform of suppression of tumorigenicity 2 (sST2) protein as model Ag, carried out “sandwich” maturation directly using the plasma samples containing the native sST2 protein and optimized a pair of antibodies with significantly improved sensitivity in the detection of the native sST2 in plasma. This method could be very useful in optimization of the diagnostic Ab pairs working in a “sandwich” manner if more antibodies were also successfully affinity‐matured with this method.
Exploring metabolic responses and pathway changes in CHO‐K1 cells under varied aeration conditions and copper supplementations using 1H NMR‐based metabolomics
Authors: Shi Yingting , Wan Yuxiang , Sun Yan , Yang Jiayu , Lu Yuting , Xie Xinyuan , Pan Jianyang , Wang Haibin , Qu Haibin
Published: February 2024

The optimization of bioprocess for CHO cell culture involves careful consideration of factors such as nutrient consumption, metabolic byproduct accumulation, cell growth, and monoclonal antibody (mAb) production. Valuable insights can be obtained by understanding cellular physiology to ensure robust and efficient bioprocess. This study aims to improve our understanding of the CHO‐K1 cell metabolism using 1H NMR‐based metabolomics. Initially, the variations in culture performance and metabolic profiles under varied aeration conditions and copper supplementations were thoroughly examined. Furthermore, a comprehensive metabolic pathway analysis was performed to assess the impact of these conditions on the implicated pathways. The results revealed substantial alterations in the pyruvate metabolism, histidine metabolism, as well as phenylalanine, tyrosine and tryptophan biosynthesis, which were especially evident in cultures subjected to copper deficiency conditions. Conclusively, significant metabolites governing cell growth and mAb titer were identified through orthogonal partial least square‐discriminant analysis (OPLS‐DA). Metabolites, including glycerol, alanine, formate, glutamate, phenylalanine, and valine, exhibited strong associations with distinct cell growth phases. Additionally, glycerol, acetate, lactate, formate, glycine, histidine, and aspartate emerged as metabolites influencing cell productivity. This study demonstrates the potential of employing 1H NMR‐based metabolomics technology in bioprocess research. It provides valuable guidance for feed medium development, feeding strategy design, bioprocess parameter adjustments, and ultimately the enhancement of cell proliferation and mAb yield.
Pipette‐free field‐deployable molecular diagnostic kit for bimodal visual detection of infectious RNA viruses
Authors: Park So Yeon , Trinh Kieu The Loan , Song Yoon‐Jae , Lee Nae Yoon
Published: February 2024

Here, we developed a field‐deployable molecular diagnostic kit for the detection of RNA viruses that operates in a pipette‐free manner. The kit is composed of acrylic sticks, PCR tubes, and palm‐sized three‐dimensional(3D)‐printed heaters operated by batteries. The kit performs RNA extraction, reverse transcriptase loop‐mediated isothermal amplification (RT‐LAMP), and visual detection in one kit. An acrylic stick was engraved with one shallow and one deep cylindrical chamber at each end for the insertion of an FTA card and ethidium homodimer‐1 (EthD‐1), respectively, to perform RNA extraction/purification and bimodal visual detection of the target amplicons. First, an intercalation of EthD‐1 into the target DNA initially produces fluorescence upon UV illumination. Next, the addition of a strong oxidant, in this case sodium (meta) periodate (NaIO4), produces intense aggregates in the presence of EthD‐1‐intercalated DNA, realized by electrostatic interaction. In the absence of the target amplicon, no fluorescence or aggregates are observed. Using this kit, two major infectious viruses—severe fever with thrombocytopenia syndrome virus (SFTSV) and severe acute respiratory syndrome coronavirus (SARS‐CoV‐2)—were successfully detected in 1 h, and the limits of detection (LOD) were approximately 1 virus μL−1 for SFTSV and 103 copies μL−1 for SARS‐CoV‐2 RNA. The introduced kit is portable, end‐user‐friendly, and can be operated in a pipette‐free manner, paving the way for simple and convenient virus detection in resource‐limited settings.
Initial cyclic‐di‐GMP upregulation triggers sporadic cellular expansion leading to improved cellular survival
Authors: Yang Yongshuai , Guo Siyu , Hong Can‐Jian , Liang Zhao‐Xun , Ho Chun Loong
Published: February 2024

Bacterial second messenger c‐di‐GMP upregulation is associated with the transition from planktonic to sessile microbial lifestyle, inhibiting cellular motility, and virulence. However, in‐depth elucidation of the cellular processes resulting from c‐di‐GMP upregulation has not been fully explored. Here, we report the role of upregulated cellular c‐di‐GMP in promoting planktonic cell growth of Escherichia coli K12 and Pseudomonas aeruginosa PAO1. We found a rapid expansion of cellular growth during initial cellular c‐di‐GMP upregulation, resulting in a larger planktonic bacterial population. The initial increase in c‐di‐GMP levels promotes bacterial swarming motility during the growth phase, which is subsequently inhibited by the continuous increase of c‐di‐GMP, and ultimately facilitates the formation of biofilms. We demonstrated that c‐di‐GMP upregulation triggers key bacterial genes linked to bacterial growth, swarming motility, and biofilm formation. These genes are mainly controlled by the master regulatory genes csgD and csrA. This study provides us a glimpse of the bacterial behavior of evading potential threats through adapting lifestyle changes via c‐di‐GMP regulation.
Synergistic application of atmospheric and room temperature plasma mutagenesis and adaptive laboratory evolution improves the tolerance of Escherichia coli to L‐cysteine
Authors: Yang Hui , Zhang Bo , Wu Zidan , Pan Jiayuan , Chen Lifeng , Xiu Xiaoling , Cai Xue , Liu Zhiqiang , Zheng Yuguo
Published: February 2024

L‐Cysteine production through fermentation stands as a promising technology. However, excessive accumulation of L‐cysteine poses a challenge due to the potential to inflict damage on cellular DNA. In this study, we employed a synergistic approach encompassing atmospheric and room temperature plasma mutagenesis (ARTP) and adaptive laboratory evolution (ALE) to improve L‐cysteine tolerance in Escherichia coli. ARTP‐treated populations obtained substantial enhancement in L‐cysteine tolerance by ALE. Whole‐genome sequencing, transcription analysis, and reverse engineering, revealed the pivotal role of an effective export mechanism mediated by gene eamB in augmenting L‐cysteine resistance. The isolated tolerant strain, 60AP03/pTrc‐cysEf, achieved a 2.2‐fold increase in L‐cysteine titer by overexpressing the critical gene cysEf during batch fermentation, underscoring its enormous potential for L‐cysteine production. The production evaluations, supplemented with L‐serine, further demonstrated the stability and superiority of tolerant strains in L‐cysteine production. Overall, our work highlighted the substantial impact of the combined ARTP and ALE strategy in increasing the tolerance of E. coli to L‐cysteine, providing valuable insights into improving L‐cysteine overproduction, and further emphasized the potential of biotechnology in industrial production.
NAT10‐mediated ac4C‐modified ANKZF1 promotes tumor progression and lymphangiogenesis in clear‐cell renal cell carcinoma by attenuating YWHAE‐driven cytoplasmic retention of YAP1
Authors: Miao Daojia , Shi Jian , Lv Qingyang , Tan Diaoyi , Zhao Chuanyi , Xiong Zhiyong , Zhang Xiaoping
Published: February 2024

Background

Lymphatic metastasis is one of the most common metastatic routes and indicates a poor prognosis in clear‐cell renal cell carcinoma (ccRCC). N‐acetyltransferase 10 (NAT10) is known to catalyze N4‐acetylcytidine (ac4C) modification of mRNA and participate in many cellular processes. However, its role in the lymphangiogenic process of ccRCC has not been reported. This study aimed to elucidate the role of NAT10 in ccRCC lymphangiogenesis, providing valuable insights into potential therapeutic targets for intervention.

Methods

ac4C modification and NAT10 expression levels in ccRCC were assessed using public databases and clinical samples. Functional investigations involved manipulating NAT10 expression in cellular and mouse models to study its role in ccRCC. Mechanistic insights were gained through a combination of RNA sequencing, mass spectrometry, co‐immunoprecipitation, RNA immunoprecipitation, immunofluorescence, and site‐specific mutation analyses.

Results

We found that ac4C modification and NAT10 expression levels increased in ccRCC. NAT10 promoted tumor progression and lymphangiogenesis of ccRCC by enhancing the nuclear import of Yes1‐associated transcriptional regulator (YAP1). Subsequently, we identified ankyrin repeat and zinc finger peptidyl tRNA hydrolase 1 (ANKZF1) as the functional target of NAT10, and its upregulation in ccRCC was caused by NAT10‐mediated ac4C modification. Mechanistic analyses demonstrated that ANKZF1 interacted with tyrosine 3‐monooxygenase/tryptophan 5‐monooxygenase activation protein epsilon (YWHAE) to competitively inhibit cytoplasmic retention of YAP1, leading to transcriptional activation of pro‐lymphangiogenic factors.

Conclusions

These results suggested a pro‐cancer role of NAT10‐mediated acetylation in ccRCC and identified the NAT10/ANKZF1/YAP1 axis as an under‐reported pathway involving tumor progression and lymphangiogenesis in ccRCC.
Maxillary first molar with two distobuccal root canals and cervical deformity: A case report
Authors: Cai Xue , Tang Rentao
Published: February 2024

Key Clinical Message

The second distobuccal canal in the maxillary first molar is often missed because of the low prevalence rate (0%–4%). The article reports this kind of variation in one case. Promising outcomes have continued up to the present (2‐year follow‐up).
Distinct distribution of subplate neuron subtypes between the sensory cortices during the early postnatal period
Authors: Chang Minzi , Nehs Sophia , Xu Zheng , Kanold Patrick O
Published: February 2024

Subplate neurons (SpNs) are a heterogeneous neuronal population actively involved in early cortical circuit formation. In rodents, many SpNs survive and form layer 6b. The molecular heterogeneity of SpNs raises the question of whether different subpopulations of SpNs survive through the early postnatal period similarly and whether such diverse SpN populations in the auditory cortex (ACtx) share a common distribution pattern with other sensory systems. To address that, we investigated the expression pattern of multiple specific SpN markers in the ACtx, as well as in the visual (VCtx) and somatosensory (SCtx) cortices as controls, using complexin 3 (Cplx3) antibodies and different SpN‐specific Cre‐driver mice, such as connective tissue growth factor (CTGF), dopamine receptor D1 (Drd1a), and neurexophilin 4 (Nxph4). We focused on two early time windows in auditory development: (1) during the second postnatal week (PNW) before ear‐canal opening and (2) during the third PNW after ear‐canal opening. We compared the expression pattern of different SpN markers in ACtx with VCtx and SCtx. At both examined timepoints, Cplx3 and Nxph4 expressing SpNs form the largest and smallest population in the ACtx, respectively. Similar distribution patterns are observable in the VCtx and SCtx during the second PNW but not during the third PNW, for a higher proportion of Drd1a expressing SpNs is detected in the VCtx and CTGF expressing SpNs in the SCtx. This study suggests that different populations of SpNs might contribute differently to the development of individual sensory circuits.
Supramolecular Synthesis of Star Polymers
Authors: Haino Takeharu , Nitta Natsumi
Published: February 2024

Supramolecular polymers, in which monomers are assembled via intermolecular interactions, have been extensively studied. The fusion of supramolecular polymers with conventional polymers has attracted the attention of many researchers. In this review article, the recent progress in the construction of supramolecular star polymers, including regular star polymers and miktoarm star polymers, is discussed. The initial sections briefly provide an overview of the conventional classification and synthesis methods for star polymers. Coordination‐driven self‐assembly was investigated for the supramolecular synthesis of star polymers. Star polymers with multiple polymer chains radiating from metal‐organic polyhedra (MOPs) have also been described. Particular focus has been placed on the synthesis of star polymers featuring supramolecular cores formed through hydrogen‐bonding‐directed self‐assembly. After describing the synthesis of star polymers based on host‐guest complexes, the construction of miktoarm star polymers based on the molecular recognition of coordination capsules is detailed.