bluuebunny/crossref_metadata_2025_split huggingface dataset:
Genetically Engineered Myeloid Cells Rebalance the Core Regulatory Immune Suppression Program in Metastasis
Authors: Kaczanowska Sabina , Beury Daniel W , Qin Haiying , Kaplan Rosandra 
Published: February 2023

Abstract
Tumor metastasis is a critical step in the progression of solid tumors that is associated with patient mortality, yet we have limited knowledge of the metastatic microenvironment to effectively target this process. The pre-metastatic niche is the tumor-promoting microenvironment that is established at distant sites in response to primary tumor growth before detectable metastatic disease. We characterized immune populations in the lungs of tumor-bearing mice by flow cytometry and RNA sequencing approaches. We identified an immune suppression gene signature in pre-metastatic niche formation that is found predominantly within myeloid cells. In addition to the increase of myeloid cells and immunosuppressive pathways, we discovered that T cell populations are reduced in pre-metastatic lungs. We hypothesized that reversing this immunosuppressive environment would restore T cell function and antitumor immunity. We designed a novel approach in which we generated Genetically-Engineered Myeloid cells (GEMys) to deliver IL-12, a potent antitumor molecule, into the pre-metastatic microenvironment. We evaluated the lungs by flow cytometry and observed that IL12-GEMy-treated mice had increased numbers of T cells and enhanced expression of activation markers, resulting in reduced metastasis and increased survival. When combined with chemotherapy pre-conditioning, IL12-GEMys cured mice of established tumors and generated long-lived T cell memory, as these mice were immune to subsequent tumor challenge. These studies demonstrate that IL12-GEMys can functionally modulate the core program of immune suppression in the pre-metastatic niche to successfully rebalance the dysregulated metastatic microenvironment in cancer.
Induction of hyperresponsivity to LPS by Porphyromonas gingivalis in gingival epithelium
Authors: Gasiorek Anna , Dobosz Ewelina , Ciaston Izabela , Potempa Jan , Koziel Joanna 
Published: February 2023

Abstract
Chronic periodontitis (CP) is an inflammatory disease of the tooth supporting tissue developed in response to formation of the dysbiotic biofilm on the subgingival tooth surface. Although exacerbated inflammation leads to the alveolar bone destruction and may cause tooth loss the molecular basis of CP initiation and progression remains elusive. The control over the inflammatory reaction and homeostasis can be efficiently restored by the negative regulators of TLRs signaling pathways, such as MCPIP-1, which is constitutively expressed in gingival keratinocytes and prevents the hyporesponsiveness in the gum. Here we found that inflammophilic periodontal species influence stability of MCPIP-1 leading to aggravated response of the epithelium to proinflammatory stimulation. We identified that among enzymes secreted by those pathogenic species, gingipains – cysteine proteases from Porphyromonas gingivalis play a crucial role. Their proteolytic activity leads to a rapid degradation of MCPIP-1 exacerbating inflammatory response induced by endotoxin. Taken together, we showed the novel mechanism of corruption of inflammatory signaling by periodontal pathogens indicating new possibilities for treatment of this chronic disease.
CARD8 is an inflammasome sensor for HIV-1 protease activity
Authors: Shan Liang , Wang Qiankun 
Published: February 2023

Abstract
Human immunodeficiency virus type 1 (HIV-1) has high mutation rates and exists as mutant swarms within the host. The adaptive immune system recognizes cognate epitopes based on amino acid sequences and associated conformations that are subject to mutation. The within-host diversity of HIV-1 allows rapid selection of antibody and T cell escape variants, leading to viral persistence. Approaches to target immutable components are needed to clear HIV-1 infection. Here, we report that the CARD8 inflammasome senses HIV-1 protease activity. HIV-1 can evade CARD8 sensing because its protease remains inactive in infected cells prior to viral budding. Induction of premature intracellular activation of the viral protease triggers CARD8 inflammasome-mediated pyroptosis of HIV-1-infected cells. The viral protease activity against CARD8 is well conserved across major HIV-1 subtypes. This strategy led to the clearance of latent HIV-1 in patient CD4+ T cells after viral reactivation. Taken together, our study identifies CARD8 as an inflammasome sensor of HIV-1 that holds promise as a strategy for clearance of persistent HIV-1 infection.
Contribution of soluble ORF2 viral protein to viral replication and host immune response during acute Hepatitis E virus infection
Authors: Ralfs Philipp , Salinas Eduardo , Ambardekar Charuta , Bremer Bill , Blasczyk Heather , Zhu Jingting , Holland Brantley , Walker Christopher M , Feng Zongdi , Grakoui Arash 
Published: February 2023

Abstract
The hepatitis E virus (HEV) is a small, positive-stranded RNA virus that is a major cause of acute viral hepatitis globally. Acute HEV infection is typically asymptomatic and resolves within 8–10 weeks. HEV encodes 2 forms of capsid protein. A cytoplasmic form (ORF2c) is essential for virion structure. A secreted glycosylated form (ORF2s) accumulates at high titer in serum and can mask anti-ORF2 neutralizing antibodies. Here, we explored the contribution of ORF2s to HEV replicative fitness in vivo, and its role in generating anti-ORF2 antibodies (Abs). Rhesus Macaques (RM) were challenged by direct hepatic injection of infectious ORF2s+ and ORF2s− RNA. The replication of an HEV mutant lacking ORF2s expression was delayed by ~2 weeks when compared with wildtype virus and peak titers were nearly 10-fold lower for ORF2s−. No reversion of the 3 ORF2s silencing mutations was detected in the ORF2s− genomes, indicating genetic stability. The delay in replication and lower peak titer was unexpected as the viruses replicate similarly in cell culture. In addition, our data demonstrated that ORF2s has a significant and unexpected impact on generation of antibodies. Specifically, serum anti-ORF2 antibodies were only transiently detected in ORF2s− infected RM. As expected, anti-ORF2 titers were high and sustained in ORF2s+ infected RM. Furthermore, anti-ORF2 Ab response primed by ORF2s− infection differed in protection against reinfection when compared to ORF2s+. The ORF2s− challenged animals were re-infected upon second exposure to HEV infection. These findings indicate ORF2s may be dispensable for viral replication in vivo but is required for long-lived antibody response to mediate protection against re-exposure.
Isolation of Circulating Plasma Cells in Multiple Myeloma on a Novel Automated System
Authors: Sa Silin , Hsu Faye , Oaks Jean , Spiteri Liz , Williams Jesse , Yu Liping 
Published: February 2023

Abstract
Multiple myeloma is a cancer of disordered plasma cells. The disease poses significant challenges for clinical management where cytogenetic analysis, such as FISH (fluorescent in situ hybridization), was commonly used for diagnosis and prognosis. Beads based enrichment of plasma cells are widely used to enhance FISH sensitivity and specificity. However conventional beads-based methods often require time-consuming sample preparation and results in low cell recovery.
Here we present an easy and automated method to positively isolated CD138+ plasma cells from bone marrow samples. The method is developed on a novel designed magnetic separation system, MARS®. MARS® is matrix-free magnetic separation platform providing high purity, high recovery, and high capacity for cell isolation. Magnetic bead labeled CD138+ cells are isolated directly from bone marrow and peripheral blood without centrifugation. In the pilot study, U266 cells were spiked into peripheral blood from 0.5% to 2.0% of white blood cells frequency range. In addition, healthy donor bone marrow cell sample with 0.04% plasma cells were processed on MARS®. Plasma cells were isolated at 1mL/min flow rate followed by automated rinsing and releasing on MARS®. 48% to 80% recovery of the plasma cells were obtained. Results show our method has advantage in fast (&lt;30 minutes), easy process with high recovery. Our next objective will be to validate the method on Multiple Myeloma patient specimens and compared to conventional beads-based enrichment methods. Enriched plasma cells will be analyzed by FISH to prove the enhanced sensitivity with better enrichment method.
Frozen CyTOF panel enables robust and consistent longitudinal assessment of the SARS-CoV-2 immune landscape
Authors: Acosta-Vega Carolina , Skibinski David , Kus Anna , Speak Cate , Long Alice , Malhotra Uma , Buckner Jane H , Wiedeman Alice 
Published: February 2023

Abstract
During the Coronavirus Disease 2019 (COVID-19) outbreak, a robust and consistent strategy was required for assessing the immune landscape. Our rapid research response to COVID-19 was enabled by use of standardized assays. The assays included a 33-marker mass cytometry (CyTOF) panel that characterized major immune cells subsets in fresh whole blood including both innate and adaptive cells. Intraassay variation, determined by staining of the same fresh whole blood sample independently by two technicians on the same day, was CV&lt;20% for populations with a frequency &gt;5%. However, technical consistency was crucial to accurately capture and compare immune changes. Thus, we tested the stability of an antibody cocktail stored at −80C to limit inherent variability in daily panel preparation. Using lyophilized whole blood and cryopreserved R848-stimulated PBMC as controls encompassing the breadth of cell subsets and markers, we assessed the panel performance 7 times (at days 0, 16, 33, 42, 56, 72 and 96 after freezing). We found stability of staining over time: variability of the frozen cocktail was comparable to repeated cocktail preparations as seen in rare plasmablasts (CV&lt;25% frozen vs 13.1% repeat fresh), but also CD4 T cells (CV&lt;13% vs 10.6%), and neutrophils (CV&lt;4% vs 9.8%). Applying the highly-consistent frozen cocktail to study of 59 COVID-19 subjects revealed changes in common subsets (CD4 central memory T cells increased &gt;30% with severe disease), but also in rare plasmablasts (increased 6% of CD19), and more subtle neutrophil changes (increased 15% of CD45). Thus, freezing a high-content CyTOF panel cocktail provides a reliable method to control variability of staining in assays where sample collection is variable.
B-cell lymphoma 6 protein modulates function of human tonsillar innate lymphoid cells
Authors: Ohayon David E , Clay Gwendolyn M. , Forney Carmy , Seelamneni Harsha , Sestito Alexandra , Duggins Angela , Fitzpatrick Sarah , Guilliams Harmon , Kottyan Leah C. , Smith David F. , Miraldi Emily R. , Waggoner Stephen N 
Published: February 2023

Abstract
Innate lymphoid cells (ILC) play a fundamental role in mucosal homeostasis and immunity via expression of cytokines such as IL-22, IL-17A and IFN-γ. However, the transcriptional network that controls ILC functional identity is incompletely defined. Previously, we demonstrated that BCL6 plays a key role in transcriptional regulation in mouse intestinal ILC1 and ILC3. Here, we performed in vitro cultures of ILCs isolated from human tonsil in the presence of cytokines that promote ILC1 or ILC3 in order to assess the role of BCL6 in functional plasticity of ILCs. ILCs were treated with IL-2 and IL-12/IL-1β or IL-23/IL-1β in the presence or absence of the BCL6 inhibitor FX-1. In the context of IL-12/IL-1β ILC1-promoting culture, FX-1 inhibition of BCL6 reduced expression of Tbet and IFN-γ. In contrast, BCL6 inhibition in the context of IL-23/IL-1β ILC3-inducing conditions had no effect on Tbet, IFN-γ, or RORγt. In either context, FX-1 resulted in reduced IL-22 but increased IL-17A expression in comparison to vehicle treated cultures. Thus, our data emphasizes the role of BCL6 as a regulator of human tonsil ILC functional identity.
Study of coronavirus disease 2019 cases at the isolation department of el-dakhla general hospital
Authors: Tageldein Mohamed A. , Mohamed Mahmoud M. , Ibrahim Mai M. 
Published: February 2023

Background
Coronavirus disease 2019 (COVID-19) infection is currently the greatest atypical pneumonia outbreak, causing widespread anxiety and fear.

Objectives
To study cases with clinical manifestations that have been identified as COVID-19 according to the Egyptian Ministry of Health’s criteria, their management protocol, and outcome data at the isolation department of El-Dakhla General Hospital.

Patients and methods
This work was performed upon 106 patients with clinical symptoms of COVID-19 at the isolation department of El-Dakhla General Hospital.

Results
Mortality was associated with older age. No patients within the died group had been vaccinated. Vaccination was significantly associated with decreased severity of chest affection, admission to the ICU, and duration of admission. Mortality was associated with severity of chest affection. All died patients were admitted to the ICU. This study also revealed that abnormal vital and laboratory data were significantly associated with mortality. On using binary regression analysis of factors to be significantly associated with mortality, lower duration of admission and not being admitted to ICU decreased the risk of mortality among the studied patients, yet was statistically nonsignificant (P>0.05), whereas higher C-reactive protein on admission increased the risk by 1.186 folds.

Conclusion
Age, vaccination state, severity of chest affection, ICU admission and duration, use of some medications, vital data, and laboratory data are important factors to assess the prognosis and severity of COVID infection.
The effect of coronavirus disease 2019 severity on male sexual functions among patients on follow-up after hospital discharge
Authors: Mousa Mohamed S. , Elshazly Mostafa , El Salam Mohamed A.A. , Ahmed Salwa H. , Tantawy Ahmed A. 
Published: February 2023

Objectives
Severe acute respiratory syndrome coronavirus 2 has infected millions of people worldwide with extensive affection and damage to body systems and organs; hence, the study of post-coronavirus disease (COVID) sequences is mandatory. Till now, reports are upcoming on the considerable effects of COVID-19 on male sexual health with no final data.

Patients and methods
Our cohort study included 76 male COVID-19-infected patients, confirmed positive via nasopharyngeal PCR swab. The rationale of this study was to estimate the influence of clinical, laboratory, and radiological severity parameters of COVID-19 on male erectile dysfunction based on erectile scores and male sex hormones (follicle-stimulating hormone, luteinizing hormone, testosterone, and estradiol).

Results
Our results have demonstrated a highly statistically significant correlation between COVID-19 severity (mild, moderate, and severe cases) and both erectile scores (erection hardness score and International Index of Erectile Dysfunction-5) and testosterone hormones at first and third month after COVID (P<0.001), except for testosterone level at third month and COVID-19 severity, which showed a statistically significant difference, with P value of 0.031.

Conclusions
The current study correlated the effect of COVID-19 severity in the terms of clinical, laboratory, and radiological presentations on male sexual dysfunction (erectile scores and testosterone hormone) at first and third month after hospital discharge, with statistical significance being highly affected in severe rather than moderate and mild cases. This strengthens the obvious effect of COVID-19 infection on male sexual dysfunction.
Reactivation of retinopathy of prematurity after intravitreal bevacizumab monotherapy in aggressive posterior retinopathy of prematurity
Authors: Verma Ritesh , Rathi Manisha , Phogat Jitender , Sodhi Amita , Lochab Sakshi 
Published: February 2023

Aims:
The aim of the study was to identify the risk factors and timing for the reactivation of retinopathy of prematurity (ROP) after intravitreal bevacizumab (IVB) in aggressive posterior retinopathy of prematurity (APROP).

Materials and Methods:
A retrospective study to analyze the outcome of 24 eyes of 12 infants treated with IVB monotherapy (0.625 mg) was done. Data were analyzed regarding the initial response to IVB, the incidence of ROP reactivation, timing, location, and stage of reactivation. Gestational age, postmenstrual age, and other neonatal comorbid risk factors were compared between the reactivation and those without reactivation after IVB.

Results:
The mean follow-up time was 50.2 ± 1.4 weeks after IVB. Reactivation was identified in 8 (33.3%) eyes. Reactivation in zone I was seen in only two eyes and zone II reactivation was present in six eyes. The mean postmenstrual age at the time of recurrence was 44.0 ± 1.15 weeks (range: 43–45). The mean time between initial treatment and treatment-requiring recurrence was 11.8 ± 0.9 weeks. Multiple pregnancy, low birth weight, sepsis, and necrotizing enterocolitis were significantly associated with ROP reactivation. None of our patients progressed to retinal detachment after laser treatment for reactivation.

Conclusions:
Although IVB treatment is effective in inducing the regression of APROP, the effect may be transient. The recurrence of ROP should be carefully watched in a long-term follow-up after IVB monotherapy, particularly in the first 12 weeks after IVB.
Correlation of the activities of antioxidant enzymes superoxide dismutase and glutathione reductase with micronuclei counts among areca nut chewers of Manipuri population using exfoliative cytology
Authors: Sanjeeta Ngairangbam , Banerjee Sumita , Mukherjee Saikat , Devi T. Premlata , Nandini D. B. , Aparnadevi P. 
Published: February 2023

Context:
Areca nut (AN) is a potent cytotoxic and genotoxic agent. Oxidative stress-induced by chewing of AN can cause DNA damage leading to nuclear anomalies such as micronuclei (MN) and also alters antioxidant defense mechanisms, leading to genomic instabilities and the development of oral cancer.

Aims:
The aim of this study is to study the correlation between the levels of glutathione reductase (GR) and superoxide dismutase (SOD) in exfoliated buccal mucosal cells and the genotoxicity levels (MN count) in chronic AN chewers.

Materials and Methods:
The present study was conducted with the approval of the Research Ethics Board in 60 individuals; 40 cases (Group I–20 raw AN chewers, Group II–20 dried areca with tobacco chewers), and 20 controls as Group III in the age group of 18–68 years who attended the outpatient department of our college. Estimation of SOD and GR and MN assessment was done using buccal exfoliated cells.

Statistical Analysis Used:
Statistical analysis was performed using the Student's t-test and one-way ANOVA.

Results:
Antioxidant levels were found to be significantly reduced in both Group I and Group II in comparison to the control group. Group II showed significantly reduced level of GR in comparison to Group I. The MN count was significantly increased in Group II in comparison to Group I. The MN counts showed an inverse correlation to the activities of the antioxidant enzymes. Greater activities of antioxidant enzymes correlated with decreased MN counts.

Conclusions:
Detection of MN in AN chewers with or without tobacco can be a useful biomarker for clinical screening procedures that may be used as a risk marker for oral cancer. It is important to increase the awareness programs to educate the public about the deleterious effects of AN chewing, emphasize on early intervention of AN chewing habit and thus prevent the development of oral cancer.
Laparoscopic-assisted surgery versus open surgery for transverse colon cancer
Authors: Tamagawa Hiroshi , Numata Masakatsu , Aoyama Toru , Kazama Keisuke , Maezawa Yukio , Atsumi Yosuke , Hara Kentaro , Kano Kazuki , Komori Keisuke , Kawahara Shinnosuke , Yukawa Norio , Sawazaki Sho , Saeki Hiroyuki , Godai Teni , Rino Yasushi , Masuda Munetaka 
Published: February 2023

Introduction:
Previous randomized controlled trials demonstrated similar oncological outcomes between laparoscopic-assisted (LA) and open (OP) colectomy; however, patients with transverse colon cancer were not analyzed. The aim of this study was to confirm the oncological safety and the advantages of the short- and long-term results of LA surgery for transverse colon cancer in comparison to OP surgery.

Materials and Methods:
The study data were retrospectively collected from the databases of four hospitals. Patients with transverse colon cancer who underwent LA or OP R0 or R1 resection were registered.

Results:
Among the 204 patients, 149 underwent OP colectomy and 55 underwent LA colectomy. The median follow-up period was 43 months. The rate of conversion to OP resection was 7.3%. The 5-year overall survival rate of the LA group was higher than that of the OP surgery group for all-stage patients (97.5% vs. 91.1%, P = 0.108), and it was similar in Stage II and Stage III patients (94.1% vs. 94.2%, P = 0.510). The LA group had significantly lower blood loss and a significantly longer operative time in comparison to the OP surgery group. Furthermore, the postoperative hospital stay was significantly shorter (9 vs. 13 days, P = 0.001) and the incidence of Grade ≥III complications was lower in the LA group (3.7% vs. 14.8%, P = 0.031).

Conclusion:
We concluded that LA surgery for transverse colon cancer is oncologically safe and yields better short-term results in comparison to OP surgery.
Formulation and Evaluation of Levofloxacin and Betamethasone Ophthalmic Emulgel
Authors: Sabry Hiba Sabah , Al-Shohani Athmar Dhahir Habeeb , Mahmood Sura Zuhair 
Published: February 2023

Objective:
Drug delivery to ocular tissues is challenging due to rapid removal of instilled drug due to low resident time in ocular tissues. The aim of the study was to formulate an ophthalmic emulgel that delivers two drugs (betamethasone sodium phosphate [BSP] and levofloxacin). The new combination will allow the simultaneous administration and extended release of the two drugs which potentially improve resident time in ocular tissues, patient compliance, and adherence to treatment.

Materials and Methods:
Formulations containing different gelling agents at different concentrations were prepared to choose the optimum combination regarding physical properties and release. The emulgel formulations F1, F2, F3, and F4 were made using gelling agent 1% and 2% xanthan gum, 1% carbopole 934, and 2% methyl cellulose, respectively. F5 was formulated using 2% methyl cellulose with double the amount of poloxamer 188 as emulsifying agent. All the formulations were examined regarding their physical appearance, pH, viscosity, drug content, and in vitro drug release. The optimum formula was also examined for antibacterial activity.

Results:
The results demonstrated that F5 was the optimum formulation having a proper physical characteristics and release profile of both drugs, 96% and 90% for BSP and levofloxacin, respectively, compared to other formulations and commercial eye drops.

Conclusion:
Simultaneous and extended release of the two drugs was achieved using one formulation of emulgel. The ability to deliver hydrophilic and hydrophobic drug through the same formulation without the need to use two drops will improve patient compliance and hence patient adherence to treatment.