bluuebunny/crossref_metadata_2025_split huggingface dataset:
Patch-clamp studies and cell viability assays suggest a distinct site for viroporin inhibitors on the E protein of SARS-CoV-2
Authors: Breitinger Ulrike , Sedky Christine Adel , Sticht Heinrich , Breitinger Hans-Georg 
Published: July 2023

Abstract
Background
SARS-CoV-2 has caused a worldwide pandemic since December 2019 and the search for pharmaceutical targets against COVID-19 remains an important challenge. Here, we studied the envelope protein E of SARS-CoV and SARS-CoV-2, a highly conserved 75–76 amino acid viroporin that is crucial for virus assembly and release. E protein channels were recombinantly expressed in HEK293 cells, a membrane-directing signal peptide ensured transfer to the plasma membrane.

Methods
Viroporin channel activity of both E proteins was investigated using patch-clamp electrophysiology in combination with a cell viability assay. We verified inhibition by classical viroporin inhibitors amantadine, rimantadine and 5-(N,N-hexamethylene)-amiloride, and tested four ivermectin derivatives.

Results
Classical inhibitors showed potent activity in patch-clamp recordings and viability assays. In contrast, ivermectin and milbemycin inhibited the E channel in patch-clamp recordings but displayed only moderate activity on the E protein in the cell viability assay, which is also sensitive to general cytotoxic activity of the tested compounds. Nemadectin and ivermectin aglycon were inactive. All ivermectin derivatives were cytotoxic at concentrations > 5 µM, i.e. below the level required for E protein inhibition.

Conclusions
This study demonstrates direct inhibition of the SARS-CoV-2 E protein by classical viroporin inhibitors. Ivermectin and milbemycin inhibit the E protein channel but their cytotoxicity argues against clinical application.
Future health expenditure in the BRICS countries: a forecasting analysis for 2035
Authors: Sahoo Pragyan Monalisa , Rout Himanshu Sekhar , Jakovljevic Mihajlo 
Published: July 2023

AbstractBackgroundAccelerated globalization especially in the late 1980s has provided opportunities for economic progress in the world of emerging economies. The BRICS nations’ economies are distinguishable from other emerging economies due to their rate of expansion and sheer size. As a result of their economic prosperity, health spending in the BRICS countries has been increasing. However, health security is still a distant dream in these countries due to low public health spending, lack of pre-paid health coverage, and heavy out-of-pocket spending. There is a need for changing the health expenditure composition to address the challenge of regressive health spending and ensure equitable access to comprehensive healthcare services.ObjectivePresent study examined the health expenditure trend among the BRICS from 2000 to 2019 and made predictions with an emphasis on public, pre-paid, and out-of-pocket expenditures for 2035.MethodsHealth expenditure data for 2000–2019 were taken from the OECD iLibrary database. The exponential smoothing model in R software (ets ()) was used for forecasting.ResultsExcept for India and Brazil, all of the BRICS countries show a long-term increase in per capita PPP health expenditure. Only India’s health expenditure is expected to decrease as a share of GDP after the completion of the SDG years. China accounts for the steepest rise in per capita expenditure until 2035, while Russia is expected to achieve the highest absolute values.ConclusionThe BRICS countries have the potential to be important leaders in a variety of social policies such as health. Each BRICS country has set a national pledge to the right to health and is working on health system reforms to achieve universal health coverage (UHC). The estimations of future health expenditures by these emerging market powers should help policymakers decide how to allocate resources to achieve this goal.
Wild food plants of Brazil: a theoretical approach to non-random selection
Authors: Gomes Lailson César Andrade , de Medeiros Patrícia Muniz , Prata Ana Paula do Nascimento 
Published: July 2023

AbstractEthnobiological investigations have focused on identifying factors that interfere with the criteria adopted for selection of plants, especially medicinal plants, by different populations, confirming the theory that plant selection is not random. However, regarding wild food plants, little effort has been made to confirm the theory in this context, especially in Brazil. Therefore, this systematic review aimed to contribute to the establishment of theoretical bases of the non-random selection of wild food plants by local populations in Brazil. For this, searches were made in 4 databases, namely, Web of Science, Scielo, Scopus and PubMed, using 8 sets of keywords in English and Portuguese in order to identify wild food plants occurring in Brazil. The steps were: application of inclusion and exclusion criteria, screening of articles, selection of studies based on risk of bias, data treatment and, finally, data analysis. Eighty articles met the inclusion criteria of this review. However, 45 of them were considered to present high risk of bias and thus 35 articles were kept for the identification of overused and underused families. The results were inferred through two different approaches (IDM and Bayesian). Annonaceae, Arecaceae, Basellaceae, Cactaceae, Capparaceae, Caryocaraceae, Myrtaceae, Passifloraceae, Rhamnaceae, Rosaceae, Sapotaceae, Talinaceae, and Typhaceae were considered overused. Eriocaulaceae, Orchidaceae, and Poaceae were considered underused. Therefore, considering that some families are more (or less) used than others, we confirm that the wild food plants occurring in Brazil, known and used by different populations, are not chosen at random.
p16INK4a and pRb expression in laryngeal squamous cell carcinoma with and without infection by EBV or different genotypes of HPV: a retrospective study
Authors: Vazquez-Guillen Jose Manuel , Palacios-Saucedo Gerardo C. , Alanis-Valdez Alondra Yamileth , Huerta-Escobedo Andrea , Zavala-Pompa Angel , Rivera-Morales Lydia Guadalupe , Martinez-Torres Ana Carolina , Gonzalez-Villasana Vianey , Serna-Hernandez Julio Cesar , Hernandez-Martinez Silvia Judith , Castelan-Maldonado Edmundo Erbey , Montalvo-Bañuelos Martha Socorro , Alonso-Tellez Cesar Alejandro , Sanchez-Fresno Ethel Corinthia , Tamez-Guerra Reyes S. , Rodriguez-Padilla Cristina 
Published: July 2023

Abstract
Background
Laryngeal squamous cell carcinoma (LSCC) represents one of the principal tumors of the head and neck. Human papillomavirus (HPV) and Epstein–Barr virus (EBV) are considered risk factors for the development and the clinical prognosis of LSCC. High levels of p16INK4a are suggested as a surrogate marker of HPV or EBV infection in some head and neck tumors but in LSCC is still controversial. Furthermore, pRb expression may be considered an additional biomarker but it has not been clearly defined. This work aimed to compare the expression of pRb and p16INK4a as possible biomarkers in tumor tissues with and without infection by EBV or different genotypes of HPV from patients with LSCC.

Methods
Tumor samples from 103 patients with LSCC were previously investigated for the presence and genotypes of HPV using the INNO-LiPA line probe assay and for the infection of EBV by qPCR. p16 INK4a and pRb expression was assessed by immunohistochemistry.

Results
Of the 103 tumor samples, expression of p16INK4a was positive in 55 (53.4%) and of this, 32 (56.1%) were positive for HPV whereas 11 (39.3%) were EBV positive but both without a significantly difference (p > 0.05). pRb expression was positive in 78 (75.7%) and a higher frequency of this expression was observed in HPV negative samples (87.0%) (p = 0.021) and in high-risk HPV negative samples (85.2%) (p = 0.010). No difference was observed when comparing pRb expression and EBV infection status (p > 0.05).

Conclusion
Our results support the suggestion that p16INK4a is not a reliable surrogate marker for identifying HPV or EBV infection in LSCC. On the other hand, most of our samples had pRb expression, which was more frequent in tumors without HPV, suggesting that pRb could indicate HPV negativity. However, more studies with a larger number of cases are required, including controls without LSCC and evaluating other molecular markers to determine the real role of p16INK4a and pRb in LSCC.
Unresolved questions in selection of therapies for treatment-naïve chronic lymphocytic leukemia
Authors: Bennett Rory , Anderson Mary Ann , Seymour John F. 
Published: July 2023

Abstract
Background
The treatment landscape for chronic lymphocytic leukemia (CLL) continues to undergo considerable evolution. Optimal selection of initial therapy from multiple effective options provides a major challenge for clinicians, who need to consider both disease and patient factors in conjunction with a view to sequencing available therapies in event of disease relapse.

Review
We explore the most topical clinically relevant unresolved questions through discussion of important available pertinent literature and propose expert opinion based on these data. (1) Shrinking role of chemoimmunotherapy (CIT); while novel therapies are generally superior, we highlight the utility of FCR for IGHV-mutated CLL. (2) Choosing between inhibitors of Bruton’s tyrosine kinase (BTKi); while efficacy between agents is likely similar there are important differences in toxicity profiles, including the incidence of cardiac arrhythmia and hypertension. (3) BTKi with or without anti-CD20 monoclonal antibodies (mAb); while obinutuzumab-acalabrutinib (AO) may confer superior progression-free survival to acalabrutinib (Acala), this is not true of rituximab (Ritux) to ibrutinib (Ib)—we highlight that potential for increased side effects should be carefully considered. (4) Continuous BTKi versus time-limited venetoclax-obinutuzumab (VenO); we propose that venetoclax (Ven)-based therapy is generally preferable to BTKi with exception of TP53 aberrant disease. (5) BTKi-Ven versus VenO as preferred time-limited therapy; we discuss comparable efficacies and the concerns about simultaneous 1L exposure to both BTKi and Ven drug classes. (6) Utility of triplet therapy (BTKi-Ven-antiCD20 mAb) versus VenO; similar rates of complete response are observed yet with greater potential for adverse events. (7) Optimal therapy for TP53 aberrant CLL; while limited data are available, there are likely effective novel therapy combinations for TP53 aberrant disease including BTKi, BTKi-Ven ± antiCD20 mAb.

Conclusion
Frontline therapy for CLL should be selected based on efficacy considering the patient specific biologic profile of their disease and potential toxicities, considering patient comorbidities and preferences. With the present paradigm of sequencing effective agents, 1L combinations of novel therapies should be used with caution in view of potential adverse events and theoretical resistance mechanism concerns in the absence of compelling randomized data to support augmented efficacy.
Lysophosphatidic acid is associated with oocyte maturation by enhancing autophagy via PI3K-AKT-mTOR signaling pathway in granulosa cells
Authors: Liu Jia , Wang Chong 
Published: July 2023

AbstractBackgroundFolliculogenesis is a complex network of interacting cellular signals between somatic cells and oocytes. Many components in ovarian follicular fluid (FF) dynamically change during folliculogenesis and play a positive role in oocyte maturation. Previous studies have reported that lysophosphatidic acid (LPA) promotes cumulus cell expansion, oocyte nuclear maturation, and in vitro maturation of oocytes.ResultsInitially, the expression of LPA was raised in matured FF significantly (P < 0.0001). Then, 10 μM LPA treated for 24 h in human granulosa cells (KGNs) aggravated cell proliferation, with increased autophagy, and reduced apoptosis. Meanwhile, we demonstrated that LPA mediated cell function through the PI3K-AKT-mTOR signaling pathway as PI3K inhibitor (LY294002) significantly prevented LPA-induced AKT, mTOR phosphorylation and autophagy activation. Such results were also verified by immunofluorescence staining and flow cytometry. In addition, an autophagy inhibitor 3 methyladenine (3MA) could also alleviate the effects of LPA, by activating apoptosis through PI3K-AKT-mTOR pathways. Finally, we found blockade with Ki16425 or knockdown LPAR1, alleviated LPA mediated autophagy activation in KGNs, suggesting that LPA enhances autophagy through activation of the LPAR1 and PI3K-AKT-mTOR signaling pathways.ConclusionThis study demonstrates that increased LPA activated PI3K-Akt-mTOR pathway through LPAR1 in granulosa cells, suppressing apoptosis by enhancing autophagy, which might play a role in oocyte maturation in vivo.
Improved language function for post-stroke aphasia in the long term following repeated repetitive transcranial magnetic stimulation and intensive speech-language-hearing therapy: a case report
Authors: Sakurai Yoshihiro , Abo Masahiro , Terajima Saki , Ikeda Saho , Ohara Kensuke , Hada Takuya 
Published: July 2023

Abstract
Background
There have been no papers reporting improvement in language function and changes in cerebral blood flow following repeated use of repetitive transcranial magnetic stimulation in combination with intensive speech-language-hearing therapy. This case report concerns the efficacy of repeated use of repetitive transcranial magnetic stimulation and intensive speech-language-hearing therapy for a certain patient suffering from aphasia following stroke, plus the findings of the cerebral blood flow measurements.

Case presentation
The patient was a 71-year-old right-handed Japanese male who developed fluent aphasia following a left middle cerebral artery stroke. He underwent repetitive transcranial magnetic stimulation and intensive speech-language-hearing therapy five times in total. The repetitive transcranial magnetic stimulation was applied to right inferior frontal gyrus at the frequency of 1 Hz plus 2 hours per day of intensive speech-language-hearing therapy. The patient’s language function was evaluated in the short term and long term. Cerebral blood flow was measured with single photon emission computed tomography scan. As a result, in the short term, the patient’s language function improved especially during the initial hospitalization. In the long term, it improved gradually and stabilized.Cerebral blood flow was increased in the right hemisphere.

Conclusions
The findings of the study indicate that the repeated use of repetitive transcranial magnetic stimulation and intensive speech-language-hearing therapy may be effective in improving and preserving language function and increasing cerebral blood flow for aphasia following stroke.
Persistent pulmonary hypertension of the newborn due to methylmalonic acidemia: a case report and review of the literature
Authors: Hemmati Fariba , Barzegar Hamide 
Published: July 2023

Abstract
Background
Persistent pulmonary hypertension of the newborn manifesting with refractory and severe cyanosis is the consequence of high pulmonary vascular resistance causing extrapulmonary right-to-left shunt. Acidosis and hypoxemia produce pulmonary vasoconstriction. Persistent pulmonary hypertension of the newborn occurs due to numerous disorders and has been rarely reported as a manifestation of methylmalonic acidemia. We report a newborn with methylmalonic acidemia who presented with persistent pulmonary hypertension of the newborn.

Case presentation
A 1-day-old Iranian girl presented with respiratory distress and refractory metabolic acidosis. She was born at 39 + 5 weeks gestational age with Apgar scores of 8 and 9 in the 1st and 5th minutes, respectively, and was in good condition up to 10 hours of life. After that, she presented with cyanosis, tachypnea, retraction, and hypotonia. Despite receiving oxygen, she had low oxygen saturation. Echocardiography revealed severe pulmonary hypertension and right-to-left shunt through patent ductus arteriosus and foramen ovale. Her acidosis worsened despite receiving full support and medical therapy. So, she was started on peritoneal dialysis. Unfortunately, she did not respond to treatment, and after she had died, biochemical tests confirmed methylmalonic acidemia.

Conclusion
Persistent pulmonary hypertension of the newborn is a very rare manifestation of methylmalonic acidemia. Severe inborn errors of metabolism may cause irreversible damage with adverse lifelong morbidity, and early diagnosis may help to prevent such complications. Furthermore, diagnosis of these disorders aids in prenatal diagnosis through the use of cultured amniocytes or chorionic villi to detect gene mutations, as well as biochemical analyses of amniotic fluid for subsequent pregnancies.
Odontoblasts release exosomes to regulate the odontoblastic differentiation of dental pulp stem cells
Authors: Luo Xinghong , Feng Weiqing , Huang Shijiang , Miao Shenghong , Jiang Tao , Lei Qian , Yin Jingyao , Zhang Sheng , Bai Xiaochun , Hao Chunbo , Li Weizhong , Ma Dandan 
Published: July 2023

Abstract
Background
Dental pulp stem cells (DPSCs) play a crucial role in dentin-pulp complex regeneration. Further understanding of the mechanism by which DPSCs remain in a quiescent state could contribute to improvements in the dentin-pulp complex and dentinogenesis.

Methods
TSC1 conditional knockout (DMP1-Cre+; TSC1f/f, hereafter CKO) mice were generated to increase the activity of mechanistic target of rapamycin complex 1 (mTORC1). H&E staining, immunofluorescence and micro-CT analysis were performed with these CKO mice and littermate controls. In vitro, exosomes were collected from the supernatants of MDPC23 cells with different levels of mTORC1 activity and then characterized by transmission electron microscopy and nanoparticle tracking analysis. DPSCs were cocultured with MDPC23 cells and MDPC23 cell-derived exosomes. Alizarin Red S staining, ALP staining, qRT‒PCR, western blotting analysis and micro-RNA sequencing were performed.

Results
Our study showed that mTORC1 activation in odontoblasts resulted in thicker dentin and higher dentin volume/tooth volume of molars, and it increased the expression levels of the exosome markers CD63 and Alix. In vitro, when DPSCs were cocultured with MDPC23 cells, odontoblastic differentiation was inhibited. However, the inhibition of odontoblastic differentiation was reversed when DPSCs were cocultured with MDPC23 cells with mTORC1 overactivation. To further study the effects of mTORC1 on exosome release from odontoblasts, MDPC23 cells were treated with rapamycin or shRNA-TSC1 to inactivate or activate mTORC1, respectively. The results revealed that exosome release from odontoblasts was negatively correlated with mTORC1 activity. Moreover, exosomes derived from MDPC23 cells with active or inactive mTORC1 inhibited the odontoblastic differentiation of DPSCs at the same concentration. miRNA sequencing analysis of exosomes that were derived from shTSC1-transfected MDPC23 cells, rapamycin-treated MDPC23 cells or nontreated MDPC23 cells revealed that the majority of the miRNAs were similar among these groups. In addition, exosomes derived from odontoblasts inhibited the odontoblastic differentiation of DPSCs, and the inhibitory effect was positively correlated with exosome concentration.

Conclusion
mTORC1 regulates exosome release from odontoblasts to inhibit the odontoblastic differentiation of DPSCs, but it does not alter exosomal contents. These findings might provide a new understanding of dental pulp complex regeneration.
Dyskerin and telomerase RNA component are sex-differentially associated with outcomes and Sunitinib response in patients with clear cell renal cell carcinoma
Authors: Yuan Huiyang , Qin Xin , Yang Qingya , Liu Li , Fang Zhiqing , Fan Yidong , Xu Dawei 
Published: July 2023

Abstract
Background
Clear cell renal cell carcinoma (ccRCC) displays sex-biased incidence, outcomes, molecular alterations and treatment efficacy; however, clinical managements are largely identical in male and female patients. Moreover, many biomarkers have been identified as predictors for ccRCC outcomes and response to therapeutic drugs, such as multitargeted tyrosine-kinase receptor (TKR) inhibitors, but little is known about their sex-specificity. Dyskerin (DKC1), encoded by the DKC1 gene within Xq28, is a telomerase co-factor stabilizing telomerase RNA component (TERC) and overexpressed in various cancers. Here, we determined whether DKC1 and/or TERC affect ccRCC sex-differentially.

Methods
DKC1 and TERC expression in primary ccRCC tumors was assessed using RNA sequencing and qPCR. DKC1 association with molecular alterations and overall or progression-free survival (OS or PFS) was analyzed in the TCGA cohort of ccRCC. The IMmotion 151 and 150 ccRCC cohorts were analyzed to evaluate impacts of DKC1 and TERC on Sunitinib response and PFS.

Results
DKC1 and TERC expression was significantly upregulated in ccRCC tumors. High DKC1 expression predicts shorter PFS independently in female but not male patients. Tumors in the female DKC1-high group exhibited more frequent alterations in PIK3CA, MYC and TP53 genes. Analyses of the IMmotion 151 ccRCC cohort treated with the TKR inhibitor Sunitinib showed that female patients in the DKC1-high group was significantly associated with lower response rates (P = 0.021) accompanied by markedly shortened PFS (6.1 vs 14.2 months, P = 0.004). DKC1 and TERC expression correlated positively with each other, and higher TERC expression predicted poor Sunitinib response (P = 0.031) and shorter PFS (P = 0.004), too. However, DKC1 rather than TERC acted as an independent predictor (P < 0.001, HR = 2.0, 95% CI 1.480–2.704). In male patients, DKC1 expression was associated with neither Sunitinib response (P = 0.131) nor PFS (P = 0.184), while higher TERC levels did not predict response rates. Similar results were obtained from the analysis of the Sunitinib-treated IMmotion 150 ccRCC patients.

Conclusions
DKC1 serves as an independent female-specific predictor for survival and Sunitinib efficacy in ccRCC, which contribute to better understanding of the sex-biased ccRCC pathogenesis and improve personalized interventions of ccRCC.
Investigation of the structure-activity relationship at the N-terminal part of minigastrin analogs
Authors: Holzleitner Nadine , Günther Thomas , Daoud-Gadieh Amira , Lapa Constantin , Wester Hans-Jürgen 
Published: July 2023

Abstract
Background
Over the last years, several strategies have been reported to improve the metabolic stability of minigastrin analogs. However, currently applied compounds still reveal limited in vitro and in vivo stability. We thus performed a glycine scan at the N-terminus of DOTA-MGS5 (DOTA-d-Glu-Ala-Tyr-Gly-Trp-(N-Me)Nle-Asp-1-Nal) to systematically analyze the peptide structure. We substituted N-terminal amino acids by simple PEG spacers and investigated in vitro stability in human serum. Furthermore, we evaluated different modifications on its tetrapeptide binding sequence (H-Trp-(N-Me)Nle-Asp-1-Nal-NH2).

Results
Affinity data of all glycine scan peptides were found to be in a low nanomolar range (4.2–8.5 nM). However, a truncated compound lacking the d-γ-Glu-Ala-Tyr sequence revealed a significant loss in CCK-2R affinity. Substitution of the d-γ-Glu-Ala-Tyr-Gly sequence of DOTA-γ-MGS5 (DOTA- d-γ-Glu-Ala-Tyr-Gly-Trp-(N-Me)Nle-Asp-1-Nal-NH2) by polyethylene glycol (PEG) spacers of different length exhibited only a minor influence on CCK-2R affinity and lipophilicity. However, in vitro stability of the PEG-containing compounds was significantly decreased. In addition, we confirmed that the tetrapeptide sequence H-Trp-Asp-(N-Me)Nle-1-Nal-NH2 is indeed sufficient for high CCK-2R affinity.

Conclusion
We could demonstrate that a substitution of d-γ-Glu-Ala-Tyr-Gly by PEG spacers simplified the peptide structure of DOTA-MGS5 while high CCK-2R affinity and favorable lipophilicity were maintained. Nevertheless, further optimization with regard to metabolic stability must be carried out for these minigastrin analogs.